Effects of combinational adenoviral vector-mediated TGF beta 3 transgene and shRNA silencing type I collagen on articular chondrogenesis of synovium-derived mesenchymal stem cells.
In this study, transgenic effects of combination of transforming growth factor (TGF) beta3 and shRNA silencing type I collagen (Col I) on chondrogenesis of synovium-derived mesenchymal stem cells (SMSCs) were evaluated. SMSCs were infected with recombinant adenoviruses encoding TGF beta 3 (Ad-TGF beta 3) and/or anti-Col I shRNA (Ad-shRNA) separately, simultaneously (Ad-combination), or conjugately (Ad-double, mediated by one vector encoding both). The transduced SMSCs were encapsulated in alginate hydrogel and cultured for 30 days in chondrogenic medium. The expression of cartilaginous extracellular matrix components was investigated by quantitative real-time RT-PCR (qRT-PCR) and histological staining. qRT-PCR showed an up-regulation in chondrocytes marker genes such as type II collagen, aggrecan, and cartilage oligomeric matrix protein (COMP) in Ad-TGF beta 3, Ad-double, and Ad-combination groups on day 30. Whereas, Ad-TGF beta 3 treatment induced significant elevation in Col I, which could be largely resisted by anti-Col I shRNA functionality. Histological and immunohistochemical staining results were consistent with our qRT-PCR data. These results demonstrate that the application of combinational adenoviral vector-mediated transgenic TGF beta 3 and shRNA targeting Col I possesses the potential in promoting the chondrogenic differentiation of SMSCs as well as inhibiting the formation of fibrocartilage.
Yao
Yongchang
Y
Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637457, Republic of Singapore.
Zhang
Feng
F
Zhou
Ruijie
R
Su
Kai
K
Fan
Jiabing
J
Wang
Dong-An
DA
D000256:Adenoviridae / Q000235:genetics; D002454:Cell Differentiation; D020219:Chondrogenesis; D024042:Collagen Type I / Q000037:antagonists & inhibitors* / Q000235:genetics; D020869:Gene Expression Profiling; D020868:Gene Silencing; D005822:Genetic Vectors; D006651:Histocytochemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013583:Synovial Membrane / Q000166:cytology; D014161:Transduction, Genetic; D053782:Transforming Growth Factor beta3 / Q000235:genetics / Q000378:metabolism*; D019076:Transgenes
D024042:Collagen Type I; D053782:Transforming Growth Factor beta3