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Federico Bech, Ignacio Alcalde, Almudena Íñigo-Portugués, Paola Braga, Enol Artime, Beatriz Garcia, Jose Alfonso, Jesus Merayo-Lloves; Induction of experimental keratoconic ectasias by Endo-β-Galactosidase extracellular matrix digestion. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):2998.
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© ARVO (1962-2015); The Authors (2016-present)
The keratoconus disease has been related with structural defects due to the overexpression of keratocan, which can alter the fibrillogenesis in the stroma. The development of pharmacological approaches to keratoconus reveals the need of reproducible experimental models. The aim of this work is to generate a mouse model of keratoconus by degradation of the proteoglycan extracellular matrix suitable to test drug or cell based therapies.
To induce a keratoconic ectasia in mice, corneas were thinned using photorefractive keratectomy (PRK). Once the epithelium was removed mice were treated with the enzyme Endo-β-Galactosidase for 5 days at two concentrations (50 mU and 100 mU). After 10 days, the eye globes were characterized by optical keratometry, hematoxylin-eosin staining and by immunohistochemistry techniques to demonstrate Keratocan, TrkA, β-Catenin and MMP9. Therapeutic treatments with adipose mesenchymal stem cells (AMSC) and Plasma Rich in Growth Factors (PRGF) were tested. AMSC were isolated from human lipoaspirates and cultured with a defined medium before dropped it directly onto the eye surface for 5 days. Other group was treated paralelly with PRGF.
Keratometric analysis reveals that 40% of the corneas treated with Endo-β-Galactosidase at 100 mU develop a cone shape fifteen days after the PRK. The hematoxylin-eosin staining reveal an epithelial and stromal thinning at the site of the keratoconic ectasia. Moreover, a relative increase ot the expression of Keratocan and β-Catenin was observed in conic corneas. Otherwise, the TrkA and MMP9 expression was virtually nil in ectasic corneas. Treatment with stem cells showed the integration of AMSC into the stroma and an increase of corneal thickness. Both AMSC and PRGF produced a partial restoration of the ectasia.
Treatment with Endo-β-Galactosidase and PRK produce a reproducible mouse model that shares morphological and molecular features with human keratoconus This model is a good platform to test new biological strategies to treat keratoconus.
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