The term angiogenesis refers to sprouting of new blood vessels from pre-existing ones. The angiogenic process involves cell migration and tubulogenesis requiring interaction between endothelial cells and the extracellular matrix. Human peroxidasin 1 (hsPxd01) is a multidomain heme peroxidase found embedded in the basement membranes. As it promotes the stabilization of extracellular matrix, we investigated its possible role in angiogenesis both in vitro and in vivo. We analysed the effects of peroxidasin 1 gene silencing and supplementation by recombinant hsPxd01 in TeloHAEC endothelial cells on cell migration, tubulogenesis in matrigel, and intracellular signal transduction as assessed by kinase phosphorylation and expression of pro-angiogenic genes as measured by qRT–PCR. We further evaluated the angiogenic potential of recombinant peroxidasin in a chicken chorioallantoic membrane model. RNA silencing of endogenous hsPxd01 significantly reduced tube formation and cell migration, whereas supplementation by the recombinant peroxidase promoted tube formation in vitro and stimulated vascularization in vivo through its catalytic activity. Moreover, recombinant hsPxd01 promoted phosphorylation of Extracellular signal-Regulated Kinases (ERK1/2), Protein kinase B (Akt), and Focal Adhesion Kinase (FAK), and induced the expression of pro-angiogenic downstream genes: Platelet Derived Growth Factor Subunit B (PDGFB), endothelial-derived Heparin Binding EGF-like growth factor (HB-EGF), CXCL-1, Hairy-Related Transcription Factor 1 (HEY-1), DNA-binding protein inhibitor (ID-2), Snail Family Zinc Finger 1 (SNAI-1), as well as endogenous hsPxd01. However, peroxidasin silencing significantly reduced Akt and FAK phosphorylation but induced ERK1/2 activation after supplementation by recombinant hsPxd01. While hsPxd01 silencing significantly reduced expression of HEY-1, ID-2, and PDGFB, it did not affect expression of SNAI-1, HB-EGF, and CXCL-1 after supplementation by recombinant hsPxd01. Our findings suggest a role of enzymatically active peroxidasin 1 as a pro-angiogenic peroxidase and a modulator of ERK1/2, Akt and FAK signalling.

Platelet spreading and retraction play a pivotal role in the platelet plugging and the thrombus formation. In routine laboratory, platelet function tests include exhaustive information about the role of the different receptors present at the platelet surface without information on the 3D structure of platelet aggregates. In this work, we develop, a method in Digital Holographic Microscopy (DHM) to characterize the platelet and aggregate 3D shapes using the quantitative phase contrast imaging. This novel method is suited to the study of platelets physiology in clinical practice as well as the development of new drugs.

It is a known fact that blood flow pattern and more specifically the pulsatile time variation of shear stress on the vascular wall play a key role in atherogenesis. The paper presents the conception, the building and the control of a new in vitro test bench that mimics the pulsatile flows behavior based on in vivo measurements. An in vitro cardiovascular simulator is alimented with in vivo constraints upstream and provided with further post-processing analysis downstream in order to mimic the pulsatile in vivo blood flow quantities. This real-time controlled system is designed to perform real pulsatile in vivo blood flow signals to study endothelial cells’ behavior under near physiological environment. The system is based on an internal model controller and a proportional-integral controller that controls a linear motor with customized piston pump, two proportional-integral controllers that control the mean flow rate and temperature of the medium. This configuration enables to mimic any resulting blood flow rate patterns between 40 and 700 ml/min. In order to feed the system with reliable periodic flow quantities in vivo measurements were performed. Data from five patients (1 female, 4 males; ages 44–63) were filtered and post-processed using the Newtonian Womersley’s solution. These resulting flow signals were compared with 2D axisymmetric, numerical simulation using a Carreau non-Newtonian model to validate the approximation of a Newtonian behavior. This in vitro test bench reproduces the measured flow rate time evolution and the complexity of in vivo hemodynamic signals within the accuracy of the relative error below 5%. This post-processing method is compatible with any real complex in vivo signal and demonstrates the heterogeneity of pulsatile patterns in coronary arteries among of different patients. The comparison between analytical and numerical solution demonstrate the fair quality of the Newtonian Womersley’s approximation. Therefore, Womersley’s solution was used to calculate input flow rate for the in vitro test bench.

Cardiovascular diseases represent a major issue in terms of morbidity and mortality for dialysis patients. This morbidity is due to the accelerated atherosclerosis observed in these patients. Atherosclerosis is a chronic inflammatory disease characterized by key players such as monocytes, macrophages, or oxidized LDLs. Monocytes-macrophages are classified into subsets of polarized cells, with M1 and M2 macrophages considered, respectively, as pro- and anti-inflammatory. (1) Methods: The monocyte subsets and phenotypes were analyzed by flow cytometry. These data were completed by the quantification of plasma M-CSF, IL-8, CRP, Mox-LDLs, Apo-B, Apo-AI, chloro-tyrosine, and homocitrulline concentrations. The statistical differences and associations between two continuous variables were assessed using the Mann–Whitney U test and Spearman’s correlation coefficient, respectively. (2) Results: Hemodialyzed patients showed a significant increase in their concentrations of CRP, M-CSF, and IL-8 (inflammation biomarkers), as well as chloro-tyrosine and homocitrulline (myeloperoxidase-associated oxidative stress biomarkers). Moreover, we observed a higher percentage of M2 monocytes in the plasma of hemodialysis patients as compared to the controls. (3) Conclusions: Our data suggest that oxidative stress and an inflammatory environment, which is amplified in hemodialysis patients, seems to favor an increase in the concentration of circulating M-CSF, therefore leading to an increase in M2 polarization among circulating monocytes.

Rationale: There are important interactions between Red Blood Cells (RBCs) and platelets in the bloodstream. These interactions lead to a phenomenon called margination. RBCs in pathological situations undergo biochemical and conformational changes leading to alterations in blood rheology. Aim: RBCs shape in volunteers (21), stable (42) and exacerbated (31) COPD patients was analyzed. We studied the effect of the RBCs spherization on the platelets transport experimentally, in vitro , and by using numerical simulations. Methods: RBC shape was estimated by the second moment of Pearson obtained through flow cytometry on fsc histogram. In vitro experiments were performed to analyze the effect of RBC shape on platelets adhesion/aggregation in dynamic conditions. Neuraminidase treatment was used to induce RBCs spherization. Numerical simulation were performed to determine the effect of RBCs spherization on platelets mean square displacement (MSD) to provide a physical explanation. Results: Significant increase of RBC sphericity was observed in COPD patients compared to volunteers (Kruskal-Wallis: p<0.0001). In vitro experiments, at shear rate of 100 s ^-1 , we observed that RBCs treated with neuraminidase mainly affect the number of platelet aggregates (p = 0.004). There was no change in the aggregates size. At a shear rate of 400 sec ^-1 neuraminidase treatment changes both the size of the aggregates (p = 0.009) and the number of platelet aggregates (p = 0.008). Numerical simulations indicated that RBCs spherization induces an increase of MSD and the effect was more pronounced when the shear rate increased. Conclusion: Our results show that the RBCs of COPD patients are more spherical than those of healthy volunteers. Experimentally we observe that the RBCs spherization induces an increase platelet transport to the wall. Additional studies are needed to better understand the possible association between the RBCs effect on the platelets transport and the increased cardiovascular events observed in COPD patients.

Cardiovascular diseases represent a major issue in terms of morbidity and mortality for dialysis patients. This morbidity is due to the accelerated atherosclerosis observed in these patients. Atherosclerosis is a chronic inflammatory disease characterized by key players such as monocytes, macrophages or oxidized LDLs. Monocytes-macrophages are classified into subsets of polarized cells, with M1 and M2 macrophages considered respectively as pro- and anti-inflammatory.The monocyte subsets and phenotypes were analyzed by flow cytometry. These data was completed by the quantification of plasma M-CSF, IL-8, CRP, Mox-LDLs, Apo-B and Apo-AI, chloro-tyrosine and homocitrulline concentrations.The statistical differences and associations between two continuous variables were assessed using the Mann-Whitney U test and Spearman’s correlation coefficient, respectively.Hemodialyzed patients showed a significant increase in the concentrations of CRP, M-CSF and IL-8 (inflammation biomarkers) as well as of chloro-tyrosine and homocitrulline (myeloperoxidase-associated oxidative stress biomarkers). Moreover we observed a higher percentage of M2 monocytes in the plasma of hemodialysis patients, as compared to the controls.Our data suggests that an oxidative stress and an inflammatory environment, amplified in hemodialysis patients, seems to favor an increase in the concentration of circulating M-CSF, therefore leading to an increase of M2 polarization among circulating monocytes.Short summary: Our data suggests that an oxidative stress and an inflammatory environment, amplified in hemodialysis patients, seems to favor an increase in the concentration of circulating M-CSF, associated with an increase of circulating M2 polarized monocytes.