Chen Fang, Wan Huiying, Xia Tian, Guo Xueqin, Wang Huan, Liu Yaowen, Li Xiaohong
Key Laboratory of Advanced Technologies of Materials, Ministry of Education of China, School of Materials Science and Engineering, Southwest Jiaotong University, Chengdu, PR China.
Eur J Pharm Biopharm. 2013 Nov;85(3 Pt A):699-710. doi: 10.1016/j.ejpb.2013.07.009. Epub 2013 Jul 24.
Vascularization is one of the capital challenges in the establishment of tissue engineering constructs and recovery of ischemic and wounded tissues. The aim of this study was to assess electrospun fibers with loadings of multiple pDNA to allow a localized delivery for an efficient regeneration of mature blood vessels. To induce sufficient protein expression, a reverse microemulsion process was adopted to load pDNA into calcium phosphate nanoparticles (CP-pDNA), which were electrospun into fibers to achieve a sustained release for 4 weeks. Compared with pDNA-infiltrated fibers, the localized and gradual release of pDNA facilitated cell proliferation, gene transfection, and extracellular matrix secretion and enhanced the generation of blood vessels after subcutaneous implantation. Compared with commonly used pDNA polyplexes with poly(ethyleneimine), CP-pDNA nanoparticles induced significantly lower cytotoxicity and less inflammation reaction after implantation into animals. Fibers with encapsulated nanoparticles containing plasmids encoding vascular endothelial growth factor (pVEGF) and basic fibroblast growth factors (pbFGF) led to significantly higher density of mature blood vessels than those containing individual plasmid. It is suggested that the integration of CP-pDNA nanoparticles with loadings of multiple plasmids into fibrous scaffolds should provide clinical relevance for therapeutic vascularization, getting fully vascularized in engineered tissues and regeneration of blood vessel substitutes.
血管化是组织工程构建体建立以及缺血和受伤组织修复中的主要挑战之一。本研究的目的是评估负载多种质粒DNA（pDNA）的电纺纤维，以实现局部递送，促进成熟血管的有效再生。为诱导足够的蛋白质表达，采用反向微乳液法将pDNA负载到磷酸钙纳米颗粒（CP-pDNA）中，然后将其电纺成纤维，以实现4周的持续释放。与pDNA浸润纤维相比，pDNA的局部和逐步释放促进了细胞增殖、基因转染和细胞外基质分泌，并增强了皮下植入后的血管生成。与常用的聚乙烯亚胺pDNA复合物相比，CP-pDNA纳米颗粒植入动物后诱导的细胞毒性显著更低，炎症反应更小。含有编码血管内皮生长因子（pVEGF）和碱性成纤维细胞生长因子（pbFGF）质粒的纳米颗粒包封纤维，比含有单个质粒的纤维导致的成熟血管密度显著更高。研究表明，将负载多种质粒的CP-pDNA纳米颗粒整合到纤维支架中，应为治疗性血管化提供临床相关性，使工程组织完全血管化并实现血管替代物的再生。
