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首页> 外文期刊>Acta biomaterialia >A self-assembling peptide matrix used to control stiffness and binding site density supports the formation of microvascular networks in three dimensions.
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A self-assembling peptide matrix used to control stiffness and binding site density supports the formation of microvascular networks in three dimensions.

机译：用于控制刚度和结合位点密度的自组装肽基质支持三维微血管网络的形成。

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A three-dimensional (3-D) cell culture system that allows control of both substrate stiffness and integrin binding density was created and characterized. This system consisted of two self-assembling peptide (SAP) sequences that were mixed in different ratios to achieve the desired gel stiffness and adhesiveness. The specific peptides used were KFE ((acetyl)-FKFEFKFE-CONH2), which has previously been reported not to support cell adhesion or MVN formation, and KFE-RGD ((acetyl)-GRGDSP-GG-FKFEFKFE-CONH2), which is a similar sequence that incorporates the RGD integrin binding site. Storage modulus for these gels ranged from ～60 to 6000Pa, depending on their composition and concentration. Atomic force microscopy revealed ECM-like fiber microarchitecture of gels consisting of both pure KFE and pure KFE-RGD as well as mixtures of the two peptides. This system was used to study the contributions of both matrix stiffness and adhesiveness on microvascular network (MVN) formation of endothelial cells and the morphology of human mesenchymal stem cells (hMSC). When endothelial cells were encapsulated within 3-D gel matrices without binding sites, little cell elongation and no network formation occurred, regardless of the stiffness. In contrast, matrices containing the RGD binding site facilitated robust MVN formation, and the extent of this MVN formation was inversely proportional to matrix stiffness. Compared with a matrix of the same stiffness with no binding sites, a matrix containing RGD-functionalized peptides resulted in a ～2.5-fold increase in the average length of network structure, which was used as a quantitative measure of MVN formation. Matrices with hMSC facilitated an increased number and length of cellular projections at higher stiffness when RGD was present, but induced a round morphology at every stiffness when RGD was absent. Taken together, these results demonstrate the ability to control both substrate stiffness and binding site density within 3-D cell-populated gels and reveal an important role for both stiffness and adhesion on cellular behavior that is cell-type specific.
机译：允许控制衬底刚度和整合蛋白结合密度的三维（3-D）细胞培养系统。该系统由两种自组装肽（SAP）序列组成，其以不同的比例混合以达到所需的凝胶刚度和粘合性。使用的特定肽是kFe（（乙酰基）-fkfefkfe-conh2），其先前报告不支持细胞粘附或mvn形成，以及Kfe-Rgd（（乙酰基）-Grgdsp-gg-Fkfefkfe-conh2），这是一种类似的序列，其包含RGD整合蛋白结合位点。这些凝胶的储存模量范围为约60至6000Pa，这取决于它们的组成和浓度。原子力显微镜显示凝胶的ECM样纤维微体系结构，包括纯KFE和纯KFE-RGD，以及两种肽的混合物。该系统用于研究基质刚度和粘合性对内皮细胞的微血管网络（MVN）形成的贡献及人间充质干细胞（HMSC）的形态学。当内皮细胞包封在3-D凝胶基质中而不结合位点时，无论刚度如何，都会发生很少的细胞伸长率并且没有网络形成。相反，含有RGD结合位点的基质促进了鲁棒MVN形成，并且该MVN形成的程度与基质刚度成反比。与没有结合位点的相同刚度的基质相比，含有RGD-官能化肽的基质导致网络结构的平均长度的〜2.5倍，其用作MVN形成的定量测量。当RGD存在时，具有HMSC的矩阵促进了在较高刚度时的细胞突起的数量和长度，但是当RGD不存在时诱导每个刚度的圆形形态。总之，这些结果证明了在3-D细胞填充的凝胶内控制衬底刚度和结合位点密度的能力，并揭示了刚度和粘合性的重要作用，其具有细胞类型的细胞类型。

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