Immature Muscular Tissue Differentiate into Cartilage Using BMP in vitro

Successful tissue engineering relies on a combination of cells, cytokines and appropriate scaffolds. Objectives: We tried to induce the formation of cartilage tissue in vitro using immature muscular tissue, BMP as a source of the cytokine, and biodegradable membranes (Bio Mend^® and GC-MEMBRANE) or a non-biodegradable membrane (GORE-TEX^®) as scaffolds and also examined which one was the most effective material as a scaffold. Methods: BMP was extracted from bovine cortical bones, dried, and dissolved in 1M urea before it was added to immature muscular tissue from the forelimbs of fetal Sprague Dawley rats at 20 days of pregnancy. The tissue was then cultured in CMRL 1066 supplemented with 15% FBS for two weeks in a carbon dioxide incubator. After finished tissue culture, H-E staining, toluidine blue staining and immunohistochemical staining of collagen typeⅡ were performed for histological observation. mRNA was extracted from cultured tissue on days 3,7,10 and 14, and reverse transcription PCR assay was performed to quantify cartilaginous extra cellular matrix gene expressions. Results: Complete cartilage tissue was observed only when GORE-TEX^® was used as a scaffold. In addition, cartilage-like tissue or calcified-like tissue formation was observed when Bio Mend^® was used, and partial cartilage tissue formation was observed for GC-MEMBRANE. Collgaen typeⅡ and aggrecan were upregulated from days 3 to 10 when GORE-TEX^® was used. These gene expression levels were significantly higher than in the case of Bio Mend^® and GC-MEMBRANE. (p<0.05) Conclusion: These results show that GORE-TEX^® is the most effective material for use as a scaffold in this model of tissue engineering.