An Analysis of the Progression of Conjunctivalisation after Transplantation of Cultivated Corneal Epithelium.

PURPOSE: To analyse the recurrence of superficial neovascularisation after previous corneal surface reconstruction with cultivated corneal epithelial cells. MATERIALS AND METHODS: Forty-eight eyes underwent autologous transplantation of cultivated corneal epithelium to treat partial or total limbal stem cell deficiency caused by chemical or thermal injury. The carrier for the epithelial sheets was a denuded amniotic membrane. Follow-up was conducted for up to 120 months. Recurrent revascularisation (measured in terms of clock hours affected) was evaluated with slit-lamp examination and the support of confocal microscopy. RESULTS: During the long-term observation, only 7 eyes had stable epithelia with no neovascularisation from the conjunctiva. Nineteen eyes developed pathologic vessels in 1 quadrant, with additional 4 eyes developing them in 2 quadrants. Twelve patients developed subtotal or total conjunctivalisation of the corneal surface. They were referred for second cultivated epithelium transplantation (3 patients), allogenic keratolimbal transplantation (7 patients), or keratoprosthesis (2 patients). Six patients withdrew consent. The use of confocal scans of up to 100 µm in resolution enabled the detection of pathologic microvasculature originating from the conjunctiva and the exclusion of stromal vascular ingrowth. CONCLUSIONS: Local ingrowth of the conjunctiva is a common complication after the transplantation of cultivated epithelial cells. Severe and progressive vascularisation inevitably leads to graft failure. However, if local ingrowth stops before reaching the central cornea, the treatment even with this complication can be considered a success.

Cultivated Oral Mucosa Epithelium in Ocular Surface Reconstruction in Aniridia Patients.

PURPOSE: Efficacy of cultivated oral mucosa epithelial transplantation (COMET) procedure in corneal epithelium restoration of aniridia patients. METHODS: Study subjects were aniridia patients (13 patients; 17 eyes) with irregular, vascular conjunctival pannus involving visual axis who underwent autologous transplantation of cultivated epithelium. For the procedure oral mucosa epithelial cells were obtained from buccal mucosa with further enzymatic treatment. Suspension of single cells was seeded on previously prepared denuded amniotic membrane. Cultures were carried on culture dishes inserts in the presence of the inactivated with Mitomycin C monolayer of 3T3 fibroblasts. Cultures were carried for seven days. Stratified oral mucosa epithelium with its amniotic membrane carrier was transplanted on the surgically denuded corneal surface of aniridia patients with total or subtotal limbal stem cell deficiency. Outcome Measures. Corneal surface, epithelial regularity, and visual acuity improvement were evaluated. RESULTS: At the end of the observation period, 76.4% of the eyes had regular transparent epithelium and 23.5% had developed epithelial defects or central corneal haze; in 88.2% of cases visual acuity had increased. VA range was from HM 0.05 before the surgery to HM up to 0.1 after surgery. CONCLUSION: Application of cultivated oral mucosa epithelium restores regular epithelium on the corneal surface with moderate improvement in quality of vision.

Hydrogel membranes based on genipin-cross-linked chitosan blends for corneal epithelium tissue engineering.

Novel polymeric hydrogel scaffolds for corneal epithelium cell culturing based on blends of chitosan with some other biopolymers such as hydroxypropylcellulose, collagen and elastin crosslinked with genipin, a natural substance, were prepared. Physicochemical and biomechanical properties of these materials were determined. The in vitro cell culture experiments with corneal epithelium cells have indicated that a membrane prepared from chitosan-collagen blend (Ch-Col) provided the regular stratified growth of the epithelium cells, good surface covering and increased number of the cell layers. Ch-Col membranes are therefore the most promising material among those studied. The performance of Ch-Col membranes is comparable with that of the amniotic membrane which is currently recommended for clinical applications.

[Regeneration of corneal epithelium using keratin modified chitosan membranes]. / Regeneracja nablonka rogówki przy zastosowaniu membran chitozanowych modyfikowanych keratyna.

The cornea is a transparent front layer of the eye. It functions like a window that controls and focuses the light entering into the eye. The cornea contributes to 65-75% of the eye's total focusing power and it acts as a physical barrier against pathogenic microorganisms, dirt and other noxious physical factors. The corneal tissue is arranged in five basic layers. The outermost layer (epithelium) is made up of highly regenerative cells that allow for quick healing of superficial injuries. Eye infections, diseases, or mechanical injury can harm corneal epithelium and cause blindness. Under certain circumstances, to prevent that, it is recommended to perform complete corneal transplantation. However, due to lack of sufficient number of donors, researchers are searching for alternative solutions.. Regeneration of epidermal tissue can restore and ensure normal functioning of cornea. For that purpose proper grafts are needed. The goal of current research was to develop the material for scaffold preparation providing optimal conditions for the epithelium cornea cell culturing and to determine its chemical, physical, and biological properties. The scaffolds, which could be applied in ophthalmology should fulfill a lot of requirements, among them such as biocompatibility, biodegradability, restorability, non-toxicity. They should also have adequate mechanical strength, flexibility and porosity. The aim of this work was to synthesize and to determine the properties of polymeric material for ophthalmic surgery applications. A hydrogel scaffold in the form of membrane was obtained from chitosan - natural, biocompatible, biologically inert, stable in the natural environmental and antibacterial polysaccharide derived from chitin. Biodegradable chitosan films containing keratin were crosslinked with genipin - a naturally occurring and nontoxic agent. In this study we present physicochemical characterization of the scaffolds. Porosity, contact angle and swelling ratio (at different pH) were determined. The optical microscope technique was used to visualize the microstructure of the scaffolds. Atomic force microscopy (AFM) measurements revealed the topography of the surfaces of membranes. The biological tests have shown that epithelial cells seeded on the membranes proliferated efficiently.