Molecularly designed alginate hydrogels susceptible to local proteolysis as three-dimensional cellular microenvironments
Molecularly designed alginate hydrogels susceptible to local proteolysis as three-dimensional cellular microenvironments
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DOI：
10.1016/j.actbio.2010.12.029
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发表时间：
2011-04-01
影响因子：
9.7
通讯作者：
Barrias, Cristina C.
中科院分区：
文献类型：
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作者：
Fonseca, Keila B.;Bidarra, Silvia J.;Barrias, Cristina C.
开发复杂的三维（3-D）细胞培养微环境，以重现自然细胞外基质（ECM）的一些复杂性，仍然是一项具有挑战性的任务。本文介绍了通过与基质金属蛋白酶（MMP）可切割肽（脯氨酸-缬氨酸-甘氨酸-亮氨酸-异亮氨酸-甘氨酸，PVGLIG）部分交联对海藻酸盐进行改性的方法，并提出了将其用于制备可注射的原位交联水凝胶样基质的方法。采用碳二亚胺化学方法合成了pvglg接枝海藻酸盐，并对其进行了表征。通过比较3d培养的间充质干细胞（MSCs）对仅含有细胞粘附肽（RGD-alginate）或两种肽（PVGLIG/RGD-alginate）的海藻酸盐水凝胶的反应来评估它们的生物学性能。1周后，细胞在rgd -海藻酸盐凝胶中基本保持圆形，而在PVGLIG/ rgd -海藻酸盐水凝胶中细胞表现出细长的形态，形成细胞网络。这表明细胞能够通过酶解PVGLIG残基在结构上重组基质，克服生物物理水凝胶抗性。明胶酶谱分析显示，MSC在被mmp敏感肽功能化的海藻酸盐中培养时，MMP-2活性更高，表明基质组成调节了细胞的蛋白水解表型。此外，PVGLIG/ rgd -海藻酸盐水凝胶在细胞培养中明显降解。综上所述，研究结果表明，在细胞粘附的rgd -海藻酸盐水凝胶中掺入mmp -不稳定肽可以提高其作为ECM类似物的性能，提供更动态和生理的三维细胞微环境。(C) 2010材料学报Elsevier Ltd.出版。版权所有。
The development of sophisticated three-dimensional (3-D) cell culture microenvironments that recreate some of the complexity of the natural extracellular matrix (ECM) remains a challenging task. Here, the modification of alginate through partial crosslinking with a matrix metalloproteinase (MMP) cleavable peptide (proline-valine-glycine-leucine-isoleucine-glycine, PVGLIG) is described, and its use in the preparation of injectable, in situ crosslinkable hydrogel-like matrices is proposed. PVGLIG-grafted alginates were synthesized by carbodiimide chemistry and characterized. Their biological performance was evaluated by comparing the response of 3-D cultured mesenchymal stem cells (MSCs) to alginate hydrogels containing only cell-adhesion peptides (RGD-alginate) or both peptides (PVGLIG/RGD-alginate). After 1 week, cells remained essentially round within RGD-alginate, while they exhibited an elongated morphology within PVGLIG/RGD-alginate hydrogels, forming cellular networks. This suggests that cells were able to structurally reorganize the matrix, through enzymatic hydrolysis of PVGLIG residues, overcoming biophysical hydrogel resistance. As shown by gelatine-zymography, MSC presented higher activity of MMP-2 when cultured within alginate functionalized with MMP-sensitive peptide, suggesting that the cell's proteolytic phenotype was modulated by the matrix composition. Additionally, PVGLIG/RGD-alginate hydrogels were clearly degraded in cell culture. Taken together, the results demonstrate that the co-incorporation of MMP-labile peptides in cell-adhesive RGD-alginate hydrogels improved their performance as ECM analogues, providing a more dynamic and physiological 3-D cellular microenvironment. (C) 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.