Three-dimensional culture of differentiating marrow stromal osteoblasts in biomimetic poly(propylene fumarate-co-ethylene glycol)-based macroporous hydrogels
Three-dimensional culture of differentiating marrow stromal osteoblasts in biomimetic poly(propylene fumarate-co-ethylene glycol)-based macroporous hydrogels
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DOI：
10.1002/jbm.a.10003
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发表时间：
2003-09-01
影响因子：
4.9
通讯作者：
Mikos, AG
中科院分区：
文献类型：
--
作者：
Behravesh, E;Mikos, AG
（T）他的研究评估了基于仿生聚（丙二醇酯-共-乙二醇）的水凝胶维持骨髓基质细胞（MSC）向成骨细胞表型分化并在体外产生矿化基质的能力。大孔水凝胶的基础上聚（丙二醇酯-共-乙二醇）与和没有共价连接的RGD细胞粘附肽的合成和接种大鼠骨髓间充质干细胞悬浮在培养基中或在I型胶原蛋白溶液。发现悬浮在培养基中的细胞粘附于RGD修饰的水凝胶，但不粘附于未修饰的水凝胶。细胞悬浮在胶原蛋白溶液中被截留后，胶原蛋白凝胶化和增殖的肽修饰的水凝胶独立。用RGD肽进行水凝胶修饰足以使MSC在成骨培养补充剂的存在下粘附和分化成成骨细胞表型。培养28天后，用胶原凝胶接种到RGD修饰的大孔水凝胶上的MSC显示出细胞数量的显著增加，从15，200 +/-2，000增加到208，600 +/-69，700（p < 0.05）。此外，与悬浮在培养基中的细胞相比，悬浮在胶原凝胶中的细胞在RGD修饰的水凝胶中培养28天后明显出现显著的钙沉积，3.47 +/-0.26相比于0.82 +/-0.20mgC2 +/支架（p < 0.05）。共聚焦显微镜显示，悬浮在胶原凝胶中并在RGD修饰的水凝胶上培养28天的MSC粘附于水凝胶的表面，而悬浮在胶原凝胶中并在未修饰的水凝胶上培养的MSC位于水凝胶表面的孔内，并且不与水凝胶表面直接接触。结果表明，这些仿生水凝胶促进粘附，并支持骨髓间充质干细胞分化成骨细胞表型的成骨培养基的存在下。
(T)his study assesses the ability of biomimetic poly(propylene fumarate-co-ethylene glycol)-based hydrogels to sustain the differentiation of marrow stromal cells (MSCs) to the osteoblastic phenotype and to produce a mineralized matrix in vitro. Macroporous hydrogels based on poly(propylene fumarate-co-ethylene glycol) with and without covalently linked RGD cell-adhesive peptide were synthesized and seeded with rat MSCs suspended in media or in a type I collagen solution. Cells suspended in media were found to adhere to RGD-modified but not to unmodified hydrogels. Cells suspended in a collagen solution were entrapped after collagen gelation and proliferated independent of the peptide modification of the hydrogel. Hydrogel modification with RGD peptide was sufficient to allow for the adhesion and differentiation of MSCs to the osteoblastic phenotype in the presence of osteogenic culture supplements. MSCs seeded with a collagen gel onto RGD-modified macroporous hydrogels after 28 days of culture showed a significant increase in cell numbers, from 15,200 +/- 2,000 to 208,600 +/- 69,700 cells (p < 0.05). Moreover, significant calcium deposition was apparent after 28 days of culture in RGD-modified hydrogels for cells suspended in a collagen gel in comparison to cells suspended in media, 3.47 +/- 0.26 compared to 0.82 +/- 0.20 mg C2+ per scaffold (p < 0.05). Confocal microscopy revealed that MSCs suspended in a collagen gel and cultured on RGD-modified hydrogels for 28 days were adhered to the surface of the hydrogel while MSCs suspended in a collagen gel and cultured on unmodified hydrogels were located within the pores of and not in direct contact with the hydrogel surface. The results demonstrate that these biomimetic hydrogels facilitate the adhesion and support the differentiation of MSCs to the osteoblastic phenotype in the presence of osteogenic culture media.