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Three-dimension Collagen-Chitosan Scaffold for Osteoblast Proliferation and Differentiation
|Authors: ||Chen-Shiu Wang|
|Contributors: ||NTOU:Department of Food Science|
|Issue Date: ||2011-06-30T08:15:22Z
|Abstract: ||本研究為利用多孔三維支架來模擬生物骨骼之環境，以人類造骨類細胞 MG-63探討於此仿生之環境對其增生與分化之影響。目前一般細胞試驗以平面培養基為主，但仍與生物內部環境有差距，故本研究乃利用高分子可塑性，以冷凍產生冰晶，再以冷凍乾燥去除冰晶，因而產生出多孔三維支架。本篇所採用的材料為幾丁聚醣，並以交聯方式使膠原蛋白與幾丁聚醣產生連接，膠原蛋白為組織中胞外基質主要成分之一，特別是結締組織，其不僅為結構上的支撐，並給予細胞其他重要的功能。來自於魚鰾之膠原蛋白經證實為第二型膠原蛋白，並且可利用來改善細胞不易黏著於幾丁聚醣表面。本篇支架採用 -20 ℃ 冷凍方式來產生孔洞，其孔徑大小為141±11 m ，由電子顯微鏡所觀測，細胞於複合支架有較好的延展性。在增生測試方面，可發現複合支架有較高的增生現象，並且持續增生至第四天，原因可能為膠原蛋白促進細胞黏著及延展。在分化測試方面，可得到複合支架於培養第一天即有較高的鹼性磷酸酶活性。實驗結果得知，複合支架相較於幾丁聚醣支架有更好的細胞黏著、延展、增生及分化，故可提升其應用性，以作為評估保健食品之研究模型。|
The purpose of this study was to fabricate a three-dimensional (3D) porous scaffold and evaluate both biofunctions for proliferation and differentiation of human osteoblast-like MG-63 cells cultured in the biomimetic scaffold. Compare to in vitro cultivation of cells on 2D dish conventional, we used polymeric materials plasticity to fabricate 3D porous scaffolds in 24 well cell culture plate by removing ice crystal through freeze-dry process. In our study, we used chtiosan and collagen as the scaffold materials, and then formed a biomimetic scaffold by cross-link reaction. Collagen is one of the key extracellular matrix of animal tissue, especially in connective tissue. Extracellular matrix usually provides structural support to the animal cells in addition to performing various other important functions. Collagen was extracted and purified from swim bladder, and indentified as type II collagen. Collagen could improve cell adhesion on chitosan surface. The porous size of scaffolds was 141±11 m through freeze process at the temperature of -20 degree Celsius. Collagen-chitosan complex scaffolds had more effect of cells spreading than chitosan scaffold by SEM detection. The result of cell proliferation showed complex scaffolds enhanced proliferation maybe via promoting cell both adhesion and spreading, and it continued for the fourth day. The result of cell differentiation showed cells cultured in complex scaffolds expressed more ALP activity at first day. In summary, complex scaffolds would be more benefic cell adhesion, spreading, proliferation and differentiation. It may be used as a desirable method for evaluation of functional foods model that keeping cell in good performance.
|Appears in Collections:||[食品科學系] 博碩士論文|
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