Effects Of Amniotic Membrane Transplantation Combined With BFGF In Promoting Healing Of Corneal Epithelium Injury In Dogs

In the common ophthalmic disease of pets, the incidence of comeal damage which results in blind with malpractice is higher than other diseases. In recent years, amniotic membrane transplantation is hot spot in ophthalmic research. As a biological material using for treating ophthalmic disease, amniotic membrane can reconstruct ocular surface by promoting cell proliferation, preventing scarring after fibrosis, and reducing inflammation and corneal neovascularization. Proper concentration of bFGF which has multiple biological functions can promote cell proliferation and speeding healing the surface layer of the cornea in our previous studies. So we treated acute corneal alkali burn by using bFGF combined with amniotic membrane transplantation to know the healing rate of corneal epithelium. In order to know the effect of amniotic membrane on corneal inflammation, we research the impact of amniotic membrane homogenate on the chemotaxis migration of neutrophils.Neutrophils were separated from peripheral blood cells of health dog, and nested with amniotic membrane homogenate with different diluted concentrations. The transwell system was used to research the chemotaxis of Neutrophils with Interleukin-8. The test was to verify that whether amniotic membrane can inhibit the chemotactic migration of neutrophils, which leaded to inflammation in the early burn alkali cornea.Corneal alkali burn model was used for destroying the corneal epithelium.10ng/mL bFGF eye drops and amniotic membrane transplantation were used on the models as combination.24hybrid dogs were randomly divided into4groups:The blank control group, the conjunctival cover group, the amniotic membrane transplantation group, and the bFGF combinated with amniotic membrane transplantation group. The dogs got surgery at0d in each group. The corneal turbidity and corneal neovascularization were observed and given clinical scores after the operations. Dogs were euthanized at10d, and the corneal was taken for pathological observation.Results:Neutrophils, nested by amniotic membrane homogenate of different diluted concentrations in vitro, had lower chemotaxis. High concentrations of amniotic membrane homogenate were significant in inhibiting neutrophils chemotaxis (P=0.022). The treatment of amniotic membrane group and the combination group were significantly better than those of the blank control group and the conjunctival cover group in terms of inflammation, tissue repairing and inhibiting corneal neovascularization. The corneal turbidity of the combined group was significantly better than that of AM group at9,10d (P=0.021;0.032). The corneal neovascularization first appears in the combination group (7d) later than amniotic membrane group (4d).Conclusion:In the dogs’corneal alkali burn treatment programs, bFGF, combined with AM transplantation in treatment of simple amniotic membrane transplantation obtained better results. The amniotic membrane, which played the anti-inflammatory effect, could inhibit inflammation in the early alkali burn phase by reducing neutrophils chemotaxis.