col v sirna engineered tenocytes for tendon tissue engineering坳v sirna工程tenocytes肌腱组织工程.pdfVIP

Col V siRNA Engineered Tenocytes for Tendon Tissue Engineering Ping Lu1,2., Guo Rong Zhang1., Xing Hui Song1,2, Xiao Hui Zou1,3, Lin Lin Wang1,2*, Hong Wei Ouyang1,2* 1 Center for Stem Cell and Tissue Engineering, School of Medicine, Zhejiang University, Hangzhou, China, 2 Institute of Cell Biology, School of Medicine, Zhejiang University, Hangzhou, China, 3 Women Hospital, School of Medicine, Zhejiang University, Hangzhou, China Abstract The presence of uniformly small collagen fibrils in tendon repair is believed to play a major role in suboptimal tendon healing. Collagen V is significantly elevated in healing tendons and plays an important role in fibrillogenesis. The objective of this study was to investigate the effect of a particular chain of collagen V on the fibrillogenesis of Sprague-Dawley rat tenocytes, as well as the efficacy of Col V siRNA engineered tenocytes for tendon tissue engineering. RNA interference gene therapy and a scaffold free tissue engineered tendon model were employed. The results showed that scaffold free tissue engineered tendon had tissue-specific tendon structure. Down regulation of collagen V a1 or a2 chains by siRNAs (Col5a1 siRNA, Col5a2 siRNA) had different effects on collagen I and decorin gene expressions. Col5a 1 siRNA treated tenocytes had smaller collagen fibrils with abnormal morphology; while those Col5a2 siRNA treated tenocytes had the same morphology as normal tenocytes. Furthermore, it was found that tendons formed by coculture of Col5a1 siRNA treated tenocytes with normal tenocytes at a proper ratio had larger collagen fibrils and relative normal contour. Conclusively, it was demonstrated that Col V siRNA engineered tenocytes improved tendon tissue regeneration. And an optimal level of collagen V is vital in regulating collagen fibrillogenesis. This may provide a basis for future development of no