| Objective:To observe corneal ultrastructure changes after using amniotic membrane(AM) suspension of rats corneal alkali bum, and to explore the effective method for eye alkaline burn.Methods:Forty eyes from40Sprague-Dawley healthy male rats without any ocular disease were randomly divided into control groups(20eyes) and experiment groups(20eyes).The model of alkaline burn of rabbit cornea were set up with1mol/L NaOH. All eyes in the control groups were received PBS four times a day, while the experiment group eyes were treated with amniotic suspension four times a day.Using slit lamp biomicroscopy, each eye’s general condition was observed postoperatively every day, such as conjunctival congestion, corneal opacity and neovascularization. The anterior segment was pictured,and the ratio of cornea neovascularization(CNV) area, epithelium defect to cornea’s were scored and calculated by computer image analysis system. Tissue ultrastructure changes of cornea were analyzed by corneal confocal microscope and the number of cells was calculated. Three rats were killed respectively at3、7、14and28days after injured in two groups. Eyeballs were taken to make tissue sections and stained with HE to observe the morphological changes of corneas.Results:The healing area of epithelium of experiment groups was more than control groups at first day to seventh day after alkaline burn of rabbit’s cornea (P<0.01). The CNV in experiment groups occurred later and grew slower than that in control groups (4.8±0.6vs3.1±0.5d, P<0.01).11d after wounding, the area of neovascularization was48.72±2.12mm2in the control group,35.12±1.77mm2in the experiment group. The area of CNV were statistical differences at each time point between control group and experiment group(all p<0.05). Inflammatory cells in corneal in control groups were3164±306cell/mm2at9day, which significant far more than that in experiment groups (2830±254cell/mm2)(P<0.01).11day after wounding, the number of fibroblast cells of homogenate amnion was less than the control group(248±84vs349±105cell/mm2, P<0.01). At28day, Cellular density of endothelial cells in experiment groups was2064±54cells/mm2, which more than that in control groups(1831±59cell/mm2),(P<0.01)。HE staining showed that the density of corneal stroma collagen fibers, inflammatory cells and the aero of CNV in the experiment groups was lower,and collagen in the anterior stromal layer was more regular than that in the control groups.Conclusions:AM suspension can enhance the proliferation of rats corneal epithelial cell effectively and accelerate the healing of rats corneal epithelial. AM suspension can inhibite inflammatory cells and significant restrain the growth of the corneal neovascularization. AM suspension can restore the corneal stroma, reduce the number of corneal stroma collagen fibers, and reduce the corneal scar. |
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