Abstract
Purpose :
It is unclear whether diabetes mellitus (DM) affects corneal stromal fibroblast (CSF) morphology, functional response, and wound healing properties. We sought to find similarities and discrepancies in cellular morphology and functional biomarkers of cell proliferation, differentiation, and migration of CSFs retrieved from Type-I Diabetic (DT1), Type-II Diabetic (DT2), and non-diabetic donor human corneas (ND).
Methods :
Experiments were done on donor human corneas received from Saving Sight, Kansas City, Missouri. Corneas were procured and processed within ten days of death, and either used to isolate human corneal stromal fibroblasts (hCSFs), mRNA for molecular studies, or corneal tissue sections for histological investigations. In vitro studies used hCSFs to determine morphological and functional differences between ND, DT1, and DT2. Ex vivo studies were also conducted, where donor human corneas were directly processed to study the differences in gene expression using qRT-PCR and protein quantification, and localization was studied using IHC staining.
Results :
The hCSFs isolated from donor ND, DT1, and DT2 human corneas showed no significant differences in cellular morphology. The functional response of hCSFs obtained from ND, DT1, and DT2 depended upon experimental conditions. In general, hCSFs of DT2 showed a slightly slow proliferation, but similar viability in vitro compared to the ND and DT1. On the other hand, hCSFs of ND, DT1, and DT2 demonstrated a notable difference in migration rate within in vitro wound scratch assays. The differentiation of hCSFs of ND, DT1, and DT2 in TGFb mimicking injury condition is under investigation. Analyses and qRT-PCR and IHC are underway.
Conclusions :
The corneal wound healing response appears to vary between diabetic and non-diabetic corneas. In-depth investigations are ongoing with animal and human experimental models.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.