In vitro & in vivo study of endothelial progenitor cell- assisted endochondral ossification LHCG platform

• Main Author: Lim, Sze Yua
• Other Authors: Wang Dongan
• Format: Final Year Project
• Language: English
• Published: 2015
• Subjects: DRNTU::Engineering::Bioengineering
• Online Access: http://hdl.handle.net/10356/64735
• Institution: Nanyang Technological University

Living hyaline cartilaginous graft (LhCG), formed by chondrocytes and their extracellular matrix, can undergo osteogenic differentiation that is similar to natural endochondral ossification. Therefore, LhCG gains more interest being used to guide bone formation in tissue engineering. However, vascularized and hypertrophic cartilaginous template should be introduced before LhCG is induced endochondral bone generation. In this study, we tend to investigate effect of endothelial progenitor cells (EPC) on promoting cartilage hypertrophy and vascularization in LhCG. Bone marrow derived EPC were seeded onto LhCG and the whole construct was cultured in osteogenic medium for 9 days before it was cultured in MCDB 131 medium. Another LhCG which was applied similar condition but without EPC seeding was established for comparison. Cultured in vitro for 18 days, both LhCGs were implanted into subcutaneous pockets of nude mice. All samples were retrieved at designated time point and analyzed by histological and immunohistochemical staining. EPC seeding LhCG showed higher degree staining of collagen X and Alizarin Red S, indicating more hypertrophic chondrocytes and mineralized extracellular matrix in LhCG, both in vitro and in vivo. The results suggested that co-culture of EPC and chondrocytes in LhCG can enhance cartilage hypertrophy and vascularization. It is recommended that various cells and mediums can be applied to LhCG to determine their co-culture effects on hypertrophic LhCG.