Purpose To explore the feasibility and efficacy of a cell delivery system using amniotic membrane (AM) fixed by a novel biomembrane-fixing device (BMFD) for corneal epithelium reconstruction in rabbits with limbal stem cell deficiency (LSCD).
Methods Sixty female rabbits with LSCD were created and randomly assigned to three groups of 20 each: LSCD rabbits without treatment (the control), LSCD rabbits treated with BMFD-fixed AM (BMFD-AM), and rabbits treated with male human limbal epithelial cells delivered with BMFD-fixed AM (BMFD-AM+cells). They were followed up with slit lamp observation and corneal fluorescein staining for 14 days. Cytokeratin K3 and K4 and mucin 5AC were used to evaluate corneal conjunctivalization. Sry gene detection was used to trace the delivered cells.
Results  The corneal re-epithelialization time was 5.60 ± 0.46 days in the BMFD-AM+cell group, significantly shorter (P < 0.05) than in the LSCD (12.45 ± 0.65 days) and the BMFD-AM (9.25 ± 0.51 days) groups. Conjunctivalization and neovascularization were observed to be severe in the LSCD group and moderate in the BMFD-AM group. The prevention of conjunctivalization in the BMFD-AM+cell group was evidenced by positive K3/K12 and negative MUC5AC and K4 observed on re-epithelialized corneal epithelium. The histologic sections at different time points and positive Sry gene expression indicated that the delivered cells adhered to the wounded corneal surface and proliferated well.
Conclusions These findings demonstrate that the BMFD with fixed AM served well as a cell delivery system for the ocular surface. The delivered limbal epithelial cells promoted corneal re-epithelialization and prevented corneas from conjunctivalization and neovascularization in rabbits with experimental LSCD.