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Hong Kyun Kim, Man-Il Huh, Kyoung Min Kang, Ji-Ahn Kim; Limbal reconstruction using decellularized human corneal lenticule with compressed collagen for treating limbal deficiency in rabbit experimental model.. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):2260.
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© ARVO (1962-2015); The Authors (2016-present)
We previously studied decellularized human cornea lenticule (HCL) generated by hypotonic enzymatic solution has great advantages as a scaffold for corneal regeneration. The aim of this study is to make a suitable bio-engineered limbal tissue for severely destructed limbal structure and then to evaluate therapeutic effect of the construct.
HCLs were decellularized using our standardized method and then gelated collagen was compressed on the decellularized human cornea. The constructs were trephined into 4 mm and examined mechanic and optical properties. Human corneal limbal epithelial cells (HCLECs) were re-seeded on compressed collagen (CC) surrounding decellularized HCL. Limbal deficient experimental model was established by alkali burn with 1N NaOH and construct with HCLECs was transplanted on the limbal deficient cornea. The operated corneas were collected at 1, 3 weeks. Limbal deficiency and invasion of conjunctival cells were examined by histology.
The construct combined CC with decellularized HCL showed better strength than CC only and decellularized HCL only, and histological results showed that it was well established. Immunologic staining showed that HCLECs were successfully reseeded expressing progenitor marker. Numerous goblet cells and several vascular were observed at inside of limbus. Histology of transplanted corneas showed that the construct with HCLECs inhibited conjunctival cell migration into center cornea compare to control, and had more transparent than the other at 2 weeks after operation.
Data in this study showed that combined construct had several advantages for limbal reconstruction ; stronger, easier for handling and and better bio-compatibility than CC only or decellularized HCL only. HCLECs on the bio-engineered limbal tissue were remained and kept its phenotype and its stemness without any infiltration of inflammatory cells. Collectively, our data suggest that combined construct with HCLECs has a great potential for the reconstruction of limbal niche in highly destructive limbal deficiency.
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