An Experimental Trial Of Non-penetration Filtering Surgery With Amniotic Membrane

Objective:To investigate and comparative the amnion membrane implanted in the rabbit non-penetrating trabeculectomy in the proliferation of anti-fiber capacity.Methods:15 adult healthy New Zealand white rabbits, both male and female, each rabbit eyes underwent non-penetrating trabeculectomy, during a randomly selected, 6mm×10mm amnion membrane implanted scleral flap and conjunctival flap for the experiment group, while the other eye does not implant amnion membrane for the control group. After the first 3 days, 1 week, 2 weeks, 3 weeks to observe intraocular eye surgery, filtering bleb morphology, complications, cornea, conjunctiva, and anterior chamber situation. After 1 week, 2 weeks, 3 weeks randomly selected 5 rabbits were killed, removal of the eye (part of a complete filtering bleb reservation), check operation of the filtering bleb eyes and organizations around the cornea and sclera, fixed with fixative for ( hematoxylin eosin HE) staining were observed in the filtration Road open, fibrous proliferation, inflammatory response and uranyl acetate, lemon lead double staining was observed under the transmission electron microscope pool filtration fibroblast proliferation and collagen fibers.Results:1) after 3 days, all the conjunctivas had functional filtering bleb and anterior chamber reaction mildly, corneal epithelial staining without abnormal, corneal endothelial cells morphous quantitative analysis no significant difference compared with the preoperative, but the intraocular pressure lower than preoperative , and differences between groups without significant. 2) after 1 week, the experimental group saw 14 diffusion filtering blebs, a slight bulge, showing small cystic cavity. The control group saw 11 filtering blebs, dispersion, and small cystic cavities, the two groups was no significant difference (p> 0.05). Corneal endothelium, intraocular pressure and fundus case no significant differences, no inflammatory components detected in aqueous humor. 3) after 2 weeks, the experimental group, 8 eyes existed bulge obvious filtering bleb, diffusion, however the control group only 3 eyes had functioning filtering bled, and filtering bleb much small and had scars adhesion in some department. Even though, There was still no significant difference (p> 0.05) between the two groups. Corneal endothelium, intraocular pressure, fundus and the aqueous was no significant change over the previous. 4) after 3 weeks, the experimental group, 4 still had filtering bleb, smaller, still micro-eminence; but in the control group filtering bleb border fuzzy, flat, functional filtering bleb disappeared. 5) HE staining biopsy showed: after 1 week, the experimental group filtration gap, and scleral fibroblasts less, non-proliferation of fibrous tissue formation, membrane began to rupture, which can be seen within a small amount of inflammatory cell infiltration; the control group exist filtering bleb space, scleral fibroblasts less and no formation of fibrous tissue hyperplasia, inflammatory cell infiltration less. After 2 weeks, the experimental group into a loose gap-like filtration, fibroblasts little less fibrous tissue hyperplasia, and inflammatory cell infiltration; the control group gradually disappear filtration gap, the proliferation of fibrous tissue was replaced more than fibroblasts. After 3 weeks, the experimental group was mesh filtering bleb, fewer fibroblasts.The surface of the membrane fragments can be seen inflammatory cell infiltration; control group, large filtering bleb was replaced by the proliferation of fibrous connective tissue, fibroblast cells. 6) electron microscopy: after 1 week, the experimental group and control group no significant difference in cell ultrastructure, majority of fibroblast proliferation actived, cell bodies and cytoplasm rich in mitochondria, rough endoplasmic reticulum, ribosomes, etc. . After 2 weeks, the control volume composed of fibroblasts, cell body, and abundant cytoplasm, nucleolus, and chromosome density, rough endoplasmic reticulum number, capsules filled with reticular and granular protein material, were active proliferative state; experimental composition of fiber volume, cell body, small, sparse cytoplasm, electron dense, rough endoplasmic reticulum the number of small, low-proliferative capacity. After 3 weeks, the number of fibroblasts were reduced, the cell body shape smaller, the density of the cytoplasm lower, the number of the rough endoplasmic reticulum decreased; experiment groug only fewer of fibroblasts, reduced cytoplasm, the cell is in a state of decomposition, the entire cells were the performance of senescent cells. 7) Corneal epithelial injury, corneal edema, conjunctival wound leakage, conjunctival epithelial defects and complications such as shallow anterior chamber were not found in any group. Conclusion:1) amnion membrane used in non-penetrating trabeculectomy, there was no obvious complications, safe and reliable. 2) amnion membrane in rabbit non-penetrating trabeculectomy in inhibiting fibroblast proliferation and promote the formation of filtering bleb and effective for clinical applications based on animal experiments...