Establishment of a 3D In Vitro Model to Accelerate the Development of Human Therapies against Corneal Diabetes

建立 3D 体外模型以加速人类角膜糖尿病疗法的开发

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作者:Shrestha Priyadarsini, Akhee Sarker-Nag, Tyler G Rowsey, Jian-Xing Ma, Dimitrios Karamichos

Conclusion

DM has significant effects on physiological and clinical aspects of the human cornea. The benefits in developing and fully characterizing our 3D in vitro model are enormous and might provide clues for the development of novel therapeutics.

Methods

Human corneal fibroblasts from Healthy (HCFs), Type 1DM (T1DM) and Type 2DM (T2DM) donors were isolated and cultured for 4 weeks with Vitamin C stimulation in order to allow for extracellular matrix (ECM) secretion and assembly.

Purpose

To establish an in vitro model that would mirror the in vivo corneal stromal environment in diabetes (DM) patients.

Results

Our data indicated altered cellular morphology, increased cellular migration, increased ECM assembly, and severe mitochondrial damage in both T1DM and T2DMs when compared to HCFs. Furthermore, we found significant downregulation of Collagen I and Collagen V expression in both T1DM and T2DMs. Furthermore, a significant up regulation of fibrotic markers was seen, including α-smooth muscle actin in T2DM and Collagen III in both T1DM and T2DMs. Metabolic analysis suggested impaired Glycolysis and Tricarboxylic acid cycle (TCA) pathway.

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