Infl ammation Induced By Cold Ischemia: A New Fast and Simple In Vivo Method to Evaluate Potential Therapeutic Tools

AMBROSI NELLA; GUERRIERI DIEGO; BOUSO CAROLINA; SALAMONE GABRIELA; MENDIBERRI JOSEFINA; INCARDONA CLAUDIO; CHULUYAN EDUARDO; CASADEI DOMINGO

San Francisco

Congreso; World Transplant Congress 2014; 2014

ASTS, TTS, AST

A prolonged cold ischemic (CI) time in kidney transplants has been associated with a high rate of DGF and reduced graft survival. The aim of this work was to develop a simple and rapid method to evaluate in vivo potential therapeutic tools, which may reduce the infl ammatory process caused by the prolonged CI. Kidney explants (Exp) derived from BALB/c (WT) were incubated ex vivo at 4 ° C for 18 h in different preservation solutions. At the end of the incubation period, explants were implanted subcutaneously (s.c.) in WT or transgenic mice expressing green fl uorescent protein (GFP). As control group and, at the same time, these animals also received explants that do not suffered CI. The recruitment of neutrophils was examined after 24 hours by measuring myeloperoxidase (MPO), as an indirect marker of neutrophils migration, and by a direct counting of the Gr1-Phe+ and GFP+ cells with a fl ow cytometer. Explants that were incubated at 4ºC for 18 h in Ringer Lactate (RL), recruited more neutrophils than control explants (p<0.001). When Wisconsin solution (WS) was used during the CI, the explants recruit less neutrophils compared to RL solution (p<0.01). Furthermore, renal medulla recruited more cells than cortex (p<0.01). Moreover, the explants that were incubated with dexamethasone decreased the recruitment of neutrophils compared to control explants (p<0.05). Finally, this model was challenged in their ability to distinguish between explants derived from healthy or damaged kidneys. For this, we obtained explants derived from mice that had suffered a period of 30 min of warm ischemia (WI) or from animals that had been previously treated with a serpine SLPI (250 ug/kg). It is important to mention that our group described that SLPI protects from the kidney damage induced by WI. After 24 h of WI, mice were sacrifi ced; the explants were obtained and incubated at 4ºC for 18 h in RL, as described above. These experiments showed that explants derived from animals with WI, recruited more cells than animals treated with SLPI (P<0.05). Conclusion: overall these results support the use of this in vivo model to study potential therapeutic tools designed to reduce infl ammation induced by CI and WI.