Tissue Engineering of Cartilage: A Chondrogenic Differentiation of Mesenchymal Stem Cells in Type I Collagen Matrix

• Main Authors: Chia-Ming Chang, 張家銘
• Other Authors: 蔡郁惠
• Format: Others
• Language: en_US
• Published: 2002
• Online Access: http://ndltd.ncl.edu.tw/handle/97666569961610467590

碩士 === 台北醫學院 === 生物醫學材料研究所碩士班 === 90 === Tissue engineering is a new strategy to produce a biocompatible and bioactive tissue for surgical implantation to solve several clinical problems. Cells, matrix (scaffolds) and growth factors, three major components of tissue engineering can be manipulated delicately into a perfect concerto for engineering a neo-tissue to promote the life quality of human beings. By using bone marrow derived stromal cells (mesenchymal stem cells, MSCs) as the cell source, transforming growth factor-beta 1 (TGF-β1) as the chondrogenic inducer and rat-tail type I collagen as the scaffold, the goal of in vitro tissue engineering of artificial cartilage (neocartilage) was attempted in this study. MSCs have been isolated, fractionated and propagated in vitro and reported to maintain their potential to differentiate into chondrocytes. Bone marrow aspiration has been performed frequently by clinical standard procedures. Consequently, MSCs are good cell source in the matter of cartilage engineering to overcome the deficiency of healthy cartilage for primary chondrocytes. On the other hand, collagen is one of the most abundant proteins in connective tissue and is a major component of extracellular matrix. Because of its triple helix structure, collagen not only as a good scaffold but also provide a good living environment for cells. Owing to its relatively low immunogenesity, collagen has been wildly used as biomaterials in wound dressing, heart valve and so on. TGF-β1, one of the most well studied growth factors in the matter of cell proliferation and differentiation, has been reported to promote chondrogenic differentiation of MSCs. Besides, cartilage engineering by using TGF-β1 and different biomaterials has also been reported in the past years. In this thesis, modified isolation protocols were used to obtain sufficient MSCs from rabbit bone marrow for tissue engineering of cartilage. During subsequent passages, the morphology and behavior of MSCs were observed. Their potential for chondrogenic differentiation after subsequent passage was also tested. Optimal MSCs seeding density within three dimensional collagen matrix was determined in this study. In order to evaluate the progression of chondrogenic differentiation, alcian blue was used to confirm the expression of cartilage proteoglycan after tissue culture in the existence of differentiation factors. This study was focused on the isolation of and the differentiating potential of MSCs after consecutive passages. The protocols for manufacturing neocartilage composed of primary chondrocytes established in Dr. Lai’s laboratory was followed to engineer a new neocartilage from MSCs. In conclusion, sufficient MSCs would be obtained easily in this laboratory. The MSCs are cultivated in rat-tail type I collagen matrix under various culture conditions. The newly formed cell-matrix constructs was induced to differentiate into cartilage-like tissue by TGF-β1. Chondrocytic morphology and expression of cartilage proteoglycan were used to evaluate the progression of chondrogenesis. Further efforts are necessary to optimize the conditions for fabricating and evaluating neo-cartilage derived from MSCs. The obtained data will provide crucial information for the development of autogenic neocartilage implantation.