1 Department of Applied Clinical Dietetics, Kitasato Junior College of Health and Hygienic Sciences, Minamiuonuma, Niigata, Japan.
2 Department of Clinical Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo, Japan.
J Biomater Appl. 2019 Feb;33(7):893-902. doi: 10.1177/0885328218823457. Epub 2019 Jan 12.
Extracellular matrix molecules are recognized by several integrin subtypes, making identification of cross-talk among different integrin subtypes difficult. Here, we evaluated the cross-talk of integrin subtypes using four different integrin-binding peptides (FIB1; integrin αvβ3/α5β1, A2G10; integrin α6β1, EF1zz; integrin α2β1, or 531; integrin α3β1) derived from extracellular matrix molecules. Various combinations of two different integrin-binding peptides were mixed and conjugated on a chitosan matrix at various molar ratios and were evaluated for cell attachment activity. FIB1/A2G10 (molar ratio 5:5; total 10 nmol/well)-chitosan matrix significantly enhanced cell attachment activity compared with sum of the cell attachment activity on FIB1 (5 nmol/well)-chitosan matrices and A2G10 (5 nmol/well)-chitosan matrices, respectively. However, none of the other peptides showed a significant activity change when they were mixed and conjugated on a chitosan matrix. We investigated the mechanisms of this enhancement. FIB1/A2G10 (8:2 or 6:4)-chitosan matrix increased the cell spreading, phosphorylation of focal adhesion kinase at Y397, and slightly decreased phosphorylation of caveolin-1 at Y14 in fibroblasts compared with FIB1-chitosan and A2G10-chitosan matrices. These results indicate that FIB1/A2G10 (8:2 or 6:4)-chitosan matrix synergistically enhances cell attachment, suggesting that integrins αvβ3/α5β1 and α6β1 are involved in a cross-talk and synergistically enhance cell attachment. These findings also suggest that the mixed peptide-chitosan matrix system can regulate the ratio of two different peptides and is useful for evaluating cellular functions through receptor-specific cross-talk. Further, FIB1/A2G10 (8:2 or 6:4)-chitosan matrix could be a useful material for tissue engineering.
细胞外基质分子被几种整合素亚基识别，这使得不同整合素亚基之间的串扰难以识别。在这里，我们使用四种不同的整合素结合肽（FIB1；整合素αvβ3/α5β1、A2G10；整合素α6β1、EF1zz；整合素α2β1 或 531；整合素α3β1）评估整合素亚基的串扰，这些肽来自细胞外基质分子。两种不同整合素结合肽的各种组合以不同的摩尔比混合并连接在壳聚糖基质上，并评估其对细胞附着活性的影响。与 FIB1（5 nmol/孔）-壳聚糖基质和 A2G10（5 nmol/孔）-壳聚糖基质的细胞附着活性之和相比，FIB1/A2G10（摩尔比 5:5；总 10 nmol/孔）-壳聚糖基质显著增强了细胞附着活性。然而，当它们混合并连接在壳聚糖基质上时，其他肽都没有显示出明显的活性变化。我们研究了这种增强的机制。与 FIB1-壳聚糖和 A2G10-壳聚糖基质相比，FIB1/A2G10（8:2 或 6:4）-壳聚糖基质增加了成纤维细胞的细胞铺展、粘着斑激酶 Y397 的磷酸化，以及 caveolin-1 Y14 的磷酸化略有减少。这些结果表明，FIB1/A2G10（8:2 或 6:4）-壳聚糖基质协同增强细胞附着，表明整合素αvβ3/α5β1 和α6β1 参与串扰并协同增强细胞附着。这些发现还表明，混合肽-壳聚糖基质系统可以调节两种不同肽的比例，并通过受体特异性串扰用于评估细胞功能。此外，FIB1/A2G10（8:2 或 6:4）-壳聚糖基质可能是组织工程的有用材料。
