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Intra-graft Coagulation Events in Clinical Renal Transplantation and Delayed Graft Function

Conditions
Reperfusion Injury
Delayed Graft Function
End Stage Renal Disease
Kidney Transplantation
Registration Number
NCT02568696
Lead Sponsor
Helsinki University Central Hospital
Brief Summary

The purpose of this study is to investigate local activation of the coagulation system in the kidney graft during organ preservation and during early reperfusion in adult kidney transplantation. Generation of thrombin and fibrin as well as activation and inhibition of fibrinolysis will be investigated. Influence of these events on delayed graft function (DGF) and acute cell-mediated rejection will be evaluated.

Detailed Description

Background

In clinical kidney transplantation organ retrieval, cold-preservation of the graft as well as restoration of the blood flow to the transplant cause tissue damage (ischemia/reperfusion injury). Clinically these events can manifest themselves as delayed graft function (DGF), which is usually defined as the need for dialysis during first week after transplantation. DGF increases the risk of developing chronic rejection and subsequently loss of the transplant.

Ischaemia/reperfusion injury is biologically characterized by local profound inflammatory response, activation of the coagulation system and endothelial dysfunction in the transplanted organ. After reperfusion activated neutrophils cause tissue damage in the graft by production of reactive oxygen species (ROS) and release of proteolytic enzymes, which lead to plugging of the capillaries by accumulation of thrombocytes and fibrin. Blood flow is further diminished by increased blood viscosity and local vasoconstriction and swelling of the endothelial cells. Disorders of the microcirculation lead to "no-reflow" phenomenon whereby locally tissues remain ischemic, despite of good blood flow in the organ artery and vein.

Coagulation is activated in the renal transplant during reperfusion, when circulating factor VIIa (FVIIa) comes into contact with the tissue factor (TF), which is expressed on the endothelium due to ischaemia. FVII-TF complex activates factor X (FX) and activated FX (FXa) cleaves thrombin (FII) from prothrombin. Thrombin activates thrombocytes, cleaves fibrin from fibrinogen and activates factor XIII( FXIII), which stabilizes fibrin clot. Fibrin has been demonstrated to accumulate in the kidney graft during reperfusion. Fibrin accumulation is aggravated by inhibition of fibrinolysis due to reperfusion.

Furthermore, the investigators conducting this current research project, have previously gained indirect evidence in a small cohort study, that accumulation of fibrin occurs even before reperfusion, during donor care and organ retrieval. Most importantly, specifically this pre-reperfusion fibrin deposition was related to DGF.

Patients and sample size

There were several limitations in investigators previous study concerning intra-graft coagulation events in DGF. It was conducted as a part of a larger trial in renal transplantation and included only 30 patients in two study arms with different immunosuppressant regimens (peri-operative basiliximab and conventional triple therapy). Therefore, a new study, with larger sample size and standardized immunosuppression is warranted.

Therefore, in this current prospective observational study surgical technique, anaesthesia and hemodynamic management, immunosuppressive medications are strictly standardized. Sample size is increased to 100. The investigators prospectively screen all adult patients receiving their first kidney transplant from cadaveric donor. Only patients scheduled to receive local standard triple immunosuppressant therapy with cyclosporine A, mycophenolate mofetil and methylprednisolone are included.

Blood samples and prospective data collection

Blood samples for assessment of intra-graft coagulation events (generation of thrombin and fibrin, activation and inhibition of fibrinolysis) are drawn peri-operatively. Predefined clinical and demographical data are collected preoperatively and prospectively during 3 months after kidney transplantation to assess the influence of these coagulation events on delayed graft function according to Halloran criteria (8) (primary outcome) and acute cell mediated graft rejection (primary outcome).

Recruitment & Eligibility

Status
UNKNOWN
Sex
All
Target Recruitment
100
Inclusion Criteria
  • adult person (over 18 years old)
  • cadaveric transplantation
  • conventional standard immunosuppression plan (methylprednisolone, cyclosporin A, mycophenolate mofetil)
Exclusion Criteria
  • previous kidney transplant
  • other than local standard immunosuppression
  • panel reactive antibodies (PRA) >30%
  • warfarin therapy
  • dual anti-platelet therapy
  • use of low molecular weight heparins (LMWH) or fondaparinux during last two weeks before surgery for other indication than hemodialysis

Study & Design

Study Type
OBSERVATIONAL
Study Design
Not specified
Primary Outcome Measures
NameTimeMethod
Trans-renal difference/ratio of plasma concentrations of coagulation measurementsTwo minutes after reperfusion of the kidney transplant

Trans-transplant difference/ratio is determined in order to correlate intra-graft coagulation events to the incidences of other primary outcomes.

Delayed Graft FunctionDuring 1 week after kidney transplantation

Delayed graft function is assessed by Halloran criteria: oliguria \< 1000ml/24h for more than 2 days after transplantation or plasma creatinine \>500 micromol/l during the first week after transplantation or more than one dialysis during the first week after transplantation (Halloran et al, Transplantation 1988;46:223-8.)

Acute cell mediated graft rejectionDuring 3 months after kidney transplantation
Secondary Outcome Measures
NameTimeMethod
Myeloperoxidase and/or lactoferrinBlood samples are taken at two timepoints: 1) immediately before the start of the surgery; 2) 2 minutes after reperfusion of the kidney transplant

Preoperative value and the trans-transplant difference and/or ratio is determined in order to assess activation of neutrophiles

Plasma urea value (mmol/L)At admission to the hospital, during the first week after transplantation and at 1 and 3 months after renal transplantation
Estimated glomerular filtration rate (ml/min/1.73 m2)At admission to the hospital, during the first week after transplantation and at 1 and 3 months after renal transplantation

Estimated glomerular filtration rate is calculated according to the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) formula

Plasma creatinine value (micromol/L)At admission to the hospital, during the first week after transplantation and at 1 and 3 months after renal transplantation
Urine output (ml/24h)Pre-operative urine output (ml/24h) and daily urine output (ml/24h) during the first week after transplantation

Urine output (ml/24h) before the surgery and during the first week after transplantation will be recorded

Renal artery and renal vein blood flow (ml/min)Immediately after blood sample retrieval during reperfusion

Renal artery and renal vein blood flow (ml/min) are measured intra-operatively immediately after blood sample retrieval using a specific probe

Fluid balance during surgery and post-anesthesia care unit stay (ml)From the start of the kidney transplantation surgery until the discharge from post-anesthesia care unit (up to 24 hours from the start of the surgery)

All fluids infused from the start of the kidney transplantation surgery until discharge from the post-anaesthesia care unit (until discharge to the ward) are recorded. All fluids losses (blood loss, urine output) during this period are recorded.

TransfusionFrom the start of the kidney transplantation surgery until the discharge from post-anesthesia care unit (up to 24 hours from the start of the surgery)

All blood products used during this time frame are recorded and reported

Prothrombin fragment 1+2 (F1+2)Blood samples are taken at two timepoints: 1) immediately before the start of the surgery; 2) 2 minutes after reperfusion of the kidney transplant

Preoperative value and the trans-transplant difference and/or ratio is determined in order to assess thrombin generation.

Fibrinopeptide ABlood samples are taken at two timepoints: 1) immediately before the start of the surgery; 2) 2 minutes after reperfusion of the kidney transplant

Preoperative value and the trans-transplant difference and/or ratio is determined in order to assess fibrin generation.

D-dimersBlood samples are taken at two timepoints: 1) immediately before the start of the surgery; 2) 2 minutes after reperfusion of the kidney transplant

Preoperative value and the trans-transplant difference and/or ratio is determined in order to assess fibrin degradation

Tissue type plasminogen activatorBlood samples are taken at two timepoints: 1) immediately before the start of the surgery; 2) 2 minutes after reperfusion of the kidney transplant

Preoperative value and the trans-transplant difference and/or ratio is determined in order to assess activation of fibrinolysis.

Plasminogen activator inhibitorBlood samples are taken at two timepoints: 1) immediately before the start of the surgery; 2) 2 minutes after reperfusion of the kidney transplant

Preoperative value and the trans-transplant difference and/or ratio is determined in order to assess inhibition of fibrinolysis.

Interleukin 6, interleukin 8, interleukin 10Blood samples are taken at two timepoints: 1) immediately before the start of the surgery; 2) 2 minutes after reperfusion of the kidney transplant

Preoperative value and the trans-transplant difference and/or ratio is determined in order to assess activation/inhibition of inflammation

Syndecan-1Blood samples are taken at two timepoints: 1) immediately before the start of the surgery; 2) 2 minutes after reperfusion of the kidney transplant

Preoperative value and the trans-transplant difference and/or ratio is determined in order to assess degradation of endothelial glycocalyx

Number of hemodialyses and their indication after surgeryFrom the start of the surgery until 3 months after surgery

All dialysis sessions will be recorded during first post-operative week. Indication for dialysis (oliguria, hyperkalemia, hypervolemia, acidosis) will be recorded during first post-operative week. At 1 and 3 months after surgery only number of dialyses/week will be recorded

Trial Locations

Locations (1)

Helsinki University and Helsinki University Hospital

🇫🇮

Helsinki, Finland

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