Enhancement of chondrogenesis of human adipose derived stem cells in a hyaluronan-enriched microenvironment
Enhancement of chondrogenesis of human adipose derived stem cells in a hyaluronan-enriched microenvironment
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DOI：
10.1016/j.biomaterials.2009.09.089
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发表时间：
2010-02-01
期刊：
影响因子：
14
通讯作者：
Ho, Mei-Ling
中科院分区：
文献类型：
--
作者：
Wu, Shun-Cheng;Chang, Je-Ken;Ho, Mei-Ling
微环境在指导干细胞分化中起着关键作用。我们研究了富含透明质酸（RA）的微环境对人脂肪源性干细胞（hADSC）关节软骨组织工程软骨形成的促进作用。hADSC从接受髋关节置换术的患者中获得。HA包被的威尔斯孔和HA修饰的聚乳酸-羟基乙酸共聚物（HA/PLGA）支架被用作HA富集的微环境。软骨形成（SOX-9、聚集蛋白聚糖和11型胶原）、纤维软骨（1型胶原）的mRNA表达。通过实时聚合酶链反应定量肥大（X型胶原）标记基因。通过阿辛蓝、番红-O染色和二甲基亚甲基蓝（DMMB）试验检测硫酸化糖胺聚糖（sGAG）沉积。免疫组化检测局部11型胶原。在HA包被的威尔斯孔（0.005-0.5 mg/cm（2））中培养的hADSC在培养24 h后显示出增强的聚集和mRNA表达（SOX-9、11型胶原和聚集蛋白聚糖），并且在培养9天后sGAG含量也显著增加。HA修饰的PLGA没有改变hADSC的细胞粘附和活力。在HA/PLGA中培养的hADSCs中软骨形成标记基因的mRNA表达在培养1、3和5天后显著增强，而不是在PLGA支架中培养的hADSCs。在HA/PLGA中培养的hADSC产生更高水平的sGAG和11型胶原，与培养4周后的PLGA支架中的那些相比。我们的研究结果表明，HA富集的微环境诱导hADSCs中的软骨形成，这可能是有益的关节软骨组织工程。(C)2009爱思唯尔有限公司保留所有权利。
Microenvironment plays a critical role in guiding stem cell differentiation. We investigated the enhancing effect of a hyaluronan (RA)-enriched microenvironment on human adipose derived stem cell (hADSC) chondrogenesis for articular cartilage tissue engineering. The hADSCs were obtained from patients undergoing hip replacement. HA-coated wells and HA-modified poly-(lactic-co-glycolic acid) (HA/PLGA) scaffolds were used as the HA-enriched microenvironment. The mRNA expressions of chondrogenic (SOX-9, aggrecan and collagen type 11), fibrocartilage (collagen type 1). and hypertrophic (collagen type X) marker genes were quantified by real-time polymerase chain reaction. Sulfated glycosaminoglycan (sGAG) deposition was detected by Alcian blue, safranin-O staining, and dimethylmethylene blue (DMMB) assays. Localized collagen type 11 was detected by immunohistochemistry. The hADSCs cultured in HA-coated wells (0.005-0.5 mg/cm(2)) showed enhanced aggregation and mRNA expressions (SOX-9, collagen type 11, and aggrecan) after 24 h, and sGAG content was also significantly increased after 9 days of culture. The HA-modified PLGA did not change the cell adherence and viability of hADSCs. The mRNA expressions of chondrogenic marker genes were significantly enhanced in hADSCs cultured in HA/PLGA rather than those cultured in the PLGA scaffold after 1, 3, and 5 days of culture. The hADSCs cultured in HA/PLGA produced higher levels of sGAG and collagen type 11, compared to those in the PLGA scaffold after 4 weeks of cultures. Our results suggest that HA-enriched microenvironment induces chondrogenesis in hADSCs, which may be beneficial in articular cartilage tissue engineering. (C) 2009 Elsevier Ltd. All rights reserved.