Effect of Vitamin D3 on Regulating Human Tenon's Fibroblasts Activity.
Jia Shuo, Chen Fushun, Wang Huogang, Kesavamoorthy Gandhervin, Lai Jimmy Shiu-Ming, Wong Ian Yat-Hing, Chiu Kin, Chan Jonathan Cheuk-Hung
AI Summary
Vitamin D3 inhibited human Tenon's fibroblast activity without toxicity, suggesting its potential to improve outcomes in pterygium and glaucoma filtering surgeries by preventing excessive scarring.
Abstract
Purpose
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.
MeSH Terms
Shields Classification
Key Concepts6
Vitamin D3 can inhibit the in vitro activity of human Tenon's fibroblasts (HTF) without compromising cellular survivability at concentrations up to 10 µM.
Vitamin D3 inhibited the proliferation and migration of pterygium and glaucoma human Tenon's fibroblasts (HTF) in a dose-dependent manner, without cytotoxicity or decrease in cellular viability with concentrations up to 10 µM.
Vitamin D3 appears to enhance CYP24A1, SHE, and KRT16 expression while suppressing CILP expression in human Tenon's fibroblasts (HTF).
The in vitro effect of vitamin D3 on the healing response of human Tenon's fibroblasts (HTF) was studied to investigate its possible role in preventing excessive postoperative subconjunctival fibrosis.
The effect of vitamin D3 on cytotoxicity and cell survival of primary cultured human Tenon's fibroblasts (HTF) was measured by lactate dehydrogenase and PrestoBlue assays, respectively.
RNA sequencing analyses were performed on control-HTF and vitamin D3-treated HTF (D3-HTF) from six subjects (three with glaucoma and long-term use of topical medications, three with primary pterygium) to identify potential biomarkers for the inhibitory effect of vitamin D3 on HTF.
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