June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Preclinical randomized controlled study comparing long-term stored human corneas in an innovative bioreactor versus standard organ-culture.
Author Affiliations & Notes
  • Thibaud GARCIN
    Ophthalmology Department, University Hospital., Laboratory “Corneal Graft Biology, Engineering and Imaging” EA2521, University Jean Monnet., SAINT ETIENNE, France
  • Fabien FOREST
    Laboratory “Corneal Graft Biology, Engineering and Imaging” EA2521, University Jean Monnet., SAINT ETIENNE, France
    Pathology department, University Hospital., SAINT ETIENNE, France
  • Paul VERHOEVEN
    Microbiology department, University Hospital., SAINT ETIENNE, France
  • Jean Lou PUGNIET
    Graft coordinator team, University Hospital., SAINT ETIENNE, France
  • Thierry PEYRAGROSSE
    Graft coordinator team, University Hospital., SAINT ETIENNE, France
  • Françoise ROGUES
    Graft coordinator team, University Hospital., SAINT ETIENNE, France
  • Pascal HERBEPIN
    Laboratory “Corneal Graft Biology, Engineering and Imaging” EA2521, University Jean Monnet., SAINT ETIENNE, France
  • Sophie ACQUART
    Eye Bank of St-Etienne, French Blood Center., SAINT ETIENNE, France
  • Fabrice COGNASSE
    Eye Bank of St-Etienne, French Blood Center., SAINT ETIENNE, France
  • Chantal PERRACHE
    Laboratory “Corneal Graft Biology, Engineering and Imaging” EA2521, University Jean Monnet., SAINT ETIENNE, France
  • Zhiguo HE
    Laboratory “Corneal Graft Biology, Engineering and Imaging” EA2521, University Jean Monnet., SAINT ETIENNE, France
  • Philippe GAIN
    Ophthalmology Department, University Hospital., Laboratory “Corneal Graft Biology, Engineering and Imaging” EA2521, University Jean Monnet., SAINT ETIENNE, France
  • Gilles Thuret
    Ophthalmology Department, University Hospital., Laboratory “Corneal Graft Biology, Engineering and Imaging” EA2521, University Jean Monnet., SAINT ETIENNE, France
    Institut Universitaire de France, Paris, France., PARIS, France
  • Footnotes
    Commercial Relationships   Thibaud GARCIN, None; Fabien FOREST, None; Paul VERHOEVEN, None; Jean Lou PUGNIET, None; Thierry PEYRAGROSSE, None; Françoise ROGUES, None; Pascal HERBEPIN, None; Sophie ACQUART, None; Fabrice COGNASSE, None; Chantal PERRACHE, None; Zhiguo HE, None; Philippe GAIN, University Jean Monnet (P); Gilles Thuret, University Jean Monnet (P)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 3793. doi:
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      Thibaud GARCIN, Fabien FOREST, Paul VERHOEVEN, Jean Lou PUGNIET, Thierry PEYRAGROSSE, Françoise ROGUES, Pascal HERBEPIN, Sophie ACQUART, Fabrice COGNASSE, Chantal PERRACHE, Zhiguo HE, Philippe GAIN, Gilles Thuret; Preclinical randomized controlled study comparing long-term stored human corneas in an innovative bioreactor versus standard organ-culture.. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3793.

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

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Abstract

Purpose : Organ-culture (OC) developed in the 70’s, is a passive corneal storage that has never been revisited. For 5 years, our lab has been developing a bioreactor (BR) that restores the corneal physiology (intra ocular pressure (IOP) and aqueous humor renewal). Aim: to compare BR versus (vs) OC for corneas included immediately at retrieval, in a real-life setting.

Methods : Preclinical study designed as a superiority controlled trial, on paired fresh scientific human corneas in parallel groups, authorized by the French Biomedicine Agency and our local IRB. Immediately at retrieval, after centralized randomization, one cornea was stored in OC and the other in the BR with 21 mmHg and 2.6 µL/min, both with the same medium (CorneaMax, Eurobio, 2% fetal calf serum). Pairs with endothelial cell density (ECD) below 2000 cells/mm2 and more than 10% difference between corneas at day (D)2 were excluded. At D26, only OC corneas were transferred to a deswelling medium with 5% Dextran (CorneaJet, Eurobio). We calculated that 49 pairs would be needed to show a difference of 500 viable cells/mm2 between groups. Assessments were performed at D2, 26 and 28. Primary endpoint: the viable ECD (vECD) determined after staining by Hoechst/Ethidium/Calcein (that we previously reported) at D28, blind of storage method. Secondary endpoints: transparency (T), central corneal thickness by OCT (CCT) and microbiological testing.

Results : Preliminary results on the first 41 pairs. At D2, ECD was 2575+/-363 in OC vs 2628+/-359 cells/mm2 in BR (p=0.087). At D28, vECD was 2186+/-394 in BR vs 1679+/-382 cells/mm2 in OC with a mean difference of 507 cells/mm2 (95%CI 423;590) (p<0.001). At D28, in BR 62% of corneas remained over 2000 viable cells/mm2 vs 21% in OC (p<0.001). At D26, T was lower in OC with 12+/-7 vs 60+/-22% in BR (p<0.001) and average CCT in OC was 1179+-156 µm. At D28, T did not differ with 44+/-10% in OC vs 40+/-12% in BR (p=0.111) and CCT was lower in OC with 604+/-56 vs 689+/-58 µm in BR (p<0.001). One cornea of each group was contaminated.

Conclusions : Restoration of the IOP and medium renewal by the BR significantly improves endothelial cell survival, resulting in dramatically higher ECD (+30%), while avoiding the deleterious deswelling step necessary in standard OC. At the end of storage, three times more corneas are suitable for transplantation and their quality is higher.

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

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