| Objective:To study and compare the antiproliferative effection of fresh amniotic membrane and preserved amniotic membrane in glaucoma filtration passage by constructing animal glaucoma filtration surgery model.Method:24 New Zealand white rabbits were randomly divided into 3 groups. Group A is operated with glaucoma filtering surgery; Group B is operated with glaucoma filtration surgery and preserved amniotic membrane implantation; Group C is operated with glaucoma filtration surgery and fresh amniotic membrane implantation. Each group consisting of 8 rabbits with 16 eyes. We observed the rabbits' filtration blebs, intraocular pressure and complications at 3 days,and at week 1,2,3, 5 after surgery. Two rabbits of every group were killed at week 1, 2, 3, 5 after surgery. Their eyes were enucleated,and their filtration blebs were kept and fixed,and then they were made paraffin slices. We observed the filtration passage and the inflammatory cells by performing HE staining; We observed the expression of proliferative cell nuclear antigen(PCNA) in the filtration area and counted the number of positive cell nuclear under microscope after performing immunohistochemical staining; We observed the collagen formation under microscope by Masson staining and made stastic analysis.Results: 1)A11 the filtration blebs in three groups were functional blebs at postoperative days 3 .There were some non-functional blebs in group A at week 1 after surgery. The filtration blebs disappeared at week 3. The area of all the functional blebs in group B and group C appeared more distinct at week 1. The percentage of the functional blebs in group C was higher than group A and B at week 2,3, 5 after surgery, and it was higher in group B than in group A. The filtration blebs in group B were disappearing at week 5. And the area of the filtration blebs in group C was diminishing.2) No injured corneal epithelium,conjunctival leaking wound, shallow anterior chamber and other complications were observed.3)There were no significant difference among the intraocular pressure of three groups before operation and postoperative days 3(P>0.05).The intraocular pressure of group A is higher than group B and group C after 3 days,and the difference is significant(P<0.05). There was no significant difference between group B and group C(P>0.05) at week 1.But the intraocular pressure of group B is higher than group C after 7 days,and the difference is significant(P<0.05).4)The results of HE:there was much loose collagen in the wound area in group A at week 1 after surgery. The filtration pathway was partly closed with the proliferation of collagen fiber and the invasion of the neovascularization after 2 weeks. The filtration pathway was closed after 3 weeks. There were many inflammatory cells appeared around the amniotic membrane in group B and group C at week 2, and the small fraction of amniotic membrane was dissolved and was imperfect in group B. The amniotic membrane in group B appeared piecely at week 3, and much collagen fiber was observed in the filtration pathway. But the amniotic membrane in group C was perfect at week 3. The amniotic membrane in group B was wholely dissolved at week 5, and the filtration pathway was displaced by the loose collagen fibre. The amniotic membrane in group C appeared piecely,and there was much collagen fibre in the filtration pathway. At different time afer surgery,the numbers of inflammatory cells in group B as well as group C were significantly increased compared to group A(P<0.05),and there was significant statistical difference between group B and group C(P<0.05). 5) The numbers of PCNA positive cell nuclear at week 1 in every group were higher than other postoperative time. The numbers of the positive cell nuclear of PCNA in group C as well as group B were significantly reduced compared to group A(P<0.05) at different time after surgery. There was significantl statistical difference in the numbers of positive cells of PCNA between group B and group C(P<0.05). 6)The numbers of newly formed collagen fiber in group B and group C were significantly decreased compared to group A(P<0.05).There was significant statistical difference between numbers of nemly formed collagen fibers in group B and group C(P<0.05).Conclusion: l)The fresh and preserved amniotic membrane in the filtration pathway of glaucoma play the role of anti-inflammation and anti-proliferation.2)The antiproliferative effection of fresh amniotic membrane is better than amniotic membrane in glaucoma filtration surgery.3)No rejection is observed in glaucoma filtration surgery with the fresh amniotic membrane implantation. It is safe and effective.
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