Peptide-Functionalized Hydrogels Modulate Integrin Expression and Stemness in Adult Human Epidermal Keratinocytes
Peptide-Functionalized Hydrogels Modulate Integrin Expression and Stemness in Adult Human Epidermal Keratinocytes
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DOI：
10.1002/adbi.201900022
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发表时间：
2019-08-23
影响因子：
4.1
通讯作者：
Magin, Chelsea M.
中科院分区：
文献类型：
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作者：
Davis-Hall, Duncan;Vy Nguyen;Magin, Chelsea M.
细胞外基质（ECM）通过β-整联蛋白信号传导控制角质形成细胞增殖、迁移和分化。伤口愈合研究需要在体外扩增细胞，同时保持复制能力;然而，传统培养条件下的早期终末分化限制了扩增。在这里，实验设计方法鉴定了具有机械性能（弹性模量，E = 20.9 +/-0.56kPa）和模拟表皮ECM的生物活性肽序列的基于聚（乙二醇）的水凝胶制剂。这些水凝胶能够系统地研究来自纤连蛋白（RGDS）、层粘连蛋白（YIGSR）和胶原IV（HepIII）的细胞结合结构域对角质形成细胞干性和β（1）整联蛋白表达的影响。14天角蛋白表达的定量显示，与标准技术相比，四种水凝胶改善了干性。三种水凝胶增加β（1）整联蛋白表达，证明干性和β（1）整联蛋白表达之间的正线性关系。多因素统计分析预测了用于维持体外干性的最佳肽组合（[RGDS] = 0.67 mm，[YIGSR] = 0.13 mm和[HepIII] = 0.02 mm）。与标准相比，表现最佳的水凝胶表现出Ki-67阳性细胞没有减少（减少15%，第7天至第14天; p < 0.05，Tukey检验）。这些数据表明，精确设计的水凝胶生物材料指导整合素表达并促进增殖，提高了培养的角质形成细胞的再生能力，用于基础科学和转化工作。
The extracellular matrix (ECM) controls keratinocyte proliferation, migration, and differentiation through beta-integrin signaling. Wound-healing research requires expanding cells in vitro while maintaining replicative capacity; however, early terminal differentiation under traditional culture conditions limits expansion. Here, a design of experiments approach identifies poly(ethylene glycol)-based hydrogel formulations with mechanical properties (elastic modulus, E = 20.9 +/- 0.56 kPa) and bioactive peptide sequences that mimic the epidermal ECM. These hydrogels enable systematic investigation of the influence of cell-binding domains from fibronectin (RGDS), laminin (YIGSR), and collagen IV (HepIII) on keratinocyte stemness and beta(1) integrin expression. Quantification of 14-day keratin protein expression shows four hydrogels improve stemness compared to standard techniques. Three hydrogels increase beta(1) integrin expression, demonstrating a positive linear relationship between stemness and beta(1) integrin expression. Multifactorial statistical analysis predicts an optimal peptide combination ([RGDS] = 0.67 mm, [YIGSR] = 0.13 mm, and [HepIII] = 0.02 mm) for maintaining stemness in vitro. Best-performing hydrogels exhibit no decrease in Ki-67-positive cells compared to standards (15% decrease, day 7 to 14; p < 0.05, Tukey Test). These data demonstrate that precisely designed hydrogel biomaterials direct integrin expression and promote proliferation, improving the regenerative capability of cultured keratinocytes for basic science and translational work.