Abstract:
Biomaterial-mediated, spatially localized gene delivery is important for the development of cell-populated scaffolds used in tissue engineering. Cells adhering to or penetrating into such a scaffold are to be transfected with a preloaded gene that induces the production of secreted proteins or cell reprogramming. In the present study, we produced silica nanoparticles-associated pDNA and electrospun scaffolds loaded with such nanoparticles, and studied the release of pDNA from scaffolds and cell-to-scaffold interactions in terms of cell viability and pDNA transfection efficacy. The pDNA-coated nanoparticles were characterized with dynamic light scattering and transmission electron microscopy. Particle sizes ranging from 56 to 78 nm were indicative of their potential for cell transfection. The scaffolds were characterized using scanning electron microscopy, X-ray photoelectron spectroscopy, stress-loading tests and interaction with HEK293T cells. It was found that the properties of materials and the pDNA released vary, depending on the scaffold\'s composition. The scaffolds loaded with pDNA-nanoparticles do not have a pronounced cytotoxic effect, and can be recommended for cell transfection. It was found that (pDNA-NPs) + PEI9-loaded scaffold demonstrates good potential for cell transfection. Thus, electrospun scaffolds suitable for the transfection of inhabiting cells are eligible for use in tissue engineering.
摘要:
生物材料介导,空间定位的基因传递对于组织工程中使用的细胞填充支架的开发很重要。粘附或穿透到这种支架中的细胞将用诱导分泌蛋白产生或细胞重编程的预加载基因转染。在本研究中,我们生产了与二氧化硅纳米颗粒相关的pDNA和载有这种纳米颗粒的静电纺丝支架,并从细胞活力和pDNA转染功效方面研究了pDNA从支架的释放和细胞与支架的相互作用。用动态光散射和透射电子显微镜表征pDNA包被的纳米颗粒。56至78nm范围内的粒度指示其用于细胞转染的潜力。使用扫描电子显微镜对支架进行表征,X射线光电子能谱,应激负荷试验和与HEK293T细胞的相互作用。发现材料的性质和释放的pDNA各不相同,取决于支架的组成。载有pDNA纳米颗粒的支架没有明显的细胞毒性作用,并可推荐用于细胞转染。发现(pDNA-NP)+负载PEI9的支架显示出良好的细胞转染潜力。因此,适用于存活细胞转染的电纺支架可用于组织工程。