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作为小干扰RNA载体生物材料合成的壳聚糖-聚(-[3-(二甲基氨基丙基)]甲基丙烯酰胺)共聚物的物理化学性质

Physico-biochemical Properties of Chitosan-poly(‑[3-(dimethylaminopropyl)] methacrylamide) Copolymers Synthesized as siRNA Carrier Biomaterials.

作者信息

Şimşek Ceyda, Demirel Erhan, Macit Semih, Yüksel Durmaz Yasemin, Erbil Candan

机构信息

Faculty of Science and Letters, Department of Chemistry, Istanbul Technical University, Istanbul, TR 34469, Turkey.

School of Engineering and Natural Sciences, Department of Biomedical Engineering, Istanbul Medipol University, Istanbul, TR 34810, Turkey.

出版信息

ACS Omega. 2025 Aug 29;10(36):42019-42038. doi: 10.1021/acsomega.5c06839. eCollection 2025 Sep 16.

Abstract

In the present study, poly-(-[3-(dimethylaminopropyl)] methacrylamide) (PDMAPMAAm) grafts/blocks and chitosan (Cs) backbone bearing tertiary and primary amine groups, respectively, were copolymerized by the conventional free-radical solution method to prepare novel nonviral cationic carriers covering a pH range between 4.0 and 9.0. All of the chain parameters calculated by experimental and theoretical methods indicated that with increasing KPS concentration, the structures of Cs-PDMAPMAAm (Cs/D) copolymers changed from graft to block copolymer. The constants of the Mark-Houwink-Sakurada (MHS) equation and were also determined to calculate the molecular weights of synthesized PDMAPMAAm and its hydrolyzed grafts. For physico-biochemical properties, FTIR, GPC, and H NMR techniques, ζ-potential, particle size, hemolytic activity, buffering capacity, and cell viability measurements were performed to discuss that at least one of the Cs/D products demonstrates an acceptable transfection efficiency as . To investigate the cytotoxicity and gene transfection properties of Cs/D copolymers and their polyplexes with small interfering RNA (siRNA) by MTS assay on L929 and MDA-MB-231, which are healthy mouse fibroblast cells and a triple-negative human breast cancer cell line, respectively, binding capacity and serum stability by agarose gel electrophoresis, cellular uptake by confocal microscopy, and transfection efficiency of siRNA on the MDA-MB-231 cell line were studied.

摘要

在本研究中,分别带有叔胺基和伯胺基的聚(- [3 - (二甲基氨基丙基)]甲基丙烯酰胺)(PDMAPMAAm)接枝/嵌段与壳聚糖(Cs)主链,通过传统自由基溶液法进行共聚,以制备覆盖pH范围在4.0至9.0之间的新型非病毒阳离子载体。通过实验和理论方法计算得到的所有链参数表明,随着过硫酸钾(KPS)浓度的增加,Cs - PDMAPMAAm(Cs / D)共聚物的结构从接枝共聚物转变为嵌段共聚物。还确定了Mark - Houwink - Sakurada(MHS)方程的常数 和 ,以计算合成的PDMAPMAAm及其水解接枝物的分子量。对于物理生化性质,采用傅里叶变换红外光谱(FTIR)、凝胶渗透色谱(GPC)和核磁共振氢谱(H NMR)技术、ζ电位、粒径、溶血活性、缓冲能力和细胞活力测量等方法来探讨至少一种Cs / D产物表现出可接受的转染效率。通过MTS法在分别为健康小鼠成纤维细胞和三阴性人乳腺癌细胞系的L929和MDA - MB - 231细胞上研究Cs / D共聚物及其与小干扰RNA(siRNA)的复合物的细胞毒性和基因转染特性,通过琼脂糖凝胶电泳研究结合能力和血清稳定性,通过共聚焦显微镜研究细胞摄取,以及研究siRNA在MDA - MB - 231细胞系上的转染效率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/64fb/12444683/74830dfac1ca/ao5c06839_0001.jpg

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