Clevenger Tracy N, Hinman Cassidy R, Ashley Rubin Rebekah K, Smither Kate, Burke Daniel J, Hawker Craig J, Messina Darin, Van Epps Dennis, Clegg Dennis O
1 Center for Stem Cell Biology and Engineering, University of California , Santa Barbara, Santa Barbara, California.
2 Department of Molecular, Cellular and Developmental Biology, University of California , Santa Barbara, Santa Barbara, California.
Tissue Eng Part A. 2016 Apr;22(7-8):597-609. doi: 10.1089/ten.TEA.2015.0550. Epub 2016 Mar 31.
Soft tissue defects are relatively common, yet currently used reconstructive treatments have varying success rates, and serious potential complications such as unpredictable volume loss and reabsorption. Human adipose-derived stem cells (ASCs), isolated from liposuction aspirate have great potential for use in soft tissue regeneration, especially when combined with a supportive scaffold. To design scaffolds that promote differentiation of these cells down an adipogenic lineage, we characterized changes in the surrounding extracellular environment during adipogenic differentiation. We found expression changes in both extracellular matrix proteins, including increases in expression of collagen-IV and vitronectin, as well as changes in the integrin expression profile, with an increase in expression of integrins such as αVβ5 and α1β1. These integrins are known to specifically interact with vitronectin and collagen-IV, respectively, through binding to an Arg-Gly-Asp (RGD) sequence. When three different short RGD-containing peptides were incorporated into three-dimensional (3D) hydrogel cultures, it was found that an RGD-containing peptide derived from vitronectin provided strong initial attachment, maintained the desired morphology, and created optimal conditions for in vitro 3D adipogenic differentiation of ASCs. These results describe a simple, nontoxic encapsulating scaffold, capable of supporting the survival and desired differentiation of ASCs for the treatment of soft tissue defects.
软组织缺损相对常见,但目前使用的重建治疗方法成功率各不相同,且存在严重的潜在并发症,如不可预测的体积损失和再吸收。从抽脂吸出物中分离出的人脂肪干细胞(ASC)在软组织再生中具有巨大的应用潜力,尤其是与支持性支架结合使用时。为了设计能促进这些细胞向脂肪生成谱系分化的支架,我们对脂肪生成分化过程中周围细胞外环境的变化进行了表征。我们发现细胞外基质蛋白的表达发生了变化,包括IV型胶原蛋白和玻连蛋白表达增加,以及整合素表达谱的变化,如αVβ5和α1β1等整合素的表达增加。已知这些整合素分别通过与精氨酸-甘氨酸-天冬氨酸(RGD)序列结合,特异性地与玻连蛋白和IV型胶原蛋白相互作用。当将三种不同的含RGD短肽掺入三维(3D)水凝胶培养物中时,发现源自玻连蛋白的含RGD肽提供了强大的初始附着,维持了所需的形态,并为ASC的体外3D脂肪生成分化创造了最佳条件。这些结果描述了一种简单、无毒的封装支架,能够支持ASC的存活和所需的分化,用于治疗软组织缺损。