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dc.contributor.authorKaramichos, D.en_US
dc.contributor.authorHutcheon, A. E. K.en_US
dc.contributor.authorRich, C. B.en_US
dc.contributor.authorTrinkaus-Randall, V.en_US
dc.contributor.authorAsara, J. M.en_US
dc.contributor.authorZieske, J. D.en_US
dc.date.accessioned2014-05-06T16:17:39Z
dc.date.issued2014en_US
dc.identifier.citationKaramichos, D., A. E. K. Hutcheon, C. B. Rich, V. Trinkaus-Randall, J. M. Asara, and J. D. Zieske. 2014. “In vitro model suggests oxidative stress involved in keratoconus disease.” Scientific Reports 4 (1): 4608. doi:10.1038/srep04608. http://dx.doi.org/10.1038/srep04608.en
dc.identifier.issn2045-2322en
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:12152919
dc.description.abstractKeratoconus (KC) affects 1:2000 people and is a disorder where cornea thins and assumes a conical shape. Advanced KC requires surgery to maintain vision. The role of oxidative stress in KC remains unclear. We aimed to identify oxidative stress levels between human corneal keratocytes (HCKs), fibroblasts (HCFs) and keratoconus cells (HKCs). Cells were cultured in 2D and 3D systems. Vitamin C (VitC) and TGF-β3 (T3) were used for 4 weeks to stimulate self-assembled extracellular matrix (ECM). No T3 used as controls. Samples were analyzed using qRT-PCR and metabolomics. qRT-PCR data showed low levels of collagen I and V, as well as keratocan for HKCs, indicating differentiation to a myofibroblast phenotype. Collagen type III, a marker for fibrosis, was up regulated in HKCs. We robustly detected more than 150 metabolites of the targeted 250 by LC-MS/MS per condition and among those metabolites several were related to oxidative stress. Lactate levels, lactate/malate and lactate/pyruvate ratios were elevated in HKCs, while arginine and glutathione/oxidized glutathione ratio were reduced. Similar patterns found in both 2D and 3D. Our data shows that fibroblasts exhibit enhanced oxidative stress compared to keratocytes. Furthermore the HKC cells exhibit the greatest level suggesting they may have a myofibroblast phenotype.en
dc.language.isoen_USen
dc.publisherNature Publishing Groupen
dc.relation.isversionofdoi:10.1038/srep04608en
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3980225/pdf/en
dash.licenseLAAen_US
dc.titleIn vitro model suggests oxidative stress involved in keratoconus diseaseen
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden
dc.relation.journalScientific Reportsen
dc.date.available2014-05-06T16:17:39Z
dc.identifier.doi10.1038/srep04608*


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