Chondrogenic differentiation of human bone marrow mesenchymal stem cells on polyhydroxyalkanoate (PHA) scaffolds coated with PHA granule binding protein PhaP fused with RGD peptide
Chondrogenic differentiation of human bone marrow mesenchymal stem cells on polyhydroxyalkanoate (PHA) scaffolds coated with PHA granule binding protein PhaP fused with RGD peptide
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DOI：
10.1016/j.biomaterials.2010.12.009
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发表时间：
2011-03-01
期刊：
影响因子：
14
通讯作者：
Chen, Guo-Qiang
中科院分区：
文献类型：
--
作者：
You, Mingliang;Peng, Gongfeng;Chen, Guo-Qiang
以3-羟基丁酸酯-羟基己酸酯共聚物（PHBHHx）为骨架材料，制备了疏水性聚羟基脂肪酸酯（PHA）支架，并将其与RGD肽融合的PHA颗粒结合蛋白PhaP（PhaP-RGD）进行了包被。将人骨髓间充质干细胞（human bone marrow mesenchymal stem cells，hBMSCs）接种于软骨组织工程支架上，诱导其向软骨细胞分化，形成关节软骨。PhaP-RGD包被的支架与未包被或包被的支架相比，细胞在支架中的分布更均匀，细胞粘附、增殖和成软骨分化更好。此外，通过扫描电子显微镜成像，软骨细胞特异性基因（包括SOX-9、聚集蛋白聚糖和II型胶原蛋白）的表达增强，证明了在PhaP-RGD涂覆的支架上/中，分化的细胞在14天的时间内产生更多的细胞外基质，表明了RGD对细胞外基质产生的积极作用。此外，软骨特异性细胞外物质硫酸化糖胺聚糖（sGAG）和总胶原蛋白含量发现上/中的PhaP-RGD涂层的支架显着更多的相比，由控制和PhaP仅涂层的支架。3周后，在PhaP-RGD包被的支架上/中观察到形成软骨样基质的均匀分布的软骨细胞样细胞。结果表明，PhaP-RGD包被的PHBHHx支架可促进hBMSCs向软骨细胞分化，为软骨组织工程提供支持。(C)2010爱思唯尔有限公司版权所有。
Hydrophobic polyhydroxyalkanoate (PHA) scaffolds made of a copolyester of 3-hydroxybutyrate-co-hydroxyhexanoate (PHBHHx) were coated with a fusion protein PHA granule binding protein PhaP fused with RGD peptide (PhaP-RGD). Human bone marrow mesenchymal stem cells (hBMSCs) were inoculated on/in the scaffolds for formation of articular cartilages derived from chondrogenic differentiation of hBMSCs for cartilage tissue engineering. PhaP-RGD coating led to more homogeneous spread of cells, better cell adhesion, proliferation and chondrogenic differentiation in the scaffolds compared with those of PhaP coated or uncoated scaffolds immerging in serum minus chondrogenic induction medium. In addition, more extracellular matrices were produced by the differentiated cells over a period of 14 days on/in the PhaP-RGD coated scaffolds evidenced by scanning electron microscopy imaging, enhanced expression of chondrocyte specific genes including SOX-9, aggrecan and type II collagen, suggesting the positive effect of RGD on extracellular matrix production. Furthermore, cartilage-specific extracellular substances sulphated glycosaminoglycans (sGAG) and total collagen content found on/in the PhaP-RGD coated scaffolds were significantly more compared with that produced by the control and PhaP only coated scaffolds. Homogeneously distributed chondrocytes-like cells forming cartilage-like matrices were observed on/in the PhaP-RGD coated scaffolds after 3 weeks. The results suggested that PhaP-RGD coated PHBHHx scaffold promoted chondrogenic differentiation of hBMSCs and could support cartilage tissue engineering. (C) 2010 Elsevier Ltd. All rights reserved.