Page 23 - 2019_03-Haematologica-web

Perspective Article
Table 1. Available models for testing blood-endothelial interactions in vitro.
Model
Static conditions
Blood unit sampling
Specific component co-incubation
Wholebloodincubation
Laminar flow conditions
Acellular planar flow model
Cellular planar flow model
Protocol
• Repeated sampling of blood product over shelf life (37-41)
• Blood products or supernatant of blood product mixed with purified cell populations from healthy volunteers
- Neutrophils (1, 17-21)
- Macrophages (3)
- Platelets (22)
- HUVECs (23)
•Bloodproductcomponentsincubated with whole blood (2, 24)
• Storage-induced RMP incubated with whole blood (43)
• Cells from blood products perfused over layer of subendothelial matrix or collagen (44)
• Endothelial cells cultured on coverslips and subjected to specific shear stress
- 0.3 dyn/cm2 (27)
- 5 dyn/cm2 (26)
- 10 dyn/cm2 (25)
- Shear stress not reported (25)
• Different mixtures of blood perfused
through system
- 10% whole blood suspension (25) - 1.5% PRBC suspension (26, 27)
• Use of activated endothelium - E. coli 0127:B8 LPS (45)
- TNF-α (25, 45)
• Human umbilical artery (up to 20 cm) and connected to circuit (28)
• Whole blood diluted 1:10 circulated with shear pressure of 0.4 dyn/cm2 • Endothelium activated with TNF-α
•Biodegradabletubularscaffold matrices (length 4 cm, inner diameter 6 mm) seeded with HUVECs (29)
Outcome measures
• Markers of platelet activation + apoptosis (37)
• Media change (38)
• Microparticle accumulation (38) • Proteomics (42)
• Procoagulant characteristics (39) • RBC anti-oxidant oxidation during
storage (40)
• Membrane deformability (41)
• Neutrophil apoptosis (17)
• Neutrophil ROS formation (17, 19, 20)
• Neutrophil activation markers (1, 18, 21) • Neutrophil phagocytosis (1)
• Macrophage cytokine elaboration (3)
• Platelet adhesion + aggregation (22)
• RBC adhesion to endothelial cells (23)
•Cytokineproductionafteraddingfresh or stored PRBC supernatant to
whole blood (2, 24)
• Thromboelastometry, microparticle characterization, cytokine concentration (43)
• Influence of PC storage duration on adhesion capacity to subendothelial matrix + collagen (44)
• Adhesion of RBCs to endothelium from PRBCs stored for different
periods (26, 27, 45)
• Tissue factor expression + adhesion to endothelial cells (25)
• Microparticle formation, endothelial ROS formation (28)
•Confocalmicroscopy,biochemical extracellular matrix analysis, endothelial nitric oxide formation (29)
Research output and clinical impact
• PC undergo deterioration of mitochondrial integrity + apoptosis (37) during storage
• PRBC storage leads to enhanced RMP formation, osmotic fragility, hemolysis, decreased deformability (38),
prolonged activated clotting time (39), increased RBC anti-oxidant oxidation (40)
• Supernatant from stored PRBCs delays neutrophil apoptosis and primes neutrophils (17)
• PMP binding to neutrophils increases CD11b expression + phagocytic activity (1)
• Non-polar lipids accumulated during ex vivo PRBC storage prime neutrophils (20)
• PRBCs stored for prolonged periods are less capable of supporting platelet adhesion + aggregation (22)
•IncubationofmicroparticlesfromPRBCs with whole blood induced host production of TNF, IL-6, IL-8 (2)
• RMPs from stored PRBCs trigger coagulation of whole blood through TF signaling (43)
• Platelet aggregation + adhesion capacity are improved in BC PC compared
to PRP PC (44)
• RBCs from stored PRBCs more adherent to HUVECs under laminar flow conditions compared to those from fresh
PRBCs (26, 27)
• Exposure to TNF-stimulated HUVECs under arterial flow conditions led to increased tissue factor expression on monocytes compared to quiescent HUVECs (25)
• RBCs more adherent to E. coli-activated endothelium compared to quiescent endothelium (45)
• Whole blood perfused through TNF-stimulated umbilical arteries demonstrated increased microparticle formation. These microparticles enhanced HUVEC ROS formation in vitro (28)
•Monocytescirculatedthroughthe TNF-stimulated model adhered to
the endothelium and transmigrated into the intima (29)
Ex vivo artery model
Syntheticarterialmodel
haematologica | 2019; 104(3)
continued on the next page
429