Collagen binding and mimetic peptide-functionalized self-assembled peptide hydrogel enhance chondrogenic differentiation of human mesenchymal stem cells.

Günnur Pulat, Oğuzhan Gökmen, Şerife Özcan, Ozan Karaman
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Abstract

The avascular structure and low cell migration to the damaged area due to the low number of cells do not allow spontaneous repair of the articular cartilage tissue. Therefore, functional scaffolds obtained from biomaterials are used for the regeneration of cartilage tissue. Here, we functionalized one of the self-assembling peptide (SAP) scaffolds KLD (KLDLKLDLKLDL) with short bioactive motifs, which are the α1 chain of type II collagen binding peptide WYRGRL (C1) and the triple helical collagen mimetic peptide GFOGER (C2) by direct coupling. Our goal was to develop injectable functional SAP hydrogels with proper mechanical characteristics that would improve chondrogenesis. Scanning electron microscopy (SEM) was used to observe the integration of peptide scaffold structure at the molecular level. To assure the stability of SAPs, the rheological characteristics and degradation profile of SAP hydrogels were assessed. The biochemical study of the DNA, glycosaminoglycan (GAG), and collagen content revealed that the developed bioactive SAP hydrogels greatly increased hMSCs proliferation compared with KLD scaffolds. Moreover, the addition of bioactive peptides to KLD dramatically increased the expression levels of important chondrogenic markers such as aggrecan, SOX-9, and collagen Type II as evaluated by real-time polymerase chain reaction (PCR). We showed that hMSC proliferation and chondrogenic differentiation were encouraged by the developed SAP scaffolds. Although the chondrogenic potentials of WYRGRL and GFOGER were previously investigated, no study compares the effect of the two peptides integrated into 3-D SAP hydrogels in chondrogenic differentiation. Our findings imply that these specifically created bioactive peptide scaffolds might help enhance cartilage tissue regeneration.

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胶原蛋白结合和模拟肽功能化的自组装肽水凝胶可增强人间质干细胞的软骨分化。
无血管结构和细胞数量少导致细胞向受损区域的迁移率低,使得关节软骨组织无法自发修复。因此,由生物材料制成的功能性支架被用于软骨组织的再生。在这里,我们通过直接偶联的方式,将一种自组装肽(SAP)支架 KLD(KLDLKLDLKLDL)与短生物活性基团(即 II 型胶原蛋白结合肽 WYRGRL 的 α1 链(C1)和三重螺旋胶原蛋白模拟肽 GFOGER(C2))功能化。我们的目标是开发具有适当机械特性的可注射功能性 SAP 水凝胶,以改善软骨生成。扫描电子显微镜(SEM)用于观察肽支架结构在分子水平上的整合。为确保 SAP 的稳定性,对 SAP 水凝胶的流变特性和降解曲线进行了评估。对 DNA、糖胺聚糖(GAG)和胶原含量的生化研究表明,与 KLD 支架相比,所开发的生物活性 SAP 水凝胶大大提高了 hMSCs 的增殖能力。此外,通过实时聚合酶链反应(PCR)评估,在 KLD 中添加生物活性肽可显著提高重要软骨生成标志物(如 aggrecan、SOX-9 和 II 型胶原)的表达水平。我们的研究表明,所开发的 SAP 支架促进了 hMSC 的增殖和软骨分化。虽然以前对 WYRGRL 和 GFOGER 的软骨生成潜能进行过研究,但还没有研究比较过这两种肽集成到三维 SAP 水凝胶中对软骨生成分化的影响。我们的研究结果表明,这些特制的生物活性肽支架可能有助于促进软骨组织再生。
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