One-year follow-up of epithelial corneal cell sheet allografts mounted on platlet poor plasma in rabbits

[摘要] Purpose: To evaluate the usefulness ofepithelial corneal sheets mounted on platelet poor plasma (PPP) forallograft transplantation of rabbits with total limbal stem celldeficiency (LSCD) and to prove its efficacy at 1 year after surgery. Methods: LSCD was induced in 21 femalerabbits by mechanical keratectomy. To configure the grafts, limbalbiopsies were taken from male rabbits and cells were cultured on afibroblast feeder layer grown on clotted autologous PPP. Afterkeratectomy, grafts were sutured over the stroma. Control groupsconsisted of no implant or an implant of clotted PPP. Rabbits wereeuthanized at 3 and 12 months. Corneas and cultured sheets wereprocessed for histopathology and immunohistochemistry (K3/12 and K19).Gender analysis was performed at 4 and 7 months. Results: One rabbit had endophthalmitis,and another died of no apparent cause. The rest of the animals treatedhad no inflammation, showed a stratified epithelium, keratin 3/12expression, and no expression of keratin 19. At 1 year, seven of eightrabbits showed no LSCD or corneal rejection signs. Y chromosomes weredetected at 4 and 7 months postoperatively. All controls showed LSCDsigns, erratic epithelium, and minimal cell differentiation; theyrevealed a slight expression of K3/12 and an expression of K19 inpatchy patterns. Conclusions: Allografts contributed torestoring a healthy eye surface without signs of graft rejection. Thistechnique seems to be a promising procedure for bilateral ocularsurface diseases and may be useful for new therapeutic strategies.

[学科分类] 生物化学/生物物理