Stefano Barabino1, Barbara De Servi2, Salvatore Aragona3, Demetrio Manenti4, Marisa Meloni2. 1. a Clinica Oculistica, Di.N.O.G.M.I. , Azienda Ospedaliera Universitaria San Martino-IST, Genoa , Italy. 2. b VitroScreen SrL-in Vitro Research Laboratories , Milan , Italy. 3. c Humanitas Mater Domini , Castellanza , Italy. 4. d Sildeha , Lugano , Switzerland.
Abstract
PURPOSE: So far tear substitutes have demonstrated a limited role in restoring ocular surface damage in dry eye syndrome (DES). The aim of this study was to assess the efficacy of a new ocular surface modulator in an in vitro model of human corneal epithelium (HCE) damaged by severe osmotic stress mirroring the features of dry eye conditions. METHODS: A reconstructed HCE model challenged by the introduction of sorbitol in the culture medium for 16 h was used to induce an inflammatory pathway and to impair the tight junctions integrity determining a severe modification of the superficial layer ultrastructure. At the end of the overnight stress period in the treated HCE series, 30 μl of the ocular surface modulator (T-LysYal, Sildeha, Switzerland) and of hyaluronic acid (HA) in the control HCE series were applied for 24 h. The following parameters were quantified: scanning electron microscopy (SEM), trans-epithelial electrical resistance (TEER), immunofluorescence analysis of integrin β1 (ITG-β1), mRNA expression of Cyclin D-1 (CCND1), and ITG-β1. RESULTS: In the positive control after the osmotic stress the HCE surface damage was visible at the ultrastructural level with loss of cell-cell interconnections, intercellular matrix destruction, and TEER reduction. After 24 h of treatment with T-LysYal, HCE showed a significant improvement of the ultrastructural morphological organization and increased expression of ITG-β1 at the tissue level when compared to positive and control series. A significant increase of mRNA expression for ITG-β1 and CCND1 was shown in the HA-treated cells compared to T-LysYal. TEER measurement showed a significant reduction in all groups after 16 h without modifications after the treatment period. CONCLUSIONS: This study has shown the possibility of a new class of agents denominated ocular surface modulators to restore corneal cells damaged by dry eye conditions. Further in vivo studies are certainly necessary to confirm these results.
PURPOSE: So far tear substitutes have demonstrated a limited role in restoring ocular surface damage in dry eye syndrome (DES). The aim of this study was to assess the efficacy of a new ocular surface modulator in an in vitro model of human corneal epithelium (HCE) damaged by severe osmotic stress mirroring the features of dry eye conditions. METHODS: A reconstructed HCE model challenged by the introduction of sorbitol in the culture medium for 16 h was used to induce an inflammatory pathway and to impair the tight junctions integrity determining a severe modification of the superficial layer ultrastructure. At the end of the overnight stress period in the treated HCE series, 30 μl of the ocular surface modulator (T-LysYal, Sildeha, Switzerland) and of hyaluronic acid (HA) in the control HCE series were applied for 24 h. The following parameters were quantified: scanning electron microscopy (SEM), trans-epithelial electrical resistance (TEER), immunofluorescence analysis of integrin β1 (ITG-β1), mRNA expression of Cyclin D-1 (CCND1), and ITG-β1. RESULTS: In the positive control after the osmotic stress the HCE surface damage was visible at the ultrastructural level with loss of cell-cell interconnections, intercellular matrix destruction, and TEER reduction. After 24 h of treatment with T-LysYal, HCE showed a significant improvement of the ultrastructural morphological organization and increased expression of ITG-β1 at the tissue level when compared to positive and control series. A significant increase of mRNA expression for ITG-β1 and CCND1 was shown in the HA-treated cells compared to T-LysYal. TEER measurement showed a significant reduction in all groups after 16 h without modifications after the treatment period. CONCLUSIONS: This study has shown the possibility of a new class of agents denominated ocular surface modulators to restore corneal cells damaged by dry eye conditions. Further in vivo studies are certainly necessary to confirm these results.
Authors: Christophe Baudouin; Murat Irkeç; Elisabeth M Messmer; José M Benítez-Del-Castillo; Stefano Bonini; Francisco C Figueiredo; Gerd Geerling; Marc Labetoulle; Michael Lemp; Maurizio Rolando; Gysbert Van Setten; Pasquale Aragona Journal: Acta Ophthalmol Date: 2017-04-08 Impact factor: 3.761
Authors: Adriana Di Benedetto; Francesca Posa; Mario Marazzi; Zamira Kalemaj; Roberta Grassi; Lorenzo Lo Muzio; Mariasevera Di Comite; Elisabetta Ada Cavalcanti-Adam; Felice Roberto Grassi; Giorgio Mori Journal: Front Endocrinol (Lausanne) Date: 2020-05-05 Impact factor: 5.555