## 3 Results
### Targeted Drug Delivery
#### 3.2.1 Active Targeting
developed several types of self-assembled chitosan nanoparticles loaded with DOX to selectively target HCC via ASGPR. Chitosan was first chemically modified with glycidol to enhance its solubility, then further functionalized with hydrophobic moieties such as vitamin E succinate (VE) or deoxychic acid (DCA) to impart amphiphilic properties, and finally galactosylated by conjugation with lactobionic acid to enable ASGPR targeting. Doxorubicin was subsequently loaded into the resulting nanoparticles using the dialysis method. In vitro drug release studies confirmed that DOX release from all nanoparticle types was pH-dependent, with faster release under acidic conditions (pH 5), which mimic the tumor microenvironment. Internalization tests conducted on HepG2 cells demonstrated superior uptake for the galactosylated nanoparticles, supporting the involvement of ASGPR in the cellular uptake mechanism. Among the variants, galactosylated nanoparticles showed superior in vitro performance and enhanced in vivo anticancer efficacy. Moreover, all tested formulations exhibited good biocompatibility, with no significant histological damage observed. However, the study also highlighted certain limitations. The exact molecular mechanisms of the interaction between the nanoparticles and ASGPR were not explored in detail. Another aspect that requires further investigation is the behavior of these nanoparticles in systemic circulation over the long term, as well as potential accumulation in other organs that weakly express ASGPR. Lastly, detailed pharmacokinetic and biodistribution studies are essential before any potential clinical applications [54]. Based on the fact that folic acid receptors are overexpressed on liver tumor cells, [PERSON] et al. employed nanomicelles functionalized with silibinin (SLB) to enhance treatment efficiency. Silibinin, a polyphenol derived from _Silbyum marianum_, is known for its anti-inflammatory, anticancer, and hepatoprotective effects, with minimal side effects. However, its poor water solubility limits clinical use [55]. To address this, the researchers synthesized FA-functionalized Pluronic F127 copolymer (F127/FA) using the Steglich esterification method and encapsulated SLB into its hydrophobic core via thin-film hydration. The resulting nanomicelles had a spherical morphology, an average size of 17.7 nm, and demonstrated 2.36% drug loading and 79.43% encapsulation efficiency. In vitro release studies showed that SLB was released more slowly and sustainably from the FA-targeted nanomicelles than in its free form. Cytotoxicity assays in HepG2 cells revealed significantly greater anticancer activity for SLB/F127/FA compared to SLB/F127 or free SLB. In summary, these findings highlight SLB/F127/FA nanomicelles as a promising targeted delivery system for liver cancer therapy, especially in tumors with overexpressed folate receptors. While the study demonstrates promising in vitro results, its translational potential remains limited due to the absence of in vivo validation. Additionally, the relatively low drug loading capacity could restrict the amount of silibinin delivered per dose, necessitating larger quantities of the carrier to achieve therapeutic effects. The formulation's long-term stability also remains uncertain, as no data were provided on potential changes in particle size, aggregation, or drug retention under physiological conditions [56].