| Objective:To Study the effects of using amniotic membrane combined with oral mucosal epithelium of rabbit transplantation to treat severe alkali burns of rabbit eyes.After transplantation,observe the growth of rabbit corneal surface,and to evaluate the feasibility of using the combination of amniotic membrane and oral mucosal epithelium transplantation in the treatment of stem cell defect,so as to explore a possible sources of rich materials and feasible methods.Methods:(1) produced rabbit alkali burn model of 18 New Zealand rabbits(2-2.5kg) after anesthesia,each elected one eye.In a sterile conditions,infiltrate the filter paper in a solution of 1mol/LNaOH and contact the cornea and comeoscleral 1min to establish animal model of corneal alkali burn;(2) Preparation of amniotic membrane Elect the pregnant women whose serum HbsAg, chlamydia,HIV and Treponema pallidum were all negative in cesarean section,remove the amniotic membrane,washed to preserve;(3)Experimental grouping limbal alkali bum model of stem cell defect of 18 rabbits were randomly divided into control group(control group),amniotic membrane transplantation alone group(amniotic membrane group,AM group) and amniotic membrane + the oral mucosa epithelium transplantation group,(AM + oral mucosal epithelium group),each group is of 6 rabbits(6)eye,do the transplantation 7d after the alkali bum.(4) transplantation①The control group After 7 d of the alkali burn,make an annular conjunctival incision 2mm in the post-limba,do the simple conjunctival incision and suture + lid margin fusion;②Simple amniotic membrane transplantation group Remove conjunctiva Group from comeal surface and the surrounding tissue of limbal conjunctiva within the scope of 2mm.Then,wash the glycerol preserved amniotic membrane under running water,immersed in Iodophor,followed,and then repeatedly washed with normal saline.Cover the ocular surface with the amniotic membrane,fix it to the cornea and sclera with 8-0 nylon suture in the margin of 5mm post limbal margin,do the fusion of lid margin and encapsulate the surgery eye;③Amniotic membrane combined with oral epithelium transplantation group At first open the mouth,shear oral mucosa from the rabbit,try to wipe out submucosal tissue and sterilize it.Remove conjunctiva Group from corneal surface and the surrounding tissue of limbal conjunctiva within the scope of 2mm.Cover the ocular surface with the amniotic membrane,fix it to the cornea and sclera with 8-0 nylon suture in the margin of 5mm post limbal margin,and then part oral mucosa into four pieces,each about 2×4mm~2,stitch them to the upper,the under,inside and outside edge of the cornea and sclera,fixed them to the amniotic membrane and sclera on shallow;do the fusion of lid margin and encapsulate the surgery eye.Results:(1) control group:under slit-lamp saw the inflammatory response,corneal perforation, epithelial defect,corneal neovascularization substantial ingrowth,corneal opacity.Histopathology examination of HE staining:corneal epithelial shedding,see a large number of inflammatory cell infiltration,goblet cell ingrowth,corneal stroma fiber swelling and degeneration,thinning,a large number of neovascular ingrowth.(2) Amniotic membrane transplantation alone group:under slit lamp saw corneal neovascularization and conjunctival epithelial ingrowth,or intrusion into the central corneal zone, corneal opacity;histopathological examination with HE staining see irregular stratified epithelial cells,a small amount of inflammatory cells and neovascularization.(3) oral epithelium amniotic membrane transplantation group:see slit-lamp corneal surface epithelium under the speed of corneal transparency,only a small limbal neovascularization,no violation of the central zone,does not affect vision;pathological examination HE staining similar to corneal epithelium can be seen,the stratified epithelial cells were arranged regularly, connecting closer,no obvious neovascularization;scanning electron microscope examination showed corneal surface cells were fiat polygonal epithelial layer of the complex,cell-defined to form a tight junction,the surface cells are micro - villi.Conelusion:(1) this experiment successfully established a complete limbal stem cell damage model in rabbits with severe ocular surface burn;(2) Simple amniotic membrane transplantation for total limbal stem cell damage do the poor treatment of burns,amniotic membrane does not have the ability to repair corneal epithelium,but the amniotic membrane transplantation can be reduced to some extent in the inflammatory response,reducing scar formation;(3) Amniotic membrane transplantation combined with oral epithelium for treatment of severe alkali burn has achieved some success in the short term.It maintain the transparency of the cornea, reducing the inflammatory response,no corneal scar,conjunctiva and the emergence of neovascularization.Amniotic membrane and oral mucosa epithelial membrane transplantation has played a common synergy;(4)The long-term results of amniotic membrane transplantation combined with oral epithelium for ocular surface reconstruction are not clear,the transfer of oral mucosa epithelium,the corneal neovascularization is increasing or decreasing,these will need to be confirmed. |
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