Effect of Vitamin D3 on Regulating Human Tenon's Fibroblasts Activity

Abstract

To study the in vitro effect of vitamin D3 on the healing response of human Tenon's fibroblasts (HTF) and its possible role in preventing excessive postoperative subconjunctival fibrosis.Methods: Effect of vitamin D3 on cytotoxicity and cell survival of primary cultured HTF was measured by lactate dehydrogenase and PrestoBlue assays, respectively. Proliferation and migration of vitamin D3-treated HTF (D3-HTF) was determined by CyQUANT proliferation and scratch assay, respectively. The mRNA expression profiles of control-HTF and D3-HTF from six subjects (three with glaucoma and long-term use of topical medications, three with primary pterygium) were assessed by RNA sequencing analyses to identify potential biomarkers for the inhibitory effect on HTF by vitamin D3. Validation of these biomarkers and their potential pathways were performed by quantitative real-time polymerase chain reaction (qRT-PCR) detection.Results: Pure monolayers of HTF from controls (retinal detachment or squint surgeries), pterygium, and glaucoma subjects were successfully prepared and passaged. Proliferation and migration of pterygium and glaucoma HTF were inhibited by vitamin D3 in a dose-dependent manner, and without cytotoxicity or decrease in cellular viability with concentrations up to 10 μM. The qRT-PCR results were consistent with the transcriptome analyses, vitamin D3 appears to enhance CYP24A1, SHE, KRT16 but suppresses CILP expression in HTF.Conclusions: Vitamin D3 can inhibit the in vitro activity of HTF without compromising cellular survivability at concentration up to 10 μM. This has potential clinical application for improving the outcome of pterygium and filtering surgeries.Translational Relevance: Vitamin D3 can suppress the in vitro proliferation, migration, and transdifferentiation of human Tenon's fibroblasts, without the cytotoxicity of mitomycin-C, the current standard antifibrotic agent in clinical use.

Digital light processing-bioprinted poly-NAGA-GelMA-based hydrogel lenticule for precise refractive errors correction

“…PNG-25%/5% tablets were transferred onto the transwell insert (0.4 µm Polyester Membrane), and then the insert was assembled with six-well plates pre-seeded with hKs. The scratch-healing assay followed our previously published protocol[42].…”

The Role of Vitamin D3 in Ocular Diseases

“…Further longitudinal studies are suggested to explore the potential impact of vitamin D supplementation on glaucoma progression. Vitamin D 3 seems capable of inhibiting the proliferation, migration, and transdifferentiation of human Tenon’s fibroblast cells, which is crucial in optimizing the outcome of trabeculectomy by maintaining aqueous filtration [ 75 ]. The levels of vitamin D found in the aqueous humor were notably lower in patients with both cataracts and glaucoma compared to those with cataracts alone, which means vit.…”