title: Establishment of a 3D In Vitro Model to Accelerate the Development of Human Therapies against Corneal Diabetes.
abstract: OBJECTIVE
  To establish an in vitro model that would mirror the in vivo corneal stromal environment in diabetes (DM) patients.
  
  
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Ophthalmology/Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States of America.
authors.forename: Shrestha
authors.initials: S
authors.lastname: Priyadarsini
authors.affiliation: Department of Ophthalmology/Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States of America.
authors.forename: Akhee
authors.initials: A
authors.lastname: Sarker-Nag
authors.affiliation: Department of Biology and Chemistry, East Central University, Ada, Oklahoma, United States of America.
authors.forename: Tyler G
authors.initials: TG
authors.lastname: Rowsey
authors.affiliation: Department of Physiology Harold Hamm Diabetes Center, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States of America.
authors.forename: Jian-Xing
authors.initials: JX
authors.lastname: Ma
authors.affiliation: Department of Ophthalmology/Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States of America.
authors.forename: Dimitrios
authors.identifier: http://orcid.org/0000-0002-8761-3824
authors.initials: D
authors.lastname: Karamichos
mesh_terms: D018929:Cell Culture Techniques; D002465:Cell Movement; D049109:Cell Proliferation; D003316:Corneal Diseases / Q000473:pathology*; D003319:Corneal Stroma / Q000473:pathology*; D048909:Diabetes Complications / Q000473:pathology*; D005347:Fibroblasts; D006801:Humans; D066298:In Vitro Techniques; D055432:Metabolomics; D046529:Microscopy, Electron, Transmission; D008928:Mitochondria / Q000648:ultrastructure