Modulation of differentiation and mineralization of marrow stromal cells cultured on biomimetic hydrogels modified with Arg-Gly-Asp containing peptides
Modulation of differentiation and mineralization of marrow stromal cells cultured on biomimetic hydrogels modified with Arg-Gly-Asp containing peptides
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DOI：
10.1002/jbm.a.30027
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发表时间：
2004-06-01
影响因子：
4.9
通讯作者：
Mikos, AG
中科院分区：
文献类型：
--
作者：
Shin, H;Zygourakis, K;Mikos, AG
合成骨桥蛋白衍生肽（ODP）修饰的仿生水凝胶，并将其作为基质体外培养骨髓基质细胞（MSCs），研究仿生表面对MSCs向成骨细胞分化的影响。16天的增殖和生物实验证明MSCs分化为成骨细胞，分泌成骨表型标志物，如碱性磷酸酶（ALP）、骨桥蛋白和矿化钙。此外，在细胞培养基中存在可溶性成骨补充剂时，细胞结合肽对MSCs的分化和矿化有附加效应。例如，第16天，肽修饰水凝胶的钙含量显著高于组织培养聚苯乙烯。观察到两个总体趋势：(1)随着分化标记物数量的增加，MSCs的增殖能力下降；(2)较高的肽浓度加速了MSCs的分化。在ODP修饰的水凝胶上，当浓度为2 μ mol/g时，ALP活性在第10天达到最大值36.7 +/- 4.2 pmol/细胞/h，而在较低浓度时，ALP活性达到最大值所需的培养时间为第13天。在同一水凝胶上，当肽浓度为2 μ mol/g时，第10天钙含量为21.4 +/- 2.3 ng/细胞；当肽浓度为1.0 μ mol/g时，钙含量为1.0 +/- 0.3 ng/细胞。我们使用Gly-Arg-Gly-Asp-Ser （GRGDS）对水凝胶进行修饰，并与ODP的结果进行比较。然而，成骨细胞的发育并没有受到结合肽序列性质的显著影响。这些结果表明，骨髓间充质干细胞的功能可以通过改变用于支架骨组织工程的仿生水凝胶中的肽浓度来调节。(C) 2004 Wiley期刊有限公司
We synthesized biomimetic hydrogels modified with an osteopontin-derived peptide (ODP) and used them as a substrate for in vitro culture of marrow stromal cells (MSCs) to investigate the effect of the biomimetic surface on differentiation of MSCs into osteoblasts. Proliferation and biological assays for 16 days proved that MSCs became differentiated into osteoblasts secreting osteogenic phenotypic markers such as alkaline phosphatase (ALP), osteopontin, and mineralized calcium. In addition, there was an additive effect of the cell-binding peptide on differentiation and mineralization of MSCs cultured in the presence of soluble osteogenic supplements in cell culture media. For example, calcium content at day 16 on peptide-modified hydrogels was significantly higher than on tissue culture polystyrene. Two general trends were observed: (1) proliferation of MSCs decreased as the amount of differentiation markers increased, and (2) higher peptide concentrations accelerated the differentiation of MSCs. On the hydrogel modified with ODP, ALP activity exhibited a maximum value of 36.7 +/- 4.2 pmol/cell/h at day 10 for the concentration of 2 mumol/g while the culture time needed for maximum ALP activity occurred on day 13 for the lower concentrations. On the same hydrogel, the calcium content at day 10 was 21.4 +/- 2.3 ng/cell for the peptide concentration of 2 mumol/g and 1.0 +/- 0.3 ng/cell for 1.0 mumol/g. We used Gly-Arg-Gly-Asp-Ser (GRGDS) for modification of the hydrogel as a comparison to the results with ODP. However, osteoblast development was not significantly affected by the nature of the binding peptide sequences. These results suggest that MSC function can be modulated by variation of the peptide concentration in biomimetic hydrogels used for scaffold-based bone tissue engineering. (C) 2004 Wiley Periodicals, Inc.