Galván-Flores Lorena E, Osorio-Trujillo Carlos, Talamás-Rohana Patricia, Gallardo-Hernández Salvador
Programa de Doctorado en Nanociencias y Nanotecnología, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Av. IPN 2508, Ciudad de Mexico C. P. 07360, Mexico.
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Av. IPN 2508, Ciudad de Mexico C. P. 07360, Mexico.
Molecules. 2025 Sep 19;30(18):3815. doi: 10.3390/molecules30183815.
Chitosan is considered an excellent biomaterial for epithelial healing treatment. However, information on its molecular interaction with cells at a molecular level is still lacking. Thus, in the present study, homemade synthesized chitosan nanoparticles (CS NPs) and their physicochemical characterization were examined; we found that NPs had average sizes of 15, 30, and 125 nanometers by modifying some variables in the synthesis protocols. It is worth noting that a crystalline structure was found on the smallest NPs, with an average size of 15 nm, as observed in high-resolution transmission electron micrographs. To study the in vitro interaction of CS NPs with Madin-Darby canine kidney (MDCK) cells, co-culturing was performed, and cell viability was assessed. We found that NPs were not toxic at concentrations of up to 400 µg/mL during the first 24 h. Additionally, quantitative mass spectrometry revealed the overexpression of several proteins induced by the co-culture of CS NPs with MDCK cells, and a proteomic analysis suggested two important things: possible clathrin-mediated endocytosis could be the interaction mechanism of CS NPS with MDCK, and proteins related to cytoskeleton formation and organization were overexpressed. Moreover, wound healing assays revealed that <125 nm> CS NPs yielded the best closure rates, where mitomycin was added to make sure that only cell migration occurred.
壳聚糖被认为是用于上皮愈合治疗的优良生物材料。然而，在分子水平上其与细胞分子相互作用的信息仍很缺乏。因此，在本研究中，对自制合成的壳聚糖纳米颗粒（CS NPs）及其理化特性进行了检测；我们发现，通过改变合成方案中的一些变量，纳米颗粒的平均尺寸分别为15、30和125纳米。值得注意的是，在高分辨率透射电子显微镜图像中观察到，平均尺寸为15纳米的最小纳米颗粒上发现了晶体结构。为了研究CS NPs与Madin-Darby犬肾（MDCK）细胞的体外相互作用，进行了共培养，并评估了细胞活力。我们发现，在前24小时内，浓度高达400 µg/mL的纳米颗粒没有毒性。此外，定量质谱分析显示，CS NPs与MDCK细胞共培养诱导了几种蛋白质的过表达，蛋白质组学分析表明了两件重要的事情：可能的网格蛋白介导的内吞作用可能是CS NPs与MDCK的相互作用机制，并且与细胞骨架形成和组织相关的蛋白质过表达。此外，伤口愈合试验显示，<125 nm>的CS NPs产生了最佳的闭合率，其中添加了丝裂霉素以确保仅发生细胞迁移。
