Human umbilical vein endothelial cells (HUVECs Lonza) were cultured in EBM-2 medium (Lonza) at 37вЂ‰вЊ in 5% CO 2/95% air by standard methodologies in 25-cm 2 tissue culture flasks (50вЂ‰ml capacity) (Falcon, Becton Dickinson Labware) in the presence of endothelial cell growth supplement. This finding, in association with modification of bioenergetics, might reflect a clear state of EC suffering. Here, we demonstrate for the first time that PTX3 induces cytoplasmic vacuolization and dilution of mitochondrial matrix. However, although it has been demonstrated that ultrastructural alteration of membrane and organelles and modification of bioenergetic parameters represent important determinants of endothelial cell malfunction, no study has focused on the possible modulation of these parameters by PTX3 in human endothelial cells (ECs). These findings clearly suggest that the endothelium represents one of the main targets of PTX3 at the vascular level.

Moreover, circulating levels of PTX3 were found to be increased in hypertensive patients, leading us to candidate PTX3 as a novel prognostic marker for arterial hypertension. Recently, we have reported that PTX3 is directly implicated in the pathogenesis of vascular endothelial dysfunction, through a P-selectin/matrix metalloproteinase-1 pathway, demonstrating that the exposure of mouse mesenteric arteries to PTX3 leads to alterations of vascular ultrastructural and impairment of nitric oxide production. Other studies have reported that PTX3 levels are higher in women with preeclampsia, speculating on its contribution to endothelial dysfunction. On this point, an elevated level of circulating Pentraxin 3 (PTX3) вЂ“ a member of a protein superfamily involved in the innate immune response вЂ“ has been described as a marker of poor prognosis in patients with stable coronary artery disease or heart failure. Moreover, the growing body of studies linking inflammation to endothelial activation and loss of nitric oxide bioactivity has promoted investigators to seek evidence on the possible correlation of acute-phase proteins with endothelial-specific alterations. In the last decade, several inflammatory mediators have been implicated in the pathogenesis of age-related cerebro- and cardio-vascular disorders.

More studies are needed to clearly demonstrate any direct correlation between these ultrastructural and bioenergetic changes with endothelial dysfunction, especially with regard to age-related cerebro- and cardio-vascular diseases. This finding can be associated with our previous demonstration of a deleterious effect of PTX3 on the endothelial layer. PTX3 has, per se, a direct action on ultrastructural and bioenergetic parameters of isolated endothelial cells. Moreover, metabolic assays revealed that PTX3 increases respiratory capacity in support of mitochondrial function, and partially sustains the glycolytic pathway. PTX3 induced cytoplasmic vacuolization and dilution of mitochondrial matrix in isolated, human endothelial cells.

Although several mechanisms of action have been identified to date in the vascular and immune systems, the direct effects of PTX3 on isolated endothelial cells at morphological and metabolic levels remain unknown. Pentraxin 3 (PTX3), an acute-phase inflammation protein produced by several cell types, has long been described as a possible biomarker for age-related cardiovascular and cerebrovascular diseases.

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