# Sm:HAp Nanopowders Present Antibacterial Activity againstEnterococcus faecalis

Authors: Carmen Steluta Ciobanu; Cristina Liana Popa; Daniela Predoi
Journal: Journal of Nanomaterials
 (2014)

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## Abstract

The synthesis of nanoparticles with inhibitory and bactericidal effects represents a great interest in development of new materials for biological applications. In this paper we present for the first time the synthesis ofCa10-xSmx(PO4)6(OH)2 nanoparticles at low temperature and primary tests concerning the adherence of Enterococcus faecalis ATCC 29212 (gram-positive bacteria). All the XRD peaks were indexed in accordance with the hexagonal HAp in P63m space group. The EDAX spectrum and elemental mapping of O, P, Ca, and Sm demonstrate that all the elements were homogeneously distributed in Ca10-xSmx(PO4)6(OH)2 with xSm=0.03. The peaks at 347.3 eV, 532.1 eV, and 133.8 eV in the XPS spectra can be attributed to the binding energy of Ca 2p, O 1s, and P 2p. The peak at 1084.4 eV observed in Ca10-xSmx(PO4)6(OH)2 was attributed to the Sm 3d5/2. Bacterial adhesion was reduced on Ca10-xSmx(PO4)6(OH)2 sample when compared to pure HAp (xSm=0) and significant differences in bacterial adhesion on pure HAp (x=0) and Sm:HAp (xSm=0.01,xSm=0.03,andxSm=0.1) were observed. The bacterial adhesion decreased when the samarium concentrations increased. Finally, we demonstrate that the Sm:HAp nanopowder with xSm>0 showed high antibacterial activity against Enterococcus faecalis ATCC 29212.


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## Body

## 1. Introduction

In the past decades scientists worldwide have tried to find new solutions for improving treatments used for different bone related diseases and injuries. Their attention has been focused to the field of biomaterials in order to create and develop new and improved materials for tissue engineering [1–3]. Among these materials, hydroxyapatite (HAp) is one of the most commonly used for bone implants [4–9] or it is a drug release system [4, 10–15]. Moreover, it has been shown that it is also a good candidate for the role of adsorbent in column chromatography, in order to separate and purify nucleic acids and proteins [3, 16, 17]. It is possible to use hydroxyapatite for a large number of applications in the biomedical field due to its remarkable properties, such as biocompatibility, bioactivity, and osteoconductivity [4, 18–21]. In live tissue, HAp is the major mineral component of human bones and teeth [4, 22, 23]. Various factors, such as grain size, morphology, surface area, and microporosity [24–26], must be taken into account when developing the biomaterial. Therefore, in order to improve the properties of HAp (for its use in different bone tissue applications), researchers have developed various methods of obtaining new compounds based on HAp doped with different rare-earth (RE) ions [24, 27]. The choice for lanthanides as doping agents has been made due to their high bioactivity and their ability to substitute calcium ions in structured molecules [24, 28–30]. It has been reported that lanthanides have been previously used in the biomedical field as contrast agents for magnetic resonance imaging and as luminescent probes for biosensors used for variousin vivo imaging applications [24, 28, 29, 31], due to their narrow emission bands and long emission lifetime [24, 32, 33]. Among the rare-earth elements, samarium (Sm) is a good candidate for different radiation therapies used for cancer treatments. For more than a decade, 153Sm isotopes have been used for cancer radiation treatments [4, 34, 35] which are now administered clinically to patients that have symptomatic bone metastases from different types of cancer, mostly prostatic and breast carcinoma, for pain alleviation [4, 34–37]. In addition, different new studies have used 153Sm-hydroxyapatite particles as radiation synovectomy agent in the treatment of chronic synovitis [38–41], especially knee synovitis [38, 39]. Radiation synovectomy is a method used to alleviate the pain of patients suffering from swelling of rheumatoid arthritis [42]. Furthermore, another major problem encountered in the medical field is the inflammation of the synovial membrane caused by prosthetic joint infections (PJI) [43] which could also be treated by radiation synovectomy. The procedure involves an intra-articular injection of the beta-emitting radiopharmaceutical into the joint space. Once the radionucleide decays, the regenerating synovium starts to be irradiated [41]. Therefore, the 153Sm-hydroxyapatite particles are better candidates for this type of treatment than other compounds based on Y-90 due to the shorter half-life of 153Sm and the lower extra-articular leakage of 153Sm-hydroxyapatite [39]. Furthermore, it has been shown that 153Sm emits β rays with a small tissue penetration (0.8 mm), more that 99% of the 153Sm being bound to the hydroxyapatitein vivo [44]. According to a clinical study made by Brazilian researchers after one year of treatment based on 153Sm-hydroxyapatite on patients suffering from haemophilic arthropathy, it was established that this treatment is effective for several types of joints. They obtained good results for 75% of elbows, 87.5% of ankles, and 40% of knees [45].The prior studies have focused on preparation and characterization of complexes of europium (III) metal ion with organic reagents [46]. On the other hand, previous studies concerning the antibacterial and antifungal activity of ligand and its metal complexes tested against gram-positive and gram-negative bacteria, yeast, and fungi show that the complexes are more potent antimicrobials than the parent ligand [47–49].The studies on the preparation, physicochemical characterization, and biological properties of the samarium doped hydroxyapatite (Sm:HAp) powders are absent in the literature. In this paper we present for the first time a method to synthesize the nanocrystalline hydroxyapatite doped with samarium at low temperature. Physicochemical characterization and antimicrobial studies on the Sm:HAp nanopowder are also presented for the first time. The results obtained suggest that the new obtained material,153Sm-hydroxyapatite, could be a good candidate for treating the inflamed and damaged synovial membrane of the joint encountered in the case of prosthetic joint infections.TheCa10-xSmx(PO4)6(OH)2 powders were synthesized by coprecipitation method at low temperature, mixing Sm(NO3)3·6H2O, Ca(NO3)2·4H2O, and (NH4)2HPO4 in deionized water. The structure, morphology, and vibrational properties of the obtained samples were studied by X-ray diffraction (XRD), scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FT-IR). In order to reveal the presence of the samarium in the Sm:HAp nanopowder with xSm=0.01, 0.03, and 0.1, the X-ray photoelectron spectroscopy (XPS) results are also presented. In addition, the preliminary antibacterial activity of the Ca10-xSmx(PO4)6(OH)2 nanopowder with xSm=0.01, 0.03, and 0.1 is reported.


## 2. Experimental Section

### 2.1. Sample Preparation

All the reagents used for the synthesis, including ammonium dihydrogen phosphate [(NH4)2HPO4], calcium nitrate tetrahydrate [Ca(NO3)2·4H2O], and Samarium(III) nitrate hexahydrate Sm(NO3)3·6H2O (Sigma-Aldrich), were purchased without further purification.Nanocrystalline hydroxyapatite doped with Sm, (Ca10-xSmx(PO4)6(OH)2, with xSm=0 to xSm=0.1 was obtained by setting the atomic ratio of Sm/[Sm+Ca] from 0% to 10% and [Ca+Sm]/P as 1.67. The Sm(NO3)3·6H2O and Ca(NO3)2·4H2O were dissolved in deionised water to obtain 300 mL [Ca+Sm] containing solution. On the other hand, the (NH4)2HPO4 was dissolved in deionised water to make 300 mL P-containing solution. The [Ca+Sm] containing solution was put into a Berzelius and stirred at 100°C for 30 minutes. Meanwhile, the pH of P-containing solution was adjusted to 10 with NH3 and stirred continuously for 30 minutes. The P-containing solution was added drop by drop into the [Ca+Sm] containing solution and stirred for 2 h, the pH being constantly adjusted and kept at 10. After the reaction, the deposited mixtures were washed several times with deionised water. The resulting material (Sm:HAp) was dried at 100°C. The scheme of the present study is presented in Figure 1.Figure 1
Schematic representation of preparation and evaluation of new antimicrobial Sm:HAp ceramics.


### 2.2. Sample Characterization

The X-ray diffraction measurements ofCa10-xSmx(PO4)6(OH)2 samples were recorded using a Bruker D8 Advance diffractometer, with nickel filtered Cu Kα (λ=1.5418Å) radiation and a high efficiency one-dimensional detector (Lynx Eye type) operated in integration mode. The diffraction patterns were collected in the 2θ range 15°–140°, with a step of 0.02° and 34 s measuring time per step.The scanning electron microscopy (SEM) study was performed on a HITACHI S2600N-type microscope equipped with an energy dispersive X-ray attachment (EDAX/2001 device). Energy dispersive X-ray analysis, referred to as EDS or EDAX, was used to identify the elemental composition of materials.The functional groups present in the prepared nanoparticles and thin films were identified by FTIR using a Spectrum BX spectrometer. In order to obtain the nanoparticles spectra, 1% of nanopowder was mixed and ground with 99% KBr. Tablets of 10 mm diameter were prepared by pressing the powder mixture at a load of 5 tons for 2 min. The spectrum was taken in the range of 500 to 4000 cm−1 with 4 cm−1 resolution.X-ray photoelectron spectroscopy (XPS) studies were conducted using a VG ESCA 3 MK II XPS installation (Ekα = 1486.7 eV). The vacuum analysis chamber pressure was P~ 3 × 10−8 torr. The XPS recorded spectrum involved an energy window w=20 eV with the resolution R = 50 eV with 256 recording channels. The XPS spectra were processed using Spectral Data Processor v 2.3 (SDP) software.In order to test thein vitro antibacterial activity, the strains of bacteria used wereEnterococcus faecalis ATCC 29212. The bacteria were grown overnight in Todd-Hewit broth supplemented with 1% yeast extract at 37°C, followed by centrifuging. The supernatants were discarded and pellets were resuspended in phosphate-buffered saline (PBS) followed by a second centrifuging and resuspension in PBS. The samples to be tested were placed in 50 mL sterilized tubs followed by the addition of 2 mL of the bacterial suspension. The tubes were incubated at 3°C for 4 h. At the end of the incubation period, the samples were gently rinsed three times with PBS. The nonadherent bacteria were eliminated. After washing, the samples were then put into a new tube containing 5 mL PBS and vigorously vortexed for 30 s to remove the adhering microorganisms. The viable organisms in the buffer were quantified by plating serial dilutions on yeast extract agar plates. Yeast extract agar plates were incubated for 24 h at 3°C and the colony forming units were counted visually.


## 2.1. Sample Preparation

All the reagents used for the synthesis, including ammonium dihydrogen phosphate [(NH4)2HPO4], calcium nitrate tetrahydrate [Ca(NO3)2·4H2O], and Samarium(III) nitrate hexahydrate Sm(NO3)3·6H2O (Sigma-Aldrich), were purchased without further purification.Nanocrystalline hydroxyapatite doped with Sm, (Ca10-xSmx(PO4)6(OH)2, with xSm=0 to xSm=0.1 was obtained by setting the atomic ratio of Sm/[Sm+Ca] from 0% to 10% and [Ca+Sm]/P as 1.67. The Sm(NO3)3·6H2O and Ca(NO3)2·4H2O were dissolved in deionised water to obtain 300 mL [Ca+Sm] containing solution. On the other hand, the (NH4)2HPO4 was dissolved in deionised water to make 300 mL P-containing solution. The [Ca+Sm] containing solution was put into a Berzelius and stirred at 100°C for 30 minutes. Meanwhile, the pH of P-containing solution was adjusted to 10 with NH3 and stirred continuously for 30 minutes. The P-containing solution was added drop by drop into the [Ca+Sm] containing solution and stirred for 2 h, the pH being constantly adjusted and kept at 10. After the reaction, the deposited mixtures were washed several times with deionised water. The resulting material (Sm:HAp) was dried at 100°C. The scheme of the present study is presented in Figure 1.Figure 1
Schematic representation of preparation and evaluation of new antimicrobial Sm:HAp ceramics.


## 2.2. Sample Characterization

The X-ray diffraction measurements ofCa10-xSmx(PO4)6(OH)2 samples were recorded using a Bruker D8 Advance diffractometer, with nickel filtered Cu Kα (λ=1.5418Å) radiation and a high efficiency one-dimensional detector (Lynx Eye type) operated in integration mode. The diffraction patterns were collected in the 2θ range 15°–140°, with a step of 0.02° and 34 s measuring time per step.The scanning electron microscopy (SEM) study was performed on a HITACHI S2600N-type microscope equipped with an energy dispersive X-ray attachment (EDAX/2001 device). Energy dispersive X-ray analysis, referred to as EDS or EDAX, was used to identify the elemental composition of materials.The functional groups present in the prepared nanoparticles and thin films were identified by FTIR using a Spectrum BX spectrometer. In order to obtain the nanoparticles spectra, 1% of nanopowder was mixed and ground with 99% KBr. Tablets of 10 mm diameter were prepared by pressing the powder mixture at a load of 5 tons for 2 min. The spectrum was taken in the range of 500 to 4000 cm−1 with 4 cm−1 resolution.X-ray photoelectron spectroscopy (XPS) studies were conducted using a VG ESCA 3 MK II XPS installation (Ekα = 1486.7 eV). The vacuum analysis chamber pressure was P~ 3 × 10−8 torr. The XPS recorded spectrum involved an energy window w=20 eV with the resolution R = 50 eV with 256 recording channels. The XPS spectra were processed using Spectral Data Processor v 2.3 (SDP) software.In order to test thein vitro antibacterial activity, the strains of bacteria used wereEnterococcus faecalis ATCC 29212. The bacteria were grown overnight in Todd-Hewit broth supplemented with 1% yeast extract at 37°C, followed by centrifuging. The supernatants were discarded and pellets were resuspended in phosphate-buffered saline (PBS) followed by a second centrifuging and resuspension in PBS. The samples to be tested were placed in 50 mL sterilized tubs followed by the addition of 2 mL of the bacterial suspension. The tubes were incubated at 3°C for 4 h. At the end of the incubation period, the samples were gently rinsed three times with PBS. The nonadherent bacteria were eliminated. After washing, the samples were then put into a new tube containing 5 mL PBS and vigorously vortexed for 30 s to remove the adhering microorganisms. The viable organisms in the buffer were quantified by plating serial dilutions on yeast extract agar plates. Yeast extract agar plates were incubated for 24 h at 3°C and the colony forming units were counted visually.


## 3. Results and Discussions

In Figure2 the XRD patterns of the synthesized Ca10-xSmx(PO4)6(OH)2 nanopowders with xSm=0.01, 0.03, and 0.1 are shown. Good pattern fit of the synthetized Sm:HAp structures was determined using the Rietveld method implemented in the software package Materials Analysis Using Diffraction (MAUD) [50] by applying the Popa approach for the anisotropic microstructure analysis [51] implemented in the MAUD as “Popa rules.”Figure 2
XRD patterns ofCa10-xSmx(PO4)6(OH)2 nanopowders with 0.01 ≤ xSm ≤ 0.1.The XRD patterns of the Sm:HAp nanopowders withxSm=0.01, 0.03, and 0.1 appeared to be identical, regardless of the Sm content in the samples. Sm doped hydroxyapatite exerted no effect on the phase composition of pure HAp. No secondary phases than those associated with HAp were detected. All the XRD peaks were indexed in accordance with the hexagonal HAp in P63m space group (JCPDS card number 9-432). On the other hand, no change took place in the symmetry of the HAp crystallographic unit cell.In the XRD spectra of the Sm:HAp powders, the diffraction peak intensities were reduced and the peak shapes were broadened when the Sm concentration increased. The broadened peaks when Sm concentration increased in the Sm:HAp samples are attributed to the small grain size. However, small variations of lattice constants and crystallite size were observed when the samarium content increases as it can be seen in Table1.Table 1
Lattice parameters of Sm doped hydroxyapatite and crystallite sized.
Samples
a-axis (Å)
c-axis (Å)
Unit cell volume (Å)
Crystallite sizeD (nm)
x
S
m
=
0.01
9.4195
6.8857
528.8875
18.14
x
S
m
=
0.03
9.4299
6.8864
529.9895
17.53
x
S
m
=
0.1
9.4398
6.8897
531.5923
12.67Typical particle morphology of the Sm:HAp (xSm=0.01, xSm=0.03 and xSm=0.1.) samples are shown in Figure 3. The surface morphology investigated by scanning electron microscopy reveals that the particles have a small, long morphology, coexisting with agglomerates. The results suggest that the doping of Sm has little influence on the morphology of Sm:HAp samples.SEM images of samarium doped hydroxyapatite (Sm:HAp) powders withxSm=0.01, xSm=0.03, and xSm=0.1.
(a)
(b)
(c)In order to investigate the uniformity of the element distribution in Sm:HAp samples, the EDX mapping technique was used. Figure4 shows the EDAX spectrum and elemental mapping of O, P, Ca, and Sm and demonstrates that all the elements were homogeneously distributed in Ca10-xSmx(PO4)6(OH)2 with xSm=0.03. On the other hand, the purity of the synthetized powders was also demonstrated. Similar results regarding the homogenous distribution of O, P, Ca, and Sm in the Sm:HAp samples were obtained for all the analysed samples.Figure 4
EDX spectrum and element maps of oxygen, phosphorus, calcium, and samarium (for the powders withxSm=0.03).FT-IR spectroscopy was performed in order to investigate the functional groups present in the nanohydroxyapatite,Ca10-xSmx(PO4)6(OH)2 (xSm=0.01, xSm=0.03, and xSm=0.1) samples. In Figure 5 the FT-IR spectra of samarium doped hydroxyapatite with various concentration are presented.Figure 5
FT-IR spectra of samarium doped hydroxyapatite (Sm:HAp) withxSm=0, xSm=0.01, xSm=0.03, and xSm=0.1.In the FT-IR spectra, the presence of [OH]− vibration peak at 632 cm−1 could be observed for all the samples [52]. This band arises from stretching vibrational mode of [OH]−. According to Markovic et al. [53], the sharpness of bands, especially sharpness of the 632 cm−1, 603 cm−1, and 567 cm−1 bands, indicates a well-crystallized HAp. The band at 470 cm−1 is assigned to the v2 [PO4]3− [54]. The vibration peaks in the regions 1600–1700 cm−1 and 3200–3600 cm−1 correspond to H–O–H bands of lattice water. The characteristic bands for carbonate are presented in the spectral regions 1400–1600 cm−1 (v3: asymmetric stretch vibration) and 873–880 cm−1 (v2: out-of-plane bend vibration). In addition, a weak band for the stretch vibration of structurally bound OH may be present in Sm:HAp samples near 3572 cm−1. Generally, it is difficult to distinguish between HPO42- and CO32- groups due to the overlapping of the characteristic peaks around 870 cm−1 [55, 56].The bands at around 1097 cm−1 and about 1037 cm−1 can be attributed to the v3 [PO4]3−, while the band at 962 arises from v1 [PO4]3−. The 603 cm−1 and 567 cm−1 bands appear from v4 [PO4]3− [57, 58]. In the FT-IR spectrum (Figure 5) the bands corresponding to the v3 vibration of C–O were observed at 1456 cm−1, characteristic of the carbonate group [59, 60].Also, in Figure5 we observed that the contribution of the area that corresponds to the phosphate bands decreases when the samarium concentration in the samples increases. The bands at 632 and 962 cm−1 progressively disappear with the increase of samarium concentration. When xSm=0.1 these bands are almost absent.Furthermore, in agreement with the results already reported in the literature, it can be observed that the use of samarium as doping agent does not induce the appearance of other peaks [61–63].The XPS analysis has been used to obtain the qualitative determination of the surface components and composition of the samples. Figure6 shows the survey of XPS narrow scan spectra of the as-prepared Ca10-xSmx(PO4)6(OH)2 with xSm=0.01 (a), xSm=0.03 (b), and xSm=0.1 (c) nanopowder samples. Concerning the binding energies in the pure HAp, the peaks at 347.3 eV, 532.1 eV and 133.8 eV can be attributed to the binding energy of Ca 2p, O 1s and P 2p, confirming the presence of apatite groups [64–66]. The peak at 1084,4 eV observed in Ca10-xSmx(PO4)6(OH)2 was attributed to the Sm 3d5/2, consistent with the normal oxidation state of Sm3+, because the binding energy of core level for Sm2O3 is generally observed at 1083.2 eV [65].XPS general spectrum ofCa10-xSmx(PO4)6(OH)2 with xSm=0.01 (a), xSm=0.03 (b), and xSm=0.1 (c) powders. XPS narrow scan spectra of Sm element are presented for all the samples.
(a)
(b)
(c)The peaks associated to Ca, P, and O states presented in Figure6 were the same in the Sm:HAp samples with different Sm3+ concentrations (xSm=0.01 (a), xSm=0.03 (b), and xSm=0.1 (c)). This showed that the increasing of the concentration of Sm did not change the binding energy assigned to Ca 2p, O 1s, and P 2p presented in Sm:HAp samples. For XPS analysis, the binding energies were calibrated with reference C 1s at 285 eV. By combination of XRD results, it can be concluded that these signals can be assigned to Sm:HAp nanopowder. XPS results provided the additional evidence for the successful doping with Sm3+ in Sm:HAp.In agreement with previous studies [52, 57, 67], the luminescence, magnetic, and nuclear properties of the ceramics can be improved by doping them with lanthanides. According to [68], the biocompatibility and valuable biological properties of samarium were proved by their use in drugs to regulate blood clotting present during the process of wound healing. Knowing that lanthanides (III) have an interesting biological role but not well known, preliminary antimicrobial tests were performed on samples of hydroxyapatite doped with samarium.Figure7 shows the results of viable bacteria adhering to the 5, 15, 25, and 50 μg/mL of Ca10-xSmx(PO4)6(OH)2 (xSm=0.01,  xSm=0.03, and xSm=0.1) when exposed toEnterococcus faecalis ATCC 29212. Bacterial adhesion was reduced on Ca10-xSmx(PO4)6(OH)2 sample when compared to pure HAp (xSm=0) and significant differences in bacterial adhesion on pure HAp (x=0) and Sm:HAp (xSm=0.01, xSm=0.03, and xSm=0.1) were observed. We observed that the bacterial adhesion decreased when the samarium concentrations increased.Figure 7
Adherence ofEnterococcus faecalis ATCC 29212 on different concentrations of Ca10-xSmx(PO4)6(OH)2 (xSm=0.01, xSm=0.03, and xSm=0.1) nanopowders compared to pure HAp (xSm=0) nanopowders.In this preliminary antimicrobial studies we observed that the Sm:HAp nanopowders have significantly lower adhesion ofEnterococcus faecalis ATCC 29212, suggesting that the Sm:HAp nanopowders are antibacterial. In the future, the effect of samarium doped hydroxyapatite on other bacterial strains will be evaluated and these strains will be selected depending on the field of applications. The influence of the size of Sm:HAp nanoparticles on bacterial strains will be also studied.The recent studies [69, 70] suggest that risk of infections in actual treatment of prosthetic infections is a major problem. In the patient treatment with hip or knee prosthetic infections it is noteworthy that infections with gram-negative bacilli or a fungus are contraindications. In this context, to optimize patient treatment with prosthetic infections the creation of new materials with antimicrobial properties is very important [71, 72]. In conclusion, it can be said that the new bioceramic Sm:HAp with antimicrobial properties may be a candidate with a good outcome in prosthetic joint infection and bone-targeting drug delivery system.


## 4. Conclusions

In this short work, the emphasis was placed on the synthesis at low temperature by a simple method of pure Sm:HAp nanopowder with efficient antimicrobial activity.The XRD studies have shown that the characteristic peaks of Sm:HAp nanopowders appeared to be identical regardless of the Sm content in the samples. All the XRD peaks were indexed in accordance with the hexagonal HAp inP63m space group (JCPDS card number 9-432) and no other secondary phases were detected. The functional groups present in nanohydroxyapatite samples, Ca10-xSmx(PO4)6(OH)2 (xSm=0.01, xSm=0.03, and xSm=0.1), were investigated by FT-IR spectroscopy. It has been observed that the use of samarium as doping agent does not induce the appearance of other peaks. FTIR spectra of the Sm:HAp samples show only the absorption bands characteristic to hydroxyapatite. A small long morphology was remarked for all the Sm:HAp nanopowder samples by SEM. The doping of Sm has little influence on the morphology of Sm:HAp samples. The presence and uniform distribution of samarium in the nanopowder samples were confirmed by elemental maps. The EDX spectrum of Sm:HAp confirms the presence of calcium (Ca), phosphorus (P), oxygen (O), and samarium (Sm) in all the synthetized samples. The successful doping of Sm3+ in Sm:HAp was also emphasized by XPS results.The inhibition ofEnterococcus faecalis ATCC 29212 bacteria increased when the samarium concentrations in Ca10-xSmx(PO4)6(OH)2 increased from xSm=0.01 to xSm=0.1. Our studies have shown that the Ca10-xSmx(PO4)6(OH)2 (xSm=0.01, xSm=0.03 and xSm=0.1) exhibited greater antimicrobial activity compared to the pure HAp (xSm=0). This will be significant for further antimicrobial studies of Ca10-xSmx(PO4)6(OH)2 on various gram-positive and gram-negative bacteria as well as fungi.In conclusion, this study on new bioceramic Sm:HAp with antimicrobial properties may present a solution in bone-targeting drug delivery system and prosthetic joint infection. On the other hand, Sm:HAp bioceramics can be used as antimicrobial coatings in various applications.



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