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Endothelial quality of eye bank-prestripped DMEK prepared form organ-cultured corneas with the Muraine technique

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Abstract

Our aim was to measure the endothelial quality of prestripped Descemet membrane endothelial keratoplasty (DMEK) 48 h after preparation in an eye bank with the Muraine technique and shipping in a distant center. Ten pairs of human corneas with similar eye bank endothelial cell density (ebECD) were stored in organ-culture (OC) for 25 days (20, 28) [median (10–90 percentiles)]. One cornea was then randomized to DMEK preparation using the Moria Muraine trephine, the other served as control. The grafts were left attached to the center of the cornea, immersed in the OC medium (without Dextran) and shipped to a distant center. After 48 h, the viable ECD (vECD) was assessed by image analysis after staining with Hoechst/Ethidium/Calcein-AM. In addition, immunostaining was performed on flat mounted tissues for structural (ZO-1, NCAM, CD166) and functional (Na+/K+ ATPase) proteins of ECs, and for collagen I. Just before stripping, ebECD was 2428 (2268–2669) cells/mm2 for DMEK and 2471 (2135–2714) for controls (P = 1). Forty-eight hours after stripping, vECD was 2057 (1829–2463) cells/mm2 for DMEK and 2119 (1496–2525) for controls (P = 0.508). The expression patterns of the 5 proteins were similar in ECs of both groups. Notably, the deep posterior folds observed in OC controls almost disappeared in prestripped DMEK due to the lack of a link between Descemet membrane and stroma. As a consequence of the elimination of mechanical stress in these zones, EC evenly covered the whole graft. In conclusion, DMEK prestripping with the Muraine technique and shipping away can be used safely by eye banks.

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Authors and Affiliations

  1. Laboratory “Biology, Engineering, and Imaging of Corneal Graft”, BiiGC, EA2521, SFR143, Faculty of Medicine, Jean Monnet University, 10, Rue de la Marandière, 42055, Saint-Étienne Cedex 2, France

    Zhiguo He, Philippe Gain & Gilles Thuret

  2. Ophthalmology Department, University Hospital, Charles Nicolles Hospital, Rouen, France

    Julie Gueudry & Marc Muraine

  3. Eye Bank of Normandy, Charles Nicolles Hospital, Rouen, France

    David Toubeau & Marc Muraine

  4. Ophthalmology Department, University Hospital, Saint-Étienne, France

    Philippe Gain & Gilles Thuret

  5. Institut Universitaire de France, Paris, France

    Gilles Thuret

Authors
  1. Zhiguo He
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  2. Julie Gueudry
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  3. David Toubeau
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  4. Philippe Gain
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  5. Gilles Thuret
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  6. Marc Muraine
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Corresponding author

Correspondence to Gilles Thuret.

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Conflict of interest

Dr. Muraine is consultant for Moria, France. Dr Gain and Dr Thuret are consultant for les Laboratoires Théa, France. The other authors have no conflicting interest to disclose.

Human participants and/or animal rights

Corneas assigned to scientific use and procured by the Normandy Eye Bank (Rouen, France) were used after informed consent of the relatives, as authorized by French bioethics laws. All procedures conformed to the tenets of the Declaration of Helsinki for biomedical research involving human subjects.

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He, Z., Gueudry, J., Toubeau, D. et al. Endothelial quality of eye bank-prestripped DMEK prepared form organ-cultured corneas with the Muraine technique. Cell Tissue Bank 19, 705–716 (2018). https://doi.org/10.1007/s10561-018-9723-0

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  • DOI: https://doi.org/10.1007/s10561-018-9723-0

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