Restoration of corneal clarity after transplantation of an engineered corneal endothelium: preliminary in vivo model

Abstract

Purpose : To determine the ability of an engineered corneal endothelium to restore corneal clarity in an animal model.

Methods : Corneal endothelial cells were isolated from three 3 months old New Zealand rabbits and harvested with a two-step approach. Collagen membranes were elaborated using bovine collagen type I (5.6 µL/mm² of the culture plate surface or 2X) dissolved in DMEM-F12 medium, 22 mM HEPES solution, 8% FBS, and 1 % antibiotics. The mixture was incubated 2h at 37 °C for gelation then placed in a 40 °C and 40% RH for 37 days. The corneal endothelial cells were seeded at a density of 1250 cells/mm² overnight over 8 mm² membranes. Five male New Zealand rabbits 20 months old were used as models. Corneal endothelium was removed under local and general anesthesia and the membrane/cells constructs were transplanted into the anterior chamber of the right eye of 4 rabbits. One rabbit was transplanted using a collagen membrane elaborated with 8.4 µL/mm² collagen (3X). The transplanted eyes were photo-documented for 30 days.

Results : The five eyes developed opacity after 5 min of corneal endothelium removal. The eyes transplanted with 2X membrane constructs (rabbits 1, 2, 3, and 4) recovered partially the peripherical clarity after 3 weeks. After 5 weeks, rabbits 1, 2, and 4 showed central corneal clarity. Central opacity remained in rabbit 3. The eye transplanted with 3X membrane construct remained with mild opacity after 5 weeks of transplantation.

Conclusions : Engineered corneal endothelium produced with corneal endothelial cells cultivated with a two-step approach and 2X collagen membranes restores the corneal clarity in old White New Zealand rabbits. The concentration of collagen in the fabrication of the membrane could affect the recovery of corneal clarity.

This is a 2021 ARVO Annual Meeting abstract.

 

Corneal clarity restoration of rabbits 1 to 5 (columns) throughout 5 weeks (rows) and comparison with the control eye (last column).