June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Ex vivo corneal wound healing model using an innovative bioreactor
Author Affiliations & Notes
  • Damien Guindolet
    Ophthalmology, Fondation Ophtalmologique Rothschild, Paris, France
    Corneal Graft Biology- Engineering and Imaging Laboratory, EA2521- Federative Institute of Research in Sciences and Health Engineering- Faculty of Medicine- Jean Monnet University, Saint Etienne, France., Saint Etienne, France
  • Emmanuel CROUZET
    Corneal Graft Biology- Engineering and Imaging Laboratory, EA2521- Federative Institute of Research in Sciences and Health Engineering- Faculty of Medicine- Jean Monnet University, Saint Etienne, France., Saint Etienne, France
  • Zhiguo HE
    Corneal Graft Biology- Engineering and Imaging Laboratory, EA2521- Federative Institute of Research in Sciences and Health Engineering- Faculty of Medicine- Jean Monnet University, Saint Etienne, France., Saint Etienne, France
  • Aurélien BERNARD
    Corneal Graft Biology- Engineering and Imaging Laboratory, EA2521- Federative Institute of Research in Sciences and Health Engineering- Faculty of Medicine- Jean Monnet University, Saint Etienne, France., Saint Etienne, France
  • Chantal PERRACHE
    Corneal Graft Biology- Engineering and Imaging Laboratory, EA2521- Federative Institute of Research in Sciences and Health Engineering- Faculty of Medicine- Jean Monnet University, Saint Etienne, France., Saint Etienne, France
  • Fabien FOREST
    Department of Pathology, University Hospital of Saint-Etienne, Saint Etienne, France
    Corneal Graft Biology- Engineering and Imaging Laboratory, EA2521- Federative Institute of Research in Sciences and Health Engineering- Faculty of Medicine- Jean Monnet University, Saint Etienne, France., Saint Etienne, France
  • Michel PEOC'H
    Department of Pathology, University Hospital of Saint-Etienne, Saint Etienne, France
    Corneal Graft Biology- Engineering and Imaging Laboratory, EA2521- Federative Institute of Research in Sciences and Health Engineering- Faculty of Medicine- Jean Monnet University, Saint Etienne, France., Saint Etienne, France
  • Philippe GAIN
    Corneal Graft Biology- Engineering and Imaging Laboratory, EA2521- Federative Institute of Research in Sciences and Health Engineering- Faculty of Medicine- Jean Monnet University, Saint Etienne, France., Saint Etienne, France
  • Gilles Thuret
    Corneal Graft Biology- Engineering and Imaging Laboratory, EA2521- Federative Institute of Research in Sciences and Health Engineering- Faculty of Medicine- Jean Monnet University, Saint Etienne, France., Saint Etienne, France
  • Eric Ernest GABISON
    Ophthalmology, Fondation Ophtalmologique Rothschild, Paris, France
  • Footnotes
    Commercial Relationships   Damien Guindolet, None; Emmanuel CROUZET, None; Zhiguo HE, None; Aurélien BERNARD, None; Chantal PERRACHE, None; Fabien FOREST, None; Michel PEOC'H, None; Philippe GAIN, university Jean Monnet (P); Gilles Thuret, university Jean Monnet (P); Eric GABISON, None
  • Footnotes
    Support  Fondation de l’Avenir, Paris, France. Study n°AP-RM- 2016-008.
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 2642. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Damien Guindolet, Emmanuel CROUZET, Zhiguo HE, Aurélien BERNARD, Chantal PERRACHE, Fabien FOREST, Michel PEOC'H, Philippe GAIN, Gilles Thuret, Eric Ernest GABISON; Ex vivo corneal wound healing model using an innovative bioreactor. Invest. Ophthalmol. Vis. Sci. 2017;58(8):2642.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Experimental models of corneal epithelial healing are critically important to study cellular mechanisms and new therapeutics. Available model generally use animal corneas and do not enable long-term storage, creation of an appropriate epithelial environment and preservation of the endothelium that influence wound healing. We designed a bioreactor (BR) for human ex-vivo corneal storage that restores a pressure equivalent to the intraocular pressure in the endothelial chamber while allowing continuous renewing of media endothelial chambers. The epithelial chamber is alternatively exposed to air and to a specific epithelial medium. Aim: to study epithelial wound healing in the BR compared to a standard ex-vivo model of human corneas.

Methods : Human corneas not suitable for graft were used. Corneas were stored either in a BR or in petri dish (immersed or airlifted with epithelial culture medium). A calibrated epithelial debridement was done with a crescent knife. Histological structure and immunohistological staining (K3-K12, Laminin-5, type IV collagen) were compared to conventionally stored corneas after 1 week. Healing rate was monitored with fluorescein staining, digital pictures, and image analysis with ImageJ. Experiments were done in triplicate.

Results : Faster epithelial healing was observed for cornea stored in bioreactor. In bioreactor corneal epithelium was differentiated (K3-K12 immunostaining) and multi-layered, whereas a 1-3 layer thick epithelium was observed in Petri Dish. Basement membrane was also restored (continuous laminin-5 and type IV collagen immunostaining) in BR whereas it was fragmented in Petri dish.

Conclusions : This innovative bioreactor could be a new ex-vivo assay to study corneal wound healing and to assess efficacy or toxicity of new therapeutics.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×