a Laboratory of Ultrasound Molecular Imaging, Department of Ultrasound Medicine , The Third Affiliated Hospital of Guangzhou Medical University , Guangzhou , China.
Drug Deliv. 2018 Nov;25(1):1740-1745. doi: 10.1080/10717544.2018.1510065.
Gene therapy is a promising strategy for treatments of various diseases. Efficient and safe introduction of therapeutic genes into targeted cells is essential to realize functions of the genes. High-molecular-weight polyethylenimines (HMW PEIs) including 25 kDa branched PEI and 22 kDa linear PEI are widely used for in vitro gene transfection. However, high-gene transfection efficiency is usually accompanied with high cytotoxicity, which hampers their further clinical study. On the contrary, low-molecular-weight polyethylenimines (LMW PEIs) such as 1.8 kDa PEI and 800 Da PEI show good biocompatibility but their applications are limited by the poor DNA condensation capability. In this study, we find that 1.8 kDa PEI, but not 800 Da PEI combined with low-dose 25 kDa PEI could significantly promote gene transfection with low cytotoxicity. Plasmids encoding enhanced green fluorescence protein (EGFP) were delivered by the combined PEI and gene transfection efficiency was evaluated by microscopic observation and flow cytometry. Parameters including concentrations of 25 kDa PEI and 1.8 kDa PEI and preparation ways were further optimized. This study presents an efficient and safe combined PEI-based non-viral gene delivery strategy with potential for in vivo applications.
基因治疗是治疗各种疾病的一种很有前途的策略。将治疗基因有效地、安全地导入靶细胞对于实现基因的功能至关重要。高分子量聚乙烯亚胺（HMW PEIs），包括 25kDa 支化聚乙烯亚胺和 22kDa 线性聚乙烯亚胺，被广泛应用于体外基因转染。然而，高基因转染效率通常伴随着高细胞毒性，这阻碍了它们进一步的临床研究。相反，低分子量聚乙烯亚胺（LMW PEIs），如 1.8kDa PEI 和 800Da PEI，具有良好的生物相容性，但由于 DNA 凝聚能力差，其应用受到限制。在本研究中，我们发现 1.8kDa PEI，而不是与低剂量 25kDa PEI 结合的 800Da PEI，可以显著提高基因转染效率而不产生细胞毒性。用编码增强型绿色荧光蛋白（EGFP）的质粒进行转染，通过显微镜观察和流式细胞术评估基因转染效率。进一步优化了 25kDa PEI 和 1.8kDa PEI 的浓度以及制备方法等参数。本研究提出了一种高效、安全的基于复合聚乙烯亚胺的非病毒基因传递策略，具有体内应用的潜力。
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