Rapid Metabolic Recovery of Donor Circulatory Death Liver Graft Using Whole Blood Perfusion: A Pig Study

Abstract Background Since autologous veins are unavailable when needed in more than 20% of cases in vascular surgery, the production of personalized biological vascular grafts for implantation has become crucial. Surface modification of decellularized xenogeneic grafts with vascular cells to achieve physiological luminal coverage and eventually thromboresistance is an important prerequisite for implantation. However, ex vivo thrombogenicity testing remains a neglected area in the field of tissue engineering of vascular grafts due to a multifold of reasons. Methods After seeding decellularized bovine carotid arteries with human endothelial progenitor cells and umbilical cord-derived mesenchymal stem cells, luminal endothelial cell coverage (LECC) was correlated with glucose and lactate levels on the cell supernatant. Then a closed loop whole blood perfusion system was designed. Recellularized grafts with a LECC > 50% and decellularized vascular grafts were perfused with human whole blood for 2 h. Hemolysis and complete blood count evaluation was performed on an hourly basis, followed by histological and immunohistochemical analysis. Results While whole blood perfusion of decellularized grafts significantly reduced platelet counts, platelet depletion from blood resulting from binding to re-endothelialized grafts was insignificant (p = 0.7284). Moreover, macroscopic evaluation revealed thrombus formation only in the lumen of unseeded grafts and histological characterization revealed lack of CD41 positive platelets in recellularized grafts, thus confirming their thromboresistance. Conclusion In the present study we were able to demonstrate the effect of surface modification of vascular grafts in their thromboresistance in an ex vivo whole blood perfusion system. To our knowledge, this is the first study to expose engineered vascular grafts to human whole blood, recirculating at high flow rates, immediately after seeding.

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Biomarkers of Early Liver Graft Damage in Circulatory Death and Brain Death Donors: A Propensity Score Matching Analysis

Transplantation Proceedings ◽

10.1016/j.transproceed.2021.07.055 ◽

2021 ◽

Author(s):

Víctor Lopez-Lopez ◽

Carlos Martínez-Caceres ◽

David Ferreras ◽

Jesus De La Peña-Moral ◽

Juan De La Cruz ◽

...

Keyword(s):

Propensity Score ◽

Brain Death ◽

Propensity Score Matching ◽

Liver Graft ◽

Propensity Score Matching Analysis ◽

Circulatory Death

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Fibulin-1 mediates platelet adhesion via a bridge of fibrinogen

Blood ◽

10.1182/blood.v88.7.2569.bloodjournal8872569 ◽

1996 ◽

Vol 88 (7) ◽

pp. 2569-2577 ◽

Cited By ~ 48

Author(s):

S Godyna ◽

M Diaz-Ricart ◽

WS Argraves

Keyword(s):

Extracellular Matrix ◽

Vascular Injury ◽

Whole Blood ◽

Platelet Adhesion ◽

Solid Phase ◽

Blood Perfusion ◽

General Mechanism ◽

Binding Assays ◽

Solid Phase Binding ◽

Coated Surfaces

Fibulin-1 is a component of the extracellular matrix that surrounds vascular smooth muscle. This observation, along with the recent finding that fibulin-1 can bind fibrinogen (J Biol Chem 270:19458, 1995), prompted investigation into the potential role of fibulin-1 as a thrombogenic agent. In perfusion chamber assays, platelets in whole blood under flow conditions attached and spread on surfaces coated with fibulin-1. This adhesion was completely blocked by fibulin-1 antibodies. Platelets free of plasma did not attach to fibulin-1 coated surfaces; however, with the addition of fibrinogen, platelet adhesion to fibulin-1 took place. When detergent extracts of platelets were subjected to fibulin-1-Sepharose affinity chromatography, the integrin alpha IIb beta 3 was selected. Solid phase binding assays using purified components showed that integrin alpha IIb beta 3 could not bind directly to fibulin-1 but in the presence of fibrinogen the integrin bound to fibulin-1-coated surfaces. Monoclonal alpha IIb beta 3 antibodies capable of blocking its interaction with fibrinogen completely blocked platelet adhesion to fibulin-1 in both whole blood perfusion and static adhesion assays. The results show that fibulin-1 can support platelet attachment via a bridge of fibrinogen to the platelet integrin alpha IIb beta 3. When fibroblast monolayers containing extracellular matrix-incorporated fibulin-1 were used as adhesion substrates, platelet adhesion in the presence of fibrinogen could be inhibited by 30% using antibodies to fibulin-1. Following vascular injury, fibulin-1 present in the extracellular matrix of the vessel wall may therefore interact with plasma fibrinogen and promote platelet adhesion, leading to the formation of a platelet plug. Thus, fibulin-1 joins the list of matrix proteins including collagens I and IV and fibronectin that mediate platelet adhesion via a plasma protein bridge. This bridging phenomenon may represent a general mechanism by which platelets interact with exposed subendothelial matrices following vascular injury.

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Comparison of Intermittent Warm and Cold Blood Perfusion During Hypothermic Myocardial Preservation on Functional and Metabolic Recovery

Echocardiography ◽

10.1111/j.1540-8175.1985.tb01420.x ◽

1985 ◽

Vol 2 (6) ◽

pp. 451-459

Author(s):

Thomas N. Masters ◽

Francis Robicsek ◽

Alexander A. Fokin ◽

Joseph W. Cook ◽

Geoffrey Gong ◽

...

Keyword(s):

Blood Perfusion ◽

Myocardial Preservation ◽

Metabolic Recovery ◽

Cold Blood

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The Influence of the Hematocrit of the Pulsatile Whole Blood Perfusion of a Stenosis in an Elastic Tube System

Cardiovascular System Dynamics ◽

10.1007/978-1-4899-6693-3_28 ◽

1982 ◽

pp. 281-287

Author(s):

P. Mönninghoff ◽

H. Rieger ◽

H. Zeller ◽

J. Reinecke

Keyword(s):

Whole Blood ◽

Blood Perfusion ◽

Elastic Tube ◽

Tube System

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Influences of Hemodilution and Anticoagulation on Antiplatelet P2Y12 Therapy: In Vitro Whole Blood Perfusion Model

Journal of Cardiothoracic and Vascular Anesthesia ◽

10.1053/j.jvca.2013.06.010 ◽

2013 ◽

Vol 27 (6) ◽

pp. e69-e71 ◽

Cited By ~ 1

Author(s):

Satoru Ogawa ◽

Kazuya Hosokawa ◽

Kenichi A. Tanaka

Keyword(s):

Whole Blood ◽

Blood Perfusion ◽

Perfusion Model

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Resin Column Perfusion with Whole Blood or Plasma Separated by the Continuous Flow Celltrifuge

Clinical Science ◽

10.1042/cs0480187 ◽

1975 ◽

Vol 48 (3) ◽

pp. 187-192

Author(s):

M. J. Weston ◽

P. J. Mellon ◽

P. G. Langley ◽

R. D. Hughes ◽

E. H. Dunlop ◽

...

Keyword(s):

Plasma Flow ◽

Whole Blood ◽

Continuous Flow ◽

Blood Perfusion ◽

Resin Column ◽

Liver Support ◽

Flow Rates ◽

Plasma Separation ◽

Exchange Resins

1. The aim of this study was to define the factors influencing plasma separation from the continuous flow celltrifuge and to evaluate plasma as an alternative to whole blood for perfusion of exchange resins as part of a system of artificial liver support. 2. Studies in vitro showed the importance of packed cell volume, centrifugal force and duration of centrifugation on the degree of plasma separation. From these data it was possible to calculate plasma flow rates likely to be obtained from the celltrifuge when used in vivo. These predicted values correlated closely with plasma flow rates obtained in twenty-six studies in dogs. 3. Comparison of whole blood perfusion with plasma perfusion of exchange resins in another series of dog experiments showed that with whole blood perfusion there was often a considerable rise in pressure across the resin column but that this did not occur with plasma perfusion. 4. Measurements of platelet losses in the same series of experiments showed a 50% reduction of arterial platelet counts over a 3 1/2 h period of perfusion when whole blood was perfused. Although the fall was lower with plasma perfusion, the difference was not statistically significant. 5. Use of the celltrifuge provides a means of resin perfusion free of the mechanical difficulties of whole blood perfusion, but platelet losses still remain a problem.

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Automated Analysis of Platelet Aggregation on Cultured Endothelium in a Microfluidic Chip Perfused with Human Whole Blood

Micromachines ◽

10.3390/mi10110781 ◽

2019 ◽

Vol 10 (11) ◽

pp. 781 ◽

Cited By ~ 1

Author(s):

Albers ◽

Passier ◽

van den Berg ◽

van der Meer

Keyword(s):

Image Analysis ◽

Whole Blood ◽

Microfluidic Channel ◽

Aggregate Size ◽

Blood Perfusion ◽

Automated Analysis ◽

Microfluidic Channels ◽

Size Distributions ◽

Platelet Aggregate ◽

Platelet Aggregates

Organ-on-a-chip models with incorporated vasculature are becoming more popular to study platelet biology. A large variety of image analysis techniques are currently used to determine platelet coverage, ranging from manually setting thresholds to scoring platelet aggregates. In this communication, an automated methodology is introduced, which corrects misalignment of a microfluidic channel, automatically defines regions of interest and utilizes a triangle threshold to determine platelet coverages and platelet aggregate size distributions. A comparison between the automated methodology and manual identification of platelet aggregates shows a high accuracy of the triangle methodology. Furthermore, the image analysis methodology can determine platelet coverages and platelet size distributions in microfluidic channels lined with either untreated or activated endothelium used for whole blood perfusion, proving the robustness of the method.

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