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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增强人间充质干细胞的软骨形成分化。

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

作者信息

Pulat Günnur, Gökmen Oğuzhan, Özcan Şerife, Karaman Ozan

机构信息

Tissue Engineering and Regenerative Medicine Laboratory, Department of Biomedical Engineering, İzmir Katip Çelebi University, İzmir, Turkey.

Bonegraft Biomaterials Co., Ege University Technopolis, İzmir, Turkey.

出版信息

J Biomed Mater Res A. 2025 Jan;113(1):e37786. doi: 10.1002/jbm.a.37786. Epub 2024 Sep 5.

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.

摘要

由于无血管结构以及细胞数量少导致细胞向受损区域的迁移能力低,关节软骨组织无法进行自发修复。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。在此,我们通过直接偶联,用短生物活性基序对自组装肽(SAP)支架之一KLD(KLDLKLDLKLDL)进行功能化,这些短生物活性基序是II型胶原结合肽WYRGRL(C1)的α1链和三螺旋胶原模拟肽GFOGER(C2)。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,以改善软骨形成。使用扫描电子显微镜(SEM)在分子水平观察肽支架结构的整合情况。为确保SAP的稳定性,评估了SAP水凝胶的流变学特性和降解情况。对DNA、糖胺聚糖(GAG)和胶原蛋白含量的生化研究表明,与KLD支架相比,所开发的生物活性SAP水凝胶极大地促进了人间充质干细胞(hMSCs)的增殖。此外,通过实时聚合酶链反应(PCR)评估,向KLD中添加生物活性肽显著提高了重要软骨生成标志物如聚集蛋白聚糖、SOX-9和II型胶原蛋白的表达水平。我们表明,所开发的SAP支架促进了hMSC的增殖和软骨生成分化。尽管之前已经研究了WYRGRL和GFOGER的软骨生成潜力,但没有研究比较这两种肽整合到三维SAP水凝胶中对软骨生成分化的影响。我们的研究结果表明,这些专门创建的生物活性肽支架可能有助于增强软骨组织再生。

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