July 2019
Volume 60, Issue 9
Free
ARVO Annual Meeting Abstract  |   July 2019
The OBSERV platform (Ophthalmic Bioreactor Specialized in Experimental Research & Valorization): simulation of a DMEK
Author Affiliations & Notes
  • Gilles Thuret
    Laboratory Biology, Engineering and Imaging of Corneal Grafts, BiiGC, EA2521, University Jean Monnet, France, France
    Institut Universitaire de France, Paris, France
  • Emmanuel CROUZET
    Laboratory Biology, Engineering and Imaging of Corneal Grafts, BiiGC, EA2521, University Jean Monnet, France, France
  • Chantal PERRACHE
    Laboratory Biology, Engineering and Imaging of Corneal Grafts, BiiGC, EA2521, University Jean Monnet, France, France
  • Thibaud GARCIN
    Laboratory Biology, Engineering and Imaging of Corneal Grafts, BiiGC, EA2521, University Jean Monnet, France, France
    Ophthalmology, University Hospital of St-Etienne, Saint-Etienne, France
  • Marie-Caroline TRONE
    Ophthalmology, University Hospital of St-Etienne, Saint-Etienne, France
    Laboratory Biology, Engineering and Imaging of Corneal Grafts, BiiGC, EA2521, University Jean Monnet, France, France
  • Fabien FOREST
    Laboratory Biology, Engineering and Imaging of Corneal Grafts, BiiGC, EA2521, University Jean Monnet, France, France
    Pathology, University Hospital, Saint-Etienne, France
  • Philippe Gain
    Laboratory Biology, Engineering and Imaging of Corneal Grafts, BiiGC, EA2521, University Jean Monnet, France, France
    Ophthalmology, University Hospital of St-Etienne, Saint-Etienne, France
  • Zhiguo HE
    Laboratory Biology, Engineering and Imaging of Corneal Grafts, BiiGC, EA2521, University Jean Monnet, France, France
  • Footnotes
    Commercial Relationships   Gilles Thuret, laboratoires Thea (C), Quantel Medical (C), Sincler (C), University Jean Monnet (P); Emmanuel CROUZET, None; Chantal PERRACHE, None; Thibaud GARCIN, None; Marie-Caroline TRONE, None; Fabien FOREST, None; Philippe Gain, Laboratoires Thea (C), Quantel Medical (C), Sincler (C), University Jean Monnet (P); Zhiguo HE, None
  • Footnotes
    Support  Agence Nationale de la Sécurité du Médicament (ANSM): research grant 2012 BANCO; EFS 2012 research grant; research grant from Jean Monnet University 2014, Fondation de France Bourse Berthe Fouassier 2016, Fondation Université Jean Monnet 2017, Fondation de l’Avenir 2017 Leg Deroche, Agence de la Biomédecine research grant 2017.
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4143. doi:
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      Gilles Thuret, Emmanuel CROUZET, Chantal PERRACHE, Thibaud GARCIN, Marie-Caroline TRONE, Fabien FOREST, Philippe Gain, Zhiguo HE; The OBSERV platform (Ophthalmic Bioreactor Specialized in Experimental Research & Valorization): simulation of a DMEK. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4143.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Our university lab BiiGC patented 2 versions of a bioreactor (BR): one for long-term eyebanking (in the process of industrialization), the other for preclinical experimentations, called OBSERV and supported by the French Agence Nationale de Sécurité du Médicament et des produits de Santé (ANSM). Aim: to reproduce a Descemet membrane endothelial keratoplasty (DMEK) inside the BR.

Methods : By restoring IOP and medium renewal, the BR maintained corneal viability over a prolonged period and its transparency allowed characterizing the tissue with existing or customized devices without compromising sterility. A 9mm diameter Descemetorhexis was performed open-sky on the recipient human cornea. An 8mm diameter endothelial grafts was dissected with a no-touch technique and transferred using a soft contact lens. The DMEK-bearing cornea was installed in the BR with an air bubble in the endothelial chamber and endothelium facing down. IOP was set to 21mmHg. The bubble was removed after 3 days. For some experiments an adherence promoting solution was used. Implanted corneas and controls (rhexis alone) were stored for 1 to 4 weeks in a custom-made medium. Monitoring: D-1, D1, W1 to 4: transparency (slit-lamp and customized transparometer), OCT corneal thickness, endothelium/epithelium (specular microscopy). Final test: triple Hoechst/Ethidium/calcein-AM staining to determine endothelial viability (Fig.) and Alizarin red staining.

Results : All DMEKs initially adhered but some of the group without adherence-promoting agents detached 48 hours after unbubbling, although their endothelial cells remained viable. The other grafts remained adherent. Implanted corneas with adherent grafts were thinner than controls. Endothelial cell survived normally and started to migrate outside the donor Descemet onto the bare recipient stroma from W1.

Conclusions : The OBSERV platform is efficient to study the biofunctionality of a DMEK. It will be useful for preclinical assessment of next generation bioengineered endothelial grafts.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

 

Triple Hoechst-Ethidium-Calcein-AM staining of a DMEK simulated in the corneal bioreactor after 2 weeks. The uniform central green staining indicated excellent endothelial cell survival on the graft.

Triple Hoechst-Ethidium-Calcein-AM staining of a DMEK simulated in the corneal bioreactor after 2 weeks. The uniform central green staining indicated excellent endothelial cell survival on the graft.

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