肝癌仍然是全世界癌症的主要原因之一。治疗方案面临着一些挑战，而纳米材料已被证明可以提高几种候选药物的生物利用度及其在纳米医学中的应用。具体来说，壳聚糖纳米粒子（CNP）具有极高的生物降解性，具有增强的生物相容性，并且被认为在医学上使用是安全的。CNP是通过离子凝胶化合成的，负载芦丁（rCNP），并通过紫外-可见光谱（UV-Vis）、傅里叶光谱进行表征。 -变换红外光谱（FTIR）、动态光散射（DLS）和透射电子显微镜（TEM）。测试了 rCNP 对人肝癌 Hep3B 细胞的细胞毒性作用，并进行实验以确定这种作用的机制。此外，在L929成纤维细胞上测试了rCNP的生物相容性，并确定了它们的血液相容性。最初，UV-vis和FTIR分析表明rCNP上可能负载有芦丁。此外，使用超高效液相色谱（UPLC）定量测量芦丁负载，0.22微米过滤的rCNP的浓度为88μg/mL。 rCNP 的载药量 (LC%) 为 13.29 ± 0.68%，包封率 (EE%) 为 19.55 ± 1.01%。药物释放具有pH响应性，在溶酶体pH 5.5下24小时后释放出88.58%的药物，而在生理pH 7.4下102小时后释放出91.44%的药物。在 5 mg/mL 芦丁前体的 0.22 微米过滤样品中，细胞毒性作用非常显着。该浓度下rCNP的粒径为144.1 nm，多分散指数（PDI）为0.244，这被认为是肿瘤靶向的理想选择。 zeta电位（ζ电位）值为16.4 mV，表明rCNPs具有良好的稳定性。 rCNP 对人肝癌 Hep3B 细胞的细胞毒性作用的 IC50 值为 9.7 ± 0.19 μg/mL 芦丁负载。此外，还观察到活性氧（ROS）产生增加和线粒体膜电位（MMP）变化。基因表达研究表明，rCNPs对Hep3B细胞的细胞毒作用除了抑制上皮细胞外，还通过激活Unc-51样自噬激活激酶（ULK1）介导的自噬和核因子κB（NF-κB）信号传导来实现。 -间质转化（EMT）。此外，与Hep3B细胞相比，rCNP对NCTC克隆929（L929）成纤维细胞的毒性较小，并且具有优异的血液相容性（溶血率低于2％）。合成的rCNP具有pH响应性，并具有适合肿瘤的理化特性瞄准。与正常细胞相比，该颗粒对 Hep3B 细胞具有有效的细胞毒性，并且具有优异的血液相容性。 rCNP 的极低溶血特性表明该药物可以静脉注射用于癌症治疗。© 2024 Wu 等人。

Liver cancer remains to be one of the leading causes of cancer worldwide. The treatment options face several challenges and nanomaterials have proven to improve the bioavailability of several drug candidates and their applications in nanomedicine. Specifically, chitosan nanoparticles (CNPs) are extremely biodegradable, pose enhanced biocompatibility and are considered safe for use in medicine.CNPs were synthesized by ionic gelation, loaded with rutin (rCNPs) and characterized by ultraviolet-visible spectroscopy (UV-Vis), Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) and transmission electron microscopy (TEM). The rCNPs were tested for their cytotoxic effects on human hepatoma Hep3B cells, and experiments were conducted to determine the mechanism of such effects. Further, the biocompatibility of the rCNPs was tested on L929 fibroblasts, and their hemocompatibility was determined.Initially, UV-vis and FTIR analyses indicated the possible loading of rutin on rCNPs. Further, the rutin load was quantitatively measured using Ultra-Performance Liquid Chromatography (UPLC) and the concentration was 88 µg/mL for 0.22 micron filtered rCNPs. The drug loading capacity (LC%) of the rCNPs was observed to be 13.29 ± 0.68%, and encapsulation efficiency (EE%) was 19.55 ± 1.01%. The drug release was pH-responsive as 88.58% of the drug was released after 24 hrs at the lysosomal pH 5.5, whereas 91.44% of the drug was released at physiological pH 7.4 after 102 hrs. The cytotoxic effects were prominent in 0.22 micron filtered samples of 5 mg/mL rutin precursor. The particle size for the rCNPs at this concentration was 144.1 nm and the polydispersity index (PDI) was 0.244, which is deemed to be ideal for tumor targeting. A zeta potential (ζ-potential) value of 16.4 mV indicated rCNPs with good stability. The IC50 value for the cytotoxic effects of rCNPs on human hepatoma Hep3B cells was 9.7 ± 0.19 μg/mL of rutin load. In addition, the increased production of reactive oxygen species (ROS) and changes in mitochondrial membrane potential (MMP) were observed. Gene expression studies indicated that the mechanism for cytotoxic effects of rCNPs on Hep3B cells was due to the activation of Unc-51-like autophagy-activating kinase (ULK1) mediated autophagy and nuclear factor kappa B (NF-κB) signaling besides inhibiting the epithelial-mesenchymal Transition (EMT). In addition, the rCNPs were less toxic on NCTC clone 929 (L929) fibroblasts in comparison to the Hep3B cells and possessed excellent hemocompatibility (less than 2% of hemolysis).The synthesized rCNPs were pH-responsive and possessed the physicochemical properties suitable for tumor targeting. The particles were effectively cytotoxic on Hep3B cells in comparison to normal cells and possessed excellent hemocompatibility. The very low hemolytic profile of rCNPs indicates that the drug could be administered intravenously for cancer therapy.© 2024 Wu et al.