Construction of recombinant adenovirus encoding both TGF-beta 3 and ShRNA targeting type I collagen gene.

• Main Author: Liu, Chengzheng.
• Other Authors: Wang Dongan
• Format: Final Year Project
• Language: English
• Published: 2009
• Subjects: DRNTU::Engineering::Chemical engineering::Biotechnology
• Online Access: http://hdl.handle.net/10356/16558
• Institution: Nanyang Technological University

The tissue engineering of Mesenchymal stem cells (MSC) are promising in treatment of cartilage defects and the repairmen of damaged cartilage. But the highly expression of type I collagen in the cells, which will result in fibrocartilage, is still a big problem to be overcome. Basing on RNA interfering (RNAi) and the idea that the TGF-β3 can promote scar-free treatment in previous finding(1), this project was designed to construct an adenoviral vector and thus an adenovirus which can secret TGF-β3 while knocking down the type I collagen expression in target cells. Here the experiments are carried out in human fetal osteoblast (hFOB) cells and human fibroblast (hFB) cells to study the performance of the constructed virus and investigate its possible clinical application. In this project, it is found that the null adenovirus can promote type I collagen expression in both cells. Results showed that the constructed recombinant virus did have suppressive effects on type I collagen expression. In addition, the RNAi by shRNA in the cells had obvious suppression on type I collagen expression, even more effective than the recombinant virus. However, the effects of TGF-β3 on type I collagen production was diversified: inhibitive in hFOB cells but promotive in hFB cells. Later experiments found that the functioning of virus in hFOB cells was very short in terms of time, while for hFB cells, it is not achieved in this project.