Type B synoviocytes induced by transforming growth factor beta 1 combined with Pluronic-F127 to construct tissue engineering cartilage in vivo
10.3969/j.issn.1673-8225.2009.46.011
- VernacularTitle:转化生长因子β1基因修饰兔B型滑膜细胞复合Pluronic-F127体内构建组织工程软骨
- Author:
Weiping LI
;
Bin SONG
;
Rui YANG
;
Jianrong HUANG
;
Yang SONG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(46):9055-9060
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:The type B synoviocytes induced by transforming growth factor β1(TGF-β1) have the potential to differentiate into chondrocyte,which can remain the phenotype in vitro. However,whether the transfected cell combined with scaffold can form cartilage tissues need further research. OBJECTIVE:Rabbit type B synoviocytes was transfected by liposome method in vitro,combined the cells with Pluronic-F127,and then implanted into nude mouse to construct tissue engineering cartilage. Additionally,to explore the feasibility of synoviocytes differentiate into chondrocytes.DESIGN,TIME AND SETTING:The randomized animal experiment of cytology observation. The experiment was performed at the Medical Research Center,the Second Affiliated Hospital of Sun Yat-sen University between April 2007 and May 2008.MATERIALS:Healthy New Zealand white rabbits,aged 3 months,and 12 BALB/c nude mice,aged 4 weeks,weighted 20 g.METHODS:The synovial membrane tissues were taken out from the rabbit knee,isolated by enzyme digestion,and then transfected. The positive cloning was screened by G418,and the expression of TGF-β1 and collagen Ⅱ was detected. The Pluronic-F127 was dissolved at 4℃ and prepared fluid with concentration of 30%. The fluid was mixed with cells. At the same time,there were 2 groups as the control group:chondrocytes with Pluronic-F127,and synoviocytes transfeced by pcDNA3.1(+)and Pluronic-F127. The cell density was 5x1010/L. Each compound (0.2 mL) was injected into the subcutaneous of the nude mouse back. Rats were sacrificed at weeks 4,6 and 8 to harvest samples.MAIN OUTCOME MEASURES:Cell growth curve;phenotype change of the cells after transfection;histological observation of the tissue in the subcutaneous of the nude mouse back. RESULTS:Cell growth curve demonstrated that the living activity of the transfected cells was temporary decreased,which returned into normal level at days 6,7 after transfection. At day 4 after transfection,TGF β1 were positive expressed,and at day 7,the collagen Ⅱ staining were positive. The compounds in the subcutaneous of the nude mouse back formed immature chondroid tissues at week 4,which turned to mature chondroid tissue at week 8,and the collagen Ⅱ staining were positive.CONCLUSION:The transfected synoviocytes can express the phenotype of chondrocytes in vitro,and form chondrocyte-likecells. The transfected synoviocytes with Pluronic-F127 can form chondroid tissues in nude mice.
