| Summary: | 碩士 === 臺北醫學大學 === 藥學研究所 === 95 === The objective of this thesis was to develop new artificial cartilage for treating cartilage disease. We used two kinds of material scaffolds, which were based on porcine cartilage matrix and collagen matrix extracted from porcine skin, followed by various treatments. In the part of porcine cartilage matrix, we used three enzymes and treated by various time and temperature for finding the best method to remove the porcine cells. Then we applied H&E stain, nested PCR, and fluorimetric DNA assay to estimate the degree of decellularization. In the other part of collagen matrix extracted from porcine skin, we used two ways of adding hyaluronic acid for adding hyaluronic acid which was advantageous to chondrocyte growth. Then we analyzed the physical properties. Finally, we seeded chondrocytes in the two kinds of material scaffolds, and observed the growth of chondrocytes. We applied MTT assay for the amount, H&E stain for the distribution, and RT-PCR for the mRNA expression. On the other hand, the acellular cartilage matrix was implanted into the lesion of Wistar rats to evaluate the biocompatibility.
The results showed that when treated with hyaluronidase and trypsin or with dispase and trypsin, and combined surfactant, for 24 hr could effectively remove the cells, and when coated with hyaluronic acid could have firm structure but the pore diameters decreased with the increase of hyaluronic acid content. Two scaffolds had porous structure for cell growth. Now we have seeded chondrocytes in the two scaffolds, but the experiments about the observation of cell growth and the biocompatibility of acellular cartilage matrix are still in progress. We expect that cell would grow well in the both two scaffolds.
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