Moradi, Ali (2015) Development of bovine cartilage extracellular matrix as a potential scaffold for chondrogenic induction of human dermal fibroblasts / Ali Moradi. PhD thesis, University of Malaya.
Abstract
Tissue engineering aims to find an ultimate solution for cartilage regeneration. Current strategies for cartilage repair using chondrocytes are limited by the usability of cell sources as observed in several contemporary techniques such as autologous cell implantations. Cartilage tissue engineering strategies involving scaffold fabrication are common. However, only few studies have described the use of scaffolds as a promoter for chondrogenic differentiation. Cartilage extracellular matrix derived scaffolds contain many bioactive factors that have chondrogenic potential of which if appropriately managed, may provide the key to successful cartilage repair outcomes. The present thesis describes a potential biomaterial derived from bovine articular cartilage that possesses chondrogenic properties, thereby being expected to promote cartilage regeneration. Bovine articular cartilage matrix derived (CMD) porous scaffolds were fabricated at three different concentrations (5%, 15% and 30%), through the use of freeze-drying method or, physically treated for cross-linking enhancement using either ultra-violet (UV) light or, dehydrothermal (DHT) treatment or a combination of both methods. The scaffolds were characterized using morphology, FESEM, FTIR, TGA, DSC, and porosimetry. UV and UVDHT treated 15% CMD scaffolds were found to be the best scaffold possessing the best possible physical properties. Human dermal fibroblast cells (HDF) isolated from redundant skin obtained through cosmoplastic surgery were used as cells for the in vitro analysis. HDF cells were cultured on UV and UVDHT treated 15% CMD scaffolds for 42 days. Resazurin reduction test and DNA quantification showed an increasing cell proliferation during the first 3 weeks. GAGs production and the up-regulation ofSOX9, COMP, COL2A1, and ACAN genes were seenthroughout6 weeks of cell-scaffold culture. The increase or decrease in HDF cell proliferation, GAGs production and gene expression appeared correlated with the scaffold shrinkage and contraction patterns. In conclusion, bovine articular CMD porous scaffolds have the potential to induce chondrogenic differentiation in human dermal fibroblasts. Whilst the present thesis did not demonstrate the precise underlying mechanisms by which chondroinduction is achieved from these CMD scaffolds, it is proposed that future studies are conducted to investigate the involved pathways to better demonstrate the chondrogenic properties of this material.
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