利用人類胎盤間葉幹細胞植入褐藻膠/奈米陶瓷複合材料結合可降解精密支架於軟骨組織工程之研究

Human placenta-derived mesenchymal cells (hPDMCs) seeded in alginate-based nanocomposites combined with biodegradable precision scaffolds for cartilage tissue engineering

鄭舜榮(Shun-Jung Cheng)

指導教授 ： 黃琮濱

共同指導教授 ： 徐善慧(Shan-Hui Hsu)

長榮大學/健康科學學院/生物科技學系(所)/碩士(2009年)

摘要

本研究著重於探討一個新穎性的組織工程細胞來源─人類胎盤間葉幹細胞(human Placenta-derived mesenchymal cells, hPDMCs )加入奈米陶瓷材料(nano-calcium deficient hydroxyapatite, nCDHA)和細胞貼附因子CBD-RGD與褐藻膠混合後再注入聚乳酸-聚甘醇酸(poly (lactide-co-glycolide), PLGA)精密支架中使用含轉形生長因子-bata 3(transforming growth factor-beta 3, TGF-beta 3)的軟骨分化劑進行誘導培養軟骨生成情形。實驗中發現經過誘導培養21天後奈米化氫氧基磷灰石(nano-calcium deficient hydroxyapatite, nCDHA)(已吸附TGF-beta 3) 結合CBD-RGD混摻褐藻膠植入支架組別其細胞生長數目、葡萄胺聚醣 (glycosaminoglycans, GAG)、第二型膠原蛋白(Type II collagens)細胞外基質(extracellular matrix, ECM)分泌含量皆比其他組別多。並且在組織染色切片上其成熟軟骨的表徵-陷窩 (lacunae)也較為明顯且完整。另外，本研究同時比較人類胎盤間葉幹細胞與骨髓間葉幹細胞在nCDHA(已吸附TGF-beta 3) 結合CBD-RGD混摻alginate植入支架其誘導培養軟骨分化情形，發現到胎盤幹細胞誘導分化軟骨生成的細胞外基質比骨髓幹細胞明顯地多。經過實驗證明幹細胞與nCDHA(已吸附TGF-beta 3) 結合CBD-RGD混摻褐藻膠植入精密支架對於幹細胞誘導進行軟骨生成效果最佳；另一方面，人類胎盤間葉幹細胞誘導生成成熟軟骨的表現優於骨髓間葉幹細胞，因此可應用於骨組織工程作為骨組織修復的來源細胞。

關鍵字

胎盤間葉幹細胞 ； 骨髓間葉幹細胞 ； 褐藻膠 ； 氫氧基磷灰石 ； 轉形生長因子-bata 3 ； 聚乳酸-聚甘醇酸 ； CBD-RGD

並列摘要

Human mesenchymal stem cells are studied extensively such as bone marrow mesenchymal stem cells (BMSCs). However, mesenchymal stem cells from bone marrow could be limited. Human term placenta, a temporary organ is discarded postpartum. In this study, we used human placenta-derived mesenchymal cells (hPDMCs) seeded into PLGA precision scaffolds for chondrogenesis. The mixture, cellulose binding domain Arginine-Glycine-
Aspartate (CBD-RGD)/nano-calcium deficient hydroxyapatite (nCDHA) which was absorbed transforming growth factor beta-3(TGF-beta 3)/Alginate, encapsulate hPDMCs to inject a scaffold. The chondrogenic differentiations of the construction are best than others. After 7 days the cell proliferation of the construction were maximum and higher than others, furthermore, it can maintain the maximum cell numbers until 21 days. On the other hand, the component of extracellular matrix (ECM) of the construction was maximum after 7 days, also it can maintain the maximum cell numbers until 21 days. Histological examination revealed the presence of lacuna formation in the ECM. In the same system, the chondrogenic differentiations of hPDMCs are better than hBMSCs. The results of this research suggested hPDMCs can be one of the cell sources for tissue engineering of cartilage.

並列關鍵字

placenta-derived mesenchymal cells (hPDMCs) ； bone marrow mesenchymal stem cells (BMSCs) ； Alginate ； nano-calcium deficient hydroxyapatite (nCDHA) ； TGF-beta 3 ； PLGA ； CBD-RGD

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