Transfection of DNA into Cochlear Cells by Functionalized Calcium Phosphate Nanoparticles
Transfection of DNA into Cochlear Cells by Functionalized Calcium Phosphate Nanoparticles
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功能化磷酸钙纳米颗粒将 DNA 转染入耳蜗细胞
DOI：
10.1055/s-0039-1686498
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发表时间：
2019
影响因子：
1
通讯作者：
S. Hansen
中科院分区：
文献类型：
--
作者：
R. Schirrmann;K. Wey;M. Epple;S. Brandau;S. Lang;S. Hansen
材料与方法：制备了平均尺寸为274nm和403nm的多层CaP-DNA CaP-DNA-CaP-PEI（聚乙烯亚胺）。使用的缀合 DNA 是绿色荧光蛋白 eGFP 或 mOrange。然后将颗粒与新生大鼠的螺旋神经节外植体以及 HeLa 细胞共培养，并在免疫组织化学染色后进行显微镜观察和评估。 结果：纳米颗粒缀合物被非常有效地摄取到螺旋神经节外植体的所有细胞中。大约 5 天后，一小部分但显着比例的细胞显示出编码染料 DNA 的稳定表达。结论：尤其是雪旺细胞，甚至成纤维细胞甚至螺旋神经节细胞都可以在 CaP-NP 的帮助下摄取 DNA，并在转染后分泌功能蛋白。因此，可以用功能化的 CaP-NP 涂覆 CI 电极，并刺激电极附近的内耳细胞分泌神经营养因子或抗炎因子。
Materials and Methods:Multi-layer CaP-DNA CaP-DNA-CaP-PEI (polyethylenimine) with an average size of 274nm and 403nm were generated. The conjugated DNA used was the green fluorescent protein eGFP or mOrange. The particles were then co-cultured with spiral ganglia explants from neonatal rats as well as with HeLa cells and, after immunohistochemical staining, were microscoped and evaluated.Results:There was a very effective uptake of the nanoparticle conjugates into all cells of the spiral ganglia explant. A smaller but significant proportion of these cells showed stable expression of dye-encoding DNA after about 5 days.Conclusion:Schwann cells in particular, but also fibroblasts and even spiral ganglion cells can take up DNA with the help of CaP-NP and secrete a functional protein after transfection. Thus, it would be possible to coat a CI electrode with functionalized CaP-NP and to stimulate inner ear cells nearby the electrode for the secretion of neurotrophic or anti-inflammatory factors.