Screening and regulation of nanozyme activity via liquid metals coined electron rearrangement and phase engineering

Domain: biology
Published: 2026-03-26
Editorial outcome: major_revision

Abstract
Liquid metals (LMs) have been used for design of advanced materials by leveraging their unique liquid and electronic properties. Here, we design a series of liquid metal-buffered amorphous molybdenum sulfide nanozymes with different gallium indium ratios, in which Ga In MoS 75.5 24.5 X nanozymes exhibit the enzyme-like activity, superior to the crystalline MoS . This has been verified and assigned to the roles of liquid metal: (i) serving as the building template, (ii) phase engineering with catalytic active sites, (iii) electron-rich microenvironment for improving catalytic activity. Besides, the amorphous Ga In MoS nanozyme possesses oxidase (OXD)-like and 75.5 24.5 X S nicotinamide adenine dinucleotide oxidase (NOX)-like activiSty to achieve multiple-enzyme-like E cascade catalytic reactions, disrupting intratumoral redox and metabolism homeostasis. As the R example of leveraging liquid metal as a well-desiPgned platform to screen high-performance N nanozyme with excellent therapeutic effects, thNis work highlights the roles of tailored electronic I structure and phase engineering for regula tion of catalytic activity of nanozymes and may also E broaden the application of liquid metaLl in catalysis and biomedical field.

Introduction
Liquid metals (LMs) aAre enigmatic materials, with simultaneous metallic and liquid nature.1, 2 The most striking features of liquid metals are the amorphous property and unusual electronic microenvironment, which originate from the electron-rich metallic cores and free delocalized electron clouds.3, 4 Such characters make liquid metals show great potential to tune the physical, chemical, and electronic properties of materials for boosting their performance in various engineering applications.5, 6, 7, 8, 9, 10 More importantly, liquid metals represent a promising technical route for manufacturing modular advanced materials.3, 11, 12, 13, 14, 15, 16, 17, 18, 19 The obtained combinatorial materials can readily maintain the typical characteristics of liquid metals and may attain better functionality.20 In particular, the intrinsic amorphous nature of liquid metals creates a facile pathway for phase engineering of materials, thus deriving more intriguing properties and innovative applications.21, 22 Interestingly, liquid metals have also shown promising prospects in catalysis by capitalizing on their unique electronic properties.23, 24, 25, 26 For example, recent studies have reported that liquid metals could create dynamic uniform catalytic sites and favorable electronic structures for catalytic metal species to remarkably enhance their catalytic capability.27, 28, 29 Similarly, liquid metals have been proposed as a unique electron mediator for temporary electron storage, and promoted the extraction and transfer of electrons, as well as charge separation, greatly improving the optoelectronic property of perovskite films.30 Furthermore, the representative Ga-based liquid metal alloys with excellent biocompatibility and attractive flexibility, have recently aroused great interest in the biological field including cancer therapy, antimicrobial treatment, and bioimaging.31, 32, 33, 34, 35, 36, 37, 38, S39, 40, 41, 42, 43 These studies indicate the exceptional advantages of liquid metals to create advanced materials for catalytic and biomedicine applications.

Nanozymes are nanomaterials with inherent Nenzyme-mimicking catalytic properties, and have ignited increasing attention due to their adv antages of high catalytic stability and durability, easy preparation, and low cost.44, 45, 46, 47 ALlthough promising, the unsatisfactory catalytic efficiency of nanozymes remains a crucial issue for future practical applications.48, 49, 50, 51, 52 This demonstrates the vital necessity of developing an appropriate strategy to optimize the nanozyme catalytic performance. From aAn in-depth perspective, the catalytic active sites and local electronic characteristics determine their catalytic performance to a large extent.49, 50, 53, 54, 55, 56, 57 Notably, phase engineering of nanomaterials has demonstrated remarkable prospects in the catalysis field because this process modulates the atomic arrangements and electronic structure of catalysts, hence providing unusual catalytic activity.58, 59 Especially, amorphous materials possess distinctive disordered atomic arrangement that enables them with abundant active sites and optimized electronic configurations, which are beneficial for enhancing their catalytic properties.60, 61 Thus, combining the ability of phase engineering and the unique electronic properties, liquid metals are expected to be a dream matrix to construct high-performance nanozymes. However, the potential of liquid metal in regulating nanozyme catalytic activity has rarely been explored mechanistically. In particular, previous research efforts have focused on the application of liquid metal-based nanozyme while overlooking the catalytic potential of liquid metals for screening and regulation of nanozyme activity. 62, 63

Here we design and screen the liquid metals-buffered nanozymes, in which the liquid metals can not only initiate phase engineering to generate an amorphous nanozyme with abundant catalytic active sites, but also afford an electron-rich microenvironment to optimize the electronic structure of nanozymes for improving the catalytic performance (Fig. 1). We first prepare a series of molybdenum sulfide nanozymes by using five liquid metals with the composition of Ga In 1-x x (with x=0, 0.125, 0.245, 0.375, and 0.50) as templates, and these nanozymes feature uniformly core-shell structure and amorphous property. We observe that tShese nanozymes exhibit peroxidase (POD)-like activities, in which the LM MoS nanozymes (LM=Ga In ) present the best X R 75.5 24.5 catalytic activity. Then, the role of liquid metals inP regulating the enzyme-mimicking catalytic activity of LM MoS nanozymes is explored. WNe demonstrate that, compared to crystalline MoS , X 2 the amorphous LM MoS X nanozymes expo se abundant active sites and defects, and show about a 10-fold increase in the POD-like catalLytic activity, together with a significantly higher glutathione (GSH) consumption ability. Meanwhile, this strategy can turn on the oxidase (OXD)-like and nicotinamide adenine dinucleotide oxidase (NOX)-like activities of LM MoS to achieve cascade X catalytic reactions, eveAntually leading to reactive oxygen species (ROS) burst. Both in vitro and in vivo experiments demonstrate that the LM MoS nanozymes with multienzyme-mimicking X catalytic activity can trigger a highly efficient cascade reaction to disrupt intratumoral redox and metabolic homeostasis, eventually killing tumor cells. In this work, we introduce liquid metal with advantages in synthesis and catalysis to screen and regulate the catalytic and therapeutic effects of nanozymes for efficient tumor inhibition. Our work not only investigates the catalytic activity of liquid metal-buffered nanozyme with different gallium indium ratios, but also elucidates the mechanism underlying the enhanced enzyme-like catalytic activity through density functional theory (DFT) calculations. This study offers promising insights into the design and regulation of nanozymes, with potential benefits for future biomedical applications of liquid metals.

Results
The design and preparation of liquid metal-buffered nanozymes
As shown in Fig. 1a, the nanozyme was synthesized through facile probe sonication. Liquid metals with different ratios of gallium and indium were prepared by dissolving a calculated amount of In pellets in liquid Ga. The resultant liquid metal alloys should retain their liquid states at close to room temperature to ensure the subsequent liquid-liquid interfacial reactions.64 According to the previous reports, the molar fraction of In should below 50 at% in order to ensure their liquid states at close to room temperature.65, 66 Specifically, the composition of Ga In (with x = 0, 0.125, 0.245, S1-x x 0.375, and 0.50) were chosen and prepared to design a series oSf liquid metal-buffered nanozymes. Molybdenum is a low-cost and biocompatible transition metal with multiple oxidation states, which endows Mo-based nanomaterials with tunablPe enzyme-like activity.67 Among Mo-based nanomaterials, molybdenum sulfide features unNique structural and electronic properties and show superior performance in various applica tions, especially for catalysis.68, 69 The generated nanozymes were denoted as Ga 50 In 5L0 MoS X , Ga 62.5 In 37.5 MoS X , Ga 75.5 In 24.5 MoS X , Ga 87.5 In 12.5 MoS , Ga MoS . During this procedure, liquid metal served as a template to trigger in situ X 100 X galvanic replacement reactions between liquid metal droplets and MoS 2- ions to produce core– 4 shell nanostructures uApon ultrasonication. The thermodynamic driving force underlying the galvanic replacement reaction originates from the difference in redox potentials between the two metal-based species.21, 64 The size distribution of nanoparticles became narrower with higher sonication intensity applied (Supplementary Fig. 1). Transmission electron microscopy (TEM) images illustrated the well-constructed core-shell structures of the synthesized five nanozymes (Fig. 2a). Then, the energy dispersive spectroscopic (EDS) mapping and X-ray photoelectron spectroscopy (XPS) results demonstrated the existence of Ga, In, Mo and S elements in these nanozymes (Supplementary Fig. 2-3). Next, high-resolution TEM (HR-TEM) images indicated the amorphous nature of five nanozymes (Fig. 2a). According to previous studies, the intrinsic amorphous nature of liquid metals and short reaction time are considered the main reasons for the formation of the amorphous structure.4, 11 Dynamic light scattering (DLS) results of five nanozymes revealed that their hydrodynamic diameters were mainly distributed around 300 nm (Fig. 2b). The surface zeta potential measurement showed that these nanozymes were negatively charged (Fig. 2c). Then, we explored the POD-like activity of five nanozymes by monitoring the oxidation of 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) at 415 nm. As revealed in Fig. 2d and 2e, obvious ABTS oxidation could be detected, indicating that all five nanozymes possessed POD-like activity. Among them, Ga In MoS exhibited the highest 75.5 24.5 X catalytic activity, while Ga In MoS showed a relatively low activity. According to previous 50 50 X studies, the Indium content in GaIn alloys significantly affects the disordered structure and the charge density distribution of alloys, thus influencing their aSmorphous structure and electrical conductivity.70 Specifically, the alloying of Ga with In causes more disordered structure. This amorphous structure is beneficial for the high catalPytic performance of catalysts. However, the electrical conductivity of GaIn alloys decreasesN with In alloying of Ga because the charge density I near the In atoms is significantly smaller, which is unfavorable to the electron transfer and the E subsequent catalytic process of catalyLsts. Thus, the catalytic activity of the resulting nanozymes C could show a trend of initially increasing and then decreasing with increasing In content in the I T GaIn alloy. These findings could explain the experimental results that the alloying effect of In with R Ga favored the catalyAtic reaction, while lower or higher In contents compromised the catalytic activity. We further employed XPS spectra to characterize the electronic properties of these nanozymes. As displayed in Fig. 2f, the binding energies of Mo 3d were positively shifted in Ga In MoS and Ga MoS nanozymes, in comparison to other nanozymes, revealing the 75.5 24.5 X 100 X occurrence of electron transfer and rearrangement. Moreover, we have analyzed the surface valence states of Ga In MoS and Ga MoS , revealing their different Mo oxidation states 75.5 24.5 X 100 X (Supplementary Fig. 4). This meant that liquid metals with varying compositions had different effects on the electronic properties and valence states of the resulting nanozyme.

Next, we took Ga In MoS (it was denoted as LM MoS in the following content unless 75.5 24.5 X X indicated otherwise) with the best catalytic activity as an example to investigate the role of liquid metal for nanozyme catalysis. First, the structure of LM MoS was thoroughly studied using X various characterization techniques. TEM image revealed that the obtained LM MoS nanozymes X exhibited well-constructed core-shell spherical structures (Fig. 3a), with a shell thickness of approximately 40 nm. The elemental mapping images demonstrated the existence of Ga, In, Mo and S elements in LM MoS nanozymes (Fig. 3b). Then, size distribution result revealed that the X Z-Average size and polydispersity index (PDI) of LM MoS were 309.3 nm and 0.188, X respectively (Fig. 3c), indicating that the LM MoS had uniform size and good dispersibility. X Moreover, the DLS results, SEM and TEM images, and XPS spectra suggested that the LM MoS X nanozymes remained monodisperse and stable over 7 days (Supplementary Fig. 5). The elements S content in LM MoS nanozymes was further analyzed by iSnductively coupled plasma optical X E emission spectrometry (ICP-OES, Supplementary Table 1). Subsequently, the amorphous nature R of LM MoS nanozymes was explored. The selectiveP area electron diffraction (SAED) pattern and X the High-resolution (HR)-TEM image revealedN that no crystal form was observable in LM MoS X I nanozymes (Fig. 3d and 3e), and the corresp onding 3D pseudo-color surface plot demonstrated the E highly disordered arrangement of atoLms (Fig. 3f). These results were consistent with the X-ray C diffraction (XRD) analysis (Fig. 3g), which showed a single broad peak, further confirming the I T amorphous structure of LM MoS nanozymes. To further characterize the detailed electronic and X R structural properties oAf amorphous LM MoS X nanozymes, crystalline MoS 2 nanospheres were prepared (Supplementary Fig. 6-9). The SAED pattern (Supplementary Fig. 7c), lattice fringes in HR-TEM image (Supplementary Fig. 7) and XRD analysis (Supplementary Fig. 8) proved the crystalline form of MoS nanospheres. Afterwards, XPS was employed to analyze the surface 2 chemical state of the LM MoS nanozymes (Fig. 3h-3k). According to these XPS results, the X amorphous LM MoS nanozymes and crystalline MoS had very different valence states and X 2 electronic properties. The LM MoS nanozymes possessed richer Mo valence states. Fig. 3i and X 3k revealed the presence of Mo6+, Mo5+, and Mo4+ oxidation states in LM MoS nanozymes, while X only Mo4+ existed in MoS (Supplementary Fig. 9). As shown in Fig. 3j, LM MoS nanozymes 2 X had many disulfide ligands in the S 2p peak as compared with MoS (Supplementary Fig. 9), which 2 were consistent with the previous reported amorphous MoS .69 These data demonstrated that the X LM MoS nanozymes exhibited abundant active sites, which were ascribed to their amorphous X structure. Meanwhile, we found that the binding energy of Mo 3d in the LM MoS nanozymes X was positively shifted when compared to MoS (Fig. 3k), which indicated the electronic 2 perturbations in the Mo 3d orbitals. Thereafter, electron spin resonance (ESR) measurement was also carried out to probe the electronic information of LM MoS nanozymes. The ESR signals in X Fig. 3l manifested the existence of S vacancies (g=2.008), Mo5+species (g=2.01, g=1.97), and sulfur chain radicals (g=2.034, g=2.004), which further confirmed the abundant active sites of LM MoS nanozymes.69 These data elucidated the successful preparation and the delicate structure of X S LM MoS nanozymes. S X

Catalytic performance of the LM MoS nanozymePs
X
The delicate structure of LM MoS nanoNzymes spurred us to investigate their catalytic X performance. As shown in Fig. 4a, a signi ficantly increased absorbance of oxidized ABTS was observed in the LM MoS X group, whiLle the MoS 2 treated group exhibited weak catalytic activity, indicating that the LM MoS nanozymes possessed high POD-like activity. We further used X steady-state kinetic assay to evaluate the catalytic performance of LM MoS and MoS . The X 2 steady-state kinetic paArameters including the Michaelis-Menten constant (K m ) and the maximum initial velocity (V ) were obtained by Michaelis–Menten saturation curves (Supplementary Fig. max 10-12) and were shown in Supplementary Table 2. The apparent K value of LM MoS with H O m X 2 2 as the substrate was about 4.33 times lower than MoS , and the V value of LM MoS was about 2 max X 3.83 times higher than MoS . The reduced K value indicated that the LM MoS had a higher 2 m X affinity and good substrate selectivity to H O , and the increased V value reflected its superior 2 2 max catalytic activity. Moreover, the apparent K value and V value of LM MoS with H O as the m max X 2 2 substrate were both better than those of MoS and many other reported molybdenum-based 2 nanozymes in Supplementary Table 3. Next, fluorescence assay was applied to assess the catalytic capability. Terephthalic acid (TA) was used as a probe for capturing •OH (Supplementary Fig. 13). According to the fluorescence spectra in Fig. 4b and Supplementary Fig. 14, the ability to generate •OH radicals of LM MoS was 10 times higher than that of MoS . As shown in Supplementary X 2 Fig. 15, the specific activity (SA) value of LM MoS (4.12 U/mg) was 7.5 times higher than that X of crystalline MoS (0.55 U/mg), showing that the designed liquid metal-buffered nanozyme had 2 higher catalytic performance. These results were consistent with the ABTS oxidation experiment, both demonstrating the excellent POD-like activity of LM MoS nanozymes. Moreover, the X nanozyme exhibited consistent POD-like catalytic performance over 7 days (Supplementary Fig. 16), confirming its good stability. To further investigate the enzymatic characteristics of LM MoS X nanozymes, we utilized 1,3-diphenylisobenzofuran (DPBF) assay to monitor the oxidase-like activity. DPBF is a classical probe for detecting the generatioSn of O •−. Upon O •− oxidation, the 2 2 intensity of the characteristic absorption of DPBF at around 426 nm would decrease (Supplementary Fig. 17a). As shown in Fig. 4c, a dPramatic decrease in DPBF absorbance was observed after LM MoS nanozymes treatmenNt, demonstrating the O •− generated by the OXD- X 2 mimicking process. By contrast, negligible D PBF degradation was observed in MoS 2 treated group under our experimental conditions (LSupplementary Fig. 17b-c). Such OXD-like activity was probably ascribed to the intervalence charge-transfer between Mo5+/Mo6+ in the LM MoS , which X was absent in MoS .71 Furthermore, various substrates were selected to explore the substrate 2 selectivity of LM MoS . For the POD-like reaction, the substrate selectivity of LM MoS X X nanozymes was examined using some classical POD-like substrates, including 3,3′,5,5′- Tetramethylbenzidine (TMB), ABTS, and 1,2-diaminobenzene (OPD). As shown in Supplementary Fig. 18, the LM MoS nanozyme could catalyze these substrates, resulting in X characteristic oxidation colors and absorption peaks, which demonstrated its good POD-like activity. For an OXD-like reaction, the substrate selectivity of LM MoS nanozymes was carried X out in O and N -saturated solutions, respectively. As shown in Supplementary Fig. 19, the 2 2 characteristic absorption of DPBF at around 426 nm substantially decreased in the presence of LM MoS nanozymes under a normal or saturated O atmosphere. However, the catalytic ability of X 2 LM MoS nanozymes was weakened in hypoxia condition, indicating that O was the typical X 2 substrate for LM MoS nanozymes. To verify the production of ROS, free radical quenching X experiment was performed. Specifically, isopropanol (IPA) and p-benzoquinone (PBQ) served as •OH quencher and O •− quencher, respectively. According to Supplementary Fig. 20, a significant 2 enhancement in fluorescence at 435 nm could be observed for LM MoS nanozymes in the X presence of H O in comparison to other control groups. However, in the presence of IPA, the 2 2 fluorescence intensity at 435 nm decreased substantially, indicating that LM MoS nanozymes X could effectively produce •OH. Then, we observed that the absorbance of DPBF at 426 nm in the LM MoS + PBQ treated group was comparable to that of the control group, proving the formation X of O •− during the catalytic process of LM MoS nanozymes (Supplementary Fig. 21). Afterward, 2 X S the Electron paramagnetic resonance (EPR) experiment was coSnducted to evaluate the generation E of radicals. As depicted in Supplementary Fig. 22, the EPR spectra using 5,5-dimethyl-1-pyrroline R N-oxide (DMPO) as probe, exhibiting the distinctivPe quartet signal (1:2:2:1) of •OH in the LM N MoS + H O group. In addition, an obvious cNharacteristic six-line pattern of O •− was observed I X 2 2 2 after LM MoS treatment. These results ind icated the brilliant POD-like and OXD-like activities E X of LM MoS X . L

In addition to the enzyme-mimicking activities mentioned above, nanozymes have shown great C prospects in regulating the biological metabolic processes. Specifically, GSH and nicotinamide I T adenine dinucleotide (ANADH) are important intracellular metabolites upregulated in cancer cells, R both serving as the essential reductive factors for maintaining cellular redox homeostasis and A modulating cellular metabolism.72 Moreover, NADH is necessary for preserving cellular bioenergetics, genomic integrity, and other important physiological pathways.73 However, these reductive metabolites play a key role in cellular oxidative stress defense mechanisms, which can efficiently scavenge cellular ROS, thereby resulting in limited therapeutic effects. In consideration of the abundant active sites and redox pairs in LM MoS nanozymes, we then explored whether X the designed nanozymes could work on these critical metabolites. As shown in Fig. 4d, the decrease in GSH was observed after treatment with LM MoS nanozymes, revealing that LM X MoS nanozymes could effectively deplete GSH. The quantitative analysis in Fig. 4e showed that X the GSH consumption efficiency of LM MoS nanozymes reached 80%, while MoS was about X 2 10%. Next, NADH oxidation in vitro was measured by absorption spectra (Fig. 4f), where the characteristic absorption peak of NADH at 340 nm gradually decreased over time under the treatment of the LM MoS nanozymes, and the NAD+ peak at 260 nm increased. In contrast, X Supplementary Fig. 23 showed that MoS treated group had negligible degradation of NADH, 2 revealing the NOX-mimicking activity of LM MoS nanozymes. In the following, the substrate X selectivity of LM MoS was explored in some biological oxidase substrates. Impressively, NADH X could be effectively consumed by LM MoS while few (less than 6%) other biomolecules were X affected (Supplementary Fig. 24). More significantly, such NOX-mimicking catalytic process was accompanied with H O production (Fig. 4g). During this proScess, NADH served as an electron 2 2 donor for the synergistic reduction of O •− by LM MoS nanozymes to generate H O , which 2 RX 2 2 further contributed to the POD-like activity of nanozyPme. In this case, LM MoS nanozymes could X efficiently induce the cascade reactions withN OXD-like, NOX-like, and POD-like catalytic I activities, thereby consuming the reductive stress and generating more oxidative stress (Fig. 4h). E Moreover, the addition of interfering iLon or protein had no obvious influence on POD-, OXD-, and C NOX-like activities of LM MoS X nanozymes, demonstrating the anti-interference ability of the I T catalytic system (Supplementary Fig. 25-27). These results elucidated that the liquid metal- R buffered nanozymes eAxhibited improved enzyme-like catalytic activity and great multienzyme- mimicking cascade catalytic activity, highlighting the key role of liquid metal in regulating catalytic performance.

DFT Calculations
To gain insights into the reasons for the enhanced enzyme-like catalytic activity of liquid metal- buffered nanozyme, density functional theory (DFT) calculations based on first principles were conducted to investigate their electronic structures. First, three computational surface models representing ideal MoS , MoS (amorphization) and LM MoS were constructed to elucidate their 2 X X microscopic electron structures. As shown in the charge density maps (Fig. 5a), the amorphous MoS exhibited an increased electron density around Mo atoms compared with the MoS model. x 2 Notably, the LM MoS model achieved the highest electron density, indicating the electron X donation from liquid metal to Mo atoms. Moreover, the differential charge density of LM MoS X in Fig. 5b revealed the electron donation from the liquid metal to Mo atoms. In addition, the Bader charge of Mo in MoS , MoS , and LM MoS was calculated. The Bader charge analysis indicated 2 X X a charge gain of 0.52 e- on the Mo sites in the MoS relative to MoS . Compared to MoS , the X 2 X electron accumulation on the Mo sites increased by an additional 0.21 e- in the LM MoS , which X further confirmed that electron donation from liquid metal to Mo atoms. Then, the density of states (DOS) of MoS , MoS and LM MoS models, as shown in Fig. 5c, revealed that LM MoS 2 X X S X exhibited a significantly higher DOS near the Fermi level and sSeveral new hybrid electronic states E appeared compared to MoS and MoS , indicating stronger interaction and charge transfer 2 X R between LM MoS and adsorbates.51, 74 FurthermorPe, the d-band center of the Mo orbital was X calculated based on the DOS of MoS , MoS Nand LM MoS . As shown in Fig. 5d, LM MoS 2 X X X I nanozymes exhibited an upward shift in d band center position (-1.686 eV), which was E significantly nearer to the Fermi levelL than those of MoS 2 (-3.619 eV) and MoS X (-2.908 eV). The C state near to the Fermi level contributed to the interaction between the adsorbed substrate and LM I T MoS nanozyme, thereby favoring the catalytic reaction.56 These results were consistent with the X R kinetic parameters stuAdy (Supplementary Table 2), in which the reduced K m value indicated that A the LM MoS had a higher affinity to H O . Based on these results, we confirmed that the pivotal X 2 2 role of liquid metal, i.e., amorphization, and electron transfer, endowing LM MoS nanozymes X with optimized enzyme-like catalytic activity. In our designed system, on the one hand, liquid metals ingeniously endowed the nanozymes with amorphous property, thus providing abundant active sites for catalysis. On the other hand, the liquid metals matrix and the amorphous character of nanozymes led to a favorable electronic perturbation, which could promote electron transfer and optimize the reaction process, thereby boosting the catalytic behavior. These data indicated the superiority of liquid metal for the construction of high-performance nanozyme.

LM MoS nanozymes-based catalytic therapy
X
The excellent catalytic capability of LM MoS nanozymes motivated us to explore the X intracellular performance. First, cytotoxicity experimental results suggested good biocompatibility of LM MoS even at the concentration of 200 μg/mL (Supplementary Fig. 28). Then, LM X MoS @RB (Rhodamin B) as a model was applied to investigate the cellular uptake behavior of X LM MoS . As shown in confocal laser scanning microscopy (CLSM) images, the red fluorescence X of RB in LM MoS @RB treated 4T1 mouse breast cancer cells increased with time and reached X the maximum after 9 h (Supplementary Fig. 29), confirming that the LM MoS @RB could be X effectively internalized by 4T1 cells. Then, we systematically studied the catalytic behavior of LM MoS within 4T1 cells. The intracellular NADH and GSH leveSls were found to gradually decrease X E with increasing concentrations of LM MoS nanozymes (Fig. 6a and 6b). These results X R demonstrated that the LM MoS nanozymes couldP efficiently diminish the reductive stress in X cancer cells. The intracellular ROS generatedN by LM MoS nanozymes was then evaluated X I utilizing the 2′,7′-dichlorofluorescein diace tate (DCFH-DA) probe. The 4T1 cells treated with E MoS 2 only presented faint fluorescencLe signals under our experimental conditions. Comparatively, C the nonfluorescent DCFH-DA could be efficiently converted into bright green fluorescent DCF I T inside LM MoS nanozymes-treated cells, confirming the burst of intracellular ROS (Fig. 6c and X R 6d). Such an elevated Alevel of ROS in cells was partly attributed to the generation of oxidative stress by the POD-mimic and OXD-mimic activities of LM MoS nanozymes, and partly caused X by the remarkable capability of LM MoS nanozymes to eliminate reductive stress, thereby X inducing severe redox imbalance within cancer cells. Furthermore, the mitochondrial membrane potential (MMP) of 4T1 cells after different treatments was determined by JC-1 assay. As seen in Fig. 6e and 6f, the JC-1 probe displayed an intense red fluorescence signal in PBS, LM NPs (pure Ga In , Supplementary Fig. 30) and MoS treated cells. In comparison, the cells exhibited 75.5 24.5 2 bright green fluorescence in LM MoS nanozymes treated group, indicating the efficient decrease X of MMP and depolarization of mitochondria. These changes were presumably ascribed to the disturbance of NADH/NAD+ cycle, because it was necessary for the mitochondrial electron transport chain (ETC). These experiment results indicated that the LM MoS nanozymes could X effectively achieve mitochondria damage and ROS burst through cascade catalytic reactions (Fig. 6g).

After assessing the catalytic efficacy of LM MoS nanozymes in cells, we evaluated the X cytotoxicity of the designed nanozymes. First, the in vitro cytotoxicity of nanozymes was measured using the methyl thiazolyl tetrazolium (MTT) method. In general, H O , GSH, and 2 2 NADH are elevated in cancer cells, whereas normal cells have comparatively low concentrations. As displayed in Fig. 6h, LM NPs, MoS and LM MoS nanozymes exhibited negligible 2 X cytotoxicity against 293T human embryonic kidney cells under our experimental conditions. For the 4T1 mouse breast cancer cells (Fig. 6i), the cell viability afSter LM MoS nanozymes treatment X substantially diminished, while the MoS treated group showed limited cytotoxicity. As shown in 2 Supplementary Fig. 31, the half maximal inhibitory cPoncentrations (IC50) value of MoS and LM 2 MoS nanozymes against 4T1 cells were 10N5.6 µg/mL and 24.26 µg/mL, respectively. The X effective cancer cell killing ability was at tributed to the multiple enzyme-mimicking catalytic activity of LM MoS X nanozymes, whLich could induce severe oxidative damage and metabolism C imbalance in cancer cells. Then, the in vitro therapy efficiency was also evaluated through the I T LIVE/DEAD cell staining assay, in which apparent cell death (red signals) had occurred in the LM R MoS X treated group (FAig. 6j). Taken together, these results indicated that the LM MoS X nanozymes A showed a significant anticancer efficacy, which was ascribed to the disruption of intracellular redox and metabolism homeostasis caused by the highly efficient enzyme-like catalytic activity of LM MoS nanozymes. X

The remarkable catalytic effect of LM MoS in cancer cell encouraged us to examine the anti- X tumor performance in vivo. We prepared hyaluronic acid (HA) coated LM MoS nanozymes X (denoted as H-LM MoS ) for anticancer therapy (Supplementary Fig. 32a-e), in which HA X modification endowed the nanoplatform with enhanced tumor targeting ability. As illustrated in Supplementary Fig. 32f, H-LM MoS exhibited negligible Mo leaching after being soaked in X complete cell culture medium for over 7 days. Subsequently, the biotoxicity of the H-LM MoS X nanozymes was systematically assessed. The hemolytic experiment was first conducted to assure its safety for intravenous injection. As shown in Supplementary Fig. 33, H-LM MoS showed X negligible hemolysis toxicity. Then, the in vivo biocompatibility of H-LM MoS was explored. X Mice were injected with H-LM MoS via their tail veins to evaluate the toxicology. The blood X biochemical test and histological analysis in the main organs of H-LM MoS -treated mice were X performed, which showed no apparent toxicity and adverse effects compared with the control group (Supplementary Fig. 34-35). Meanwhile, the mice growth was not impacted within 28 days (Supplementary Fig. 36). These data demonstrated the favorable biocompatibility of H-LM MoS X nanozymes for bioapplication at an injection dose of 10 mg/kg. The half-life of H-LM MoS was X determined to be 1.97 h, as shown in Supplementary Fig. 37S. Subsequently, 4T1 tumor-bearing mice were established to investigate the in vivo anticancer ability (Fig. 7a). First, the tissue distribution of H-LM MoS was evaluated by ex vivoP fluorescence. Supplementary Fig. 38 showed X that a stronger fluorescence signal was detectedN in the tumor treated by H-LM MoS @Cy3, which X I revealed its good tumor accumulation capability. The inductively coupled plasma mass E spectrometry (ICP-MS) results revealLed the enhanced accumulation of H-LM MoS X in the tumor C region (Supplementary Fig. 39). The tumor-targeting capability of H-LM MoS was also evaluated X I T through in vivo fluorescence imaging system. As shown in Supplementary Fig. 40, strong red R fluorescence signals wAere observed within the tumor, confirming the effective tumor accumulation A of nanoparticles. Then, the overall anti-tumor performance of nanozymes was evaluated. The mice were randomly divided into four groups: (I) Control, (II) H-LM, (III) H-MoS , and (IV) H-LM 2 MoS . Different formulations were intravenously administered to the mice. As shown in Fig. 7c- X 7e, the tumor growth in H-LM MoS dosed group was significantly inhibited, while H-LM and H- X MoS treated groups had little inhibitory effect under our experimental conditions. We further 2 characterized the pathological damage to tumors through the Hematoxylin and eosin (H&E) and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) method (Fig. 7f). These results indicated that H-LM MoS treatment could induce apparent tumor histological X damage and cancer cell apoptosis. Notably, negligible adverse effects on mice were found during the whole therapy, as verified by the body weight analysis and histological analysis of key organs (Fig. 7b and Supplementary Fig. 41). All the results revealed that the designed H-LM MoS X nanozymes could attain highly efficient cascade catalysis, thereby presenting remarkable antitumor therapeutic effects with satisfactory biocompatibility.

In summary, we leverage liquid metal as a well-designed platform to screen high-performance nanozyme with excellent therapeutic effects. First, we have designed and synthesized a series of liquid metal-buffered nanozymes and screened the most favorable nanozyme to achieve efficient tumor catalytic therapy. In this strategy, the liquid metals possess inherent unique advantages: (i) serving as the building template of nanozymes, (ii) initiatingS phase engineering to generate an E amorphous nanozyme with abundant catalytic active sites, (iii) featuring electron-rich R microenvironment to affect electronic structure of naPnozymes, which are beneficial for regulating nanozyme catalytic activity. The amorphous LMN MoS nanozymes exhibit superior multienzyme- X I mimicking cascade catalytic activity, whic h can induce ROS burst and metabolic imbalance to E drive the efficient catalytic therapy Lof tumors with great biocompatibility. Overall, this work C provides insights into understanding the roles of tailored electronic structure and phase engineering I T for regulation of catalytic activity of nanozymes. Meanwhile, it may open prospects for future R biomedical applicationAs of liquid metals.

Methods
Ethical statement. All studies used healthy female Balb/c mice (Jilin University Laboratory Animal Center, China), which were 7-8 weeks old and weighed 18-20 g. All animal procedures were approved by the Ethics Committee of Science and Technology of Northeast Normal University (202502088). According to this protocol, the maximal tumor burden is limited to 2000 mm3. In our study, all tumor sizes were kept below 2000 mm³. Sex was not taken into consideration in the study design. Specific pathogen-free conditions were maintained for mouse housing: 26 ± 1 °C, 50 ± 5% humidity, with a 12 h light-dark cycle.

Cell lines. 4T1 cell line (Catalog no. CRL-2539, American TEype Culture Collection, ATCC), 293T cell line (HEK293T, GDC0067, the China Center for Type Culture Collection, CCTCC).

Preparations of LM MoS . Typically, 0.1 g liqNuid metal alloy was placed in 50 mL (NH ) MoS X 4 2 4 solution (20 mM). Subsequently, the mixtu re was sonicated with a tip ultrasonicator at 1000 W and 60% amplitude for 10 minutes. ThLe ultrasound was applied in 5-second pulses with a 3-second interval. An ice bath was used to keep the mixture temperature constant during ultrasonication. Following the reaction, the largest particles precipitated within seconds. To remove large particles, the remaining nanosphAeres were subjected to centrifugation (95 × g, 5 min). The product was centrifuged (13708 × g, 8 min) and washed repeatedly with Milli-Q water. To optimize the ultrasonication power, the reaction system was exposed to different power levels.

ICP-OES measurement. The sample (1 mg) was digested with 1 mL of aqua regia (HCl:HNO 3 =3:1, v/v). After standing at room temperature for 30 min, the mixture was heated (80 °C) until a clear solution was obtained. Elemental analysis was performed using ICP-OES (Agilent 5800).

Peroxidase-like Catalytic Activity Characterization and Kinetic measurements. The experiment was conducted in NaAc-HAc buffer solution (pH =4.0, 25 mM) with LM MoS (10 X µg/mL) and ABTS (2.5 mM) in the presence of H O (0.25, 0.5, 1, 2.5, 5, and 10 mM). The 2 2 catalytic activity was assessed by monitoring the absorbance of ABTS at 415 nm over time. According to the Michaelis–Menten model, =V [S]/(K + [S]), where  represent the initial max m velocity, V is the maximal reaction velocity, and [S] is substrate concentration. max

EPR Detection of •OH and O •−. For the EPR assay, •OH and O •− were detected using DMPO 2 2 as the trapping probe. In a typical process, samples were dispersed in HAc-NaAc buffer (pH =4.0, 25 mM) (solvent for •OH) and methanol (solvent for O •−), respectively. (10 mM H O , 100 µg/mL 2 2 2 LM MoS , 100 mM DMPO) X

Trapping experiments. To determine the active species generated by LM MoS nanozymes, IPA X (isopropanol alcohol, 10 μL) and PBQ (1,4-Benzoquinone,E saturated PBQ solution) were used as scavengers (10 μL) of •OH and O •−, respectively. 2

Anti-interference study. NaCl, KCl, CaCl , Na SO , NaNO and NaBr were employed to study 2 2 4 3 the interference of anions, such as Na+, K+, Ca2+ Cl-, SO 2-, NO - and Br-, and the concentration of 4 3 each anion was set at 10 mM. For Lprotein interference study, 10 μg/mL BSA (bovine serum albumin) was employed.

Assessment of Catalytic Stability of LM MoS Nanozymes. The nanozyme was dispersed in X PBS and stored at room temperature for a certain period of time, and its POD-like activity was measured at certain time. All catalytic stability experiments were evaluated in the NaAc-HAc buffer solution (pH 4.0, 25 mM), which contained LM MoS (10 µg/mL), ABTS (2.5 mM), and X H O (10 mM). 2 2

O •- generation. DPBF (1 mg/mL, 100 µL) solution was added to LM MoS solution (50 µg/mL, 2 X 600 µL). Absorbance changes were monitored with a UV–Vis spectrophotometer.

Detection of hydroxyl radical (•OH). •OH generation was probed by the fluorescence of TAOH (E =435 nm). Experiments were conducted in PBS (25 mM, pH 7.4) with the following mixtures: m TA (0.5 mM); TA and H O (1 mM); TA and nanozymes (50 μg/mL); only nanozymes; TA, 2 2 nanozymes and H O . After 12 h, samples were centrifuged and measured by fluorescence spectra. 2 2

In vitro GSH consumption. DTNB (2.0 mg/mL, 100 µL) and GSH (1×10−3 M, 200 µL) solutions were added into LM MoS solutions (1.7 mL) with varying concentrations (0, 20, 40, 60, 80, and X 100 μg/mL). After 6-h magnetic stirring (37 °C), the residual GSH was determined by absorbance spectroscopy.

Catalytic reactions of LM MoS with NADH. LM MoS solution (50 µg/mL, 2.5 mL) was X X incubated with NADH solution (50 µg/mL) under magnetic stirring at 25 °C. The amount of NADH was determined by the absorbance spectroscopy.

Detection of H O formation during NADH oxidation. NADH (800 μL, 5 mM) was incubated 2 2 with LM MoS solution (1 mL, 100 µg/mL) and reacted for 5 min. Then, the mixture was X transferred to a solution containing Amplex Red (40 μM) and HRP (10 U/mL). Absorbance changes were monitored with a UV–Vis spectrophotometer.

Evaluation of substrate selectivity of LM MoS . To study the selectivity of LM MoS towards X X NADH, uric acid (UA,A 100 μM), xanthine (XA, 100 μM), L-Tyrosine (100 μM) and NADPH (100 μM) were mixed with LM MoS (final concentration: 50 μg/mL), respectively. The uric acid X content was detected (λ 290 nm) by UV-vis spectroscopy. The xanthine content was monitored max (λ 270 nm) by UV-vis spectroscopy. To confirm the dityrosine formation (for L-Tyrosine), max fluorescence spectra were recorded (E =300 nm). NADPH content was measured at wavelength X 340 nm by UV-vis spectroscopy.

Computational Method. All DFT calculations employed the Vienna Ab initio Simulation Package (VASP 4.6) software. Amorphous models were obtained by annealing through ab initio molecular dynamics (AIMD) method. The simulations employed the canonical (NVT) ensemble with a Nosé-Hoover thermostat. The system was initially heated to and maintained at 2000 K for 10 ps using a 1 fs time step, and was finally quenched to 298.15 K. The generalized gradient approximation of Perdew-Burke-Ernzerhof (GGA-PBE) was used to describe the exchange- correlation potential. The ion cores–valence electrons were described using the projector augmented-wave (PAW) method. A plane-wave cutoff energy of 450 eV was used. The structural relaxation was performed with convergence thresholds set to -0.02 eV/Å for the Hellmann– Feynman forces and 10-5 eV for the energy change. Charge density analysis was obtained from CHGCAR, and all atomic/electronic structures were illustrated with VESTA software.

Definition of d-Band Center. The d-band center is defined as the energy center of the d-orbital electron density of states (DOS) for transition metal atoms, which is the weighted average energy value of all electronic states in the d-orbital according to their DOS. The algorithm is as follows: ∫𝜀 ∙𝑛 (P𝜀)𝑑𝜀 d 𝜀 = d ∫𝑛 (𝜀)𝑑𝜀 Nd 𝑛 (𝜀) is the d-orbital electron density of sItates, 𝜀 is the energy of electronic states, and the d integration range covers all electronic states of the d orbital (usually from the binding state below the Fermi level to the antibonding Cstate above).

Cytotoxicity assay. 4T1 cRells or 293T cells were plated in a 96-well plate (1×104 cells/well) and cultured for 24 h, followed by a 24 h treatment with varying concentrations of LM NPs, MoS and 2 LM MoS X (0, 25, 50, 75, and 100 µg/mL). The MTT solution (0.5 mg/mL) was incubated with cells for 4 h at 37 °C and then replaced with 100 µL of DMSO to dissolve the formazan product. Subsequently, cell viability was analyzed using a microplate reader.

Cellular uptake. 4T1 cells were treated with LM MoS @RB (40 μg/mL). Afterward, cells were X incubated with Hoechst (5 µg/mL) for 15 minutes and visualized using CLSM.

Determination of ROS generation in vitro. 4T1 cells were treated with LM NPs, MoS , LM 2 MoS (50 μg/mL) for 24 h. Afterward, the cells were incubated with DCFH-DA (10 μM, 30 min), X followed by Hoechst staining. The cells were imaged using CLSM.

Detection of intracellular GSH/ NADH level. 4T1 cells were seeded in 6-well plates at 5×105 cells per well and cultured for 24 h, followed by a 12 h treatment with LM MoS X at concentrations of 0, 25, 50, 75, and 100 µg/mL. The corresponding GSH contents were quantified using Ellman’s reagent, and the corresponding NADH contents were quantified using NAD+/NADH assay kit.

Live/Dead assay. 4T1 cells were planted in 6-well plate (5×105 cells/well) and incubated for 24 h. 4T1 cells were incubated with various nanoparticles (50 μg/mL) for 24 h. The cells were co-stained with calcein AM (5 µg/mL) and PI (10 µg/mL) for 30 minutes prior to visualization by CLSM.

Synthesis of H-LM MoS . The as-prepare LM MoS X X suspension (2 mg/mL) was mixed with hyaluronic acid (0.1 mg/mL) and stirred at room temperature for 4 h in the dark, followed by harvest and washing with Milli-Q water. H-LM, and H-MoS were obtained using the same method. 2

The biocompatibility of the H-LM MoS in mice. Mice were randomly assigned to two groups, X Control (PBS), H-LM MoS (10 mg/kg, intravenous injection). Body weight was monitored every X 4 days for 28 days following injIection. After 28 days, mice were euthanized for blood collection (for complete blood panelR analysis and serum biochemistry) and organ harvesting. The harvested organs were subjected to H&E staining for microscopic examination.

In vivo pharmacokinetic of H-LM MoS . H-LM MoS @Cy3 (10 mg/kg) was intravenously X X administered to healthy mice. At various time points post-injection (0.25, 0.5, 1, 2, 4, 8, 12, and 24 h), mice were anesthetized for blood collection. The collected blood was centrifuged (95 × g, 10 min), and then detected by a fluorescence spectrophotometer.

In vivo anti-tumor study. When tumor volume reached approximately 50 mm³ following subcutaneous inoculation of 4T1 cells, mice were randomly allocated into four experimental groups (n=4) for intravenous administration of the respective treatments. PBS (I), H-LM (II), H- MoS (III), H-LM MoS (10 mg/kg) (IV). Body weight and tumor volume (width2×length/2) were 2 X monitored. At the study endpoint, mice were euthanized. Following H&E or TUNEL staining, the harvested tumors and major organs were examined under an optical microscope.

In vivo biodistribution of H-LM MoS and LM MoS after injection. 4T1 tumor-bearing mice X X received intravenous injections of the various formulations (10 mg/kg, with equal Mo). At the 12 h time point, organs and tumors were harvested and weighed. The samples were digested with 1 mL of aqua regia (HCl:HNO =3:1, v/v) under heat treatment (80 °C) for 3 h, then cooled and left 3 overnight at room temperature. Subsequently, the digested mixtures were centrifuged (595 × g, 10 min) and filtered. The Mo content was quantified by ICP-MS.

Statistical analysis. All data were expressed in this articlEe as mean result ± standard deviation (SD). Statistical analysis was conducted using OriginPro 2020, GraphPad Prism 8, and Microsoft Excel software. Statistical evaluation was performe d using the Student’s t-test analysis.

Reporting Summary. Further informatiEon on research design is available in the Nature Research Reporting Summary linked to thisC article.

Peer review rounds
[{"round": 1, "reviewer_comments": {"reviewer_1": "This manuscript reports the design of LM-buffered amorphous MoSx nanozymes using Ga-In alloys as templates. The\nauthors synthesize a series of Ga1-x Inx MoSx nanozymes and identify Ga75.5 In24.5 MoSx as exhibiting superior POD-like\nactivity compared to crystalline MoS2. They attribute this enhancement to LM’s roles in templating, inducing amorphization,\nand providing an electron-rich microenvironment. The optimized nanozyme demonstrates multi-enzyme activities (POD-,\nOXD-, and NOX-like), enabling cascade reactions that disrupt redox/metabolic homeostasis in tumor cells. In vitro and in\nvivo studies show efficient ROS generation, GSH/NADH depletion, mitochondrial damage, and tumor growth inhibition. DFT\ncalculations suggest electron transfer from LM optimizes the electronic structure for catalysis. The work applies established\nmethodologies without demonstrating a significant conceptual or practical leap beyond existing literature. The absence of\ncomparative analysis with relevant systems and recurring presentation issues further diminishes its suitability for Nature\nCommunications. Should the authors address these gaps (e.g., by rigorously benchmarking against competitors and\nclarifying the innovation boundary), this study may find a better fit in a specialized journal. Therefore, I recommend rejection\nin its current form.\nMajor Weaknesses Reinforcing Rejection:\n1. Combines two well-explored materials (LM + MoS2) without novel design principles or mechanistic insights. The concept\nof using LM (Ga-In alloys) for catalysis is well-established (e.g., Refs 23–30, 62–63), as is MoS2-based nanozyme design\n(Ref 55). The work combines these existing strategies without introducing a transformative mechanism, material, or\napplication.\n2. Claims of LM’s unique roles lack substantial evidence, except for citing prior literatures\n3. Catalytic performance lacks comparison to state-of-the-art LM-based catalysts or high-activity MoS2-based nanozymes.\nAssertions of \"superior activity\" remain unsupported without context.\n4. No optimization data for critical parameters (e.g., sonication power/temperature effects).\nOther concerns:\n1. Explain POD, ABTS, ……\n2. Do not use “liquid metal” and “LM” casually.\n3. In Fig. 2D, what is the meaning of the curve named control?\n4. “Among them, Ga75.5In24.5 MoSX presented the highest catalytic activity”. Why? The explanation below is insufficient.\n5. The LM MoSx model in Fig. 5 is too coarse. MoSx is an amorphous structure and the LM is wrapped by it, as shown in\nFig. 2, which is almost irrelevant with the model in Fig. 5.\n6. “the amorphous MoSx exhibited an increased electron the amorphous exhibited an increased electron”, but I can only see\nan opposite trend. Also, “revealing the electron donation from LM to Mo atoms” is questionable.\n7. What’s the meaning of “d-band center” and what’s the algorithm to determine its center?", "reviewer_2": "In this manuscript, Zhang et al. designed a series of liquid metal-buffered nanozymes and screened the most favorable\nnanozyme to achieve efficient tumor catalytic therapy. The findings highlight the roles of tailored electronic structure and\nphase engineering for regulating the catalytic activity of nanozymes. Authors demonstrate that this is the first report of\nscreening and regulating the catalytic and therapeutic effects of LM nanozymes to achieve efficient tumor therapy. The\noverall work is well-designed, and the application of LM in catalysis and the biomedical field is interesting. However, at the\ncurrent stage, the characterization of the structure-activity relationship and the anti-cancer mechanism of LM-based\nnanozyme is not solid. Also, other issues need to be addressed.\n1)The most significant concern of utilizing LM-based nanozymes for catalytic cancer therapy lies in the intrinsic property of\nLM. As is known, amorphous LM nanoparticles could gradually transform into crystalline material, especially in aqueous\nsolution. Due to such a dynamic process, the composition of LM-based nanozyme could hardly be maintained during in vitro\nand in vivo experiments or characterizations. Also, it is necessary to ensure that the MoSx content in a series of LM-buffered\nnanozymes is the same. Or else, the differences in nanozyme activity could also be influenced by different MoSx contents.\n2)In a recently published paper (Small 2025, 2502752), FeOx was used to hybridize LM to generate nanozymes. Therefore,\nthe reason for choosing molybdenum sulfide should be clarified to distinguish the innovation of this work from previous\nworks, and the advantages of molybdenum sulfide should be explained as well.\n3)It was demonstrated that Ga75.5In24.5 (commercial Gallium-Indium eutectic) MoSX performed the best catalytic activity,\nbut the possible reason was not given. With subtle variations of the compositions, the melting points and the fluidities of\nresultant LM-MoSX nanozymes would alter significantly. In this context, the discussion of the catalytic activity of LM-MoSX\nnanozymes should be careful. Additionally, the authors did not provide detailed synthesis methods and characterization\ntechniques. It is suggested to supplement relevant details.\n4)Although decorating with HA, it remains challenging for LM-MoSX nanozyme (ca. 300 nm) to achieve efficient cellular\nuptake and high accumulation in the stiff 4T1 tumor via intravenous injection (Figure S35). And what is the half-life of LM-\nMoSX nanozyme?\n5)In Figure S2B, D, the In 3d curves were poorly fitted.\n6)In Figure S20, the curve shape of EPR result of O2•− seems abnormal. Please confirm the result and the discussion.\n7)In Figure S27, why did the cellular fluorescence of DAPI present large deviations at variable time points?\n8)The changes in particle size and potential of LM MoSx should be measured after the modification of HA.\n9)In Figure S33, there is a difference in H&E staining of the lung between the H-LM MoSx group and the control group. Were\nthe images captured at the same magnification?\n10)In Figure S35, quantitative data on in vivo biodistribution should be supplemented.\n11)The stability test of the LM MoSx nanozymes should be provided.\n12)In the article, POD-like activity was used to screen for suitable ratios of GaIn. Why not use other nanozyme activities for\nscreening?\n13)In Figure 6J, the live and dead cells co-staining was not well performed, and the cell morphology could hardly be\ndistinguished.Please provide fluorescence images under high magnification.\n14)The grammar and language of the manuscript should be carefully polished.", "reviewer_3": "The authors present a comprehensive and well-executed study on liquid metal-buffered MoSₓ nanozymes, systematically\nsynthesized using Ga–In alloys to optimize catalytic performance. The Ga₇₅.₅In₂₄.₅-based nanozyme exhibits enhanced\nperoxidase-, oxidase-, and NADH oxidase-like activities, enabling a catalytic cascade that disrupts redox homeostasis in\ncancer cells and induces selective cytotoxicity both in vitro and in vivo. The combination of multi-enzyme functionality,\neffective tumor targeting, and minimal toxicity underscores the therapeutic potential of this platform. The study’s originality\nlies in leveraging liquid metal templating to precisely control nanozyme composition, structure, and electronic properties,\nrepresenting a clear advancement over previous LM-based nanozyme approaches. The conclusions are well-supported by\ncomprehensive materials characterization, catalytic assays, theoretical modeling, and biological validation. The\nexperimental methodology is appropriate and rigorously executed. Overall, this work constitutes a significant contribution to\nnanozyme research and catalytic cancer therapy.\nTo further enhance the manuscript, several clarifications and suggestions are\nproposed as detailed in the following comments (also provided in the attached file).\n1. Particle Size and Delivery: A potential concern is the relatively large hydrodynamic size of the nanozyme (~300 nm\ndiameter). Particles of this size may have limited tumor penetration and faster clearance by the mononuclear\nphagocyte system. The in vivo success after HA coating suggests good tumor accumulation capability, but the authors might comment\non how the size could influence distribution. Could smaller LM-templated nanozymes be produced (e.g., by adjusting\nsonication conditions) to further improve tissue penetration?\n2. Comparison to Other Approaches: The introduction could better distinguish this work from prior related studies. Although\nthe manuscript cites previous reports on LM-based nanozymes (Wu et al., 2023; Xu et al., 2024), it does not sufficiently\narticulate how the present study advances beyond these earlier efforts. It would be valuable for the authors to explicitly\nhighlight how their approach advances beyond previous reports, particularly in terms of the systematic composition tuning\nand realization of a multi-enzyme cascade function with therapeutic efficacy. This clarification would better establish the\noriginality and significance of the current work.\n3. Mechanistic Detail – Role of Indium vs Gallium: The study identifies an optimal Ga/In ratio; however, the mechanistic\nreasoning for why both excessive and insufficient indium content are detrimental could be further elaborated. The authors\nrefer to prior work on charge distribution within Ga–In alloys (ref 67), indicating that indium incorporation increases structural\ndisorder and enhances electron density around gallium. They conclude that lower or higher indium levels compromise\ncatalytic activity. This is a key insight and perhaps a more comprehensive discussion here would strengthen the\nunderstanding of how composition ties to structure and catalytic activity.\n4. Undefined Terms and Abbreviations: Several key terms and abbreviations that are central to the manuscript are not\ndefined upon first use:\n- POD-like activity: The manuscript repeatedly refers to \"POD-like activity,\" but fails to define \"POD.\"\n- ABTS: Used as a substrate in peroxidase-mimicking activity assays but not defined.\n- ROS: The term \"ROS\" is frequently used but never defined.\n- NADH: Although common in biochemical studies, NADH is not defined.\n- The MTT method is mentioned as the method for cytotoxicity evaluation, but the manuscript does not define MTT.\n5. Missing Figure Legends in Fig. 4C and Fig. 4F: Figures 4C (DPBF assay for O₂•⁻ detection) and 4F (NADH oxidation\nspectra) lack in-figure legends. The data series in the plots are not labeled. The authors should include proper legends\ndirectly in the figure.", "reviewer_4": "This manuscript outlined the synthesis, characterisation and application of novel amorphous liquid Ga/MoSx structures. The\nauthors show that these materials, coined as nanozymes, showcase some impressive catalytic activity, and can be used in\nbiological contexts where their redox activity may confer therapeutic effects (e.g. for tumour cells). Overall, I am highly\nimpressed by the work contained in this manuscript. I note that the uncovering of cascade catalysis that relies on the liquid\nmetal system is a particularly interesting finding. Therefore, I would consider the work of interest to the readership, and\nof good quality for Nat. Commun. However, I believe there are several areas requiring clarification or amendment before I can\nrecommend the manuscript for publication.\nMajor comments:\n1. The manuscript could benefit from clarifying the physical state of the LM/MoSx systems, especially during catalysis\ntesting. The LMs were presumably melted and in liquid form during synthesis of the LM/MoSx structures, but can they still be\nconsidered to be in the liquid state during application? This is perhaps especially relevant in the biological context, where\ntemperature ranges are limited and must be carefully controlled.\n2. The authors discuss the dynamic nature of liquid metals as key to enhancing their catalytic activity in the introduction, but\nlater suggest the activity of their LM/MoSx systems may be due to an abundance of active sites and interesting electronic\ncharacters in the amorphous structure. Several groups are now releasing atomistic modelling suggesting that the dynamic\nmotion of liquid metal systems still plays a key role as well:\na. https://doi.org/10.1038/s41467-025-56222-0\nb. https://doi.org/10.1039/D3SC04416E\nc. https://doi.org/10.1002/ange.202407124\nThe manuscript would benefit from addressing this discussion point in the context of their results. In particular, perhaps the\nongoing dynamics of a liquid metal based system may have links to true enzymatic character?\n3. I strongly suggest the authors move the computational details of the density functional theory calculations to the main text\n(i.e. in the methods). A reader should ideally be able to find the software used and exchange-correlation functional in the\nmain text.\n4. More detail should be included on how the MoSx and LM/MoSx structural models were developed/selected for DFT. Did\nthe authors perform at-temperature molecular dynamics simulations? Were these structures adapted from other work? As is,\nit is impossible to identify whether these models are representative or accurate for the system they are trying to describe. This is\nespecially true of the LM system, where a single snapshot is often not sufficient.\n5. Can the authors clarify whether HSE06 was used for all DOS calculations as well as determination of the band centres?\nWas it also used for the charge density analyses? This needs to be clearly stated in the methods or results.\n6. It was unclear to me exactly which quantity is plotted in Figure 5A on the charge density analyses. Does this plot contain\nthe simple charge density obtained form the CHGCAR in VASP? Or is this a charge density difference relative to some\nreference state (e.g. MoSx and LM separate)? I would suggest the later analysis is more meaningful for showing charge\naccumulation (e.g. at Mo atoms, as the authors identify in the LM/MoSx system).\n7. The DOS plots in 5B should likely be enlarged to make them more clear. It is difficult to read detailed information from\nthem.\n8. The analysis of a d-band centre in Mo is interesting. I recommend the following paper for its interesting discussion on how\nit is the gap between Mo and S states in MoS2 that determines redox activity for hydrogen evolution, rather than the d-band\ncentre: https://doi.org/10.1002/anie.202003091. Could the authors comment on whether they think a similar analysis would\nbe of benefit here?\nMinor corrections:\n1. In the introduction the authors begin to use abbreviations without defining them in the text. (e.g. POD-like activities, LM,\nOXD- and NOX-like are defined in the abstract but not the main text). These should be defined in their first mention in the\nmain text.\n2. Should have \"an\" added in two places in the following sentence: \"Here we design and screen the liquid metals-buffered\nnanozymes, in which the liquid metals can not only initiate phase engineering to generate AN amorphous nanozyme with\nabundant catalytic active sites, but also afford AN electron-rich microenvironment to optimize the electronic structure of\nnanozymes for improving the catalytic performance (Fig. 1). \"\n3. \"... free delocalized electrons cloud\" should likely read \"... free delocalized electron clouds\""}, "author_response": "Next is the summarization of our point-by-point response to the comments raised by\nthe reviewers.\nReviewer #1 (Remarks to the Author):\nThis manuscript reports the design of LM-buffered amorphous MoSx\nnanozymes using Ga-In alloys as templates. The authors synthesize a series\nof Ga1-x Inx MoSx nanozymes and identify Ga75.5 In24.5 MoSx as exhibiting\nsuperior POD-like activity compared to crystalline MoS2. They attribute this\nenhancement to LM’s roles in templating, inducing amorphization, and\nproviding an electron-rich microenvironment. The optimized nanozyme\ndemonstrates multi-enzyme activities (POD-, OXD-, and NOX-like), enabling\ncascade reactions that disrupt redox/metabolic homeostasis in tumor cells. In\nvitro and in vivo studies show efficient ROS generation, GSH/NADH depletion,\nmitochondrial damage, and tumor growth inhibition. DFT calculations suggest\nelectron transfer from LM optimizes the electronic structure for catalysis. The\nwork applies established methodologies without demonstrating a significant\nconceptual or practical leap beyond existing literature. The absence of\ncomparative analysis with relevant systems and recurring presentation issues\nfurther diminishes its suitability for Nature Communications. Should the\nauthors address these gaps (e.g., by rigorously benchmarking against\ncompetitors and clarifying the innovation boundary), this study may find a\nbetter fit in a specialized journal. Therefore, I recommend rejection\nin its current form.\nMajor Weaknesses Reinforcing Rejection:\n1. Combines two well-explored materials (LM + MoS2) without novel design\nprinciples or mechanistic insights. The concept of using LM (Ga-In alloys) for\ncatalysis is well-established (e.g., Refs 23–30, 62–63), as is MoS2-based\nnanozyme design (Ref 55). The work combines these existing strategies\nwithout introducing a transformative mechanism, material, or application.\nResponse: We are grateful for the time and effort the reviewer has spent in\nreviewing our manuscript. The review comments are very helpful for further\nstrengthening the manuscript.\nAs for the previous works mentioned by the reviewer, references 23-30\nfocused on the catalytic performance of liquid metal on propane\ndehydrogenation or electrocatalytic reaction. References 62 reported on\nengineering liquid metal-based nanozyme for enhancing microwave dynamic\ntherapy, and their study focused on solving the problem of poor delivery of\nmicrowave sensitizer and the obstacle of tumor hypoxic microenvironment in\nmicrowave dynamic therapy (MDT). However, their nanozyme catalytic activity\nwas attributed to the Co2+ center in ZIF-67 rather than the liquid metal.\nReferences 63 reported the galvanic replacement synthesis of\nVOx@EGaIn−PEG for high-performance biosensors, and their study focused\non solving the limitations in biocompatibility, regenerability and recyclability of\nb of\nbiosensors. References 55 reported that positive and negative charges,\nproduced on the surface of piezoelectric material under ultrasound (US), could\nenhance the POD-like activity of MoS . These studies laid the key foundation\n2\nfor our work, which revealed the great potential of liquid metal for nanozyme\ncatalysis. However, these studies are mainly focus on the various application\nof LM-based catalysts, while overlooking the potential of liquid metal with\nintrinsic amorphous property and unusual electronic microenvironment in\nregulating nanozyme catalytic activity.\nOur work highlighted the catalytic potential of liquid metals for screening and\nregulation of nanozyme activity. We have not only investigated the catalytic\nactivity of liquid metal-buffered nanozyme with different gallium indium ratios,\nbut also explored the role of liquid metal in regulating catalytic performance\nand the reasons for the enhanced enzyme-like catalytic activity of liquid\nmetal-buffered nanozyme. In particular, we have performed the density\nfunctional theory (DFT) calculations to investigate the electronic structure of\nLM-based nanozymes in the manuscript (Fig. 5). The calculation results\nrevealed that, compared to MoS , the LM MoS model not only achieved the\nincreased electron density of Mo, but also appeared a significantly higher DOS\nnear the Fermi level and several new hybrid electronic states. Moreover, the\nLM MoS nanozymes exhibited an upward shift d band center position, which\nX\nwas significantly nearer to the Fermi level than MoS . These changes\n2\ncontributed to the electron transfer and the interaction between the adsorbed\nsubstrate and LM MoS nanozyme, thereby favoring the catalytic reaction. In\nX\naddition, our designed LM-based nanozyme exhibited multienzyme mimetic\nactivity, i.e., POD-like activity, OXD-like activity, NOX-like activity, which could\nwork together to achieve multiple-enzyme-like cascade catalytic reactions to\ndisrupt intracellular redox and metabolic homeostasis. Our work provides not\nonly the first example of leveraging LM as a well-designed platform to screen\nhigh-performance nanozyme with excellent therapeutic effects, but also\nhighlights the roles of tailored electronic structure and phase engineering for\nregulation of catalytic activity of nanozymes.\nIn the revision, we have performed additional experiments and provided more\ndata to revise the manuscript, which further strengthens our catalyst design\nand the understanding of the role of liquid metal in nanozyme catalysis.\nHopefully, it is more rigorous and integrated than the original version.\nIn the revised manuscript, we emphasized in the introduction section:\n“However, the potential of liquid metal in regulating nanozyme catalytic activity\nhas rarely been explored mechanistically. In particular, previous research\nefforts have focused on the application of liquid metal-based nanozyme while\noverlooking the catalytic potential of liquid metals for screening and regulation\nof nanozyme activity.”\nFig. 5. DFT Calculations. a The computational surface models and charge density maps\nof MoS 2 , MoS X and LM MoS X . b The differential charge density analysis at the Mo and Ga\ninterfaces of LM MoS . The yellow area indicates electron gain and the blue area\nX\nindicates electron loss. c Calculated DOS of the MoS 2 , MoS X and LM MoS X models. d\nCalculated Mo 3d DOS of the MoS , MoS and LM MoS models.\n2. Claims of LM’s unique roles lack substantial evidence, except for citing prior\nliteratures.\nResponse: Thanks for the reviewer’s important question. Liquid metals have\ndemonstrated their potential in the catalysis field in recent years, as the\nprevious literatures reported. However, the potential of liquid metal in\nregulating nanozyme catalytic activity has rarely been explored. In our work,\nthe main contribution is focused on revealing the catalytic potential of liquid\nmetals for screening and regulation of nanozyme activity. Liquid metals can not\nonly initiate phase engineering to generate amorphous nanozyme with\nabundant catalytic active sites, but also afford electron-rich microenvironment\nto optimize the electronic structure of nanozymes for improving the catalytic\nperformance. Specifically, we systematically studied the amorphous\ncharacteristics of liquid metal-buffered nanozyme through TEM, XRD, and\nSAED (refer to Fig. 3). More importantly, to gain insights into the reasons for\nthe enhanced enzyme-like catalytic activity of liquid metal-buffered nanozyme,\ndensity functional theory (DFT) calculations were conducted based on first\nprinciples to investigate their electronic structures (refer to Fig. 5). Based on\nthe experimental and calculation results, we unveiled that the pivotal role of\nliquid metal, i.e., amorphization, and electron transfer, endowing LM MoS\nX\nnanozymes with optimized enzyme-like catalytic activity. In our designed\nsystem, on the one hand, liquid metals ingeniously endowed the nanozymes\nwith amorphous property, thus providing abundant active sites for catalysis. On\nthe other hand, the liquid metals matrix and the amorphous character of\nnanozymes led to a favorable electronic perturbation, which could promote\nelectron transfer and optimize the reaction process, thereby boosting the\ncatalytic behavior. These data indicated the unique roles of liquid metal for\nnanozyme catalysis.\nIn the manuscript, we stated in the DFT Calculations section:\n“We confirmed that the pivotal role of liquid metal, i.e., amorphization, and\nelectron transfer, endowing LM MoS nanozymes with optimized enzyme-like\nX\ncatalytic activity. In our designed system, on the one hand, liquid metals\ningeniously endowed the nanozymes with amorphous property, thus providing\nabundant active sites for catalysis. On the other hand, the liquid metals matrix\nand the amorphous character of nanozymes led to a favorable electronic\nperturbation, which could promote electron transfer and optimize the reaction\nprocess, thereby boosting the catalytic behavior.”\nFig. 3. Characterization of LM MoS X nanozymes. a TEM image of LM MoS X\nnanozymes. b HAADF image and elemental mapping of LM MoS X nanozymes. c Size\ndistribution result of LM MoS X . d SAED pattern of LM MoS X nanozymes. e HR-TEM image\nof LM MoS\nX\nnanozymes and (f) the corresponding 3D pseudo-color surface plot of the\nselected domain. g XRD spectra of LM MoS X and MoS 2 . h XPS spectra of LM MoS X . i The\nMo 3d and (j) S 2p XPS patterns of LM MoS X . k The Mo 3d XPS analysis of LM MoS X and\nMoS 2 . l The ESR spectra of LM MoS X and MoS 2 .\nFig. 5. DFT Calculations. a The computational surface models and charge density maps\nof MoS 2 , MoS X and LM MoS X . b The differential charge density analysis at the Mo and Ga\ninterfaces of LM MoS . The yellow area indicates electron gain and the blue area\nX\nindicates electron loss. c Calculated DOS of the MoS 2 , MoS X and LM MoS X models. d\nCalculated Mo 3d DOS of the MoS , MoS and LM MoS models.\n3. Catalytic performance lacks comparison to state-of-the-art LM-based\ncatalysts or high-activity MoS2-based nanozymes. Assertions of \"superior\nactivity\" remain unsupported without context.\nResponse: Thanks for the helpful comments. Currently, the state-of-the-art\nLM-based catalysts are mainly applied in hydrocarbon dehydrogenation\nreactions or electrocatalytic reactions (Nat. Catal. 2023, 6, 1131, Nat.\nNanotechnol. 2024, 19, 306). Given the different types of reactions, they are\nnot suitable for comparison here. We strongly agree with the reviewer’s\nopinion that the catalytic performance of nanozymes needs comparison. As\nthe catalytic performance of nanozymes is largely determined by their\ncomponent elements, so we collected catalytic performance of different\nmolybdenum-based nanozymes and make the comparison in Supplementary\nTable 3. The catalytic performance of the nanozymes is dependent on their\nkinetics (Nat. Cata. 2021, 4, 407). So, the steady-state kinetic parameters\nincluding the Michaelis–Menten constant (K ) and the maximum initial velocity\nm\n(V ) of LM MoS with H O as the substrate were measured in the\nmax X 2 2\nmanuscript. The reduced Km value indicated that the LM MoS had a higher\nX\naffinity to H O , and the increased V value reflected its superior catalytic\n2 2 max\nactivity. The apparent K value and V value of LM MoS with H O as the\nm max X 2 2\nsubstrate were both better than those of MoS , natural horseradish peroxide\n2\nand many other reported molybdenum-based nanozymes.\nSupplementary Table 3. The kinetic parameters of LM MoS X and MoS 2\nnanozymes and reported nanozymes for POD-like catalysis.\nSubstrate\nNanozymes H 2 O 2 Reference\nK (mM) V (10-8 M/s)\nm max\nLM MoS 2.033 17.04 This work\nX\nMoS 8.805 4.439 This work\nH-Mo-CDs 2.65 3.27 [3]\nMo N NPs 12.8 10.4\n[4]\nMo-R2 163 2.38\nMo-R3 181 2.40\nNH-MoO 2.41 13 [5]\n3-x\nMoS /CoS 20.50 8.928 [6]\n2 2\nPEG-MoS 2.812 8.01 [7]\n3 4\nHRP 3.7 8.71 [10]\n*References can be found in the supplementary information\n4. No optimization data for critical parameters (e.g., sonication\npower/temperature effects).\nResponse: Thanks for your professional comments. We have complemented\nthe experiment on the influence of sonication power on the synthesized\nmaterials. These different LM MoS nanoparticles were obtained by sonicating\nX\nGa In bulk in 20 mM (NH ) MoS solution at various power of\n75.5 24.5 4 2 4\nultrasonication. The SEM images and particle size distribution results were\nshown in Supplementary Fig. 1. It was found that increasing the applied\npower of ultrasonication from 50 W to 500 W results in a decrease in the\noverall size of the LM MoS nanostructures from micrometer to hundreds of\nX\nnanometers. The size distribution also becomes narrower with higher\nsonication intensity applied. The enhancing oscillating shear force from high\npower sonication is regarded as the main factor to separate small droplets\nfrom the bulk liquid metal in a short time. As for the temperature effects, the\ntemperature of the mixture during ultrasonication was controlled by using an\nice bath. We have supplemented these experiment results and description in\nthe revised manuscript. Thanks again for our manuscript.\nIn the revised manuscript, we added:\n“The size distribution of nanoparticles became narrower with higher sonication\nintensity applied (Supplementary Fig. 1).”\nIn the methods section, we added:\n“An ice bath was used to keep the mixture temperature constant during\nultrasonication.”\n“To optimize the ultrasonication power, the reaction system was exposed to\ndifferent power levels.”\nSupplementary Figure 1. SEM images and particle size distributions of LM MoS X\nobtained by sonicating Ga In bulk in 20 mM (NH ) MoS solution at various power of\n75.5 24.5 4 2 4\nultrasonication: (a) 50 W, (b) 100 W, (c) 200 W, (d) 300 W, (e) 400 W, (f) 500 W. It was\nfound that increasing the applied power of ultrasonication from 50 W to 500 W results in a\ndecrease in the overall size of the LM MoS nanostructures from micrometer to hundreds\nX\nof nanometers. The size distribution also becomes narrower with higher sonication\nintensity applied. The enhancing oscillating shear force from high power sonication is\nregarded as the main factor to separate small droplets from the bulk liquid metal in a short\ntime.\nOther concerns:\n1. Explain POD, ABTS, ……\nResponse: Thanks for your careful reading and professional advice. We have\ndefined these abbreviations in the revised manuscript, and reviewed the entire\nmanuscript to avoid similar issues.\nIn the revised manuscript, we modified:\n“We observe that these nanozymes exhibit peroxidase (POD)-like activities.”\n“We explored the POD-like activity of five nanozymes by monitoring the\noxidation of 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) at\n415 nm.”\n2. Do not use “liquid metal” and “LM” casually.\nResponse: Thanks for your important suggestions. We have paid particular\nattention to this question, and amended the expression in the revised\nmanuscript. The abbreviation LM is used only for the name of nanozyme (LM\nMoS ), and the full name of liquid metal is used elsewhere.\nX\n3. In Fig. 2D, what is the meaning of the curve named control?\nResponse: Thanks for your valuable questions. The control group refers to\nABTS+H O , and the reaction condition are the same as other groups except\n2 2\nnanozyme. To have a better understanding, the relevant explanation has been\nsupplemented in the figure caption.\nFig. 2. (d) The POD-like activity of five liquid metal-buffered MoS X nanozymes, and the\ncontrol group refers to ABTS+H O .\n2 2\n4. “Among them, Ga75.5In24.5 MoS presented the highest catalytic activity”.\nX\nWhy? The explanation below is insufficient.\nResponse: Thanks for your valuable questions. In order to eliminate\nreviewer’s concern, we have added a comprehensive discussion to further\nstrengthen the understanding of composition and catalytic activity. Yu and\ncoworkers investigated the effects of alloying Ga with In on the atomic\nstructure and electronic transport properties of liquid EGaIn using the AIMD\nsimulations (J. Chem. Phys. 2014, 140, 064303). They found that with In\nalloying of Ga, their larger atomic volume leads to the disordered structure as\nwell as weak inter-atomic interactions by expanding the cell volume and\nbroadening their inter-atomic distance. Moreover, the altered free-electron\ndensity is higher between Ga atoms, while the charge density near the In\natoms is significantly smaller. This feature of the In atoms explain the dominant\nrole of In in reducing the electrical transport property and electrical conductivity.\nAs mentioned in the introduction section, the catalytic active sites and local\nelectronic characteristics determine their catalytic performance to a large\nextent. According to previous studies, the higher electrical conductivity favors\nefficient electron transport, which is beneficial to improve the catalytic\nperformance of nanozyme (Nat. Commun. 2022, 13, 5365; Adv. Mater. 2025,\n37, 2502804). Meanwhile, amorphous materials possess distinctive disordered\natomic arrangement that enables them with abundant active sites and\noptimized electronic configurations, which are beneficial for enhancing their\ncatalytic properties (Nat. Commun. 2019, 10, 4855; Chem. Rev. 2023, 123,\n13489). Thereafter, we mentioned that “the alloying of Ga with In results in a\nmore disordered structure, and the charge density is higher between Ga atoms\nwhile significantly smaller in the vicinity of In atoms” in the manuscript. To\nfurther strengthen the understanding of composition and catalytic activity, we\nsupplemented that the Indium content in GaIn alloys significantly affects the\ndisordered structure and the charge density distribution of alloys, thus\ninfluencing their amorphous structure and electrical conductivity. Specifically,\nthe alloying of Ga with In causes more disordered structure. This amorphous\nstructure is beneficial for the high catalytic performance of catalysts. However,\nthe electrical conductivity of GaIn alloys decreases with In alloying of Ga\nbecause the charge density near the In atoms is significantly smaller, which is\nunfavorable to the electron transfer and the subsequent catalytic process.\nThus, the catalytic activity of the resulting nanozymes could show a trend of\ninitially increasing and then decreasing with increasing In content in the GaIn\nalloy. We sincerely thank the reviewer for the insightful and constructive\nsuggestion, and we have added the corresponding discussion into the revised\nmanuscript.\nIn the revised manuscripts, we added:\n“The Indium content in GaIn alloys significantly affects the disordered structure\nand the charge density distribution of alloys, thus influencing their amorphous\nstructure and electrical conductivity. Specifically, the alloying of Ga with In\ncauses more disordered structure. This amorphous structure is beneficial for\nthe high catalytic performance of catalysts. However, the electrical conductivity\nof GaIn alloys decreases with In alloying of Ga because the charge density\nnear the In atoms is significantly smaller, which is unfavorable to the electron\ntransfer and the subsequent catalytic process of catalysts. Thus, the catalytic\nactivity of the resulting nanozymes shows a trend of initially increasing and\nthen decreasing with increasing In content in the GaIn alloy.”\n5. The LM MoSx model in Fig. 5 is too coarse. MoSx is an amorphous\nstructure and the LM is wrapped by it, as shown in Fig. 2, which is almost\nirrelevant with the model in Fig. 5.\nResponse: Thanks for your professional and careful comments. Typically, it is\nchallenging to achieve perfect consistency between simulated and actual\nstructures. The TEM image in Fig. 2 was observed at the macroscopic scale.\nHowever, the LM MoS X model in Fig. 5 was simulated at the microscopic scale,\nwhich was closer to planar configuration, unless they represented small-sized\ncluster structures. We fully understand the reviewer’s concerns. Actually, we\nopted for this simplified model primarily due to considerations of computational\nfeasibility. Accurately simulating large-scale systems with complex\nthree-dimensional morphologies via density functional theory (DFT) remains\nextremely challenging. Although simplified, our model is constructed based on\nkey experimental characterizations. Moreover, in line with common practice in\nmany computational studies (Nat Commun 2024, 15, 6888, Angew. Chem. Int.\nEd. 2024, 63, e202405641), we constructed a planar periodic model\nrepresenting the crystal facet, which is the most stable and has the largest\nexposed surface area in the materials. We hope that these explanations can\nalleviate your concerns regarding the simulation model.\n6. “the amorphous MoSx exhibited an increased electron the amorphous\nexhibited an increased electron”, but I can only see an opposite trend. Also,\n“revealing the electron donation from LM to Mo atoms” is questionable.\nResponse: Thanks for your valuable questions. We used density functional\ntheory (DFT) calculations in Fig.5 to investigate the electronic structure of\nnanozyme. As revealed by the charge density maps in Fig. 5a, the amorphous\nMoS exhibited an increased electron density of Mo compared with the MoS\nx 2\nmodel. The LM MoS model achieved the highest electron density, revealing\nX\nthe electron donation from liquid metal to Mo atoms. In order to response\nreviewer’s concerns as well as get a convincing demonstration, we have\nsupplemented the differential charge density and Bader charge analysis of LM\nMoS X . As shown in Fig. 5b, the differential charge density of LM MoS X\nrevealed the electron donation from the liquid metal to Mo atoms. In addition,\nthe Bader charge of Mo in MoS , MoS , and LM MoS was calculated. The\nBader charge analysis indicated a charge gain of 0.52 e- on the Mo sites in the\nMoS relative to MoS . Compared to MoS , the electron accumulation on the\nX 2 X\nMo sites increased by an additional 0.21 e- in the LM MoS , which further\nX\nconfirmed that electron donation from liquid metal to Mo atoms. We sincerely\nthank the reviewer for the valuable question, and we have added the following\nfigure and corresponding description into the revised manuscript.\nIn the revised manuscript, we added:\n“Moreover, the differential charge density of LM MoS in Fig. 5b revealed the\nX\nelectron donation from the liquid metal to Mo atoms. In addition, the Bader\ncharge of Mo in MoS , MoS , and LM MoS was calculated. The Bader charge\nanalysis indicated a charge gain of 0.52 e- on the Mo sites in the MoS relative\nX\nto MoS . Compared to MoS , the electron accumulation on the Mo sites\nincreased by an additional 0.21 e- in the LM MoS , which further confirmed that\nX\nelectron donation from liquid metal to Mo atoms.”\nFig. 5. DFT Calculations. a The computational surface models and charge density maps\nof MoS 2 , MoS X and LM MoS X . b The differential charge density analysis at the Mo and Ga\ninterfaces of LM MoS . The yellow area indicates electron gain and the blue area\nX\nindicates electron loss.\n7. What’s the meaning of “d-band center” and what’s the algorithm to\ndetermine its center?\nResponse: Thanks for your professional questions. D-band center refers to\nthe energy center of the d-orbital electron density of states (DOS) of transition\nmetal atoms, which is the weighted average energy value of all electronic\nstates in the d-orbital according to their DOS.\nThe algorithm to determine the d-band center is as follows:\nis the d-orbital electron density of states, is the energy of electronic\nstates, and the integration range covers all electronic states of the d orbital\n(usually from the binding state below the Fermi level to the antibonding state\nabove). We have added these discussions in the revised supplementary\ninformation.\nIn the revised supplementary information, we added:\nDefinition of d-Band Center\nd-band center refers to the energy center of the d-orbital electron density of\nstates (DOS) of transition metal atoms, which is the weighted average energy\nvalue of all electronic states in the d-orbital according to their DOS.\nThe algorithm to determine the d-band center is as follows:\nis the d-orbital electron density of states, is the energy of electronic\nstates, and the integration range covers all electronic states of the d orbital\n(usually from the binding state below the Fermi level to the antibonding state\nabove).\nReviewer #2 (Remarks to the Author):\nIn this manuscript, Zhang et al. designed a series of liquid metal-buffered\nnanozymes and screened the most favorable nanozyme to achieve efficient\ntumor catalytic therapy. The findings highlight the roles of tailored electronic\nstructure and phase engineering for regulating the catalytic activity of\nnanozymes. Authors demonstrate that this is the first report of screening and\nregulating the catalytic and therapeutic effects of LM nanozymes to achieve\nefficient tumor therapy. The overall work is well-designed, and the application\nof LM in catalysis and the biomedical field is interesting. However, at the\ncurrent stage, the characterization of the structure-activity relationship and the\nanti-cancer mechanism of LM-based nanozyme is not solid. Also, other issues\nneed to be addressed.\n1) The most significant concern of utilizing LM-based nanozymes for catalytic\ncancer therapy lies in the intrinsic property of LM. As is known, amorphous LM\nnanoparticles could gradually transform into crystalline material, especially in\naqueous solution. Due to such a dynamic process, the composition of\nLM-based nanozyme could hardly be maintained during in vitro and in vivo\nexperiments or characterizations. Also, it is necessary to ensure that the MoSx\ncontent in a series of LM-buffered nanozymes is the same. Or else, the\ndifferences in nanozyme activity could also be influenced by different MoSx\ncontents.\nResponse: Thank you for your precious advice. Your insightful suggestion\nwas very helpful for improving our study. We strongly agree with the reviewer’s\nopinion that “the stability and sustained catalytic activity are crucial”. We have\nconducted time-dependent stability assessments of LM MoS under simulated\nX\nphysiological conditions. As shown in supplementary Fig. 5, the LM MoS X\nnanozymes remained monodisperse and stable in PBS over 7 days.\nMeanwhile, the nanozyme exhibited consistent POD-like catalytic performance,\nconfirming its good stability (Supplementary Fig. 16). In addition, we have\nmonitored the Mo leakage rate of H-LM MoS after being soaked in complete\nX\ncell culture medium through ICP-MS. As illustrated in Supplementary Fig. 32f,\nH-LM MoS exhibited negligible Mo leaching over 7 days. These results\nX\ndemonstrated the sufficient stability of nanozyme. Especially, according to\nprevious studies, hyaluronic acid withabundant hydroxyl and carbonyl groups\nthat can stabilize metal-based nanoparticles and thuspreventing degradation\nor oxidation (Trends Biotechnol. 2021, 39, 624, Mater. Horiz., 2021, 8, 56).\nMoreover, many reported amorphous nanomaterial demonstrated excellent\nperformance in vivo (Nat Commun 2022, 13, 5088, Angew. Chem. Int. Ed.\n2025, 64, e202412031, J. Am. Chem. Soc. 2025, 147, 26060). We hope that\nthese explanations can address your concern.\nIn addition, we are completely in agreement with the reviewer’s opinion that “it\nis necessary to ensure that the MoSx content in a series of LM-buffered\nnanozymes is the same”. When comparing the POD-like activities of\nLM-buffered nanozymes, the Mo content was kept consistent. We sincerely\nthank the reviewer for the valuable question, and we have added the following\nfigure and corresponding description into the revised manuscript.\nIn the revised manuscript, we added:\n“Moreover, the SEM images and DLS results suggested that the LM MoS\nX\nnanozymes remained monodisperse and stable over 7 days (Supplementary\nFig. 5).”\n“Moreover, the nanozyme exhibited consistent POD-like catalytic performance\nover 7 days (Supplementary Fig. 16), confirming its good stability.”\n“As illustrated in Supplementary Fig. 32f, H-LM MoS exhibited negligible Mo\nX\nleaching after being soaked in complete cell culture medium for over 7 days.”\nSupplementary Figure 5. The stability study of LM MoS X . Dynamic light scattering (DLS)\nanalysis (a) and SEM images (b) of LM MoS\nX\nafter incubation with PBS for various times.\nSupplementary Figure 16. The POD-like activity of LM MoS X after incubation with PBS\nfor various times.\nSupplementary Figure 32. f The Mo leakage rate of H-LM MoS X after being soaked in\ncomplete cell culture medium (1640+10%FBS) for 0, 3, and 7 days.\n2)In a recently published paper (Small 2025, 2502752), FeOx was used to\nhybridize LM to generate nanozymes. Therefore, the reason for choosing\nmolybdenum sulfide should be clarified to distinguish the innovation of this\nwork from previous works, and the advantages of molybdenum sulfide should\nbe explained as well.\nResponse: Thank you for your professional suggestion. As an essential trace\nelement for the survival of living organisms and a vital cofactor for\nmolybdenum (Mo)-based natural enzymes, Mo is a low-cost transition metal\nwith desirable biosafety (Nat. Commun. 2024, 15, 8720). Notably, due to the\nabundance of easily lost single electrons in the electronic configuration, Mo\nhas a variety of oxidation states, and the reversible transition between Mo (IV),\nMo (V), and Mo (VI) imparts tunable enzyme-like activity to Mo-based\nnanomaterials (Nat. Commun. 2024, 15, 8720; VIEW. 2021, 2, 20200188).\nAmong Mo-based nanomaterials, molybdenum disulfide (MoS ) is a kind of\n2\nprototypical transition-metal dichalcogenide (TMD) and has a graphene-like\n2D layered structure, which is constituted by S-Mo-S layers that combined\ntogether by van der Waals force, always shows unique electronic properties\n(Nat Commun 2022, 13, 5954; Nat Electron 2022, 5, 849; Nat Commun 2024\n15, 10290). More importantly, amorphous MoS materials with specific\nX\nstructure have shown superior performance in various applications, especially\nin catalysis (Nat. Mater. 2016, 15, 640; Nat Catal 2022, 5, 397). Thus, we are\ninterested about the catalytic performance of liquid metal-buffered MoSx\nnanozyme.\nIn the revised manuscript, we added:\n“Molybdenum is a low-cost and biocompatible transition metal with multiple\noxidation states, which endows Mo-based nanomaterials with tunable\nenzyme-like activity.67 Among Mo-based nanomaterials, molybdenum sulfide\nfeatures unique structural and electronic properties and shows superior\nperformance in various applications, especially for catalysis.68, 69”\nIn reference, we added:\n67. Li, LY. et al. Valence-engineered catalysis-selectivity regulation of\nmolybdenum oxide nanozyme for acute kidney injury therapy and\npost-cure assessment. Nat. Commun. 15, 8720 (2024).\n68. Guo, X. et al. Charge self-regulation in 1T'''-MoS structure with rich S\n2\nvacancies for enhanced hydrogen evolution activity. Nat. Commun. 13,\n5954 (2022).\n69. Tran, PD. et al. Coordination polymer structure and revisited hydrogen\nevolution catalytic mechanism for amorphous molybdenum sulfide. Nat.\nMater. 15, 640-646 (2016).\n3)It was demonstrated that Ga75.5In24.5 (commercial Gallium-Indium eutectic)\nMoS performed the best catalytic activity, but the possible reason was not\nX\ngiven. With subtle variations of the compositions, the melting points and the\nfluidities of resultant LM-MoS nanozymes would alter significantly. In this\nX\ncontext, the discussion of the catalytic activity of LM-MoS nanozymes should\nX\nbe careful. Additionally, the authors did not provide detailed synthesis methods\nand characterization techniques. It is suggested to supplement relevant\ndetails.\nResponse: Thank you for your valuable advice. In order to response\nreviewer’s concern, we have added a comprehensive discussion to further\nstrengthen the understanding of composition and catalytic activity. Yu and\ncoworkers investigated the effects of alloying Ga with In on the atomic\nstructure and electronic transport properties of liquid EGaIn using the AIMD\nsimulations. They found that with In alloying of Ga, their larger atomic volume\nleads to the disordered structure as well as weak inter-atomic interactions by\nexpanding the cell volume and broadening their inter-atomic distance.\nMoreover, the altered free-electron density is higher between Ga atoms, while the charge\ndensity near the In atoms is significantly smaller. This feature of the In atoms\nexplain the dominant role of In in reducing the electrical transport property and\nelectrical conductivity (J. Chem. Phys. 2014, 140, 064303). As mentioned in\nthe introduction section, the catalytic active sites and local electronic\ncharacteristics determine their catalytic performance to a large extent.\nAccording to previous studies, the higher electrical conductivity favors\nefficient electron transport, which is beneficial to improve the catalytic\nperformance of nanozyme (Nat. Commun. 2022, 13, 5365; Adv. Mater. 2025,\n37, 2502804). Meanwhile, amorphous materials possess distinctive disordered\natomic arrangement that enables them with abundant active sites and\noptimized electronic configurations, which are beneficial for enhancing their\ncatalytic properties (Nat. Commun. 2019, 10, 4855; Chem. Rev. 2023, 123,\n13489). Thereafter, we mentioned that “the alloying of Ga with In results in a\nmore disordered structure, and the charge density is higher between Ga atoms\nwhile significantly smaller in the vicinity of In atoms” in the manuscript. To\nfurther strengthen the understanding of composition and catalytic activity, we\nsupplemented that the indium content in GaIn alloys significantly affects the\ndisordered structure and the charge density distribution of alloys, thus\ninfluencing their amorphous structure and electrical conductivity. Specifically,\nthe alloying of Ga with In causes more disordered structure. This amorphous\nstructure is beneficial for the high catalytic performance of catalysts. However,\nthe electrical conductivity of GaIn alloys decreases with In alloying of Ga\nbecause the charge density near the In atoms is significantly smaller, which is\nunfavorable to the electron transfer and the subsequent catalytic process.\nThus, the catalytic activity of the resulting nanozymes could show a trend of\ninitially increasing and then decreasing with increasing In content in the GaIn\nalloy. We sincerely thank the reviewer for the insightful and constructive\nsuggestion, and we have added the corresponding discussion into the revised\nmanuscript.\nMoreover, we thank the review’s suggestion that the discussion of the catalytic\nactivity of LM MoS nanozymes should be careful. These liquid metal-based\nX\nnanozymes were synthesized under identical conditions, and the POD-like\nactivities of LM-buffered nanozymes were compared under the same\nexperimental conditions. In addition, we have supplemented some details\nabout synthesis methods and characterization techniques, and the relevant\ncontent was marked in red in the revised manuscript and supplementary\ninformation.\nIn the revised manuscripts, we added:\n“The Indium content in GaIn alloys significantly affects the disordered structure\nand the charge density distribution of alloys, thus influencing their amorphous\nstructure and electrical conductivity. Specifically, the alloying of Ga with In\ncauses more disordered structure. This amorphous structure is beneficial for\nthe high catalytic performance of catalysts. However, the electrical conductivity\nof GaIn alloys decreases with In alloying of Ga because the charge density\nnear the In atoms is significantly smaller, which is unfavorable to the electron\ntransfer and the subsequent catalytic process of catalysts. Thus, the catalytic\nactivity of the resulting nanozymes could show a trend of initially increasing\nand then decreasing with increasing In content in the GaIn alloy.”\n4)Although decorating with HA, it remains challenging for LM-MoS nanozyme\nX\n(ca. 300 nm) to achieve efficient cellular uptake and high accumulation in the\nstiff 4T1 tumor via intravenous injection (Figure S35). And what is the half-life\nof LM-MoSX nanozyme?\nResponse: Thank you for your professional suggestion. We have\nsupplemented relevant experiment to study the half-life of LM MoS nanozyme\nX\nin the revised SI. As revealed in Supplementary Fig. 37, the half-life of H-LM\nMoS was 1.97 h. We have added these discussion and figure into the further\nX\nrevised manuscript.\nIn the revised manuscript, we added:\n“Meanwhile, the half-life of H-LM MoS was determined to be 1.97 h, as shown\nX\nin Supplementary Fig. 37.”\nSupplementary Figure 37. Blood circulation curve of H-LM MoS X nanoparticles\nquantified by fluorescence spectroscopy at different time points after injection. The blood\ncirculation curve showed that the pharmacokinetic profile of H-LM MoS nanoparticles\nX\nwas fitted by a two-compartment model with a half-life of 1.97 h.\nWith regard to the tumor accumulation of nanoparticles, according to the\npredominant belief in the nanotechnology community, nanoparticles cross the\ntumor vascular barrier through intercellular gaps between adjacent endothelial\ncells and are retained in the tumor owing to pressure created by poor\nlymphatic drainage — a process termed ‘enhanced permeability and retention’\n(EPR) (Nat. Rev. Mater. 2016, 1, 16014). For tumor blood vessels, adjacent\nendothelial cells adhere to each other less tightly, forming interendothelial cell\ngaps across the vessel wall. These gaps were found to have a size range up to\n2,000 nm (Nat. Mater. 2020, 19, 566). Nanoparticles should be designed with\nsizes smaller than the size of interendothelial gaps to accumulate inside the\ntumor. Researchers have found that tumor vessels were permeable to\nliposomes of up to 400 nm in actual diameter (Cancer Res. 1995, 55, 3752).\nMoreover, Du’s research group have synthesized a cerium single-atom\nnanozyme (Ce SAs@NC) with a hydrodynamic size of 370.2 nm (J. Am. Chem.\nSoc. 2024, 146, 34092). Fluorescence images of mouse organs 24 h after\ninjection showed nanozyme accumulation in the liver and tumor. Ning’s\nresearch group have constructed a dual-atom sites nanozyme (FeCu-DA), and\nthe hydrodynamic size was determined to be approximately 300 nm (Adv. Sci.\n2025, 12, 2414734). They have investigated the biodistribution of FeCu-DA\nnanozyme, and found that the fluorescence intensity was notably high in the\ntumor, liver, and kidney, suggesting that nanozyme effectively accumulated in\ntumor tissue and was primarily cleared through the hepatic and renal pathways.\nTheir data also confirmed that nanoparticles with a hydrodynamic size larger\nthan 300 nm exhibited selective uptake by tumor.\nMoreover, the size measured by DLS is usually larger than the actual size of\nnanoparticle observed under TEM. This discrepancy is likely caused by\nvariations in the degree of coating expansion, which can be attributed to the\nrelatively weak electrostatic interactions under different conditions (Angew.\nChem. Int. Ed. 2023, 62, e202310741). More importantly, the tumor targeting\neffect of HA could enhance the enrichment at the tumor site (Nat Commun\n2023, 14, 4647; Nat. Nanotechnol. 2025, 20, 935). In addition, we have\nsupplemented the quantitative data on in vivo biodistribution. ICP analysis in\nSupplementary Fig. 39 demonstrated that H-LM MoS X could be efficiently\naccumulated at tumor sites, and HA coating promoted the enrichment of\nnanoparticles in the tumor. We hope that these explanations can eliminate\nyour concerns regarding the delivery of nanoparticle.\nIn the revised manuscript, we added:\n“The inductively coupled plasma mass spectrometry (ICP-MS) results revealed\nthe enhanced accumulation of H-LM MoS in the tumor region (Supplementary\nX\nFig. 39).”\nSupplementary Figure 39. The biodistribution of LM MoS X and H-LM MoS X in 4T1\ntumor-bearing mice at 12 h using ICP-MS analysis.\n5)In Figure S2B, D, the In 3d curves were poorly fitted.\nResponse: Thank you for your important comments. We have re-fitted the In\n3d curves in these figures in the revised supplementary information. We really\nappreciated the helpful suggestions.\nSupplementary Figure 3. The In 3d XPS spectra of (a) Ga 50 In 50 MoS X , (b) Ga 62.5 In 37.5\nMoS\nX\n, (c) Ga\n75.5\nIn\n24.5\nMoS\nX\nand (d) Ga\n87.5\nIn\n12.5\nMoS\nX\n.\n6)In Figure S20, the curve shape of EPR result of O2•− seems abnormal.\nPlease confirm the result and the discussion.\nResponse: Thank you for your precious comments. We have checked the\nEPR result of O •−, and the anomalous curve shape was primarily attributed to\n2\nthe partial oxidation of DMPO. We have purchased a new DMPO for\nexperimental use. As shown in Supplementary Fig. 22, an obvious\ncharacteristic six-line pattern of O •− was observed after LM MoS treatment,\nwhich indicated the OXD-like activity of LM MoS . We modified this figure in\nX\nthe revised supplementary information.\nSupplementary Figure 22. EPR spectra of the O 2 •− spin adduct generated by LM MoS X .\n7)In Figure S27, why did the cellular fluorescence of DAPI present large\ndeviations at variable time points?\nResponse: Thanks for your professional question. We are sorry for causing\nthis concern. The difference of Hoechst fluorescence may be due to the weak\nsignal from Hoechst. In order to eliminate reviewer’s concerns as well as get a\nconvincing demonstration, we have reperformed this experiment. As shown in\nSupplementary Fig. 29, the red fluorescence of RB in LM MoS X @RB treated\n4T1 mouse breast cancer cells increased with time and reached the maximum\nafter 9 h, confirming that the LM MoS @RB could be effectively internalized by\nX\n4T1 cells. The experimental results are consistent with previous data. We\nmodified this fluorescence images in the revised supplementary information.\nSupplementary Figure 29. CLSM images of 4T1 cells upon incubation with LM\nMoS @RB. The nuclei were stained with Hoechst (blue). The red fluorescence was from\nX\nRB.\n8)The changes in particle size and potential of LM MoSx should be measured\nafter the modification of HA.\nResponse: Thanks for your important advice. We measured the change in\nparticle size of LM MoS after the modification of HA and have provided the\nX\nresults in Supplementary Fig. 32b. In addition, the potential change of LM\nMoS X after the modification of HA was shown in Supplementary Fig. 32d. The\nparticle size of LM MoS is slightly increased after the modification of HA, and\nX\nthe potential of H-LM MoS was more negative than LM MoS .\nX X\nSupplementary Figure 32. b Size distribution result of LM MoS X and H-LM MoS X . d\nSurface potential of LM MoS and H-LM MoS .\nX X\n9)In Figure S33, there is a difference in H&E staining of the lung between the\nH-LM MoSx group and the control group. Were the images captured at the\nsame magnification?\nResponse: Thanks for your question. We have carefully checked that the\nimages were captured at the same magnification. Both of them were captured\nunder a 20X lens, and the original images with scale bar have been exported\n(Figure R1). Pulmonary alveolus are irregular spherical or hemispherical\nvesicles with varying sizes, and the alveolus structure in these groups was\nnormal. Actually, it is common for Balb/c mice to exhibit certain variations in\nalveolar size within lung tissue under identical rearing conditions. Even within\nthe same lung tissue, there is natural variation in the size and shape of\npulmonary alveolus. Meanwhile, several previous reports have also described\nsimilar phenomena (Figure R2-R3) (Nat Commun 2024, 15, 1626, Nat\nCommun 2024, 115, 8097). Thanks for pointing out this important issue, and\nwe hope that these explanations have addressed your question.\nFigure R1. The original H&E images obtained from the lung of the PBS, H-LM MoS X\ntreated mice at 28 days’ post-injection. Scale bar: 50 µm.\nFigure R2. HE staining images of the tissue sections of heart, liver, spleen, lung, and\nkidney obtained from mice after treatment with Au Pd -FA nanozymes and PBS at 28\n1 3\ndays post-injection (Nat Commun 2024, 15, 1626).\nFigure R3. HE staining images of the tissue sections of heart, liver, spleen, lung, and\nkidney obtained from mice after treatment with Ru1000 nanozymes at 14 days\npost-injection (Nat Commun 2024, 115, 8097).\n10)In Figure S35, quantitative data on in vivo biodistribution should be\nsupplemented.\nResponse: Thanks for your professional advice. In the revision, we have\nsupplemented the quantitative data on in vivo biodistribution. As shown in\nSupplementary Fig. 39, the inductively coupled plasma mass spectrometry\n(ICP-MS) results revealed that H-LM MoS showed enhanced tumor\nX\naccumulation in the tumor region. The corresponding analysis and discussion\nhave been changed or added in the revised manuscript.\nIn the revised manuscript, we added:\n“The inductively coupled plasma mass spectrometry (ICP-MS) results revealed\nthe enhanced accumulation of H-LM MoS in the tumor region (Supplementary\nX\nFig. 39).”\nSupplementary Figure 39. The biodistribution of LM MoS X and H-LM MoS X in 4T1\ntumor-bearing mice at 12 h using ICP-MS analysis.\n11)The stability test of the LM MoSx nanozymes should be provided.\nResponse: Thanks for your valuable suggestion. We have supplemented the\nstability test of the LM MoS nanozymes in the revised SI. As shown in\nX\nSupplementary Fig. 5, the LM MoS X nanozymes remained monodisperse and\nstable over 7 days. We added the following figure and corresponding\ndescription into the revised version.\nIn the revised manuscript, we added:\n“Moreover, the SEM images and DLS results suggested that the LM MoS\nX\nnanozymes remained monodisperse and stable over 7 days (Supplementary\nFig. 5).”\nSupplementary Figure 5. The stability study of LM MoS X . Dynamic light scattering (DLS)\nanalysis (a) and SEM images (b) of LM MoS\nX\nafter incubation with PBS for various times.\n12)In the article, POD-like activity was used to screen for suitable ratios of\nGaIn. Why not use other nanozyme activities for screening?\nResponse: Thanks for your instructive question. From the perspective of\ncatalytic mechanism, the use of POD (Peroxidase)-like activity to screen the\ncatalytic activity of nanozyme is mainly attributed to the extensively studied\ncatalytic mechanism. When comparing the POD activity of nanozymes, we are\ninterested in whether the nanozymes can efficiently decompose H O and the\n2 2\nelectron transfer pathway in the catalytic reaction, which are conducive to\nunderstanding and comparing the electronic properties of nanozymes.\nElectron transfer is essential for catalysis, and catalytic activity is the external\nperformance of electronic properties for nanozyme. Therefore, POD-like\nactivity is informative to screen the suitable ratios of GaIn in the construction of\nnanozymes, and may contribute to clarify the difference of GaIn alloys with\nvarious ratio. From an experimental perspective, the POD-like catalytic system\nis advantageous because it not only uses H O , a readily available substrate,\n2 2\nbut also produces intense and easily detectable color changes with various\nchromogenic substrates. In contrast, OXD (Oxidase)-like activity is dependent\non unstable O in the environment, leading to unstable signals and significant\n2\nbackground interference. The product of CAT (Catalase)-like activity is O ,\n2\nwhich are not easy to quantify (typically requiring a dissolved oxygen electrode)\nand are not as intuitive as color changes. More importantly, there are\nstandardized assays for determining the catalytic activity and kinetics of\nperoxidase-like nanozymes (Nat Protoc 2018, 13, 1506; Nat Catal 2021, 4,\n407; Nat Protoc 2024, 19, 3470). From the perspective of bioapplication, the\nsubstrate and product of POD-like catalytic reaction are important\nbiomolecules and signaling molecules in biological systems, which can be\nused for disease treatment and diagnosis. Therefore, we use POD-like activity\nto screen for suitable ratios of GaIn.\n13)In Figure 6J, the live and dead cells co-staining was not well performed,\nand the cell morphology could hardly be distinguished. Please provide\nfluorescence images under high magnification.\nResponse: Thanks for your important advice. In the revision, we have\nperformed the live/dead staining analysis and provided fluorescence images\nunder high magnification. As can be seen in Fig. 6j, the cell morphology can\nbe clearly distinguished. Green signals represent live 4T1 cells, while red\nsignals represent dead 4T1 cells. We modified this fluorescence images in the\nrevised manuscript.\nFigure 6. j Live and dead co-staining of 4T1 cells with different treatments. Scale bar: 100\nµm.\n14)The grammar and language of the manuscript should be carefully polished.\nResponse: Thanks for the kindness and advice. The whole manuscript has\nbeen thoroughly examined, and the grammar and language have been\npolished to improve its overall clarity and readability.\nReviewer #3 (Remarks to the Author):\nThe authors present a comprehensive and well-executed study on liquid\nmetal-buffered MoSₓ nanozymes, systematically synthesized using Ga–In\nalloys to optimize catalytic performance. The Ga . In . -based nanozyme\n₇₅ ₅ ₂₄ ₅\nexhibits enhanced peroxidase-, oxidase-, and NADH oxidase-like activities,\nenabling a catalytic cascade that disrupts redox homeostasis in cancer cells\nand induces selective cytotoxicity both in vitro and in vivo. The combination of\nmulti-enzyme functionality, effective tumor targeting, and minimal toxicity\nunderscores the therapeutic potential of this platform. The study’s originality\nlies in leveraging liquid metal templating to precisely control nanozyme\ncomposition, structure, and electronic properties, representing a clear\nadvancement over previous LM-based nanozyme approaches. The\nconclusions are well-supported by comprehensive materials characterization,\ncatalytic assays, theoretical modeling, and biological validation. The\nexperimental methodology is appropriate and rigorously executed. Overall, this\nwork constitutes a significant contribution to nanozyme research and catalytic\ncancer therapy.\nTo further enhance the manuscript, several clarifications and suggestions are\nproposed as detailed in the following comments (also provided in the attached\nfile).\n1. Particle Size and Delivery: A potential concern is the relatively large\nhydrodynamic size of the nanozyme (~300 nm diameter). Particles of this size\nmay have limited tumor penetration and faster clearance by the mononuclear\nphagocyte system. The in vivo success after HA coating suggests good tumor\naccumulation capability, but the authors might comment on how the size could\ninfluence distribution. Could smaller LM-templated nanozymes be produced\n(e.g., by adjusting sonication conditions) to further improve tissue penetration?\nResponse: Thanks for your precious questions. We have complemented the\nexperiment on the influence of sonication power on the size of synthesized\nmaterials. These different LM MoS nanoparticles were obtained by sonicating\nX\nGa In bulk in 20 mM (NH ) MoS solution at various power of\n75.5 24.5 4 2 4\nultrasonication. The SEM images and particle size distribution results were\nshown in Supplementary Fig. 1. It was found that increasing the applied\npower of ultrasonication from 50 W to 500 W results in a decrease in the\noverall size of the LM MoS nanostructures from micrometer to hundreds of\nX\nnanometers. The size distribution also becomes narrower with higher\nsonication intensity applied. When the highest power (1000 W) was employed,\nthe hydrodynamic size of the materials was about 300 nm. This represents the\nsmallest size achievable for the nanozyme under this synthesis conditions. We\nhave supplemented these experiment results and description in the revised\nmanuscript.\nSupplementary Figure 1. SEM images and particle size distributions of LM MoS X\nobtained by sonicating Ga In bulk in 20 mM (NH ) MoS solution at various power of\n75.5 24.5 4 2 4\nultrasonication: (a) 50 W, (b) 100 W, (c) 200 W, (d) 300 W, (e) 400 W, (f) 500 W. It was\nfound that increasing the applied power of ultrasonication from 50 W to 500 W results in a\ndecrease in the overall size of the LM MoS nanostructures from micrometer to hundreds\nX\nof nanometers. The size distribution also becomes narrower with higher sonication\nintensity applied. The enhancing oscillating shear force from high power sonication is\nregarded as the main factor to separate small droplets from the bulk liquid metal in a short\ntime.\nWith regard to the tumor accumulation of nanoparticles, according to the\npredominant belief in the nanotechnology community, nanoparticles cross the\ntumor vascular barrier through intercellular gaps between adjacent endothelial\ncells and are retained in the tumor owing to pressure created by poor\nlymphatic drainage — a process termed ‘enhanced permeability and retention’\n(EPR) (Nat. Rev. Mater. 2016, 1, 16014). For tumor blood vessels, adjacent\nendothelial cells adhere to each other less tightly, forming interendothelial cell\ngaps across the vessel wall. These gaps were found to have a size range up to\n2,000 nm (Nat. Mater. 2020, 19, 566). Nanoparticles should be designed with\nsizes smaller than the size of interendothelial gaps to accumulate inside the\ntumor. Researchers have found that tumor vessels were permeable to\nliposomes of up to 400 nm in actual diameter (Cancer Res. 1995, 55, 3752).\nMoreover, Du’s research group have synthesized a cerium single-atom\nnanozyme (Ce SAs@NC) with a hydrodynamic size of 370.2 nm (J. Am. Chem.\nSoc. 2024, 146, 34092). Fluorescence images of mouse organs 24 h after\ninjection showed nanozyme accumulation in the liver and tumor. Ning’s\nresearch group have constructed a dual-atom sites nanozyme (FeCu-DA), and\nthe hydrodynamic size was determined to be approximately 300 nm (Adv. Sci.\n2025, 12, 2414734). They have investigated the biodistribution of FeCu-DA\nnanozyme, and found that the fluorescence intensity was notably high in the\ntumor, liver, and kidney, suggesting that nanozyme effectively accumulated in\ntumor tissue and was primarily cleared through the hepatic and renal pathways.\nTheir data also confirmed that nanoparticles with a hydrodynamic size larger\nthan 300 nm exhibited selective uptake by tumor.\nMoreover, the size measured by DLS is usually larger than the actual size of\nnanoparticle observed under TEM. This discrepancy is likely caused by\nvariations in the degree of coating expansion, which can be attributed to the\nrelatively weak electrostatic interactions under different conditions (Angew.\nChem. Int. Ed. 2023, 62, e202310741). More importantly, the tumor targeting\neffect of HA could enhance the enrichment at the tumor site (Nat Commun\n2023, 14, 4647; Nat. Nanotechnol. 2025, 20, 935). In addition, we have\nsupplemented the quantitative data on in vivo biodistribution. ICP analysis in\nSupplementary Fig. 39 demonstrated that H-LM MoS X could be efficiently\naccumulated at tumor sites, and HA coating promoted the enrichment of\nnanoparticles in the tumor. We hope that these explanations can eliminate\nyour concerns regarding the particle size and delivery.\nSupplementary Figure 39. The biodistribution of LM MoS X and H-LM MoS X in 4T1\ntumor-bearing mice at 12 h using ICP-MS analysis.\n2. Comparison to Other Approaches: The introduction could better distinguish\nthis work from prior related studies. Although the manuscript cites previous\nreports on LM-based nanozymes (Wu et al., 2023; Xu et al., 2024), it does not\nsufficiently articulate how the present study advances beyond these earlier\nefforts. It would be valuable for the authors to explicitly highlight how their\napproach advances beyond previous reports, particularly in terms of the\nsystematic composition tuning and realization of a multi-enzyme cascade\nfunction with therapeutic efficacy. This clarification would better establish the\noriginality and significance of the current work.\nResponse: Thank you for your important question. Wu and coworkers\nsynthesized a liquid metal-based nanozyme LM@ZIF@HA (LZH) with eutectic\nGallium Indium (EGaIn) as the core, which was coated with CoNi-bimetallic\nzeolite imidazole framework (ZIF) and hyaluronic acid (HA) (J. Nanobiotechnol.\n2023, 21, 399). However, their nanozyme catalytic activity was attributed to the\nCo2+ center in ZIF-67 rather than the liquid metal. Their study focused on\nsolving the problem of poor delivery of microwave sensitizer and the obstacle\nof tumor hypoxic microenvironment in microwave dynamic therapy (MDT). Xu\nand coworkers reported the galvanic replacement synthesis of\nVOx@EGaIn−PEG, and the designed nanohybrids demonstrated excellent\nperoxidase-like activity (ACS Appl. Mater. Interfaces 2024, 16, 21975). Their\nstudy focused on solving the limitations in biocompatibility, regenerability and\nrecyclability of biosensors. They showed that the combination of gallium\noxides and V O led to structural changes and alterations in band gap energy,\n2 5\nresulting in enhanced catalytic activity of the VOx@EGaInPEG nanozyme.\nMoreover, the designed nanozyme demonstrated catalytic stability and\nregeneration ability due to the self-healing and regenerative properties of LM.\nThey further developed a nanozyme colorimetric sensor for the convenient and\nsensitive detection of glucose in physiological environments.\nThese studies are mainly focus on the application of LM-based nanozyme\nwhile overlooking the potential of liquid metal with intrinsic amorphous property\nand unusual electronic microenvironment in regulating nanozyme catalytic\nactivity. Moreover, the reported LM-based nanozyme only displayed a single\nenzyme-mimic activity. Our work highlighted the catalytic potential of liquid\nmetals for screening and regulation of nanozyme activity. We have not only\ninvestigated the catalytic activity of liquid metal-buffered nanozyme with\ndifferent gallium indium ratios, but also explored the role of liquid metal in\nregulating catalytic performance and the reasons for the enhanced\nenzyme-like catalytic activity of liquid metal-buffered nanozyme. In addition,\nour designed LM-based nanozyme exhibited multienzyme mimetic activity, i.e.,\nPOD-like activity, OXD-like activity, NOX-like activity, which could work\ntogether to achieve multiple-enzyme-like cascade catalytic reactions to disrupt\nintracellular redox and metabolic homeostasis.\nIn the revised manuscripts, we added:\n“However, the potential of liquid metal in regulating nanozyme catalytic activity\nhas rarely been explored mechanistically. In particular, previous research\nefforts have focused on the application of liquid metal-based nanozyme while\noverlooking the catalytic potential of liquid metals for screening and regulation\nof nanozyme activity. 62, 63”\n“Both in vitro and in vivo experiments demonstrate that the LM MoS\nX\nnanozymes with multienzyme-mimicking catalytic activity can trigger a highly\nefficient cascade reaction to disrupt intratumoral redox and metabolic\nhomeostasis, eventually killing tumor cells.”\n“Our work not only investigates the catalytic activity of liquid metal-buffered\nnanozyme with different gallium indium ratios, but also elucidates the\nmechanism underlying the enhanced enzyme-like catalytic activity through\ndensity functional theory (DFT) calculations.”\n3. Mechanistic Detail – Role of Indium vs Gallium: The study identifies an\noptimal Ga/In ratio; however, the mechanistic reasoning for why both\nexcessive and insufficient indium content are detrimental could be further\nelaborated. The authors refer to prior work on charge distribution within Ga–In\nalloys (ref 67), indicating that indium incorporation increases structural\ndisorder and enhances electron density around gallium. They conclude that\nlower or higher indium levels compromise catalytic activity. This is a key insight\nand perhaps a more comprehensive discussion here would strengthen the\nunderstanding of how composition ties to structure and catalytic activity.\nResponse: Thanks for your professional suggestion. Yu and coworkers\ninvestigated the effects of alloying Ga with In on the atomic structure and\nelectronic transport properties of liquid EGaIn using the AIMD simulations.\nThey found that with In alloying of Ga, their larger atomic volume leads to the\ndisordered structure as well as weak inter-atomic interactions by expanding\nthe cell volume and broadening their inter-atomic distance. Moreover, the\naltered free-electron density is higher between Ga atoms, while the charge\ndensity near the In atoms is significantly smaller. This feature of the In atoms\nexplain the dominant role of In in reducing the electrical transport property and\nelectrical conductivity (J. Chem. Phys. 2014, 140, 064303). As mentioned in\nthe introduction section, the catalytic active sites and local electronic\ncharacteristics determine their catalytic performance to a large extent.\nAccording to previous studies, the higher electrical conductivity favors efficient\nelectron transport, which is beneficial to improve the catalytic performance of\nnanozyme (Nat. Commun. 2022, 13, 5365; Adv. Mater. 2025, 37, 2502804).\nMeanwhile, amorphous materials possess distinctive disordered atomic\narrangement that enables them with abundant active sites and optimized\nelectronic configurations, which are beneficial for enhancing their catalytic\nproperties (Nat. Commun. 2019, 10, 4855; Chem. Rev. 2023, 123, 13489).\nThereafter, we mentioned that “the alloying of Ga with In results in a more\ndisordered structure, and the charge density is higher between Ga atoms while\nsignificantly smaller in the vicinity of In atoms” in the manuscript. To further\nstrengthen the understanding of composition and catalytic activity, we\nsupplemented that the Indium content in GaIn alloys significantly affects the\ndisordered structure and the charge density distribution of alloys, thus\ninfluencing their amorphous structure and electrical conductivity. Specifically,\nthe alloying of Ga with In causes more disordered structure. This amorphous\nstructure is beneficial for the high catalytic performance of catalysts. However,\nthe electrical conductivity of GaIn alloys decreases with In alloying of Ga\nbecause the charge density near the In atoms is significantly smaller, which is\nunfavorable to the electron transfer and the subsequent catalytic process.\nThus, the catalytic activity of the resulting nanozymes could show a trend of\ninitially increasing and then decreasing with increasing In content in the GaIn\nalloy. We sincerely thank the reviewer for the insightful and constructive\nsuggestion, and we have added the corresponding discussion into the revised\nmanuscript.\nIn the revised manuscripts, we added:\n“The Indium content in GaIn alloys significantly affects the disordered structure\nand the charge density distribution of alloys, thus influencing their amorphous\nstructure and electrical conductivity. Specifically, the alloying of Ga with In\ncauses more disordered structure. This amorphous structure is beneficial for\nthe high catalytic performance of catalysts. However, the electrical conductivity\nof GaIn alloys decreases with In alloying of Ga because the charge density\nnear the In atoms is significantly smaller, which is unfavorable to the electron\ntransfer and the subsequent catalytic process of catalysts. Thus, the catalytic\nactivity of the resulting nanozymes could show a trend of initially increasing\nand then decreasing with increasing In content in the GaIn alloy.”\nIn the introduction section, we mentioned:\n“From an in-depth perspective, the catalytic active sites and local electronic\ncharacteristics determine their catalytic performance to a large extent.”\n“Amorphous materials possess distinctive disordered atomic arrangement that\nenables them with abundant active sites and optimized electronic\nconfigurations, which are beneficial for enhancing their catalytic properties.”\n4. Undefined Terms and Abbreviations: Several key terms and abbreviations\nthat are central to the manuscript are not defined upon first use:\n- POD-like activity: The manuscript repeatedly refers to \"POD-like activity,\" but\nfails to define \"POD.\"\n- ABTS: Used as a substrate in peroxidase-mimicking activity assays but not\ndefined.\n- ROS: The term \"ROS\" is frequently used but never defined.\n- NADH: Although common in biochemical studies, NADH is not defined.\n- The MTT method is mentioned as the method for cytotoxicity evaluation, but\nthe manuscript does not define MTT.\nResponse: Thanks for your suggestions. We have defined these\nabbreviations in the revised manuscript, and reviewed the entire manuscript to\navoid similar issues.\n“We observe that these nanozymes exhibit peroxidase (POD)-like activities.”\n“We explored the POD-like activity of five nanozymes by monitoring the\noxidation of 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) at\n415 nm.”\n“eventually leading to reactive oxygen species (ROS) burst”\n“GSH and nicotinamide adenine dinucleotide (NADH) are important\nintracellular metabolites upregulated in cancer cells”\n“the in vitro cytotoxicity of nanozymes was measured by methyl thiazolyl\ntetrazolium (MTT) method”\n5. Missing Figure Legends in Fig. 4C and Fig. 4F: Figures 4C (DPBF assay for\nO • detection) and 4F (NADH oxidation spectra) lack in-figure legends. The\n₂ ⁻\ndata series in the plots are not labeled. The authors should include proper\nlegends directly in the figure.\nResponse: Thanks for your attention and professional comments. We have\nsupplemented the figure legends in Fig. 4c and Fig. 4f in the revised\nmanuscript. We have re-examined the whole article and corrected the similar\nissues in the revised manuscript.\nFig. 4. c The time-dependent generation of O 2 •− monitored by DPBF probe. f The\nNOX-like activity of LM MoS nanozymes.\nX\nSupplementary Figure 23. a The NADH consumption ability of MoS 2 (0-80 min).\nReviewer #4 (Remarks to the Author):\nThis manuscript outlined the synthesis, characterisation and application of\nnovel amorphous liquid Ga/MoSx structures. The authors show that these\nmaterials, coined as nanozymes, showcase some impressive catalytic activity,\nand can be used in biological contexts where their redox activity may confer\ntherapeutic effects (e.g. for tumour cells). Overall, I am highly impressed by the\nwork contained in this manuscript. I note that the uncovering of cascade\ncatalysis that relies on the liquid metal system is a particularly interesting\nfinding. Therefore, I would consider the work of interest to the readership, and\nof good quality for Nat. Commun. However, I believe there are several areas\nrequiring clarification or amendment before I can recommend the manuscript\nfor publication.\nMajor comments:\n1. The manuscript could benefit from clarifying the physical state of the\nLM/MoSx systems, especially during catalysis testing. The LMs were\npresumably melted and in liquid form during synthesis of the LM/MoSx\nstructures, but can they still be considered to be in the liquid state during\napplication? This is perhaps especially relevant in the biological context, where\ntemperature ranges are limited and must be carefully controlled.\nResponse: Thank you for your important question. For Ga-based liquid metal,\nthe native gallium oxide that forms on these metals lowers the interfacial\ntension of liquid metal while adding a thin, mechanical “skin” to the surface\n(Acc. Mater. Res. 2021, 2, 1093). The oxide layers allow the liquid metal to\nsustain non-spherical structures and facilitate their dispersion (ACS Nano\n2019, 13, 7388). In our work, the liquid metal droplets served as templates,\ntriggering in situ galvanic replacement reactions between liquid metal droplets\nand MoS 2- ions to produce core-shell nanostructures upon ultrasonication.\nX\nmetal from leaking out and improve the colloidal stability of nanoparticles\n(Chem. Soc. Rev., 2018, 47, 2518). This structure can be regarded as a\ncore-shell configuration consisting of a solid shell and a liquid inner core (ACS\nNano 2019, 13, 7388-7395, Adv. Mater. 2025, 37, 2419060). In addition, we\nhave also attached great importance to the reviewer's comments regarding the\nbiological applications. In fact, liquid metal has shown great potential in\nbiomedical field. For example, Park and coworkers reported liquid metal-based\nthree-dimensional microelectrode arrays that were integrated with implantable\nultrathin retinal prosthesis, which led to the restoration of vision (Nat.\nNanotechnol. 2024, 19, 688). Owing to their liquid form, liquid metal could\nminimize damage to the retina. Their group also reported liquid metal-based\nsoft neural probes that can monitor the single-unit activities of neurons with\nlong-term stability (Nat Commun 2024, 15, 1772). These studies demonstrated\nthat liquid state does not limit the application of liquid metals in biological\nsystems. Gu and coworkers have reported that doxorubicin (DOX) fragments\nwere good ligands to stabilize aqueous liquid metal nanodroplet, and the\nresulted liquid metal nanomedicine demonstrated excellent tumor penetration\nand anticancer efficacy (Adv. Mater. 2024, 36, 2307817). In our work, the\ndesigned LM MoS systems possess a MoS shell that protects the inner liquid\nX X\nmetal, demonstrating sufficient stability for biological applications. In order to\nresponse reviewer’s concern, we have conducted time-dependent stability\nassessments of LM MoS under simulated physiological conditions. As shown\nX\nin Supplementary Fig. 5, the LM MoS X nanozymes remained monodisperse\nand stable in PBS over 7 days. Meanwhile, the nanozyme exhibited consistent\nPOD-like catalytic performance, confirming its good stability (Supplementary\nFig. 16). Thanks again for the reviewer’s support and valuable comments for\nour manuscript.\nIn the revised manuscript, we added:\n“Moreover, the SEM images and DLS results suggested that the LM MoS\nX\nnanozymes remained monodisperse and stable over 7 days (Supplementary\nFig. 5).” “Moreover, the nanozyme exhibited consistent POD-like catalytic\nperformance over 7 days (Supplementary Fig. 16), confirming its good\nstability.”\nSupplementary Figure 5. The stability study of LM MoS X . Dynamic light scattering (DLS)\nanalysis (a) and SEM images (b) of LM MoS\nX\nafter incubation with PBS for various times.\nSupplementary Figure 16. The POD-like activity of LM MoS X after incubation with PBS\nfor various times.\n2. The authors discuss the dynamic nature of liquid metals as key to enhancing\ntheir catalytic activity in the introduction, but later suggest the activity of their\nLM/MoSx systems may be due to an abundance of active sites and interesting\nelectronic characters in the amorphous structure. Several groups are now\nreleasing atomistic modelling suggesting that the dynamic motion of liquid\nmetal systems still plays a key role as well:\na. https://doi.org/10.1038/s41467-025-56222-0\nb. https://doi.org/10.1039/D3SC04416E\nc. https://doi.org/10.1002/ange.202407124\nThe manuscript would benefit from addressing this discussion point in the\ncontext of their results. In particular, perhaps the ongoing dynamics of a liquid\nmetal based system may have links to true enzymatic character?\nResponse: Thanks for your careful reading and professional advice. We\nsincerely appreciate your recommendation of these important papers. We\nhave thoroughly read these literatures. In the first article, Tang and coworkers\nreported that Sn solvent, as a low-cost commodity, supports unique atomic\nassemblies at the interface of molten SnIn Cu which are highly\n0.1034 0.0094,\nselective for H synthesis from hydrocarbons. Atomistic simulations reveal that\n2\ndistinctive adsorption patterns with hexadecane can be established with Cu\ntransiently reaching the interfacial layer, ensuring an energy-favorable route for\nH generation. In the second paper, Ruffman and coworkers proposed a new\n2\napproach based on ab initio molecular dynamics sampling of an adsorbate on\na liquid catalyst. They described time-resolved structures for formate adsorbed\non liquid Ga–In, and for all intermediates in the methanol oxidation pathway on\nGa-Pt. This yields a range of accessible adsorption energies that take into\naccount the at-temperature motion of the liquid metal. Their research suggests\nthat liquid metals have the ability to more readily adsorb species by\nspontaneously extending under-coordinated atoms from the surface as a part\nof their dynamic motion at temperature. In the third article, Ruffman and\ncoworkers found that the constant rearrangement of the liquid structure indeed\nforced the ongoing motion of H . This mobility allows H to drift adjacent to\nads ads\ndopant sites on the liquid surface, enabling low-barrier CO reduction in a way\n2\nthat could not readily occur on the solid. These findings underscore the opinion\nthat dynamic flexibility and constant atomic motion are critical to the unique\nactivity of liquid metal catalysts. It breaks the paradigm of traditional solid\ncatalysts by providing a constantly evolving, self-healing, flexible, and highly\nactive interface, thus conferring optimal selectivity towards specific products at\nthe molecular level. These unique features encourage further exploration of\nliquid metal catalysts for various other reactions.\nAs the reviewer pointed out, we have discussed in the introduction that the\nliquid metal could create dynamic uniform catalytic sites and favorable\nelectronic structures for catalytic metal species, which remarkably enhanced\ntheir catalytic capability (Nat. Chem. 2017, 9, 862; Nat. Chem. 2022, 14, 935;\nNat. Nanotechnol. 2024, 19, 306). These studies were similar to the\nabove-mentioned papers. Nevertheless, our current study was mainly focused\non leveraging the unusual electronic structure and phase engineering ability of\nliquid metal to regulate nanozyme catalytic activity. Their liquid nature plays a\nkey role in the liquid-liquid interfacial reactions during the synthesis process\nand is critical for the amorphous structure of the resulting nanozyme (Adv.\nMater. 2023, 35, e2303533; Chem. Soc. Rev. 2024, 53, 7158). After detailed\ncharacterization and DFT calculations, the current experimental results\nconfirmed the unique role of liquid metals in nanozyme catalysis.\nMoreover, in the above-mentioned literature, the catalytic active sites of\ncatalysts are atomically dispersed within the liquid metal, leading to a dynamic\ncoordination environment. However, in our work, the designed nanozyme are\nmore like a core-shell heterogeneous nanostructures. The MoS shell is\nX\ngenerated by using liquid metal as a template, which maintains the typical\namorphous characteristics of liquid metal. Meanwhile, the liquid metal within\nthe nanozyme affords an electron-rich microenvironment to optimize the\nelectronic structure of nanozymes. Based on their different microstructures,\nthe dynamic nature of liquid metals and their relevance to enzyme-like\ncharacteristics should be carefully considered in the catalytic process of\nnanozyme. To clarify this relevance, more complicated modeling and\ncalculations are needed. Dynamic sampling simulations of substrates and\ncatalysts are highly necessary. However, there are obstacles including the\nmulti-scale character of the processes, the spatial and chemical heterogeneity,\nand the high-dimensional phase space, which are difficult for sufficient\nstatistical sampling. We thank the reviewer for this insightful and constructive\nsuggestion, which is highly instructive. We will focus on the relationship\nbetween the dynamic motion of liquid metal and the catalytic activity of\nnanozymes in future work.\n3. I strongly suggest the authors move the computational details of the density\nfunctional theory calculations to the main text (i.e. in the methods). A reader\nshould ideally be able to find the software used and exchange-correlation\nfunctional in the main text.\nResponse: Thank you for your careful review and precious advice. We have\nmoved the computational details of the density functional theory calculations to\nthe main text in the revised manuscript (method section), and the relevant\nsoftware has been supplemented. This is a very good suggestion.\nIn the method section of revised manuscript, we updated as following:\n“We used the DFT as implemented in the Vienna Ab initio simulation package\n(VASP) in all calculations. The amorphous models were obtained by annealing\nthrough ab initio molecular dynamics (AIMD) calculation method. The\ncanonical (NVT) ensemble and Nosé-Hoover thermostats were used with the\ntemperature set to 2000 K and maintained over 10 ps while the time step is 1\nfs, and finally the temperature was returned to 298.15 K. The\nexchange-correlation potential is described by using the generalized gradient\napproximation of Perdew-Burke-Ernzerhof (GGA-PBE). The projector\naugmented-wave (PAW) method is employed to treat interactions between ion\ncores and valence electrons. The plane-wave cutoff energy was fixed to 450\neV. Given structural models were relaxed until the Hellmann-Feynman forces\nsmaller than -0.02 eV/Å and the change in energy smaller than 10-5 eV was\nattained. The charge density analysis was obtained from CHGCAR. All the\natomic and electronic structures were illustrated with VESTA software.”\n4. More detail should be included on how the MoSx and LM/MoSx structural\nmodels were developed/selected for DFT. Did the authors perform\nat-temperature molecular dynamics simulations? Were these structures\nadapted from other work? As is, it is impossible to identify whether these\nmodels are representative or accurate for the system they are trying to\ndescribe. This is especially true of the LM system, where a single snapshot is\noften not sufficient.\nResponse: Thanks for your valuable questions. We have supplemented more\ndetail in the revised manuscript. The MoS and LM/MoS structural models\nX X\nwere obtained by annealing through ab initio molecular dynamics (AIMD)\ncalculation method, and were not adapted from other work. The canonical\n(NVT) ensemble and Nosé-Hoover thermostats were used with the\ntemperature set to 2000 K and maintained over 10 ps while the time step was\n1 fs, and finally the temperature was returned to 298.15 K. Actually, the AIMD\nmethod is usually used in the simulation of the amorphous structure. There\nhave been some related reports using AIMD method to simulate amorphous\nstructures for performance study (Angew. Chem. Int. Ed. 2024, 63,\ne202405641, Nat. Mater. 2025, 24, 1186-1194). The basic principle of AIMD\nfor constructing amorphous structures is based on the dynamic and\nthermodynamic processes of amorphous formation. When constructing\namorphous structures using AIMD, it is first necessary to establish an\nappropriate initial model. After the initial model is created, the system is heated\nto a high-temperature state to provide atoms with sufficient kinetic energy for\nextensive movement and diffusion within the system. Once thermal equilibrium\nis reached, rapid cooling is initiated. These processes require enough time to\nobtain optimized structural information, so the resulting structure is not based\non a single snapshot. Thanks again for the reviewer's valuable suggestions.\nIn the method section of revised manuscript, we updated as following:\n“We used the DFT as implemented in the Vienna Ab initio simulation package\n(VASP) in all calculations. The amorphous models were obtained by annealing\nthrough ab initio molecular dynamics (AIMD) calculation method. The\ncanonical (NVT) ensemble and Nosé-Hoover thermostats were used with the\ntemperature set to 2000 K and maintained over 10 ps while the time step is 1\nfs, and finally the temperature was returned to 298.15 K. The\nexchange-correlation potential is described by using the generalized gradient\napproximation of Perdew-Burke-Ernzerhof (GGA-PBE). The projector\naugmented-wave (PAW) method is employed to treat interactions between ion\ncores and valence electrons. The plane-wave cutoff energy was fixed to 450\neV. Given structural models were relaxed until the Hellmann-Feynman forces\nsmaller than -0.02 eV/Å and the change in energy smaller than 10-5 eV was\nattained. The charge density analysis was obtained from CHGCAR. All the\natomic and electronic structures were illustrated with VESTA software.”\n5. Can the authors clarify whether HSE06 was used for all DOS calculations as\nwell as determination of the band centres? Was it also used for the charge\ndensity analyses? This needs to be clearly stated in the methods or results.\nResponse: Thanks for your precious questions. The DOS calculations and\ncharge density analyses were based on the Perdew-Burke-Ernzerhof (PBE)\nmethod. The charge density analysis was obtained from CHGCAR. We have\ndetailed this in the method section. We also supplemented the definition of\nd-band center in the revised supplementary information. The d band center\nrefers to the energy center of the d-orbital electron density of states (DOS) of\ntransition metal atoms, which is the weighted average energy value of all\nelectronic states in the d-orbital according to their DOS.\nIn the method section of revised manuscript, we updated as following:\n“We used the DFT as implemented in the Vienna Ab initio simulation package\n(VASP) in all calculations. The amorphous models were obtained by annealing\nthrough ab initio molecular dynamics (AIMD) calculation method. The\ncanonical (NVT) ensemble and Nosé-Hoover thermostats were used with the\ntemperature set to 2000 K and maintained over 10 ps while the time step is 1\nfs, and finally the temperature was returned to 298.15 K. The\nexchange-correlation potential is described by using the generalized gradient\napproximation of Perdew-Burke-Ernzerhof (GGA-PBE). The projector\naugmented-wave (PAW) method is employed to treat interactions between ion\ncores and valence electrons. The plane-wave cutoff energy was fixed to 450\neV. Given structural models were relaxed until the Hellmann-Feynman forces\nsmaller than -0.02 eV/Å and the change in energy smaller than 10-5 eV was\nattained. The charge density analysis was obtained from CHGCAR. All the\natomic and electronic structures were illustrated with VESTA software.”\nIn the revised supplementary information, we added:\nDefinition of d-Band Center\nd-band center refers to the energy center of the d-orbital electron density of\nstates (DOS) of transition metal atoms, which is the weighted average energy\nvalue of all electronic states in the d-orbital according to their DOS.\nThe algorithm to determine the d-band center is as follows:\nis the d-orbital electron density of states, is the energy of electronic\nstates, and the integration range covers all electronic states of the d orbital\n(usually from the binding state below the Fermi level to the antibonding state\nabove).\n6. It was unclear to me exactly which quantity is plotted in Figure 5A on the\ncharge density analyses. Does this plot contain the simple charge density\nobtained form the CHGCAR in VASP? Or is this a charge density difference\nrelative to some reference state (e.g. MoSx and LM separate)? I would\nsuggest the later analysis is more meaningful for showing charge\naccumulation (e.g. at Mo atoms, as the authors identify in the LM/MoSx\nsystem).\nResponse: Thank you for your important comments. The charge density\nanalyses in Fig 5a were refer to the distribution of electrons in real space,\nwhich provided a visual depiction of the probability density distribution of\nelectrons around the atomic nucleus. We have clarified that the charge density\nanalysis was obtained from CHGCAR in the revised manuscript. According to\nthe reviewer’s suggestions, we have supplemented the differential charge\ndensity and Bader charge analysis of LM MoS X . As shown in Fig 5b, the\ndifferential charge density of LM MoS revealed the electron donation from the\nX\nliquid metal to Mo atoms. In addition, the Bader charge of Mo in MoS , MoS ,\nand LM MoS was calculated. The Bader charge analysis indicated a charge\nX\ngain of 0.52 e- on the Mo sites in the MoS relative to MoS . Compared to\nX 2\nMoS , the electron accumulation on the Mo sites increased by an additional\nX\n0.21 e- in the LM MoS , which further confirmed that electron donation from\nX\nliquid metal to Mo atoms. We sincerely thank the reviewer for the valuable\nquestion, and we have added the following figure and corresponding\ndescription into the revised manuscript.\nIn the revised manuscript, we added:\n“The charge density analysis was obtained from CHGCAR.”\n“Moreover, the differential charge density of LM MoS in Fig. 5b revealed the\nX\nelectron donation from the liquid metal to Mo atoms. In addition, the Bader\ncharge of Mo in MoS , MoS , and LM MoS was calculated. The Bader charge\nanalysis indicated a charge gain of 0.52 e- on the Mo sites in the MoS relative\nX\nto MoS . Compared to MoS , the electron accumulation on the Mo sites\nincreased by an additional 0.21 e- in the LM MoS , which further confirmed that\nX\nelectron donation from liquid metal to Mo atoms.”\nFig. 5. DFT Calculations. a The computational surface models and charge density maps\nof MoS 2 , MoS X and LM MoS X . b The differential charge density analysis at the Mo and Ga\ninterfaces of LM MoS . The yellow area indicates electron gain and the blue area\nX\nindicates electron loss.\n7. The DOS plots in 5B should likely be enlarged to make them more clear. It is\ndifficult to read detailed information from them.\nResponse: Thanks for your important advice. The DOS plots in 5B-C have\nbeen enlarged to make them clearer in the revised manuscript (now was Fig.\n5c-d). Thanks again for your attention and professional comments.\nFig. 5. DFT Calculations. c Calculated DOS of the MoS 2 , MoS X and LM MoS X models. d\nCalculated Mo 3d DOS of the MoS , MoS and LM MoS models.\n8. The analysis of a d-band centre in Mo is interesting. I recommend the\nfollowing paper for its interesting discussion on how it is the gap between Mo\nand S states in MoS2 that determines redox activity for hydrogen evolution,\nrather than the d-band centre: https://doi.org/10.1002/anie.202003091. Could\nthe authors comment on whether they think a similar analysis would be of\nbenefit here?\nResponse: Thanks for your professional advice. We sincerely appreciate your\nrecommendation of this important paper. We have thoroughly read this\nliterature. Liu and coworkers proposed a hydrogen-binding mechanism for\nbasal-plane sulfur atoms in MoS -based materials and validated the model\n2\nusing extensive DFT calculations. The activity of an HER catalyst can be\nstudied through its binding energy of hydrogen atoms. Their proposed model\nwas based on the electronic promotion energy required to partially open the\nfull valence shell of a local S atom and therefore enable it to bond with a H\natom. They hypothesized that the two-step H-S binding process introduced.\nThe first step is an electronic-promotion process, where an electron in the\nvalence shell of the S atom is pushed back to Mo atoms, thus leaving the S\natom with an open valence shell. The valence shell of this S atom will be filled\nagain after it forms a single bond with the H atom in the second step. The extra\nelectron remains localized on nearby Mo sites. They have demonstrated a\nspecific, electronic-structure based descriptor for the hydrogen-binding\nstrength: Δ , the local interband energy separation between the lowest empty\ndp\nd-states on the dopant metal atoms and occupied p-states on S. They used\nthis model to explore the degree of S activation for hydrogen binding across\nnearly 30 different 3d-metal-doped MoS systems, confirming that the\n2\nproposed descriptor, Δ , was quantitatively effective in predicting the\ndp\nhydrogen binding energy. In this paper, the author demonstrated the\nrelationship between hydrogen-binding strength and HER activity of catalysts.\nTaking the POD-like catalytic process of nanozymes as an example, H O was\n2 2\ninitially adsorbed on top of the metal site within nanozyme (Nat Catal 2021, 4,\n407; Angew. Chem. Int. Ed. 2025, 64, e202505742). Subsequently, H O was\n2 2\ncleaved heterogeneously on the metal site, generating an H O molecule and\n2 2\nH O). Immediately, H O departure occurred to give free H O and hydroxyl\n2 2 2\nadsorbed on the metal site (*OH). The *OH species were subjected to further\nprotonation, forming H O adsorbed on the metal site (*H O). Finally, the entire\n2 2\ncatalytic reaction was completed with H O removal and metal-site\n2\nregeneration. The proposed reaction pathways were shown in Figure R4.\nFigure R4. Schematic illustration of the proposed reaction pathways for FeSA during the\nPOD-mimicking process (Angew. Chem. Int. Ed. 2025, 64, e202505742).\nIn fact, the d-band center in our study was employed to describe the interaction\nbetween the adsorbed substrate and LM MoS nanozyme. Although different\nX\nsubstrates and catalytic mechanisms are involved, both the d-band center and\nthe above-mentioned descriptor Δ are used to describe the interaction\ndp\nbetween substrates and catalysts. However, Δ is a highly localized\ndp\nparameter specific to a particular atom pair, which represents the inter-band\nenergy separation between occupied S p-states and unoccupied neighboring\nmetal d-states. The activated basal plane S atoms in transition-metal-doped\nMoS are the main active sites for hydrogen evolution reaction, while the metal\n2\natoms are employed to accept electrons from the valence shell of the S atom.\nIn contrast, the metal sites are the main active sites for the adsorption and\nactivation of substrates in the catalytic process of nanozyme. Notably, the\nneighboring sulfur atoms and metal atoms can influence the electronic\nproperties of metal active sites, thereby affecting their d band center. Therefore,\nwe speculate the similar analysis of Δ may have beneficial effect on the\ndp\ncatalytic activity of nanozyme. However, considering the complicated\nstructures and catalytic mechanism of nanozymes, theformula for Δ should\ndp\nbe carefully redefined according to different catalytic systems. For example, it\nis necessary to carefully consider the active sites of catalysts, the electronic\nproperties of dopants, the overall catalytic mechanism, and the electron\ntransfer direction.\nAs mentioned by the reviewer, it is imperative to develop activity descriptors for\nthe rational design of nanozymes. In fact, researchers have made significant\nefforts in this regard. Guo and coworkers have reported a strategy for precisely\ntuning the d-band center of dual-atomic sites to enhance the affinity of metal\natomic sites and substrates on a class of edge-rich N-doped porous carbon\ndual-atomic sites Fe-Mn for greatly boosting multiple-enzyme-like catalytic\nactivities (J. Am. Chem. Soc. 2024, 146, 10023-10031). Tang’s and Zhang’s\ngroup also reported that tuning the d-band center position of metal sites could\nimprove the adsorption and activation of substrates, thereby further optimizing\nthe catalytic activity of nanozyme (Angew. Chem. Int. Ed. 2025, 64,\ne202414625; Adv. Mater. 2024, 36, 2412368). These studies indicated that the\nd band center correlated with the catalytic activity of nanozymes. In addition,\nWei’s group has successively proposed several activity descriptors to predict\nthe peroxidase-like activities of nanozymes. They have investigated the\noccupancy of e orbitals as a possible descriptor for the peroxidase-like activity\ng\nof transition metal oxide (including perovskite oxide) nanozymes (Nat Commun\n2019, 10, 704). They also used the descriptor hydroxyl adsorption energy\n(E ,OH) to predict the POD activities of the nanosurfaces of nonnoble metals,\nads\nnoble metals, monometallic oxides, perovskites, and carbon materials (ACS\nCatal. 2020, 10, 12657). Recently, they identified the effectiveness and\npredictive capacity of t occupancy as a descriptor of spinel oxide nanozymes\n2\nfor the tetrahedral site (Angew. Chem. Int. Ed. 2025, 64, e202421790). These\nstudies have significant impact on the rational design of catalysts. Finally, we\nsincerely thank the reviewer for recommending this valuable literature, which\nhas given us great inspiration. We will pay close attention to the future\ndevelopments in this area.\nMinor corrections:\n1. In the introduction the authors begin to use abbreviations without defining\nthem in the text. (e.g. POD-like activities, LM, OXD- and NOX-like are defined\nin the abstract but not the main text). These should be defined in their first\nmention in the main text.\nResponse: Thanks for the reviewer’s kindly suggestions. We have defined\nthese abbreviations in the abstract and main text, and re-examined the whole\nmanuscript to avoid similar problems.\n“We observe that these nanozymes exhibit peroxidase (POD)-like activities.”\n“this strategy can turn on the oxidase (OXD)-like and nicotinamide adenine\ndinucleotide oxidase (NOX)-like activities of LM MoS to achieve cascade\nX\ncatalytic reactions, eventually leading to reactive oxygen species (ROS) burst.”\n2. Should have \"an\" added in two places in the following sentence: \"Here we\ndesign and screen the liquid metals-buffered nanozymes, in which the liquid\nmetals can not only initiate phase engineering to generate AN amorphous\nnanozyme with abundant catalytic active sites, but also afford AN electron-rich\nmicroenvironment to optimize the electronic structure of nanozymes for\nimproving the catalytic performance (Fig. 1). \"\nResponse: Thanks for your careful and important advice. We have added “an”\nin these sentences and similar places. This is a great suggestion for\nimprovement.\n“Here we design and screen the liquid metals-buffered nanozymes, in which\nthe liquid metals can not only initiate phase engineering to generate an\namorphous nanozyme with abundant catalytic active sites, but also afford an\nelectron-rich microenvironment to optimize the electronic structure of\nnanozymes for improving the catalytic performance.”\n“(ii) initiating phase engineering to generate an amorphous nanozyme with\nabundant catalytic active sites”\n3. \"... free delocalized electrons cloud\" should likely read \"... free delocalized\nelectron clouds\"\nResponse: Thanks for your correction. We have changed this expression and\nre-examine the whole article to revise similar mistake.\n“The most striking features of liquid metals are the amorphous property and\nunusual electronic microenvironment, which originate from the electron-rich\nmetallic cores and free delocalized electron clouds.”"}, {"round": 2, "reviewer_comments": {"reviewer_1": "All my questions are answered.", "reviewer_2": "After the revision, the quality of the manuscript has significantly improved, and it has largely resolved most of my concerns.\nSeveral minors still need amendment.\n1. Applying DLS and SEM measurements can only demonstrate the dimensional stability rather than the composition\nstability. For instance, the low Mo leakage rate cannot guarantee the absence of LM-based phase separation. Therefore,\ncharacterizations such as TEM or XPS are recommended to verify the stability of LM-based nanozymes.\n2. The response to excellent tumor penetration of LM-MoSX nanozyme (ca. 300 nm) is not convincing. Referring to Fig. S38,\nthe fluorescence image showed that the LM-MoSX@Cy3 was located in the center of the liver rather than at the boundary\n(what if Cy3 leakage occurs), so the 3D reconstruction image in IVIS should be provided. As Reviewer 3 suggested, an\ninvestigation into the correlation between size and penetrability LM-MoSX nanozyme could help. Moreover, I wonder if other\nimaging techniques (e.g., CT, MRI or PET) could be employed to trace the real-time and dynamic distributions of LM-MoSX\nnanozyme.\n3. I strongly agree with Reviewer 4 that whether the models used in theoretical calculations are representative or accurate\nfor LM-based nanoparticles, since they are hybrid nanomaterials (with varying and dynamic compositions) rather than\nmonodispersed nanomaterials reported previously, such as metal oxides and single-atom catalysts.", "reviewer_3": "The revision and point-by-point responses are clear and complete. The manuscript now addresses all of my earlier\nconcerns, including size considerations, positioning relative to prior LM-based nanozyme studies, mechanistic rationale for\nthe Ga/In ratio, definition of terms and abbreviations, and missing figure legends. I have no additional comments and\nrecommend acceptance.", "reviewer_4": "I have previously reviewed this manuscript, and left several substantive comments on the computational section in particular.\nIt is my belief that the authors have now satisfactorily addressed the methodological questions which I raised. The changes\nmade to the manuscript significantly enhance its clarity. Therefore, I can now recommend the manuscript for publication."}, "author_response": "Next is the summarization of our point-by-point response to the comments raised by\nthe reviewers.\nReviewer #1 (Remarks to the Author):\nAll my questions are answered.\nResponse: We sincerely thank the reviewer for the support of publication and the\ncomments are really helpful to improve the quality of this article.\nReviewer #2 (Remarks to the Author):\nAfter the revision, the quality of the manuscript has significantly improved, and it has\nlargely resolved most of my concerns. Several minors still need amendment.\n1. Applying DLS and SEM measurements can only demonstrate the dimensional\nstability rather than the composition stability. For instance, the low Mo leakage\nrate cannot guarantee the absence of LM-based phase separation. Therefore,\ncharacterizations such as TEM or XPS are recommended to verify the stability of\nLM-based nanozymes.\nResponse: Thank you for your professional suggestion. Your insightful suggestion\nwas very helpful for improving our study. We have supplemented the TEM images\nand XPS spectra to verify the stability of LM MoS nanozymes. As shown in\nX\nSupplementary Figure 5c-f, the LM MoS nanozymes remained stable in PBS over 7\nX\ndays. We have added the following figures and corresponding description into the\nrevised manuscript.\nSupplementary Figure 5. The stability study of LM MoS . XPS spectra and TEM\nX\nimages (c-f) of LM MoS after incubation with PBS for various times.\nX\nIn the revised manuscript, we added:\nMoreover, the DLS results, SEM and TEM images, and XPS spectra suggested that\nthe LM MoS nanozymes remained monodisperse and stable over 7 days\nX\n(Supplementary Fig. 5).\n2. The response to excellent tumor penetration of LM-MoSX nanozyme (ca. 300 nm)\nis not convincing. Referring to Fig. S38, the fluorescence image showed that the\nLM-MoSX@Cy3 was located in the center of the liver rather than at the boundary\n(what if Cy3 leakage occurs), so the 3D reconstruction image in IVIS should be\nprovided. As Reviewer 3 suggested, an investigation into the correlation between size\nand penetrability LM-MoSX nanozyme could help. Moreover, I wonder if other\nimaging techniques (e.g., CT, MRI or PET) could be employed to trace the real-time\nand dynamic distributions of LM-MoSX nanozyme.\nResponse: Thank you for your valuable advice. Accordingly, next is our\npoint-by-point response.\n(1) We fully understand the reviewer’s concerns. In Supplementary Figure 38,\nfluorescence image showed that the LM-MoS @Cy3 was located in the center of\nX\nthe liver rather than at the boundary. These images were obtained through ex vivo\nfluorescence imaging system. This time, the reviewer suggested providing the 3D\nreconstruction image in IVIS. However, this system has yet to add the 3D\nscanning function into their apps. So, it is difficult to provide the 3D\nreconstruction image in IVIS. However, the fluorescence signals in the figure\nwere shown as relative intensity. Therefore, the fluorescence intensity range was\nrefined, and the resulting images were provided in Figure R1. The fluorescence\nsignals were actually distributed throughout the liver, with relatively higher\nintensity in some areas. Notably, several previous reports have also described\nsimilar phenomena that the nanoparticles tend to accumulate more in the central\nregions of the liver (Figure R2-R5) (Nat. Biomed. Eng 2023, 7, 298; Nat.\nNanotechnol. 2024, 19, 1723; Nat. Commun. 2024, 15, 7824; Nat. Nanotechnol.\n2025, 20, 1131). Thanks for pointing out this important issue, and we hope that\nthese explanations have addressed your concern.\nFigure R1. In vivo biodistribution of H-LM MoS in different intensity range.\nX\nFigure R2. Ex vivo fluorescence images of tumors and main organs at 48 h post\ninjection of SNAP-M (Nat. Biomed. Eng 2023, 7, 298).\nFigure R3. Ex vivo NIRF images of tumors and major organs resected at 8 and 24 h\npost-injection of P-ZWNIR NPs into orthotopic HT-29 tumor-bearing mice.\nAbbreviations: He, heart; Lu, lungs; Li, liver; Sp, spleen; Ki, kidney; and Tu, tumor\n(Nat. Nanotechnol. 2024, 19, 1723).\nFigure R4. Images of major organs from 4T1 tumour-bearing mice at 48 h\npost-injection of iP5B5 ICG NPs (Nat. Commun. 2024, 15, 7824).\nFigure R5. In vivo biodistribution of DV1@O-Colase nanogels detected by IVIS\n(Nat. Nanotechnol. 2025, 20, 1131).\n(2) As for the correlation between size and penetrability of nanoparticles, researchers\nhave investigated the accumulation and effectiveness of different sizes of\nnanoparticles in tumors. For example, Yuan and coworkers found that tumor vessels\nare less permselective than normal vessels, presumably due to large pores in the\nvessel wall (Cancer Res. 1995, 55, 3752). They showed that tumor vessels were\npermeable to liposomes of up to 400 nm in diameter, suggesting that the cutoff size of\nthe pores is between 400 and 600 nm in diameter. Bawendi and coworkers developed\na size series of luminescent, water-soluble particles within the 10-150 nm size range\nfor the purpose of studying the size-dependent properties of particle distributions in\nsolid tumors (Angew. Chem. Int. Ed. 2010, 49: 8649-8652). They found that the 12 nm\nnanoparticles extravasate easily, though heterogeneously, and diffuse away from the\nvessels with minimal hindrance. In contrast, the 60 nm nanoparticles do extravasate\nbut do not leave the immediate perivascular space (they remain within 10 mm of the\nvessel walls), whilst the 125 nm particles do not appreciably extravasate. Moreover,\nKataoka and coworkers have demonstrated that a variety of sub-100 nm polymer\nmicelles of different sizes (30 nm, 50 nm, 70 nm and 100 nm) penetrate well within\nhighly permeable tumors (Nat. Nanotechnol. 2011, 6, 815). However, in a poorly\npermeable human pancreatic adenocarcinoma, characterized by low vascularity and\ndense fibrosis, only small size nanoparticles <50 nm in diameter are able to\naccumulate in tumors. The extent of nanoparticle accumulation also varies depending\non tumor type due to the interplay of various microenvironmental factors (Nat.\nBiotechnol. 2015, 33, 941). In general, nanoparticles with large particle size tend to\nhave elevated blood circulation time and prefer to accumulate surrounding tumor\ntissues through enhanced permeability and retention (EPR) phenomenon.\nComparatively, small-sized nanoparticles are more advantageous in penetrating into\ndeep tumor tissues, yet are frequently restricted by their rapid clearance from the body\nand insufficient accumulation within tumor region (ACS Nano 2023, 17, 20825).\nApart from these, recent studies showed that nanoparticles enter the tumor through\nactive endothelial transport processes, are retained in the tumor due to interactions\nwith tumor components (Nat. Rev. Bioeng. 2024, 2, 201). Researchers found that\nnanoparticles cross tumor endothelial cells predominantly using the\nnon-receptor-based macropinocytosis pathway. They discovered that tumor\nendothelial cell membrane ruffles capture circulating nanoparticles, internalize them\nin intracellular vesicles and release them into the tumor interstitium. Tumor\nendothelial cells have a higher membrane ruffle density than healthy endothelium,\nwhich may partially explain the elevated nanoparticle tumor accumulation. They\nfound the nanoparticle-releasing exocytosis pockets ranged from 89 nm to 841 nm in\ndiameter, suggesting that nanoparticles in this size range could accumulate more in\nthe tumor than in other non-MPS (mononuclear phagocytic system) organs through\nthis pathway. Moreover, this pathway has been verified in 4T1 tumor-bearing mice\n(Nat. Nanotechnol. 2025, 20, 672). Therefore, we chose nanoparticles with relatively\nsmaller sizes to achieve better tumor therapeutic effect in our work.\n(3) Gallium-based liquid metal particles have been used in various therapeutic\napplications, including X-ray computerized tomography (CT) imaging, photoacoustic\n(PA) imaging, and MRI (Trends Biotechnol. 2021, 39, 624). Because of the high\ndensity of metal Ga (6.08 kg/m3), Ga particles are radiopaque to some extent for\nsimultaneously serving as contrast agents under X rays (Matter 2022, 5, 219; Adv.\nMater. 2023, 35, 2203391). Moreover, liquid metal exhibits the large absorption at\nNIR wavelength and a high coefficient of thermal expansion, so it can be used as a\nhigh-quality contrast agent for photoacoustic (PA) imaging (Adv. Mater. 2022, 34,\n2201956; J. Am. Chem. Soc. 2022, 144, 6779). For example, Liu and coworkers\nreported that gallium microparticles are demonstrated to enhance the T2 magnetic\nresonance imaging (MRI) effect and mediate the dual-mode imaging of CT and MRI\n(Adv. Funct. Mater. 2020, 30, 2003359). Miyako and coworkers reported liquid metal\nnanocapsules could be used for photoacoustic and X-ray imaging (Nat Commun 2017,\n8, 15432). In addition, Mo-based nanomaterials can be applied to contrast agents in\nCT imaging because Mo atom with high absorption coefficient and atomic number\nhas stronger X-ray attenuation than the components of the body (ACS Nano 2014, 8, 7,\n6922; Adv. Sci. 2021, 8, 2100165). Furthermore, molybdenum sulfide nanoparticles\nhave outstanding photothermal conversion efficiency and excellent NIR absorption\ncapacity, which can be used for high-quality PA imaging (Theranostics, 2015, 5, 931,\nAdv. Sci. 2021, 8, 2100165). Therefore, the designed LM-MoS nanozymes are\nX\nexpected to achieve CT, MRI or PA imaging to trace their real-time and dynamic\ndistributions. However, due to the limited experimental conditions, the real-time and\ndynamic distributions of LM-MoS nanozyme could not be systematically\nX\ninvestigated through CT, MRI and PET. We thank the reviewer for this insightful and\nconstructive suggestion, in order to eliminate reviewer’s concerns as well as get a\nconvincing demonstration, we evaluated the tumor-targeting capability of H-LM\nMoS through in vivo fluorescence imaging system. As shown in Supplementary\nX\nFigure 40, strong red fluorescence signals were observed within the tumor, confirming\nthe effective tumor accumulation of nanoparticles. We have added the following\nfigure and corresponding description into the revised manuscript.\nIn the revised manuscript, we added:\nThe tumor-targeting capability of H-LM MoS was also evaluated through in vivo\nX\nfluorescence imaging system. As shown in Supplementary Figure 40, strong red\nfluorescence signals were observed within the tumor, confirming the effective tumor\naccumulation of nanoparticles.\nSupplementary Figure 40. In vivo fluorescence images of tumor-bearing mice after\nbeing injected with Cy5-labeled H-LM MoS .\nX\n3. I strongly agree with Reviewer 4 that whether the models used in theoretical\ncalculations are representative or accurate for LM-based nanoparticles, since they are\nhybrid nanomaterials (with varying and dynamic compositions) rather than\nmonodispersed nanomaterials reported previously, such as metal oxides and\nsingle-atom catalysts.\nResponse: Thank you for your important question. The LM/MoS structural model\nX\nwas obtained by annealing through ab initio molecular dynamics (AIMD) calculation\nmethod. AIMD is a powerful dynamic sampling tool that integrates first-principles\naccuracy with the time-evolution capability of molecular dynamics and dynamically\ncouples atomic motion with electronic structure calculations based on quantum\nmechanics. When using AIMD modeling, it is necessary to establish an appropriate\ninitial model. After the initial model is created, the system is heated to a\nhigh-temperature state to provide atoms with sufficient kinetic energy for extensive\nmovement and diffusion within the system. Once thermal equilibrium is reached,\nrapid cooling is initiated. These processes require enough time to obtain optimized\nstructural information. In our work, the canonical (NVT) ensemble and Nosé-Hoover\nthermostats were used with the temperature set to 2000 K and maintained over 10 ps\nwhile the time step was 1 fs, and finally the temperature was returned to 298.15 K.\nActually, the AIMD method is usually used in the simulation of the liquid metal-based\nstructures (Nat. Chem. 2022, 14, 935; Angew. Chem. Int. Ed. 2023, 62, e202219009;\nNat. Nanotechnol. 2024, 19, 306). Arifur Rahim and coworkers employ AIMD\nsimulations to obtain atomistic insights into the configuration of Pt in Ga in the bulk\nand at an interface (Nat. Chem. 2022, 14, 935). Lambie and coworkers employ AIMD\nsimulations to probe the dynamics, initially, of GaPt systems in isolation and then in\nthe presence of adsorbates (Angew. Chem. Int. Ed. 2023, 62, e202219009). Tang and\ncoworkers employ AIMD simulations to investigate the status of the dispersed atoms\nin the Ga matrix (Nat. Nanotechnol. 2024, 19, 306).\nThese studies demonstrate the effectiveness of AIMD simulations for constructing\nliquid metal-based hybrid models, and Reviewer 4 agreed with our last round\nresponse. Thank you again for the support and valuable comments on our manuscript.\nReviewer #3 (Remarks to the Author):\nThe revision and point-by-point responses are clear and complete. The manuscript\nnow addresses all of my earlier concerns, including size considerations, positioning\nrelative to prior LM-based nanozyme studies, mechanistic rationale for the Ga/In ratio,\ndefinition of terms and abbreviations, and missing figure legends. I have no additional\ncomments and recommend acceptance.\nResponse: We sincerely thank the reviewer for the support of publication and the\ncomments are really helpful to improve the quality of this article.\nReviewer #4 (Remarks to the Author):\nI have previously reviewed this manuscript, and left several substantive comments on\nthe computational section in particular. It is my belief that the authors have now\nsatisfactorily addressed the methodological questions which I raised. The changes\nmade to the manuscript significantly enhance its clarity. Therefore, I can now\nrecommend the manuscript for publication.\nResponse: We are grateful to the reviewer’s comments and really appreciate the\nagreement of publication.\nNext is the summarization of our point-by-point response to the comments raised by\nthe reviewer.\nReviewer #2 (Remarks to the Author):\nThe authors have well addressed my concerns, and the manuscript can be accepted now.\nResponse: We sincerely thank the reviewer for the support of publication and the\ncomments are very helpful to improve the quality of the manuscript."}, {"round": 3, "reviewer_comments": {"reviewer_2": "The authors have well addressed my concerns,and the manuscript can be accepted now."}, "author_response": "Response: We sincerely thank the reviewer for the support of publication and the\ncomments are very helpful to improve the quality of the manuscript."}]

Decision letter