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|標題:||Fabrication and Evaluation of Biodegradable Polymer Scaffolds for Chondrocyte Seeding
In this study, immortalized rat chondrocytes (IRC) were used to evaluate the polymer scaffolds for cartilage repair. Fibroblasts (L929), rabbit chondrocytes, porcine chondrocytes and IRC were seeded onto the surface of biodegradable polymers. The cell adhesion, growth and morphology of chondrocytes were similar to those of IRC. Therefore, IRC can be used as a tool to evaluate the materials as scaffolds. In all experiments, PLGA (polylactide-co-glycolide) 50:50 had significantly better cytocompatibility than PLLA and PLGA 85:15. SEM observations of the surface and cross-section of the substrates revealed that the pore size was 40-60μm (except for samples made by the freeze drying-liquid nitrogen fast quench method). So the pore size should not account for the different cell growth. The cells were fully spread after 96 hours of culture in scaffolds with morphology similar to that grown in petri dish. Cell migration into the porous substrates was examined by a laser confocal microscope. The amount of cells at a fixed depth for different materials was consistent with that described earlier.
Also in this study, the structures of scaffolds made by the solvent casting-salt leaching technique or by the freeze drying method were compared with each other. The pore size of scaffolds was found to be similar to that of the substrates examined under SEM. The cell growth and the amount of matrix produced in the scaffolds were evaluated by histological sectioning and biochemical analyses. The cells in the scaffolds increased in number and distributed within the scaffolds with time. The amount of glycosaminoglycan (GAG) components in matrix was determined. Although the amount was little at an early stage, the GAG content in the scaffolds also increased with time.
In summary, the optimum biodegradable scaffolds for chondrocyte seeding used in repairing the damaged articular cartilage should have a pore size close to 50 μm or up and a stable three-dimensional structure with good cytocompatibility. From our study, the optimum scaffolds were PLGA50: 50 by solvent-casting salt leaching, PLLA and blend of PLLA and PLGA50: 50 (Blend) by freeze drying (quenched in refrigerator). In the future, it is hoped that scaffolds developed here can be combined with natural materials such as collagen to modify the surface of scaffolds in order to further enhance the cell migration and to retain the cell morphology. Once the tissue regeneration in scaffolds is improved, clinical applications could thereafter be expected.
本研究中採取組織工程的方法，首次以軟骨細胞株取代動物細胞之初代培養軟骨細胞方面進行支架材料篩選，研究中將纖維母細胞、紐西蘭白兔軟骨細胞、小豬關節軟骨細胞、IRC細胞植覆於可降解材料表面，經培養後經計數細胞貼附及生長的數量，由結果可以確認軟骨細胞株在貼附及生長的狀況與正常軟骨細胞類似，且能更迅速判斷。而在多孔性基材上，細胞經植覆後96小時，細胞貼附於多孔性數量以PLGA 50：50與PLLA、PLGA 85:15有顯著的差異。由SEM觀察基材的表面及截面，不同種類材料間所產生的孔洞主要分佈在40~60 m之間（除冷凍乾燥-液態氮驟冷製備方法以外），因此可以排除孔洞尺寸對細胞生長的效應。將細胞植覆於支架上經96小時，細胞已經完全貼附，並且呈現完全伸展的型態，和在培養皿培養的型態類似。使用共軛焦雷射顯微鏡觀察多孔性基材在一定深度下，可以看出與薄膜測試結果相同的趨勢之活細胞的分佈。
|Appears in Collections:||化學工程學系所|
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