Chernonosova Vera, Khlebnikova Marianna, Popova Victoriya, Starostina Ekaterina, Kiseleva Elena, Chelobanov Boris, Kvon Ren, Dmitrienko Elena, Laktionov Pavel
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, 630090 Novosibirsk, Russia.
State Research Center of Virology and Biotechnology VECTOR, Rospotrebnadzor, 630559 Koltsovo, Russia.
Polymers (Basel). 2023 Jul 27;15(15):3202. doi: 10.3390/polym15153202.
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.
生物材料介导的空间定位基因递送对于组织工程中使用的细胞填充支架的开发至关重要。附着于或穿透此类支架的细胞要用预先加载的基因进行转染,该基因可诱导分泌蛋白的产生或细胞重编程。在本研究中,我们制备了与二氧化硅纳米颗粒相关的质粒DNA(pDNA)以及负载此类纳米颗粒的电纺支架,并从细胞活力和pDNA转染效率方面研究了pDNA从支架中的释放以及细胞与支架的相互作用。用动态光散射和透射电子显微镜对包被pDNA的纳米颗粒进行了表征。56至78纳米的粒径表明其具有细胞转染的潜力。使用扫描电子显微镜、X射线光电子能谱、应力加载测试以及与HEK293T细胞的相互作用对支架进行了表征。发现材料的性质和释放的pDNA会因支架的组成而异。负载pDNA纳米颗粒的支架没有明显的细胞毒性作用,可推荐用于细胞转染。发现负载(pDNA-纳米颗粒)+聚乙烯亚胺9的支架具有良好的细胞转染潜力。因此,适用于转染驻留细胞的电纺支架有资格用于组织工程。