Investigation of the inhibitory effect of trehalose on fibroblasts in pterygium tissue

Pterygium is a degenerative ocular surface disorder, the etiology of which has not been fully elucidated by studies, many hypotheses have been put forward, but in the pathogenesis of fibroblast proliferation. Surgical excision is generally preferred for its treatment. In our study, we aimed to examine the inhibitory effect of trehalose on fibroblasts in vitro and we aimed to analyze which cellular pathways and through which genes it does this. We included a total of 6 patients with primary pterygium, 3 male and 3 female, who applied to our clinic. First, the pterygium and normal conjunctiva of these patients were excised, and primary cell culture was performed. Trehalose(100, 200, 400, 600 mM) and mitomycin C(0.5, 1, 2.5, 5, 10 and 20 µl) were treated and incubated for 48 hours at 37oC in 5% CO2 and 95% humidity was done. At this stage, we observed that trehalose inhibited the proliferation of fibroblasts. Total RNA isolation of drug-free control and trehalose, mitomycin C-treated pterygium and conjunctival cells were performed using Geneall brand Hybrid-R total RNA isolation kit. Two of these samples were then sent for microarray analysis. We detected VIM, FGF7, FGF10, TGFBR2, TGBR3 MSMO1, Tenascin C, RANBP3L, NR4A1, IGFBP4, CLCA2, SERPIN2, ASPN genes is change in pterygium compared to normal conjunctiva. Changes in these genes were striking when trehalose was given. These genes are generally involved in events such as vasculogenesis, migration, wound healing, adhesion, fibroblast proliferation, positive regulation of fibroblast growth factor, and when trehalose was given, the expression of these genes changed to block the formation of pterygium. However, since the number of patients was limited in our study, larger patient groups are needed. Key Words: pterygium, fibroblast, microarray analysis, trehalose, mitomycin C