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Preclinical study in a corneal bioreactor of the effectiveness of amniotic membrane of umbilical cord extract (AMUCE) for the treatment of corneal deep stromal ulcerations
Author(s) -
Crouzet Emmanuel,
Peyret Benjamin,
Caroline Trone Marie,
Poinard Sylvain,
Garcin Thibaud,
Dumollard JeanMarc,
Barnouin Laurence,
Gain Philippe,
Thuret Gilles
Publication year - 2021
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2020.0101
Subject(s) - cornea , wound healing , umbilical cord , medicine , ex vivo , stromal cell , epithelium , histology , corneal epithelium , amnion , ophthalmology , staining , pathology , anatomy , in vivo , surgery , biology , fetus , pregnancy , genetics , microbiology and biotechnology
Purpose Our patented corneal bioreactor (BR) improves the long‐term survival of human corneas ex vivo by restoring intraocular pressure and renewal of storage medium. Aim To use the BR to test the efficacy of eye drops containing devitalized ground amniotic membrane of umbilical cord extract (AMUCE) to treat deep stromal ulcerations. AMUCE contains high level of proteoglycans, collagen and physiological growth factors. Methods Only pairs of human corneas (with ECD > 800 cells/mm 2 ) were used to obtain best possible controls. They were first stored for 2 weeks into the BR filled with CorneaMax (Eurobio, France), with 21 mmHg in the endothelial chamber to allow regeneration of a normal epithelium. Then, an 8.0 mm diameter calibrated epithelial ulcer was made with a spatula and a 6 mm diameter × 100 µ m in depth was made with Excimer laser in the stroma. Using a specific BR lid allowing sterile handling, each cornea was treated topically 4 times daily either by the AMUCE eye drops (TBF, France) or by the vehicle (0.9% NaCl). Closure of the ulceration was monitored daily using fluorescein staining quantified by image analysis (FIJI) of macro‐zoom microscopy. After wound healing corneas were fixed for histology (HES) and immunolabeling of epithelial markers. Results Three pairs were used. Epithelium wound healing was obtained at D3, D9, D10 with AMUCE and D3, D9, D19 with the vehicle. Histology performed in ulcerations centre showed 3‐6 epithelial layers with AMUCE and 1‐4 with the vehicle. At D21, significant epithelial detachment was observed only for 2 corneas treated with the vehicle. For one pair, immunolabelling for E‐cadherin and Cytokeratin K3/K12 showed better epithelial differentiation with AMUCE than with vehicle. No evidence of stromal regeneration was observed in the 6 corneas. Conclusions AMUCE has potential for a faster and lasting epithelium regeneration (stratification and maturation). Nevertheless, it has no impact on stromal regeneration in this model.