Co-culture with corneal epithelial cells decreases transformation of tenon`s capsule fibroblasts into myofibroblasts by benzalkonium chloride.

Abstract

Purpose : Long-term use of anti-glaucoma eye drops containing benzalkonium chloride (BAC) is a risk factor of bleb dysfunction after trabeculectomy. The influence of BAC on the transformation of tenon’s capsule fibroblasts into myofibroblasts, considering interactions with corneal epithelium, has not been elucidated yet. In this study we examined the transformation of human tenon`s capsule fibroblast (HTFs) into myofibroblasts by BAC as a single-culture of HTFs or as a co-culture with corneal epithelial cells.

Methods : Primary HTFs were cultured alone or co-cultured with human corneal epithelial cells (HCEs) using hanging mesh inserts. After reaching subconfluence, cells were incubated with serum-free medium for 24h, and subsequently exposed to serum-free medium containing BAC for 24h. BAC at non-toxic concentrations was applied in the study, which were determined by measuring lactate dehydrogenase in the supernatant of the incubating medium. Transformations of myofibroblasts of HTFs were evaluated by immunostaining and immunoblotting for α-smooth muscle actin (α-SMA). The supernatants of the incubating medium were also examined with immunoassays for inflammatory cytokines such as Interleukin-6.

Results : BAC induced transformation of HTF into myofibroblasts and promoted the expression of α-SMA even at non-toxic concentrations. When co-cultured with HCEs, the transformation of HTFs into myofibroblasts was reduced, compared with HTFs when cultured alone. The levels of inflammatory cytokines in the supernatant were also significantly inhibited in co-cultures of HTFs with HCEs compared with those of HTFs alone.</h2>

Conclusions : Transformation of HTFs into myofibroblasts by BAC was suppressed when co-cultured with HCEs. Maintenance of a healthy ocular surface may aid in avoiding postoperative bleb failure due to HTF fibrosis.</h2>

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.