Sustained delivery of therapeutic genes in vitro and in vivo has a wide range of applications in studying biology and in developing therapies for treating disease or repairing tissue. Nonviral vectors such as cationic polymers still present promising approaches; however, bolus transfection methods with polyethylenimine-based DNA polyplexes suffer from considerable levels of cytotoxicity and short-lived transgene expression levels. Here, we designed and characterized a hyaluronic acid-based porous hydrogel system for nonviral gene delivery by loading with surface-associated DNA polyplexes. With this, we observed tunable, enhanced, and sustained transgene expression over 30 days of cell culture with better cell viability and marked improvements over comparable bolus transfection techniques. Finally, we investigated mechanisms thought to be responsible for the sustained expression profile, finding that multiple transfection events are likely responsible for the observed sustained expression.

中文翻译：

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通过多次转染事件的水凝胶介导的基因转移结果持续转基因表达

在体外和体内持续递送治疗性基因在研究生物学和开发治疗疾病或修复组织的疗法方面具有广泛的应用。阳离子聚合物等非病毒载体仍然是很有前途的方法；然而，使用基于聚乙烯亚胺的 DNA 复合物的推注转染方法存在相当大的细胞毒性和短暂的转基因表达水平。在这里，我们设计并表征了一种基于透明质酸的多孔水凝胶系统，用于通过加载表面相关的 DNA 复合物来传递非病毒基因。有了这个，我们在 30 天的细胞培养中观察到可调谐、增强和持续的转基因表达，具有更好的细胞活力，并且比可比的推注转染技术有显着改进。最后，