The Efficacy Of Topical Immunosuppressive Agents On The Survival Of Cultivated Conjunctival Equivalents

Abstract

Purpose: : To investigate the efficacy of topical immunosuppressive agents on the survival of conjunctival equivalents with cultivated rabbit epithelial cells on amniotic membrane

Methods: : 26 eyes of the rabbits were allocated to two groups: cultivated conjunctival equivalent transplantation group with rabbit allo–conjunctival epithelial cells (A) and allo– conjunctival graft transplantation group (B). Conjunctival equivalents were constructed by 2 weeks expansion of conjunctival epithelial cells on human amniotic membranes (HAM) in DMEM/F10 (1:1). Temporal conjunctiva was trephined in diameter of 7.5mm, cultivated conjunctival equivalent or allo–conjunctival graft were transplanted in that place. Each eye was treated with topical four times application of 1% steroid (n=7), 1% mycophenolate mofetil (n=4), 0.5% cyclosporine (n=6), 0.03% tacrolimus (n=3), and 0.01% rapamycin (n=2) and control (n=6) in both group. Epithelial defect, and graft edema were evaluated clinically for 7 days. All the rabbits were scarified at 7 days and the expression of pkh26, keratin–4, and –7, as well as MUC5AC were evaluated.

Results: : Epithelial defects were reduced in both groups and were more persistent in group A (p=0.017, Mann–Whitney U test at day 7). Steroid– and rapamycin– applied eyes showed more pkh26 staining cells, less inflammatory cells, more positive staining for keratin–4, –7, and MUC5AC mucin irrespective of the groups. Goblet cell density in group A tended to be higher than that in group B, which was insignificant.

Conclusions: : Topical application of steroid and rapamycin were most effective to enhance graft survival and globlet cell function. Conjunctival equivalents seem to have an equal function compared with allograft.

Keywords: conjunctiva • cell survival • immunomodulation/immunoregulation