Liu Shao Qiong, Tian Quan, Wang Lei, Hedrick James L, Hui James Hoi Po, Yang Yi Yan, Ee Pui Lai Rachel
Department of Pharmacy, National University of Singapore, 18 Science Drive 4, Singapore 117543.
Macromol Rapid Commun. 2010 Jul 1;31(13):1148-54. doi: 10.1002/marc.200900818. Epub 2010 May 25.
In this study, an injectable and biodegradable poly(ethylene glycol) (PEG)/arginine-glycine-aspartic (RGD) peptide hybrid hydrogel has been synthesized and used as a biomimetic scaffold for encapsulation of human mesenchymal stem cells (hMSCs). Tetrahydroxyl PEG was functionalized with acrylate, and then reacted with thiol-containing peptide (RGD). Gelation occurred within 30 min with the addition of cells and PEG-dithiol via Michael addition. The hydrogels synthesized with a peptide concentration of 1.0-5.0 mM achieved significantly greater cell viability when compared to the hydrogels without the RGD peptide. However, the effect of RGD on chondrogenesis was found to be dose-dependent. Immunohistology studies demonstrated that hMSCs encapsulated in the hydrogel matrix with 1.0 mM RGD and TGF-β3 showed enhanced positive staining for aggrecan and type II collagen as compared to that with 5.0 mM RGD and unmodified PEG hydrogels. RT-PCR results further revealed that the cells in hydrogels with 1 mM RGD expressed significantly higher levels of type II collagen than those in PEG hydogels without RGD peptide. These findings have demonstrated that the PEG-RGD hydrogels can be a promising scaffold to deliver hMSCs for cartilage repair.
在本研究中,一种可注射且可生物降解的聚乙二醇(PEG)/精氨酸-甘氨酸-天冬氨酸(RGD)肽杂化水凝胶已被合成,并用作封装人间充质干细胞(hMSCs)的仿生支架。四羟基聚乙二醇用丙烯酸酯官能化,然后与含硫醇的肽(RGD)反应。通过迈克尔加成反应,在加入细胞和聚乙二醇二硫醇后30分钟内发生凝胶化。与不含RGD肽的水凝胶相比,肽浓度为1.0 - 5.0 mM合成的水凝胶具有显著更高的细胞活力。然而,发现RGD对软骨形成的影响是剂量依赖性的。免疫组织学研究表明,与含5.0 mM RGD的水凝胶和未修饰的聚乙二醇水凝胶相比,封装在含1.0 mM RGD和转化生长因子-β3(TGF-β3)的水凝胶基质中的hMSCs对聚集蛋白聚糖和II型胶原的阳性染色增强。逆转录-聚合酶链反应(RT-PCR)结果进一步显示,含1 mM RGD的水凝胶中的细胞比不含RGD肽的聚乙二醇水凝胶中的细胞表达显著更高水平的II型胶原。这些发现表明,聚乙二醇-RGD水凝胶可能是一种有前景的支架,用于递送hMSCs进行软骨修复。