Communication Dans Un Congrès Année : 2009

Understand the machine: mechanistic analysis of the benefits of machine perfusion on diseased after cardiac arrest donors grafts

Résumé

Introduction and aims: Diseased after cardiac arrest donors (DACAD) represent a source of organs to attempt to fill the gap between number of patients waiting for a graft and the number of donation. Due to the period of warm ischemia undergone by the graft before cold preservation, European law states that organs from DACAD must be preserved using machine perfusion (MP) prior to be transplanted, as this technique demonstrated improvement of graft early function and some beneficial effects on the long term. However, to date no careful analysis of the mechanistic impact of MP versus static cold storage (CS) has not been established.
Methods: We propose a deep evaluation of the mechanisms present in the graft 3 month after transplant, using RT-PCR for markers of inflammation, immune cell invasion, oxidative stress, epithelial to mesenchymal transition and fibrosis. Analysis were performed on grafts performed in large white pigs, for which warm ischemia was induced 60 minutes before graft CS in either static or MP conditions, using KPS as a conservation solution.
Results: We determined that after 3 month, kidney function was improved in grafts preserved with MP, as GFR was increased (125+/-25 vs. 83+/-11mL/min in CS) and proteinuria decreased (0,56+/- 0,2 vs. 1,72+/-0,3 g/24h). Moreover, kidney fibrosis at 3 month was improved by MP (Red Sirus Staining 4.16±0.08% vs 19.67±1.50 in CS, p<0.001).MP of the graft favored a decrease in pro-inflammatory factors TLR4 (5.21±2.57 vs. 17.38±7.23, folds to Control, p<0.05), TLR 2 (7.14±2.51 vs. 46.07±15.18 folds, p<0.01), Thrombospondin (6.58±1.66 vs. 11.35±1.65 folds, p<0.05) and TNFα (106.03±53.3 vs. 357.19±135.01 folds, p<0.05), as well as pro-fibrosis markers TGFß (7.20±2.13 vs. 20.18±5.99 folds, p<0.05), MMP9 (6.58±3.87 vs. 16.83±1.89 folds, p<0.05) and Timp2 (0.55±0.09 vs. 1.86±0.52 folds, p<0.01). We also defined a protective effect of MP against oxidative stress, as the expression of Phoxp47 was decreased (10.96±3.05 vs. 39.03±11.78 folds, p<0.01). Moreover, we detected a decrease in macrophage invasion evidenced by a decrease in CD11b expression (0.96±0.25 vs. 5.50±2.05 folds, p<0.01). Furthermore, MP permitted an increase in the expression of anti-fribrotic markers MMP2 (1.87±0.51 vs. 0.71±0.26 folds, p<0.05).
Conclusions: In conclusion, we defined the impact of machine perfusion on kidney graft on the mechanistic level, highlighting beneficial alterations in pro-inflammatory, oxidative stress and fibrosis markers. There is however still room for improvement, as several pro-injury factors were unchanged. We thus confirm the benefits of MP for the conservation of grafts from DACAD on the mechanistic levels. Our data suggests that improvement to the technique can and should be made, to further protect the grafts and use this donor pool to its full potential.

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hal-05099384 , version 1 (05-06-2025)

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  • HAL Id : hal-05099384 , version 1

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Raphaël Thuillier, Frédéric Favreau, Olivier Celhay, Thierry Hauet. Understand the machine: mechanistic analysis of the benefits of machine perfusion on diseased after cardiac arrest donors grafts. World Congress of Nephrology 2009, European Renal Association–European Dialysis and Transplant Association (ERA-EDTA); International Society of Nephrology (ISN), May 2009, Milan, Italy. ⟨hal-05099384⟩
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