Trypsin versus Triton X-100 for decellularization of porcine articular cartilage
10.3969/j.issn.1673-8225.2009.46.016
- VernacularTitle:胰蛋白酶法和Trilon X-100法脱猪软骨细胞的比较
- Author:
Xiaoping CUI
;
Yonghong ZHANG
;
Jun XUE
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(46):9080-9083
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:As a material to repair osteochondral defect,the acellular cartilage matdx has attracted more and more researchers'attention,however,there is no comparison between two ways of decelluladzation.OBJECTIVE:To test trypsin and Triton X-100 methods for their potential of cell removal.DESIGN,TIME AND SETTING:Controlled observation was performed at the Central Laboratory of Orthopaedics in the Second Hospital of Shanxi Medical University between March and July in 2008.MATERIALS:Fresh knee and hip articular cartilages of pig were purchased from the market.METHODS:Porcine articular cartilages were treated with either 0.25%trypsin or 0.25% Triton X-100 respectively for decelluladzation,while fresh untreated articular cartilage served as control.MAIN OUTCOME MEASURES:①General observation on the morphology of acellular articular cartilage.②Immunohistochemistry and scanning electron microscopy were used to determine the effect of two decellularization methods on the extracellular matrix collagen.③Ten articular cartilage leaflets in each group were measured for breaking strength and percentage elongation through loading,unloading and breaking tests.RESULTS:①By use of trypsin and Triton X-100 methods,the acellular matrix had no significance difference in the morphology compared with untreated cartilage,only presenting slightly white color and feeling less soft.②lmmunohistochemistry and scanning electron microscopy results showed that,trypsin and Triton X-100 methods achieved complete decelluladzation.Trypsin had no significant influence in extracellular matrix and cartilage leaflets microstructure;Triton X-100 caused some tiny structural alterations,such as more collagen and disordered structure.It is possibly that Triton X-100 damaged extracellular matrix.③No significant difference was identified between untreated and trypsin,Triton X-100 groups in breaking strength and percentage elongation.CONCLUSION:Trypsin and Triton X-100 all achieve complete decellularization.but trypsin uses short time and low costs with well preservation,while Triton X-100 with long procedure and slight damage to collagen structure.They all have no significance influence to the mechanics characteristics of articular cartilage.
