Extracellular histone release by renal cells after warm and cold ischemic kidney injury: Studies in an ex-vivo porcine kidney perfusion model.

Extracellular histones are cytotoxic molecules involved in experimental acute kidney injury. In patients receiving a renal transplant from donors after circulatory death, who suffer from additional warm ischemia, worse graft outcome is associated with higher machine perfusate extracellular histone H3 concentrations. We now investigated temperature-dependent extracellular histone release in an ex vivo porcine renal perfusion model, and subsequently studied histone release in the absence and presence of non-anticoagulant heparin. Seven pairs of ischemically damaged porcine kidneys were machine perfused at 4°C (cold ischemia) or 28°C (warm ischemia). Perfusate histone H3 concentration was higher after warm as compared to cold ischemia (median (IQR) = 0.48 (0.20-0.83) µg/mL vs. 0.02 (0.00-0.06) µg/mL; p = .045, respectively). Employing immune-electron microscopy (EM), histone containing cytoplasmic protrusions of tubular and endothelial cells were found after warm ischemic injury. Furthermore, abundant histone localization was detected in debris surrounding severely damaged glomerular cells, in a "buck shot" pattern. In vitro, histones were cytotoxic to endothelial and kidney epithelial cells in a temperature-dependent manner. In a separate ex vivo experiment, addition of heparin did not change the total histone H3 levels observed in the perfusate but revealed a continuous increase in the level of a lower molecular weight histone H3 variant. Our findings show that ischemically damaged kidneys release more extracellular histones in warm ischemia, which by EM was due to histone release by renal cells. Blocking of histone-mediated damage during transplantation may be beneficial in prevention of renal injury.

The Benefits of Hypothermic Machine Preservation and Short Cold Ischemia Times in Deceased Donor Kidneys.

BACKGROUND: Hypothermic machine perfusion (HMP) of deceased donor kidneys is associated with better outcome when compared to static cold storage (CS). Nevertheless, there is little evidence whether kidneys with short cold ischemia time (CIT) also benefit from HMP and whether HMP can safely extend CIT. METHODS: We analyzed prospectively collected data from the Machine Preservation Trial, an international randomized controlled trial. Seven hundred fifty-two consecutive renal transplants were included: 1 kidney of each of the 376 donors was preserved by HMP, the contralateral organ was preserved by CS. RESULTS: The mean CIT was 3:05 PM (SD, 4:58 AM). A subgroup analysis was performed, groups were based on CIT duration: 0 to 10 hours, 10 to 15 hours, 15 to 20 hours, or 20 hours or longer. Delayed graft function (DGF) incidence in the subgroup with up to 10 hours CIT was 6.0% (N = 3/50) in the HMP arm and 28.1% (N = 18/64) in the CS arm (univariable P = 0.002; multivariable odds ratio [OR], 0.02; P = 0.007). Cold ischemia time remained an independent risk factor for DGF for machine perfused kidneys recovered from donation after brain death donors (OR, 1.06; 95% confidence interval [CI], 1.017-1.117; P = 0.008), donation after circulatory death donors (OR, 1.13; 95% CI, 1.035-1.233; P = 0.006) and expanded criteria donors (OR, 1.14; 95% CI, 1.057-1.236; P = 0.001). CONCLUSIONS: In conclusion, HMP resulted in remarkably lower rates of DGF in renal grafts that were transplanted after a short CIT. Also, CIT remained an independent risk factor for DGF in HMP-preserved kidneys.

Capsulotomy of Ischemically Damaged Donor Kidneys: A Pig Study.

BACKGROUND: Ischemia-reperfusion injury of donor kidneys may worsen transplant outcome. Kidneys with severe injury, such as kidneys of donors after circulatory death, develop edema, which may lead to renal compartment syndrome with reduced tissue perfusion. OBJECTIVE: We studied the effect of capsulotomy during hypothermic machine perfusion (HMP) of ischemically damaged porcine kidneys. METHODS: Eight pairs of kidneys from slaughterhouse pigs were assigned to two groups (20 and 45 min of warm ischemia). After 21 h of HMP, capsulotomy was performed, and perfusion was continued for 2 h. During perfusion, machine flow (Q), renal resistance (RR), renovascular circulating volume (RCV), intraparenchymal pressure (IPP) and weight were recorded. Parenchymal injury was examined with methylene blue infusion. RESULTS: Mean Q and RCV increased directly after capsulotomy [percentage increase (95% confidence interval): x0394;Q = 32% (17, 47), p = 0.001, and x0394;RCV = 19% (3, 35), p = 0.023]. Mean RR decreased [x0394;RR = -23% (-31, -15), p < 0.001]. Subanalysis comparing both warm ischemia groups showed no significantly different effect of capsulotomy between groups. There was no methylene blue leakage after capsulotomy in any kidney. CONCLUSIONS: Renovascular perfusion can be improved with capsulotomy during HMP, without damaging the renal parenchyma. Follow-up studies need to determine which donor kidneys may benefit from capsulotomy.