本論文利用脂肪組織中取得具有分化能力的脂肪幹細胞(hASCs)結合生
物相容材料支架來誘導人體脂肪幹細胞形成組織工程軟骨。本實驗中評估
三種以明膠為主的細胞載體材料，應用在幹細胞分化成軟骨的可行性。本
實驗的主要支架明膠(gelatin)是一種具有良好的生物相容性材料，另外選擇
加入第ㄧ型膠原蛋白 (collagen I) 、第二型膠原蛋白(collagen II)，並且加
入聚己內酯(PCL)，形成gelatin/collagen II:PCL(GCII), gelatin:PCL(GP),
gelatin/collagen I:PCL(GCI) 能有效控制支架在細胞培養過程中，產生高度
的降解，維持其穩定性。並探討幹細胞在不同支架的生長及分化的情形。
在掃描式電子顯微鏡及螢光顯微鏡和細胞生長曲線鑑定下, 證明材料具
有良好的生物相容性。在脂肪幹細胞的誘導分化能力測定，免疫螢光染色
的結果顯示脂肪幹細胞在人工支架的微環境下，有被誘導走向軟骨細胞分
化的情形，且細胞表現量隨著時間而增加。另外，利用Alcian blue stain 和
DMMB assay 測定葡萄糖糖胺聚合糖分析，在GP 和GCII 有較多的表現
量。藉由Sirius red stain 來分析誘導後的軟骨中膠原蛋白的種類，使用可
見光/紫外光光譜儀測定其吸收值(波長:350nm; 540nm)，結果顯示誘導後的
軟骨所分泌的膠原蛋白以第ㄧ型膠原蛋白為主。並利用Von Kossa stain 來
探討長時間後的軟骨基質鈣化(chondralcalcification）的情形，在誘導後第
三十天，鈣化出現在軟骨細胞周圍區。

This study investigated the induction of human adipose tissue-derived stem
cells (hASCs) into chondrocytes, which made a possible source for cartilage
tissue engineering in clinical therapy. The hASCs were cultured on three
different biocompatible biomaterial groups including gelatin/collagen I: PCL
(GCI), gelatin/collagen II:PCL (GCII) and gelatin:PCL (GP). Considering the
high degradability of natural biomaterials, the addition of PCL for formulation
of scaffolds is necessary to resist absorption in vitro. In addition, we
characterized the attachment, growth and differentiation abilities of hASCs in
biomaterials by SEM, fluorescence microscopy and cell growth curve in vitro.
The immunofluorescence data showed that the hASCs were differentiated with
the increasing time on biocomposite membrane groups. For investigation of
cartilage differentiation, alcian blue staining and dimethylmethylene blue
(DMMB) assay were used for both quality and quantity evaluation of
glycosaminoglycans. The results showed that the groups of gelatin and GCII
contained more glycosaminoglycans than GCI. For further distinguishing
distinct collagen types produced by chondrocytes based on different
biocomposite membranes, sirius red staining was used and type I collagen
showed the absorbance at 540 nm and type II collagen at 350nm with
UV-visble spectrophotometer. We demonstrated that the chondrocytes release
mainly type I collagen on all biocomposite membranes. In detecting the long
term expression of calcium accumulation in differentiated chondrocytes using
von kossa stain, chondral calcification represented after induction of
chondrocyte differentiation on biocomposite membranes for 30 days

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