Chondrogenesis using mesenchymal stem cells and PCL scaffolds
Chondrogenesis using mesenchymal stem cells and PCL scaffolds
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DOI：
10.1002/jbm.a.32414
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发表时间：
2010-02-01
影响因子：
4.9
通讯作者：
Im, Gun-Il
中科院分区：
文献类型：
--
作者：
Kim, Hye-Joung;Lee, Jin-Ho;Im, Gun-Il
我们检测了多孔PCL(聚己内酯)作为骨髓间充质干细胞(MSCs)软骨组织工程支架材料的体外可行性，并测定了不同表面处理的效果。研究了三种多孔PCL支架的修饰：(1)PCL/Pluronic F127，(2)PCL/胶原，(3)PCL/Pluronic F127/胶原，(4)仅PCL。将MSCs(5×105)接种于孔径为100~150微米的PCL支架上，体外培养3周后，观察MSC支架的大体形态、DNA含量、糖胺聚糖(GAG)含量、成软骨基因表达及组织学改变。大体上，细胞-支架复合体在培养3周后变得更坚硬，更容易用钳子操作。经表面处理的三种支架的DNA含量均高于单纯PCL支架，PCL/胶原和PCL/F127/胶原支架中的GAG含量高于单纯PCL支架。实时定量聚合酶链式反应显示，与单纯PCL支架相比，PCL/胶原和PCL/F127/胶原支架中SOX-9和COL2A1的表达水平显著升高。另一方面，三种修饰的PCL支架中Col1A1和Col10A1的mRNA水平低于仅PCL支架。组织学结果与GAG和RT-PCR结果基本一致，证实了PCL支架对MSCs的亲和力以及这些支架诱导软骨分化的潜力。PCL/胶原和PCL/胶原/F127支架细胞分化程度较高，细胞外基质形成较丰富。我们的研究结果表明，基于PCL的多孔支架可能是软骨组织工程领域中MSC移植的有用载体，而基于胶原的表面修饰进一步促进了MSCs的软骨分化。(C)2009年Wiley期刊，Inc.《生物医学杂志》92A：659-666,2010
We tested the in vitro feasibility of porous PCL (poly(epsilon-caprolactone)) as a scaffold for cartilage tissue engineering from mesenchyrnal stem cells (MSCs) and determined the effects of various surface treatments. Three porous PCL scaffold modifications were examined: (1) PCL/Pluronic F127, (2) PCL/collagen, and (3) PCL/Pluronic F127/collagen, in addition to (4) PCL-only. MSCs (5 X 105) were seeded in PCL scaffolds of pore size 100-150 mu m, and after 3 weeks of in vitro culture, MSC-scaffolds were investigated for gross appearance, DNA amount, glycosaminoglycan (GAG) content, chondrogenic gene expression, aid histology. Grossly, the cell-scaffold complexes became harder, and were more easily manipulated with a forceps after 3 weeks of culture. The three surface-treated scaffolds had higher DNA contents than did the PCL-only scaffold, and the GAG contents in PCL/collagen and PCL/F127/collagen scaffolds were higher than those seen in the PCL-only scaffold. Real-time PCR showed that Sox-9 and COL2A1 mRNA levels were remarkably elevated in PCL/collagen and PCL/F127/collagen scaffolds versus the PCL-only scaffold. On the other hand, Col1A1 and Col10A1 mRNA levels were lower in the three modified PCL scaffolds than in the PCL-only scaffold. Histological findings generally concurred with GAG and RT-PCR findings, and demonstrated the affinity of PCL-based scaffolds for MSCs and the potentials of these scaffold to induce chondrogenic differentiation. Cells showed more differentiated appearance and more abundant extracellular matrix formation in PCL/collagen and PCL/collagen/F127 scaffolds. Our findings suggest that PCL-based porous scaffolds may be useful carriers for MSC transplantation in the cartilage tissue engineering field, and that collagen-based surface modifications further enhance the chondrogenic differentiation of MSCs. (C) 2009 Wiley Periodicals, Inc. J Biomed Mater Res 92A: 659-666, 2010