Cellular Contractility Profiles of Human Diabetic Corneal Stromal Cells
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...
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| Format: | Article |
| Language: | English |
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Wiley
2021-01-01
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| Series: | Analytical Cellular Pathology |
| Online Access: | http://dx.doi.org/10.1155/2021/9913210 |
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| author | Thi N. Lam Sarah E. Nicholas Alexander Choi Jian-Xing Ma Dimitrios Karamichos |
| author_facet | Thi N. Lam Sarah E. Nicholas Alexander Choi Jian-Xing Ma Dimitrios Karamichos |
| author_sort | Thi N. Lam |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-0315c89e88164973b454f3162e8cd4ef |
| institution | Kabale University |
| issn | 2210-7177 2210-7185 |
| language | English |
| publishDate | 2021-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Analytical Cellular Pathology |
| spelling | doaj-art-0315c89e88164973b454f3162e8cd4ef2025-02-03T01:08:52ZengWileyAnalytical Cellular Pathology2210-71772210-71852021-01-01202110.1155/2021/99132109913210Cellular Contractility Profiles of Human Diabetic Corneal Stromal CellsThi N. Lam0Sarah E. Nicholas1Alexander Choi2Jian-Xing Ma3Dimitrios Karamichos4Dean McGee Eye Institute, Oklahoma University Health Sciences Center, 608 Stanton L Young Blvd, Oklahoma City, OK 73104, USANorth Texas Eye Research Institute, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, Fort Worth, TX 76107, USADean McGee Eye Institute, Oklahoma University Health Sciences Center, 608 Stanton L Young Blvd, Oklahoma City, OK 73104, USADepartment of Physiology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young, Oklahoma City, OK, USANorth Texas Eye Research Institute, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, Fort Worth, TX 76107, USADiabetic 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.http://dx.doi.org/10.1155/2021/9913210 |
| spellingShingle | Thi N. Lam Sarah E. Nicholas Alexander Choi Jian-Xing Ma Dimitrios Karamichos Cellular Contractility Profiles of Human Diabetic Corneal Stromal Cells Analytical Cellular Pathology |
| title | Cellular Contractility Profiles of Human Diabetic Corneal Stromal Cells |
| title_full | Cellular Contractility Profiles of Human Diabetic Corneal Stromal Cells |
| title_fullStr | Cellular Contractility Profiles of Human Diabetic Corneal Stromal Cells |
| title_full_unstemmed | Cellular Contractility Profiles of Human Diabetic Corneal Stromal Cells |
| title_short | Cellular Contractility Profiles of Human Diabetic Corneal Stromal Cells |
| title_sort | cellular contractility profiles of human diabetic corneal stromal cells |
| url | http://dx.doi.org/10.1155/2021/9913210 |
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