Department of Biomedical Science, College of Life Science, CHA University, 6F, CHA Biocomplex, Sampyeong-Dong, Bundang-gu, Seongnam-si, 13488, Republic of Korea.
CHA Advanced Research Institute, 7F, CHA Biocomplex, Sampyeong-Dong, Bundang-gu, Seongnam-si, 13488, Republic of Korea.
Theranostics. 2019 Jul 9;9(17):5009-5019. doi: 10.7150/thno.34487. eCollection 2019.
Development of a stable and prolonged gene delivery system is a key goal in the gene therapy field. To this end, we designed and fabricated a gene delivery system based on multiply-clustered gold particles that could achieve prolonged gene delivery in stem cells, leading to improved induction of differentiation. : Inorganic gold nanoparticles (AuNPs) underwent three rounds of complexation with catechol-functionalized polyethyleneimine (CPEI) and plasmid DNAs (pDNAs), in that order, with addition of heparin (HP) between rounds, yielding multiply-clustered gold-based nanoparticles (mCGNPs). Via metal-catechol group interactions, the AuNP surface was easily coordinated with positively charged CPEIs, which in turn allowed binding of pDNAs. : Negatively charged HP was encapsulated with the positive charge of CPEIs via electrostatic interactions, making the NPs more compact. Repeating the complexation process yielded mCGNPs with improved transfection efficiency in human mesenchymal stem cells (hMSCs); moreover, these particles exhibited lower cytotoxicity and longer expression of pDNAs than conventional NPs. This design was applied to induction of chondrogenesis in hMSCs using pDNA harboring SOX9, an important chondrogenic transcription factor. Prolonged expression of SOX9 induced by mCGNPs triggered expression of chondrocyte extracellular matrix (ECM) protein after 14 days, leading to more efficient chondrogenic differentiation and .
开发稳定且长效的基因传递系统是基因治疗领域的一个关键目标。为此，我们设计并制造了一种基于多聚金纳米颗粒的基因传递系统，该系统可实现干细胞内的长效基因传递，从而提高分化诱导效果。无机金纳米颗粒（AuNPs）经过三轮与儿茶酚功能化的聚乙烯亚胺（CPEI）和质粒 DNA（pDNA）的络合，顺序为在轮间添加肝素（HP），得到多聚金基纳米颗粒（mCGNPs）。通过金属-儿茶酚基团相互作用，AuNP 表面很容易与带正电荷的 CPEIs 配位，从而允许 pDNAs 的结合。带负电荷的 HP 通过静电相互作用与 CPEIs 的正电荷结合，使 NPs 更加紧凑。重复络合过程可提高人骨髓间充质干细胞（hMSCs）中的转染效率；此外，与常规 NPs 相比，这些颗粒表现出更低的细胞毒性和更长的 pDNA 表达。该设计应用于通过携带 SOX9 的 pDNA 诱导 hMSCs 软骨生成，SOX9 是一种重要的软骨生成转录因子。mCGNPs 诱导的 SOX9 持续表达可在 14 天后触发软骨细胞细胞外基质（ECM）蛋白的表达，从而更有效地诱导软骨分化。
