fibrin and Cadaver

Topics: Angiography; Animals; Anuria; Cadaver; Diuresis; Dogs; Fibrin; Glomerular Filtration Rate; Graft Rejection; Humans; Hypertension, Renal; Kidney; Kidney Glomerulus; Kidney Transplantation; Kidney Tubules; Postoperative Complications; Proteinuria; Thrombosis; Time Factors; Tissue Preservation; Transplantation Immunology; Transplantation, Autologous; Transplantation, Homologous

The aim of this study was to determine the clinical and histologic outcomes related to transplanting kidneys from deceased donors with glomerular fibrin thrombi (GFT). We included all cases transplanted between October 2003 and October 2014 that had either a preimplantation biopsy or an immediate postreperfusion biopsy showing GFT. The study cohort included 61 recipients (9.9%) with GFT and 557 in the control group without GFT. Delayed graft function occurred in 49% of the GFT group and 39% in the control group (p = 0.14). Serum creatinine at 1, 4, and 12 months and estimated GFR at 12 months were similar in the two groups. Estimated 1-year graft survival was 93.2% in the GFT group and 95.1% in the control group (p = 0.22 by log-rank). Fifty-two of the 61 patients in the GFT group (85%) had a 1-month protocol biopsy, and only two biopsies (4%) showed residual focal glomerular thrombi. At the 1-year protocol biopsy, the prevalence of moderate to severe interstitial fibrosis and tubular atrophy was 24% in the GFT group and 30% in the control group (p = 0.42). We concluded that GFT resolves rapidly after transplantation and that transplanting selected kidneys from deceased donors with GFT is a safe practice.

Topics: Adult; Cadaver; Female; Fibrin; Follow-Up Studies; Glomerular Filtration Rate; Graft Rejection; Graft Survival; Humans; Kidney Failure, Chronic; Kidney Function Tests; Kidney Glomerulus; Kidney Transplantation; Male; Middle Aged; Prognosis; Retrospective Studies; Risk Factors; Thrombosis; Tissue Donors

Biological materials derived from extracellular matrix (ECM) proteins have garnered interest as their composition is very similar to that of native tissue. Herein, we report the use of human cornea derived decellularized ECM (dECM) microparticles dispersed in human fibrin sealant as an accessible therapeutic alternative for corneal anterior stromal reconstruction. dECM microparticles had good particle size distribution (≤10 µm) and retained the majority of corneal ECM components found in native tissue. Fibrin-dECM hydrogels exhibited compressive modulus of 70.83 ± 9.17 kPa matching that of native tissue, maximum burst pressure of 34.3 ± 3.7 kPa, and demonstrated a short crosslinking time of ~17 min. The fibrin-dECM hydrogels were found to be biodegradable, cytocompatible, non-mutagenic, non-sensitive, non-irritant, and supported the growth and maintained the phenotype of encapsulated human corneal stem cells (hCSCs) in vitro. In a rabbit model of anterior lamellar keratectomy, fibrin-dECM bio-adhesives promoted corneal re-epithelialization within 14 days, induced stromal tissue repair, and displayed integration with corneal tissues in vivo. Overall, our results suggest that the incorporation of cornea tissue-derived ECM microparticles in fibrin hydrogels is non-toxic, safe, and shows tremendous promise as a minimally invasive therapeutic approach for the treatment of superficial corneal epithelial wounds and anterior stromal injuries.

Topics: Animals; Cadaver; Cell Proliferation; Cornea; Corneal Diseases; Extracellular Matrix; Fibrin; Humans; Hydrogels; Rabbits; Stem Cell Transplantation; Stem Cells; Tissue Engineering; Wound Healing

De novo generation of axially vascularized tissue with clinically relevant dimensions in a large animal model and implementation of clinically established imaging modalities for in vivo evaluation of vascularization. To be used for reconstruction of tissue defects, engineered grafts need to be axially vascularized to enable transplantation without graft loss due to hypoxia. Limitations to dimensions in small animal models had not yet been overcome, which is necessary to yield clinical relevance. Anatomical studies of groin and axillary regions in eight merino sheep were followed by microsurgical creation of an arteriovenous loop (AV-loop), embedded in an isolation chamber filled with fibrin matrix. Constructs were implanted in the groin of six sheep for up to 6 weeks. Course of vascularization in de novo forming tissue was assessed by sequential computed tomography angiography (CTA) and magnetic resonance angiography (MRA) in vivo, as well as by postexplantational micro-computed tomography and histology. A vascular axis was constantly found epifascially at the medial aspect of all sheep's thighs, which was used for AV-loop creation. Patency of AV-loop could be visualized by CTA and MRA scans during 1-6 weeks. Complex 3D-vessel-reconstruction revealed increasing axial vascularization of the fibrin matrix and growing connective tissue within the isolation chamber, which was confirmed by micro-computed tomography and histology postexplantation. De novo formation of axially vascularized tissue was demonstrated for the first time ever in a large animal model, paving the way for the first application of tissue engineering vascularized grafts with clinically relevant dimensions.

Topics: Animals; Arteries; Arteriovenous Shunt, Surgical; Axilla; Cadaver; Disease Models, Animal; Female; Fibrin; Groin; Magnetic Resonance Angiography; Microsurgery; Neovascularization, Physiologic; Saphenous Vein; Tissue Engineering; Tomography, X-Ray Computed

A thrombotic/inflammatory reaction is elicited when isolated islets of Langerhans come in contact with ABO-compatible blood. The detrimental effects of this instant blood-mediated inflammatory reaction (IBMIR) provide a reasonable explanation for the observation that an unexpectedly high number of islets, from several donors, are needed to produce normoglycemia in transplant patients with type 1 diabetes. In this study, the hypothesis that a specific thrombin inhibitor, Melagatran, could reduce IBMIR in an in vitro model in which human islets are exposed to ABO-compatible blood was tested. The administration of Melagatran abrogated IBMIR dose-dependently. Islets exposed to blood, in the absence or presence of 0.4 micromol/l Melagatran, exhibited a loss of integrity and were found to be trapped in macroscopic clots containing platelets and CD11b(+) leukocytes. At concentrations from 1 to 10 micromol/l, Melagatran inhibited both coagulation and complement activation. Also, platelet and leukocyte activation and consumption were decreased. Islet morphology was maintained with almost no platelets adhering to the surface, and infiltration by CD11b(+) leukocytes was considerably reduced. In conclusion, Melagatran significantly reduced IBMIR in this model system. This protective effect indicates that thrombin plays a pivotal role in IBMIR and suggests that thrombin inhibition can improve the outcome of clinical islet transplantation.

Topics: ABO Blood-Group System; Aged; Aged, 80 and over; Anticoagulants; Azetidines; Benzylamines; Blood Platelets; Cadaver; CD11 Antigens; Female; Fibrin; Glycine; Humans; Immunohistochemistry; Inflammation; Islets of Langerhans; Islets of Langerhans Transplantation; Male; Middle Aged; Neutrophils; Platelet Glycoprotein GPIIb-IIIa Complex; Thrombin; Thrombosis

Although FK 506 has been shown to effectively reverse refractory renal allograft rejection, its ability to reverse accelerated renal allograft rejection as a primary agent has not been specifically addressed. Herein evidence of the ability of FK 506 to reverse accelerated renal allograft rejection is presented. A 16-yr-old highly sensitized (PRA 75%) male underwent a second cadaveric renal transplant procedure. Despite induction immunosuppression with ATGAM, cyclosporine, azathioprine, and corticosteroids, a marked elevation in serum creatinine (1.6-->2.1 ng/dl) and reduction in urine output (4000 ml/d-->1000 ml/d) were observed on the sixth post-transplant day. Renal allograft biopsy performed at that time revealed typical features of accelerated rejection including neutrophil margination in glomerular and interstitial capillaries, and C3, IgG, and fibrin deposition in glomerular and interstitial capillaries (by immunofluorescence). FK 506 therapy was promptly instituted and ATGAM therapy discontinued. Serum creatinine peaked within 3 d of FK 506 therapy (2.5 mg/dl) and subsequently progressively dropped to 1.2 mg/dl. Repeat biopsy on FK 506 treatment day 12 revealed marked histologic improvement. Renal function remains excellent (1.3 mg/dl) 18 months after initiation of FK 506 therapy, and recurrent rejection has not been observed. This experience provides evidence that FK 506 therapy may effectively reverse accelerated renal allograft rejection, and that it provides a means for treating antibody-mediated mechanisms of allograft rejection.

Topics: Adolescent; Anti-Inflammatory Agents; Antibodies; Antilymphocyte Serum; Azathioprine; Biopsy; Cadaver; Capillaries; Complement C3; Creatinine; Cyclosporine; Fibrin; Glucocorticoids; Graft Rejection; Humans; Immunoglobulin G; Immunosuppressive Agents; Kidney Glomerulus; Kidney Transplantation; Male; Methylprednisolone Hemisuccinate; Neutrophils; Reoperation; Tacrolimus; Urinary Retention

The implantation and one hour post-transplant renal biopsies from three types of allograft recipients were compared with a blind grading system: (1) 25 cadaver kidneys preserved by pulsatile perfusion, (2) seven cadaver kidneys preserved by simple hypothermia following electrolyte solution flush, (3) 18 kidneys from living-related donors. Significant lesions were found only in cadaver kidneys which had received pulsatile preservation. Microscopic findings were correlated with perfusing agent, length of perfusion and its characteristics, and subsequent clinical course of the patient. Perfusion-related injury was found to be morphologically identical to hyperacute rejection, although the lesion is produced by quite different mechanisms. Pulsatile preservation appears to be associated with a spectrum of mechanical endothelial injury ranging from minute breaks visible only ultrastructurally to areas of complete denudation baring the basement membrane. The exposed collagen activates the clotting sequence resulting in platelet and fibrin deposition, whereas in classical hyperacute rejection the triggering mechanism is cytotoxic recipient antibody. The extent of perfusion-related injury correlates well with length of preservation, quantity of fibrin deposited, and, most importantly, with both the immediate and long-term post-transplant failure rate. In some patients the injury appears to be produced by cytotoxic antibodies in the plasma perfusate, which combine with antigens in the kidney ex vivo. The Ag-Ab complex activates complement and coagulation sequences in vivo after reimplantation. Early results with albumin or purified plasma fraction perfusates suggest this portion of perfusion-related injury can be eliminated. Comparison of pre- and postimplantation biopsies of the kidneys preserved by simple hypothermia or by pulsatile preservation suggests that perfusion-related injury is much more common than is true hyperacute rejection mediated by recipient cytotoxic antibodies. We suggest that the term "hyperacute rejection" be reserved for situations where significant endothelial drainage has been excluded by preimplantation biopsy and where recipient cytotoxic antibodies can be proved.

Topics: Cadaver; Fibrin; Fluorescent Antibody Technique; Humans; Kidney Diseases; Kidney Glomerulus; Kidney Transplantation; Organ Preservation; Perfusion; Postoperative Complications; Tissue Survival; Transplantation, Homologous

The effect of a cadaver-derived vascular plasminogen activator (VA) on the degradation of fibrinogen, soluble fibrin monomer, and fibrin was studied and compared with the effect of equivalent fibrinolytic potencies of streptokinase (SK), urokinase (UK), and plasmin. The proteolytic activity of the three activators and plasmin was determined by a standard fibrin plate assay and was expressed in CTA units from a UK reference curve. Fibrinogen degradation was measured by clottable protein determinations and by an electrophoretic technique sensitive to small changes in the molecular weight of fibrinogen. When VA was incubated in plasma, no degradation of fibrinogen occurred, whereas rapid fibrinolysis took place after the plasma was clotted. By contrast, equivalent potencies of SK, UK, and plasmin caused extensive fibrinogenolysis. Since the plasmin added and that formed by the three activators had equivalent fibrinolytic activity, the failure of VA to induce fibrinogen degradation was attributed to antiactivators rather than antiplasmins. VA activity in plasma was consumed by clotting, whereas the antiactivator activity remained in the serum, suggesting dissociation of the VA-antiactivator complex on the fibrin clot. Fibrinogen and its soluble derivatives resisted degradation by VA in plasma because a solid phase appeared necessary for the complex to dissociate. The findings indicated that the degradation of fibrinogen or soluble fibrin in blood as a result of plasminogen activation by VA was unlikely to occur due to a large excess of antiactivator activity. Alternative pathways for their catabolism are discussed.

Topics: Cadaver; Electrophoresis, Polyacrylamide Gel; Extremities; Femoral Artery; Femoral Vein; Fibrin; Fibrinogen; Fibrinolysin; Fibrinolysis; Humans; Plasminogen Activators; Streptokinase; Urokinase-Type Plasminogen Activator

Acute renal failure developed in a 55-year-old man 6 days after he had received a cadaver renal allograft. This was associated with thrombocytopenia. Extensive intraglomerular fibrin deposition was seen in a renal biopsy specimen. He was treated with corticosteroids, azathioprine, cyclophosphamide and hemodialysis with regional heparinization but not with systemic anticoagulation. This was followed by complete recovery of both renal function and histologic damage despite the fact that he did not receive anticoagulant therapy. This suggests that treatment with anticoagulants may not be necessary for all patients with intraglomerular deposits of fibrin.

Topics: Acute Kidney Injury; Azathioprine; Biopsy, Needle; Cadaver; Cyclophosphamide; Fibrin; Graft Rejection; Humans; Kidney Glomerulus; Kidney Transplantation; Male; Methylprednisolone; Middle Aged; Renal Dialysis; Thrombocytopenia; Transplantation, Homologous

Topics: Azathioprine; Blood Coagulation; Cadaver; Creatinine; Drug Evaluation; Fibrin; Graft Rejection; Hemoglobins; Humans; Immunosuppression Therapy; Kidney Function Tests; Kidney Transplantation; Prednisone; Proteinuria; Transplantation, Homologous; Urea; Warfarin

Topics: Aged; Autopsy; Blood Coagulation; Cadaver; Erythrocytes; Fibrin; Hemorrhage; Humans; Male; Microscopy, Electron, Scanning; Middle Aged; Postmortem Changes; Skin; Streptokinase; Thrombosis; Time Factors

Topics: Ammonium Sulfate; Blood Vessels; Cadaver; Caseins; Chemical Precipitation; Chloromercuribenzoates; Chromatography; Chromatography, DEAE-Cellulose; Chromatography, Gel; Dithiothreitol; Electrophoresis, Polyacrylamide Gel; Ethanol; Fibrin; Fibrinolytic Agents; Hemoglobins; Humans; Hydrogen-Ion Concentration; Immune Sera; Molecular Weight; Peptide Hydrolases; Plasminogen; Zinc

The urinary excretion of fibrin/fibrinogen degradation products (F.D.P.) of 81 human cadaver kidney transplants has been measured serially by the techniques of tanned red cell haemagglutination inhibition immunoassay and immunonephelometry. Acute rejection episodes in functioning transplants have been associated with increased F.D.P. excretion which in 80% of cases has preceded clinical diagnosis by periods of one to seven days. Recovery from these episodes has been associated with a rapid fall of F.D.P. excretion to undetectable levels. The level of F.D.P. excretion during a rejection episode is a guide to its ultimate outcome. Irreversibly rejected kidneys excrete high levels of F.D.P. for long periods. Viable kidney transplants with prolonged oliguric phases can be distinguished, while still oliguric, from rejected kidneys by their low F.D.P. excretion. F.D.P. cannot usually be detected in the urine of well-functioning transplants. Episodes of raised F.D.P. excretion in the absence of acute clinical rejection, however, occur occasionally and may be associated with permanent impairment of renal function.

Topics: Anuria; Cadaver; Creatinine; Fibrin; Fibrinogen; Graft Rejection; Hemagglutination Inhibition Tests; Humans; Hypertension; Kidney Transplantation; Transplantation, Homologous; Urea

Topics: Blood Urea Nitrogen; Cadaver; Creatinine; Fibrin; Graft Rejection; Humans; Kidney Transplantation; Methods; Tissue Donors; Transplantation Immunology; Transplantation, Homologous

Topics: Animals; Aorta, Abdominal; Arteries; Blood Platelets; Cadaver; Endothelium; Fibrin; Forensic Medicine; Humans; Microscopy, Electron, Scanning; Postmortem Changes; Rabbits; Rats; Time Factors; Wounds and Injuries

Topics: Blood Coagulation; Cadaver; Death, Sudden; Fibrin; Fibrinogen; Fibrinolysis; Fibrinolytic Agents; Forensic Medicine; Hot Temperature; Humans; Immunoelectrophoresis; Postmortem Changes; Streptokinase

Topics: Biopsy; Blood Platelets; Cadaver; Complement System Proteins; Cytotoxicity Tests, Immunologic; Fibrin; Fluorescent Antibody Technique; Graft Rejection; Humans; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Kidney; Kidney Glomerulus; Kidney Transplantation; Leukocytes; Microscopy, Electron; Renal Artery; Serum Albumin; Thrombosis; Transplantation Immunology; Transplantation, Homologous

Topics: Adolescent; Adult; Cadaver; Female; Fibrin; Humans; Immunosuppressive Agents; Kidney; Kidney Glomerulus; Kidney Transplantation; Male; Renal Dialysis; Sepsis; Shwartzman Phenomenon; Toxemia; Transplantation Immunology; Transplantation, Homologous

Topics: Autopsy; Cadaver; Death; Fibrin; Fibrinolysis; Humans