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Investigation of the Internalization of Fluorescently Labeled Lipophilic siRNA into Cultured Tumor Cells
摘要： The attachment of lipophilic molecules of natural origin, which have natural means for cell internalization, to small interfering RNA (siRNA) is an effective way of delivering siRNA to cells for biomedical purposes in vitro and in vivo. Earlier, we showed that the attachment of cholesterol to the 5'-end of the sense strand of nuclease-resistant siRNA through the optimized linker allows it to penetrate the cells and suppress the expression of the target gene. However, the effectiveness of the conjugates is different for cells of different origin, and in hematopoietic cells, they are not active, despite effective accumulation. In this work, we investigated the accumulation of fluorescently labeled cholesterol conjugates of siRNA using endocytosis inhibitors and showed that fluorescently labeled 5'-cholesterol conjugate of siRNAs penetrate KB-3-1 and K562 cells in several ways whose contribution differs depending on cell type and the presence of serum. In a serum-free medium, it was found that macropinocytosis and clathrin-dependent endocytosis contribute to the accumulation of the conjugate in KB-3-1 cells, while clathrin-dependent endocytosis makes the main contribution in K562 cells, while inhibitors of different types of endocytosis do not reduce the biological activity of the conjugate without a fluorescent label.
关键词： cell delivery mechanism,inhibitors of endocytosis,siRNA,cholesterol conjugate,chemical modifications
更新于2025-09-19 17:13:59
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Layer-by-Layer-Assembled Antifouling Films with Surface Microtopography Inspired by Laminaria japonica
摘要： Marine biofouling is an important obstacle to the development of marine resources. Problems such as huge economic losses caused by biofouling and environmental damage caused by toxic antifouling agents have not been resolved. Therefore, it is of great significance to develop a novel, environmentally friendly antifouling (AF) materials. However, in the marine environment, Laminaria japonica still has excellent antifouling ability in a relatively static state compared to those parade creatures. Inspired by this, this study reports a synergistic effect between surface topography and chemical modification to inhibit marine biofouling. Firstly, the surface of the Laminaria japonica was analyzed and its morphology was reproduced using a simple moulding process. Additionally, the polyelectrolyte layer composed of sodium alginate and (guanidine-hexamethylenediamine-PEI) (poly(GHPEI)) was chemically modified on the isotropic microstructure surface of PDMS replicas by layer-by-layer assembly method. The anti-adhesion ability of the biomimetic material was tested with Nitzschia closterium (N. closterium) (9±5 diatoms mm-2 of N. closterium adhered). Meanwhile, the antifouling performance of the modified films were evaluated by Escherichia coli (E. coli), and its antibacterial ability were as high as 96.2±1.3%. The combination of microtopography and (GHPEI/ALG) * n films endow the coating with excellent antifouling ability.
关键词： marine antifouling,surface microtopography,layer-by-layer assembly,alginate,poly(GHPEI),chemical modifications
更新于2025-09-16 10:30:52