The Study On Corneal Cell Sheet Engineering Of Fibrin Glue And Its Adhesive Role For The Conjunctival Epithelium

Objective: (1) To research the characteristics of comeal epithelial cells, keratocytes andendothelial cells of rabbit and lay a foundation for tissue engineering of cornea. (2) To study thestatus of three rabbit corneal cells growing on fibrin glue (FG) and investigate the feasibility ofFG for cell sheet engineering. (3) To study the cohesion of FG on the conjunctival epitheliumand investigate the practicability of sutureless conjunctival autograft transplantation for thetreatment of pterygium with FG.Methods: (1) Using tissue inoculation or digestion methods, rabbit corneal epithelial cells,keratocytes and endothelial cells were cultured. The cells were subcultured after they becameconfluent. The growing characteristics of cells were observed every day. (2) Three comeal cellswere seeded on FG. After confluence, the corneal cell sheets were detached from the plate with acell scraper, examined by inverted microscope, histologically by hematoxylin and eosin, and byscanning electron microscopy. (3) The cohesive surgery of ocular surface of 20 rabbits wascarried, which were applied into three operating styles: exposed group, implanted group,pterygial group. The postoperative conglutinant effect, inflammatory reaction, reepithlializationand scar formation of conjunctival epithelium were observed.Results: (1) The epithelial cells, keratocytes and endothelial cells adhered and grew within 24-48hours and formed a confluent monolayer in 6-9 days. The shape of the primary isolated cells wastypical, and the passage ceils still maintained the characteristics of the primary cells. The cellsgrew very well and could be cultured for a long time. (2) The prepared membranes of FG weresmooth and transparent. With cell growth, FG could degrade partly, and obtain cell sheetengineering with a small amount of FG. Corneal cells could grow well on the FG in vitro andmaintain the physiological state of cells. Corneal epithelial cells formed unilaminar or stratifiedlayers with cellular joins. Corneal endothelial cells with almost same size formed round orpolygon and lined up tightly. Corneal keratocytes formed triangle and arborization, cell-celljunctions were obvious, and formed network link. (3) Inflammatory reaction of implanted groupand pterygial group were relatively slight three days after operation. In two weeks, they werecovered with epithelial layer completely, histological section showed that the reconstruction of the damaged conjunctival tissue with less scar formation.Conclusions: (1) The research established a simple, highly efficient method for culture of rabbitcorneal epithelial cells, keratocytes and endothelial cells in vitro. (2) FG is well compatible withthree corneal cells, which can construct tissue engineered cell sheet of cornea, so as toreconstruct ocular surface. (3) FG is a safe and effective cohesive for the conjunctival grafts. Theuse of FG results in shorter operating times, less postoperative inflammatory reaction and scarformation.