| ObjectiveThe ocular surface disorder with severe alkali burns is one of common diseases and difficult to treat in the clinic. The corneal graft is usually failed to survival because of immune rejection. The purposes of this studies are: (1) To investigate the pharmacological and toxicological effect of mitomycin C (MMC) and angiostatin (AS) to the vascular endothelial cells, corneal epithelials cell and fibloblast, and to detere the best concentration of MMC and AS which would be applied in animal experimental and clinical studies. (2) To investigate the curative effect of the therapeutics treated with corneal graft, limbal graft, limbal graft and MMC applied intraoperatively, limbal graft and AS applied post-operatively, and limbal graft + MMC + AS for severe ocular surface alkali burns in rabbits. Consequently, to evaluate the effects of corneal limbal stem cells, mitomycin C and angistatin treating the immune rejection after allograft transplantation on ocular surface alkali burns. (3) To estimate the clinical curative effects of combining with allogenic limbal stem cells graft, MMC and AS on treating the severe ocular surface alkali bums in clinic, and compare with the effect of the corneal allograft transplantation.Materials and Methods(1) The experimental studies of cells culture in vitro: After the vascular endothelial cells, corneal epithelial cells and fibloblast were separated, all these cells were cultured and transferred in the incubation(37℃ -, 5%CCO). When transferred for the second generation, cells were inoculated in different culture pore. Each kind of cells were transferred in 96 wells toculture respectively, and were divided into seven groups: No. 1-the control group, No.2, 3, 4-MMC grops, No. 5, 6, 7 AS groups; each group had 12 wells.When cells anchoring, 12 wells of No. 1 group were added with 200μl DMEM culture medium, MMC grops were added with 0.1 > 0.3 -. 0.5mg/ml MMC-DMEM respectively, for 5 utes,then removed the MMC-DMEM and added new DMEM; and 0.K0.3>0.5μg/ml AS-DMEM culture medium were adding to AS groups, respectively. The conditions of cellular growth were observed in 24 and 48 hours, and the OD values of all wells were exaated by MTT mothods.The rate of the cellular inhibition=[(the control OD-the experimental OD) the control OD] X100%. This experiment was repeated for three times.(2) The experimental studies of treatments to resume vision for ocular surface alkali burns in rabbits: Ocular surface alkali burned models were made (with I.ON NaOH liquid) on bilateral eyes of 25 rabbits (serial number: No.1 to No.25). One day latter after burned, lamellar corneal allograft transplantation were performed on the left eyes of No. 1-10 rabbits(the first group, corneal graft group), the right eyes underwent lamellar limbal allograft transplantation(the second group, limbal graft.group); all eyes of No. 11-15 were treated with lamellar limbus allograft and mitomycin-C(0.25mg/ml) were applied intraoperatively for 5 utes on limbus of all eyes.(the third group, MMC group); each eye of No. 15-20 rabbits was managed with lamellar limbus allograft and angiostatin(0.3μg/ml) were locally adistered to these eyes by quarter in day for 3 months(the fourth group, AS group); eyes of No.20-25 rabbits were treated with lamellar limbal allograft transplantation combining with mitomycin-C applied intraoperatively and angiostatin eye-drops adistered locally post-operatively(the fifth group, the united therapeutic group).All eyes Of 25 rabbits were adistered locally with FK506 eye-drops and Tobramycin Co eye-drops post-operatively (tid, for 3 months), and were observed for six months after operation: the neovascularizatio, turbidity, edema and the rejection time of cornea were exaed, and worked out the RI(rejection index),the corneal livability were estimated by the RI, the comeal organization was exaed with immunohistochemistry.(3) The clinical applied studies of treatments for severe ocular surface alkali burns: Of eight patients (eight eyes) with serious ocular surface... |
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