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W. Wang, Y. Feng, Y. Xu, X. Chen, C. Zhang; Investigation of Host Response of Xenogenic Bioengineered Corneal Matrix in Lamellar Corneal Transplantation in Rabbit. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1898. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To explore the host response of xenogenic bioengineered corneal matrix for cornea transplantation substitute
Fresh dog corneas were de-cellularized by serial digestion confirmed by light microscope (LM) and transmission electron microscope (TEM). Deep lamellar corneal transplantation (DLCT) were performed by using the xenogeneic de-cellularized corneal matrix in 8 rabbit eyes and observed by slit-lamp microscope (SLM). The rabbit were sacrificed at 1day, 7days, 1month and 3 months after DLCT and morphology structure of the cornea grafts were studied by LM and scanning electron microscope (SEM). The cells in grafts were identified by immunocytochemical staining for specific markers such as keratin 3, Vimentin, MUC5AC and P63. Proliferative cells were identified by BrdU staining. Apoptotic cells were detected by TUNEL
(1) Under LM and TEM, de-cellularized dog corneas after showed three-dimensional reticular structure without any cells detected. It was transparent with well-organized matrix fibers. (2) After DLCT, the grafts kept transparency until 1 month by SLM. The neovascularization appeared from 5 to 6 weeks after DLCT and recessed after the sutures removal. Cornea fluorescein staining was positive in 3weeks after DLCT. (3) The donor cornea grafts was intact and new vessels were found under the epithelium layer under LM. Cells showed cobblestone form with expression of K3 on surface of the grafts. Spindlelike cells expressed Vimentin were noted in the cornea stroma. Both of them didn’t express MUC5AC, expressed limbal stem cell marker P63 slightly. (4) TUNEL labeling showed scattered positive staining located in grafts stroma. (5) BrdU positive cells were noted in the matrix
The xenogenic corneal matrix after de-cellularized can keep transparent for some time after DLCT. Host cells can survive and proliferate into grafts. The grafts were merged with host tissue 3 months after DLCT. The grafts were epithelization and vasculization 3 months after DLCT. Both proliferative cells and apoptosis cells were noted in the grafts. Although extensive testing and further development are still required, this kind of bioengineering cornea shows a great potential for being a substitute of human donor cornea
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