Effects of docosahexaenoic acid on the viability of human tenon's fibroblasts

Abstract

PurposeExcessive scarring of the conjunctiva is the major cause of failure of the surgical treatment for glaucoma. The activation of human Tenon's fibroblast (HTFs) is supposed to be responsible for wound healing and scar formation. Antimetabolites are used to inhibit subconjunctival fibrosis but these molecules may lead to serious complication. Previous studies have shown that omega‐3 polyunsaturated fatty acids (PUFAs) such as docosahexaenoic acid (DHA) have anti‐inflammatory, anti‐proliferative and anti‐angiogenic properties. The aims of this study were to evaluate DHA as a potential inhibitor of human Tenon's fibroblasts.MethodsPrimary HTF cells were obtained from patients during glaucoma surgery and were cultured in DMEM medium. After stimulation with TGF‐β2, HTFs were treated with MMC (0.01 mg/mL) and different concentrations of DHA (50, 100 and 200 μM) for 48 hours. Lipid profile of HTFs was determined by gas chromatography, cell proliferation by Ki67 assay, cell migration was evaluated by videomicroscopy and cell toxicity by MTT assay. The expression α‐SMactin and MAPK p38, ERK, JNK in their non‐phosphorylated and phosphorylated forms was determined by western blotting.ResultsMMC reduced proliferation, migration compared to control but showed important toxicity suggested by the increased respiratory rate of mitochondrie in cytotoxicity assay and confirmed by a G2 arrest in cell cycle. DHA had no significant impact on cellular viability. DHA at 200 μM showed an efficacity similar to MMC. A reduction of the ability of cells to migrate was observed with DHA in a dose‐dependent manner. DHA efficiently dicreased α‐SMactin expression of myofibroblasts. Proteins from the Smad signaling pathway could not be observed.ConclusionsThese findings indicate that DHA inhibits migration and differentiation of HTFs to myofibroblats.