Diabetic keratopathy is a corneal complication of diabetes mellitus (DM). Patients with diabetic keratopathy are prone to developing corneal haze, scarring, recurrent erosions, and significant wound healing defects/delays. The purpose of this study was to determine the contractility profiles in the diabetic human corneal stromal cells and characterize their molecular signatures. Primary human corneal fibroblasts from healthy, Type 1 DM (T1DM), and Type 2 DM (T2DM) donors were cultured using an established 3D collagen gel model. We tracked, measured, and quantified the contractile footprint over 9 days and quantified the modulation of specific corneal/diabetes markers in the conditional media and cell lysates using western blot analysis. Human corneal fibroblasts (HCFs) exhibited delayed and decreased contractility compared to that from T1DMs and T2DMs. Compared to HCFs, T2DMs demonstrated an initial downregulation of collagen I (day 3), followed by a significant upregulation by day 9. Collagen V was significantly upregulated in both T1DMs and T2DMs based on basal secretion, when compared to HCFs. Cell lysates were upregulated in the myofibroblast-associated marker, α-smooth muscle actin, in T2DMs on day 9, corresponding to the significant increase in contractility rate observed at the same time point. Furthermore, our data demonstrated a significant upregulation in IGF-1 expression in T2DMs, when compared to HCFs and T1DMs, at day 9. T1DMs demonstrated significant downregulation of IGF-1 expression, when compared to HCFs. Overall, both T1DMs and T2DMs exhibited increased contractility associated with fibrotic phenotypes. These findings, and future studies, may contribute to better understanding of the pathobiology of diabetic keratopathy and ultimately the development of new therapeutic approaches.

中文翻译：

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人糖尿病角膜基质细胞的细胞收缩性特征

糖尿病性角膜病是糖尿病（DM）的角膜并发症。糖尿病性角膜病患者容易出现角膜混浊、疤痕、反复糜烂和严重的伤口愈合缺陷/延迟。本研究的目的是确定糖尿病人角膜基质细胞的收缩性并表征其分子特征。使用已建立的 3D 胶原凝胶模型培养来自健康、1 型糖尿病 (T1DM) 和 2 型糖尿病 (T2DM) 供体的原代人角膜成纤维细胞。我们跟踪、测量和量化了 9 天的收缩足迹，并使用蛋白质印迹分析量化了条件培养基和细胞裂解物中特定角膜/糖尿病标记物的调节。与 T1DM 和 T2DM 相比，人角膜成纤维细胞 (HCF) 表现出延迟和降低的收缩性。与 HCF 相比，T2DM 最初表现出 I 型胶原蛋白下调（第 3 天），随后在第 9 天显着上调。与 HCF 相比，基于基础分泌，T1DM 和 T2DM 中的胶原蛋白 V 显着上调。第 9 天，T2DM 中细胞裂解物中肌成纤维细胞相关标记物α-平滑肌肌动蛋白的表达上调，这与同一时间点观察到的收缩率显着增加相对应。此外，我们的数据表明，与 HCF 和 T1DM 相比，第 9 天时 T2DM 中 IGF-1 表达显着上调。与 HCF 相比，T1DM 表现出 IGF-1 表达显着下调。总体而言，T1DM 和 T2DM 均表现出与纤维化表型相关的收缩性增加。这些发现和未来的研究可能有助于更好地了解糖尿病角膜病的病理学，并最终开发新的治疗方法。