Farjadian Fatemeh, Behzad-Behbahani Abbas, Mohammadi-Samani Soliman, Ghasemi Soheila
Pharmaceutical Sciences Research Center, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran.
Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Prog Biomater. 2022 Jun;11(2):219-227. doi: 10.1007/s40204-022-00187-6. Epub 2022 May 9.
Nanohydrogels (NHs) with the benefits of both nanomaterials and hydrogels unlock novel opportunities and applications in biomedicine. Nowadays, cationic NHs have attracted attention in the delivery of genetic materials into cells. Herein, by using reversible addition-fragmentation chain transfer method, an NH-based poly(hydroxyethyl methacrylate-co-N,N-dimethylaminoethyl methacrylate) and cross-linked by poly(ethylene glycol)diacrylate with pH responsiveness character was developed. Several techniques including nuclear magnetic resonance, Fourier-transform infrared spectroscopy, and gel permeation chromatography confirmed the success in the synthesis. The pH responsiveness of the developed NH was shown by transmission electron microscopy and dynamic light scattering technique. The average sizes of NHs in the normal (7.4) and acidic pH (5.5) were 180 and 390 nm, respectively. The ability of the developed NH to condense genetic materials was checked using gel retardation assay with different ratios of NH and pCMV6-IRES-AcGFP, as a plasmid encoding green fluorescence protein. Results of gel retardation assay showed a decreasing trend in plasmid electrophoretic mobility with the increase in the NH concentration. The NH/plasmid complexes were stopped completely at the ratio of 5 and the plasmid band vanished at the ratio of 10. The quantitative and qualitative results of the cell transfection experiment using different ratios of NH/plasmid showed the ability of NH to carry plasmid molecules into the cancerous cells. The best transfection efficiency was observed by nanohydrogel/plasmid weight ratio of 10, while other ratios including 2, 5 and 20 showed 0.8, 10 and 12% of transfection efficiency, respectively. All the assessed factors showed that NH has the potential to be considered as an efficient gene delivery vehicle.
兼具纳米材料和水凝胶优点的纳米水凝胶（NHs）为生物医学带来了新的机遇和应用。如今，阳离子纳米水凝胶在将遗传物质递送至细胞方面受到了关注。在此，通过可逆加成-断裂链转移法，开发了一种基于NH的聚（甲基丙烯酸羟乙酯-co-N，N-二甲基氨基乙基甲基丙烯酸酯），并通过聚（乙二醇）二丙烯酸酯交联，具有pH响应特性。核磁共振、傅里叶变换红外光谱和凝胶渗透色谱等多种技术证实了合成的成功。通过透射电子显微镜和动态光散射技术展示了所开发NH的pH响应性。正常pH（7.4）和酸性pH（5.5）下NHs的平均尺寸分别为180和390nm。使用凝胶阻滞试验，以不同比例的NH和pCMV6-IRES-AcGFP（一种编码绿色荧光蛋白的质粒）检测所开发NH浓缩遗传物质的能力。凝胶阻滞试验结果表明，随着NH浓度的增加，质粒电泳迁移率呈下降趋势。NH/质粒复合物在比例为5时完全停止，在比例为10时质粒条带消失。使用不同比例的NH/质粒进行细胞转染实验的定量和定性结果表明，NH有能力将质粒分子携带到癌细胞中。纳米水凝胶/质粒重量比为10时观察到最佳转染效率，而其他比例（包括2、5和20）的转染效率分别为0.8%、10%和12%。所有评估因素表明，NH有潜力被视为一种高效的基因递送载体。
