Endothelial Progenitor Cells Counts Research Articles

Change of endothelial progenitor cells in the bone marrow and peripheral blood of patients with acute leukemia and its clinical significance

Objectives To evaluate the count of endothelial progenitor cells (EPCs) in peripheral blood (PB) and bone marrow (BM) of acute leukemia (AL) patients and explore its clinical significance. Methods EPCs were detected by flow cytometry procedures in 43 AL patients and in 10 benign hematologic patients as control group. Results The absolute counts of EPCs in AL patients before the treatment [(119.46±72.23)/μl in BM and (13.69±8.26)/μl in PB] were significantly higher than those in control group [(23.21 ±12.59)/μl in BM and (1.86±1.18)/μl in PB] (P 0.05). After the treatment, the absolute counts of EPCs both in BM and in BP of CR group [(26.32±17.44)/μl and (2.54±2.12)/μl, respectively] were significantly lower than those before treatment [(113.18±69.22)/μl and (14.45±10.76)/μl, respectively] (P 0.05). The absolute counts of EPCs whether in PB or in BM were no significant difference between acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL) (P > 0.05). The absolute counts of EPCs in PB of AL had a positive correlation with β2-MG and LDH (P < 0.05). Conclusions EPC levels are significantly increased in BM and BP of AL patients and may correlate with disease status , response to treatment and prognosis. Key words: Acute leukemia; Endothelial progenitor cells; Flow cytometry

Rosuvastatin for enhancement of aneurysm neck endothelialization after coil embolization: promotion of endothelial progenitor cells in a rodent model.

OBJECT Coil embolization is a safe, efficient, and minimally invasive technique for the treatment of intracranial aneurysms. However, coil embolization is associated with a higher risk of recurrence than clip ligation. In this study, the authors explore a new approach through the promotion of endothelial progenitor cells (EPCs) to optimize endothelialization of the aneurysm neck and reduce the risk of recurrence. METHODS A coiled aneurysm model was created in 48 adult male Sprague-Dawley rats via microsurgery. Half of these animals were treated with rosuvastatin (20 mg/kg) in saline via gavage for 10, 20, or 30 days. The other half were administered saline without rosuvastatin. An additional 15 rats underwent "mock surgery" (identical anesthesia and saline gavage but no surgery). The endothelial repair process in the coiled aneurysms was evaluated via flow cytometry, im-munostaining, and electronic microscopy. The mock surgery group was used for comparison in flow cytometry studies. The effects of rosuvastatin on viability and functioning of Sprague-Dawley rat bone marrow-derived EPCs were also explored via MTT, migration, and tube formation assays. RESULTS The aneurysm neck repair score was significantly higher in the rosuvastatin-treated rats than in the untreated rats (p < 0.05). The circulating EPC count was increased and maintained at a higher level in rosuvastatin-treated rats compared with the aneurysm rats that did not receive rosuvastatin (p < 0.05). Immunostaining showed that the aneurysm neck endothelium was more integrated and the number of kinase insert domain receptor-positive cells was increased in the rosuvastatin-treated rats. Further study demonstrated that rosuvastatin promoted EPC proliferation, migration, and tube formation. CONCLUSIONS Rosuvastatin promoted endothelialization of the coiled aneurysm neck via induction of EPCs, suggesting that promoting endothelialization provides an additional therapeutic opportunity during vascular endothelium repair.

Endothelial Progenitor Cells Predict Cardiovascular Events after Atherothrombotic Stroke and Acute Myocardial Infarction. A PROCELL Substudy.

IntroductionThe aim of this study was to determine prognostic factors for the risk of new vascular events during the first 6 months after acute myocardial infarction (AMI) or atherothrombotic stroke (AS). We were interested in the prognostic role of endothelial progenitor cells (EPC) and circulating endothelial cells (CEC)MethodsBetween February 2009 and July 2012, 100 AMI and 50 AS patients were consecutively studied in three Spanish centres. Patients with previously documented coronary artery disease or ischemic strokes were excluded. Samples were collected within 24h of onset of symptoms. EPC and CEC were studied using flow cytometry and categorized by quartiles. Patients were followed for up to 6 months. NVE was defined as new acute coronary syndrome, transient ischemic attack (TIA), stroke, or any hospitalization or death from cardiovascular causes. The variables included in the analysis included: vascular risk factors, carotid intima-media thickness (IMT), atherosclerotic burden and basal EPC and CEC count. Multivariate survival analysis was performed using Cox regression analysis.ResultsDuring follow-up, 19 patients (12.66%) had a new vascular event (5 strokes; 3 TIAs; 4 AMI; 6 hospitalizations; 1 death). Vascular events were associated with age (P = 0.039), carotid IMT≥0.9 (P = 0.044), and EPC count (P = 0.041) in the univariate analysis. Multivariate Cox regression analysis showed an independent association with EPC in the lowest quartile (HR: 10.33, 95%CI (1.22–87.34), P = 0.032] and IMT≥0.9 [HR: 4.12, 95%CI (1.21–13.95), P = 0.023].ConclusionsBasal EPC and IMT≥0.9 can predict future vascular events in patients with AMI and AS, but CEC count does not affect cardiovascular risk.

Circulating endothelial progenitor cells in obese children and adolescents

ObjectiveThis study aimed to investigate the relationship between circulating endothelial progenitor cell count and endothelial activation in a pediatric population with obesity. MethodsObservational and transversal study, including 120 children and adolescents with primary obesity of both sexes, aged 6–17 years, who were recruited at this Cardiovascular Risk Clinic. The control group was made up of 41 children and adolescents with normal body mass index. The variables analyzed were: age, gender, body mass index, systolic and diastolic blood pressure, high-sensitivity C-reactive protein, lipid profile, leptin, adiponectin, homeostasis model assessment-insulin resistance, monocyte chemoattractant protein-1, E-selectin, asymmetric dimethylarginine and circulating progenitor endothelial cell count. ResultsInsulin resistance was correlated to asymmetric dimethylarginine (ρ=0.340; p=0.003), which was directly, but weakly correlated to E-selectin (ρ=0.252; p=0.046). High sensitivity C-reactive protein was not found to be correlated to markers of endothelial activation. Systolic blood pressure was directly correlated to body mass index (ρ=0.471; p<0.001) and the homeostasis model assessment-insulin resistance (ρ=0.230; p=0.012), and inversely correlated to adiponectin (ρ=−0.331; p<0.001) and high-density lipoprotein cholesterol (ρ=−0.319; p<0.001). Circulating endothelial progenitor cell count was directly, but weakly correlated, to body mass index (r=0.211; p=0.016), leptin (ρ=0.245; p=0.006), triglyceride levels (r=0.241; p=0.031), and E-selectin (ρ=0.297; p=0.004). ConclusionCirculating endothelial progenitor cell count is elevated in obese children and adolescents with evidence of endothelial activation, suggesting that, during infancy, endothelial repairing mechanisms are present in the context of endothelial activation.

Effect of onion peel extract on endothelial function and endothelial progenitor cells in overweight and obese individuals

ObjectivesAcute or chronic intake of polyphenol-rich foods has been reported to improve endothelial function. Quercetin, found abundantly in onion, is a potent antioxidant flavonoid. The aim of this study was to investigate whether consumption of onion peel extract (OPE) improves endothelial function in healthy overweight and obese individuals. MethodsThis was a randomized double-blind, placebo-controlled study. Seventy-two healthy overweight and obese participants were randomly assigned to receive a red, soft capsule of OPE (100 mg quercetin/d, 50 mg quercetin twice daily; n = 36 participants) or an identical placebo capsule (n = 36) for 12 wk. Endothelial function, defined by flow-mediated dilation (FMD), circulating endothelial progenitor cells (EPCs) by flow cytometry, and laboratory test were determined at baseline and after treatment. ResultsBaseline characteristics and laboratory findings did not significantly differ between the two groups. Compared with baseline values, the OPE group showed significantly improved FMD at 12 wk (from 12.5 ± 5.2 to 15.2 ± 6.1; P = 0.002), whereas the placebo group showed no difference. Nitroglycerin-mediated dilation did not change in either group. EPC counts (44.2 ± 25.6 versus 52.3 ± 18.6; P = 0.005) and the percentage of EPCs were significantly increased in the OPE group. When FMD was divided into quartiles, rate of patients with endothelial dysfunction defined as lowest quartile (cutoff value, 8.6%) of FMD improved from 26% to 9% by OPE. ConclusionMedium-term administration of OPE an improvement in FMD and circulating EPCs.

Oral uracil and tegafur as postoperative adjuvant metronomic chemotherapy in patients with advanced oral squamous cell carcinoma

Background/purposeIn this study, we evaluated the effectiveness of oral uracil–tegafur (UFUR) as a postoperative adjuvant treatment for patients with advanced oral squamous cell carcinoma (OSCC). Materials and methodsThe study cohort consisted 80 patients with advance cancer treated between January 2003 and December 2007. Half of the patients received oral UFUR as postoperative metronomic adjuvant chemotherapy, while the other half received no treatment. No patients received postoperative radiotherapy or other systemic chemotherapy. Disease-free survival and toxicity were evaluated in these two groups. Twenty-two patients were assessed pre- and postoperatively for viable circulating endothelial progenitor cells using flow cytometry. ResultsThe disease-free survival rates at 4 years were 84.6% with oral UFUR treatment and 60.9% without UFUR therapy (P = 0.02). The toxicity and disease progression profiles did not differ significantly between the two groups, but viable circulating endothelial progenitor cell counts decreased after the administration of oral UFUR. Advanced OSCC patients who received oral UFUR had a better prognosis than those who did not. ConclusionOral UFUR is a promising postoperative metronomic adjuvant chemotherapy regimen for advanced OSCC.

Exercise-induced mobilisation of endothelial progenitor cells in patients with premature coronary heart disease.

Endothelial progenitor cells (EPC) derive from bone marrow and participate in both endothelial regeneration and development of new blood vessels. EPC also play a role in the atherosclerotic process, and their number correlates negatively with the presence of classical risk factors. To evaluate circulating EPC count and their exercise-induced mobilisation in patients with premature coronary artery disease (CAD). The study group included 60 patients with stable CAD diagnosed before 45 years of age. The control group consisted of 33 healthy age- and gender-matched volunteers. Venous blood was sampled 3 times in order to assess circulating EPC count immediately before an exercise test (EPC 0) and at 15 min (EPC 15) and 60 min (EPC 60) after the exercise test. Circulating EPC count in the study group at rest and at 15 min after exercise was comparable (2.1 vs. 2.1 cell/μL, p = 0.35) and increased significantly at 60 min after exercise in comparison to resting values (2.1 vs. 3.2 cell/μL, p < 0.00001). In the control group, circulating EPC count increased significantly at 15 min after exercise (2.0 vs. 3.5 cell/μL, p < 0.0001) but later decreased at 60 min after exercise, although it remained greater than at rest (2.7 vs. 2.0 cell/μL, p < 0.0002). Circulating EPC count at rest and at 60 min after exercise was comparable in the two groups (2.1 vs. 2.0 cell/μL, p = 0.96; and 3.2 vs. 2.7 cell/μL, p = 0.13, respectively) but it was significantly lower in the study group compared to the control group at 15 min after exercise (2.1 vs. 3.5 cell/μL, p < 0.00001). Circulating EPC count at rest and at 15 min after exercise did not correlate with the number of stenosed coronary arteries but at 60 min after exercise it was greater in patients with one-vessel disease compared to those with two- or three-vessel disease (4.2 vs. 3.4 cell/μL, p = 0.01; and 4.2 vs. 2.3 cell/μL, p = 0.00003). However, no difference in circulating EPC count was seen at 60 min after exercise between patients with two- or three-vessel disease (3.4 vs. 2.3 cell/μL, p = 0.3). 1. Circulating EPC count at rest is comparable between subjects with premature atherosclerosis and healthy volunteers. 2. A single bout of physical exercise causes a significant increase in circulating EPC count in both groups, but the dynamics of exercise-induced EPC mobilisation is different, with delayed exercise-induced EPC mobilisation in subjects with premature CAD. 3. The extent of atherosclerotic coronary lesions does not influence circulating EPC count at rest.

Circulating Endothelial Progenitor Cell Levels Predict Cardiovascular Events in End-Stage Renal Disease Patients on Maintenance Hemodialysis

Background: The number of circulating endothelial progenitor cells (EPCs) has been identified as a surrogate biologic marker for vascular function and cumulative cardiovascular (CV) risk in the general population. Patients with end-stage renal disease (ESRD) on hemodialysis (HD) have markedly decreased EPC counts and function. We hypothesized that the number of circulating EPCs predicts death from all causes and CV events in patients with ESRD on HD. Methods: We quantified the EPCs in blood samples from 70 patients with ESRD on HD. Circulating EPCs were counted by flow cytometry as the number of CD45^lowCD34⁺VEGFR2⁺ cells. Death from all causes and CV events served as outcome variables over a median follow-up period of 20 months. Results: It has been postulated that the number of circulating EPCs at baseline ranged from 1 to 350 cells/200 μl, with a mean of ± standard deviation (SD) of 26.0 ± 48.2 cells/200 μl. The median, lowest and highest tertiles of EPC counts were 11.0, 9.0, and 17.0 cells/200 μl, respectively. Patients with the lowest tertile EPC counts had significantly higher rates of CV events, but mortality was similar between the two groups. After adjusting for these risk factors, HbA1c and the lowest tertile EPC count remained as independent predictors of CV events. A cutoff value of 9.5 cells/200 μl maximized the power of the EPC count to predict future CV events as determined by ROC curve analysis. Conclusions: Reduced circulating EPC counts independently predicted CV events in 70 patients with ESRD on maintenance HD. Circulating EPCs may play a role in vascular repair, thereby affecting the clinical course of CV events.

Association between mobilization of circulating endothelial progenitor cells and time or degree of injury from angioplasty in patients with exertional angina: A prospective study.

The aim of the present study was to investigate the effect of coronary artery angioplasty on the recruitment of circulating endothelial progenitor cells (EPCs) in patients with angina pectoris. A total of 66 patients treated by coronary stenting were enrolled in the PCI group and 17 patients that underwent angiography alone were enrolled in the control group. The EPC count in the blood was measured by flow cytometry prior to and at 1, 3, 5, 7 and 24 h following angioplasty in the percutaneous coronary intervention (PCI) group, and at three time-points following angiography in the control group. Differences between the two groups included the characteristics of the coronary artery lesions, the incidence of diabetes and family history of coronary heart disease. The mean surface area of the stent deployed was 335.59±234.99 mm2. No significant change in EPC count was measured in the control group. In the PCI group, a moderate and delayed increase in the number of cluster of differentiation (CD)34+/kinase domain receptor (KDR)+ EPCs occurred at 24 h post-balloon inflation compared with the baseline level. The CD133-/CD34+/KDR+ subpopulations showed undulating changes at 3, 7 and 24 h post-PCI (P=0.016, P=0.01 and P=0.032, respectively). An arch shape was displayed in CD133+/KDR+ cells; initially, a reduction occurred at 3 h and was maintained constantly until 7 h (P=0.003, P=0.013 and P=0.033 at 3, 5 and 7 h, respectively), after which a slight increase to the baseline level occurred at 24 h (P=0.084). The CD133+/CD34+ cells increased in stepwise manner until 24 h. The CD34+/KDR+ EPC change magnitude correlated significantly with a global damage index by partial correlation analysis (P<0.001). The results suggested that a time-dependent mobilization of EPCs may be initiated by PCI; the change magnitude of the CD34+/KDR+ cells was associated particularly with endothelial injury degree from the PCI procedure.

Assessment of left ventricular function in systemic lupus erythematosus patients by speckle tracking echocardiography: Relation to circulating endothelial progenitor cells

BackgroundSystemic lupus erythematosus is an autoimmune disease associated with reduced number and impaired function of endothelial progenitor cells (EPCs) responsible for vascular regeneration. Aim of the workto assess left ventricular (LV) function of SLE patients using relatively new speckle tracking echocardiography (STE) and examine the relation of any detected abnormalities with peripheral circulating EPC level. Patients and methodsFifty SLE patients and 25 healthy controls were subjected to quantification of peripheral circulating Cluster of differentiation133+/vascular endothelial growth factor receptor2+(CD133+/VEGFR2+) and CD34+/VEGFR2+ EPCs, transthoracic echocardiography (TTE), tissue Doppler imaging (TDI) and STE. ResultsPatients showed a significantly lower CD133+/VEGFR2+ EPCs (p=0.009) and CD34+/VEGFR2+ EPC counts (p=0.0001) compared to controls. TTE/TDI revealed a significantly lower LV ejection fraction (EF) (p=0.007), higher LV end systolic dimensions (p=0.02), myocardial performance index (MPI) (p=0.0001) and mitral flow E/lateral annulus E′ ratio (p=0.002) in patients compared to controls. STE showed a significantly lower global longitudinal strain (GLS) (p<0.001), global circumferential strain (GCS) (p<0.001) and global strain rate during isovolumic relaxation period (GSRivr) (p=0.01) in patients compared to controls. By multiple logistic regression analysis, the independent variables affecting GCS and GSRivr were the prednisolone dose and the LVEF respectively. (95% CI=−0.46 to −0.03; p=0.03 and 95% CI=0.001–0.01; p=0.021; respectively). There was no significant correlation of the GLS, GCS or GSRivr with the EPCs. ConclusionSTE detected subclinical systolic and diastolic abnormalities of LV function in SLE patients. These abnormalities of LV function did not show however any relation with the significantly lower EPC count detected in patients.

Long-term, regular remote ischemic preconditioning improves endothelial function in patients with coronary heart disease.

Remote ischemic preconditioning (RIPre) can prevent myocardial injury. The purpose of this study was to assess the beneficial effects of long-term regular RIPre on human arteries. Forty patients scheduled for coronary artery bypass graft (CABG) surgery were assigned randomly to a RIPre group (n=20) or coronary heart disease (CHD) group (n=20). Twenty patients scheduled for mastectomy were enrolled as a control group. RIPre was achieved by occluding arterial blood flow 5 min with a mercury sphygmomanometer followed by a 5-min reperfusion period, and this was repeated 4 times. The RIPre procedure was repeated 3 times a day for 20 days. In all patients, arterial fragments discarded during surgery were collected to evaluate endothelial function by flow-mediated dilation (FMD), CD34+ monocyte count, and endothelial nitric oxide synthase (eNOS expression). Phosphorylation levels of STAT-3 and Akt were also assayed to explore the underlying mechanisms. Compared with the CHD group, long-term regular RIPre significantly improved FMD after 20 days (8.5±2.4 vs 4.9±4.2%, P<0.05) and significantly reduced troponin after CABG surgery (0.72±0.31 and 1.64±0.19, P<0.05). RIPre activated STAT-3 and increased CD34+ endothelial progenitor cell counts found in arteries. Long-term, regular RIPre improved endothelial function in patients with CHD, possibly due to STAT-3 activation, and this may have led to an increase in endothelial progenitor cells.

Maintenance chemotherapy in children with ALL exerts metronomic-like thrombospondin-1 associated anti-endothelial effect.

Maintenance chemotherapy is an important part of the treatment of ALL in children. It relies on the long-term oral administration of daily low-dose mercaptopurin and weekly low-dose methotrexate. Although it has been used in the clinic for decades, its mechanisms of action remain unclear. Here, we investigated different angiogenic and immune biomarkers to gain insights into the mechanisms of action of maintenance therapy in children with ALL. We thus monitored circulating endothelial cells (CEC), endothelial progenitor cells (EPC) and endothelial microparticles (EMP), pro-angiogenic factors (VEGF, VEGFR-1 and Ang-2), anti-angiogenic factor thrombospondin-1 (THBS1) and regulatory T lymphocytes (Treg) in 47 children with ALL during the maintenance phase of their treatment (at treatment initiation and after 6, 12 and 18 months). We observed a statistically significant decrease in EPC and EMP counts throughout the maintenance phase associated with a significant increase in THBS1 levels. No significant change was detected in other angiogenic markers or in Treg numbers.The results presented here indicate that maintenance therapy in children with ALL exerts its antitumor activity at least in part through anti-angiogenic effects, similar to those induced by metronomic chemotherapy. Larger studies are now warranted to validate these findings and determine their clinical implications.

Poor Outcome of Experimental Ischemic Stroke in Type 2 Diabetic Rats: Impaired Circulating Endothelial Progenitor Cells Mobilization

It is well accepted that type 2 diabetic mellitus (T2DM) results in the poor outcome of ischemic stroke. However, the mechanisms by which T2DM causes aggravated cerebral ischemic/reperfusion (I/R) injury are not clear. Recently, endothelial progenitor cells (EPCs) are considered to be related with the outcome of ischemic stroke. More importantly, T2DM can affect the function of circulating EPCs. This study tried to investigate whether T2DM worsens the cerebral I/R injury via affecting circulating EPCs. We used high-fat diet-fed and low-dose streptozotocin-treated male rats receiving middle cerebral artery occlusion surgery as animal model of focal cerebral I/R injury with T2DM (diabetic operated). And the rats were divided into 4 groups: normal sham, diabetic sham, normal operated, and diabetic operated. We measured the circulating EPCs counts and the levels of vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS) in peripheral plasma of 4 groups. We found that diabetic rats subjected to I/R exhibited significantly severe deterioration in neurologic deficits compared with nondiabetic counterparts, which manifested higher infarct volume and cell apoptosis as well as lower neurologic defective score. There was no significant difference on the plasma glucose of groups before cerebral I/R injury compared with that of the groups posterior to cerebral I/R injury despite cerebral I/R injury had the tendency to increase the plasma glucose no matter in the presence or the absence of T2DM. In addition, there were the marked downregulation of circulating EPCs counts and the levels of VEGF and eNOS in diabetic rats before the cerebral I/R injury. Despite I/R injury without T2DM, there was a significant increase in the circulating EPCs counts, the circulating EPCs counts in I/R injury with T2DM group were significantly decreased compared with those in the other 3 groups. We also observed that the level of eNOS was significantly improved by I/R injury without considering the presence of T2DM. Thus, our present study suggested that it might be the impaired EPCs mobilization into theblood that contributed to the worse outcome of cerebral I/R injury with T2DM.

Red cell distribution width is associated with endothelial progenitor cell depletion and vascular-related mediators in rheumatoid arthritis

ObjectivesThe role of Red Cell Distribution Width (RDW) as a predictor of cardiovascular (CV) events has been proposed in a variety of conditions, including Rheumatoid Arthritis (RA). However, the mechanisms underlying this effect are still unknown. Since inflammation and Endothelial Progenitor Cells (EPCs) imbalance have been reported in RA patients to be related to CV disease, we wondered whether RDW could be linked to endothelial repair failure in RA. MethodsEPCs (CD34+VEGFR2+CD133+) were quantified by flow cytometry in peripheral blood samples from 194 RA patients. IFNα, TNFα, IFNγ, IL-8, VEGF, GM-CSF, MCP-1, ICAM-1, EGF, Leptin and Resistin serum levels were quantified by immunoassays. Clinical and immunological parameters as well as history of traditional CV risk factors and CV events were registered from medical records. RDW was measured in complete blood cell count analyses. ResultsRDW was negatively related to EPC counts in patients with established disease (>1 year, n = 125) (r = −0.306, p < 0.001). Moreover, RDW was independently associated to an EPC depletion in the whole group (β [95% CI]: −3.537 [−6.162, −0.911], p = 0.009) after adjusting for clinical parameters, disease duration, treatments and traditional CV risk factors. Additionally, RDW was positively correlated with IFNα serum levels, a cytokine related to endothelial damage, and with IL-8, VEGF and neutrophil to lymphocyte ratio, thus supporting the association with inflammation and vascular remodelling. ConclusionsRDW was associated to EPC depletion and increased levels of different mediators linked to endothelial damage and vascular repair failure, thereby shedding new light on the nature of RDW as CV-predictor.

Myeloproliferative neoplasms and endothelial progenitor cells

JAK2V617F is a novel oncogene involved in myeloproliferative neoplasms (MPN) and closely related to its vascular complications. Recent researches found the vascular endothelial injury in patients with MPN. It is increasingly being recognized that endothelial cells and endothelial progenitor cells (EPC) play an important role in MPN. Here, the pathogenesis of the EPC JAK2V617F mutation and peripheral blood EPC counts in patients with MPN or with vascular complications are reviewed.

Elevated levels of pentraxin 3 in systemic sclerosis: associations with vascular manifestations and defective vasculogenesis.

To clarify the role of pentraxin 3 (PTX3), a multifunctional pattern recognition protein that can suppress fibroblast growth factor 2 (FGF-2), in systemic sclerosis (SSc)-related vasculopathy. We assessed 171 SSc patients and 19 age- and sex-matched healthy control subjects. Circulating PTX3 and FGF-2 levels were measured by enzyme immunoassay, and CD34+CD133+CD309+ endothelial progenitor cells (EPCs) were counted by flow cytometry. Correlations between PTX3 and FGF-2 and the presence or future development of vascular manifestations, including digital ulcers and pulmonary arterial hypertension (PAH), were identified by univariate and multivariate analysis. The effect of PTX3 on EPC differentiation was evaluated in proangiogenic cultures of mouse bone marrow cells in combination with colony formation assay. Circulating PTX3 and FGF-2 levels were significantly higher in SSc patients than in healthy control subjects. PTX3 was elevated in SSc patients who had digital ulcers or PAH, while FGF-2 was reduced in SSc patients with PAH. Multivariate analysis identified elevated PTX3 as an independent parameter associated with the presence of digital ulcers and PAH, and PTX3 levels were a useful predictor of future occurrences of digital ulcers. Reduced FGF-2 was independently associated with the presence of PAH. EPC counts were significantly lower in patients with digital ulcers or PAH and correlated negatively with circulating PTX3 concentrations. Finally, PTX3 inhibited EPC differentiation in vitro. In SSc patients, exposure to high concentrations of PTX3 may suppress EPC-mediated vasculogenesis and promote vascular manifestations such as digital ulcers and PAH.

Mobilization of endothelial progenitor cells in acute cardiovascular events in the PROCELL study: Time-course after acute myocardial infarction and stroke

The mobilization pattern and functionality of endothelial progenitor cells after an acute ischemic event remain largely unknown. The aim of our study was to characterize and compare the short- and long-term mobilization of endothelial progenitor cells and circulating endothelial cells after acute myocardial infarction or atherothrombotic stroke, and to determine the relationship between these cell counts and plasma concentrations of vascular cell adhesion molecule (VCAM-1) and Von Willebrand factor (VWF) as surrogate markers of endothelial damage and inflammation. In addition, we assessed whether endothelial progenitor cells behave like functional endothelial cells. We included 150 patients with acute myocardial infarction or atherothrombotic stroke and 145 controls. Endothelial progenitor cells [CD45−, CD34+, KDR+, CD133+], circulating endothelial cells [CD45−, CD146+, CD31+], VWF, and VCAM-1 levels were measured in controls (baseline only) and in patients within 24h (baseline) and at 7, 30, and 180days after the event. Myocardial infarction patients had higher counts of endothelial progenitor cells and circulating endothelial cells than the controls (201.0/mL vs. 57.0/mL; p<0.01 and 181.0/mL vs. 62.0/mL; p<0.01). Endothelial progenitor cells peaked at 30days post-infarction (201.0/mL vs. 369.5/mL; p<0.01), as did VCAM-1 (573.7ng/mL vs. 701.8ng/mL; p<0.01). At 180days post-infarction, circulating endothelial cells and VWF decreased, compared to baseline. In stroke patients, the number of endothelial progenitor cells — but not circulating endothelial cells — was higher than in controls (90.0/mL vs. 37.0/mL; p=0.01; 105.0/mL vs. 71.0/mL; p=0.11). At 30days after stroke, however, VCAM-1 peaked (628.1/mL vs. 869.1/mL; p<0.01) but there was no significant change in endothelial progenitor cells (90/mL vs. 78/mL; p<0.34). At 180days after stroke, circulating endothelial cells and VWF decreased, compared to baseline. Cultured endothelial progenitor cells from controls and myocardial infarction patients had endothelial phenotype characteristics and exhibited functional differences in adhesion and Ca2+ influx, but not in proliferation and vasculogenesis. In myocardial infarction patients, VCAM-1 levels and mobilization of endothelial progenitor cells peaked at 30days after the ischemic event. Although a similar VCAM-1 kinetic was observed in stroke patients, endothelial progenitor cells did not increase. Endothelial progenitor cells had mature endothelial capabilities in vitro.

Tumor necrosis factor-alpha antagonism with etanercept improves endothelial progenitor cell counts in patients with psoriasis: Etanercept, vascular function and endothelial progenitor cells in psoriasis

a Division of Cardiology, Emory University School of Medicine, Atlanta, GA, United States b Division of Pulmonary Medicine, Emory University School of Medicine, Atlanta, GA, United States c Department of Biostatistics and Bioinformatics, Rollins School of Public Health, Emory University, Atlanta, GA, United States d Department of Dermatology, Atlanta VA Medical Center and Emory University School of Medicine, Atlanta, GA, United States e Department of Hematology and Oncology, Winship Cancer Institute, Emory University, Atlanta, GA, United States

Abstract 19820: Heterogeneity of Disease Progression Rates in Patients With Bicuspid Aortic Valve: Is There a Suitable Bio-marker?

Introduction: Bicuspid aortic valve (BAV) is associated with inflammatory activation and endothelial dysfunction. Worsening of aortic valve stenosis/regurgitation represents the most common complication of BAV, but clinical and biochemical markers of disease progression are not currently available. Objective: The objective of the current study was to (1) evaluate rates of progression of aortopathy and valve dysfunction, and (2) identify correlates of accelerated progression among BAV patients. Methods: 43 BAV patients aged 45 ± 16 (SD) years were evaluated clinically and with echo/MRI. Inflammatory activation was assessed via hs-CRP and myeloperoxidase (MPO) levels, endothelial function/NO responsiveness via flow-mediated dilatation (FMD), plasma asymmetric dimethylarginine (ADMA) levels, inhibition of platelet aggregation by SNP, and endothelial progenitor cell counts. Follow-up was undertaken after 47 ±16 months to determine rate of progression and of need for valve replacement; some patients did not undergo repeat echo (Figure). Biochemical/physiological correlates of valve dysfunction and aortic dimensions were sought via univariate and multiple linear regression analyses. Results: AV max increased from 2.1 ± 0.6 to 2.5 ± 1.1(m/s), p &lt; 0.05, while ascending aorta (Asc Ao) dimensions increased from 18.2 ± 6.1 to 21.2 ± 4.9 (mm/m 2 ), p &lt; 0.05 over the monitoring period. Although there was a direct correlation (p &lt; 0.01) between baseline AV max and MPO levels on multivariate analysis, this correlation did not extend to progression rates. Indeed neither clinical nor biochemical parameters predicted accelerated progression. Conclusions: (1) Progression of valve disease and aortopathy in BAV is often rapid, with annual mean rate of 0.1 m/s for AV max and 0.4 mm/m 2 for Asc Ao. (2) Despite physiological evidence linking BAV with inflammation and endothelial dysfunction, these biochemical factors did not appear to predict rapid disease progression.

Markers of subclinical atherosclerosis in premenopausal women with vitamin D deficiency and effect of vitamin D replacement

Background: Recent studies have revealed a relationship between vitamin D deficiency and atherosclerosis. This study aims to investigate the impact of vitamin D deficiency and replacement on markers of subclinical atherosclerosis in young premenopausal women in whom vitamin D deficiency is prevalent. Methods: Thirty-one premenopausal vitamin D deficient women and 27 age and gender-matched control subjects were enrolled in this study. Markers of subclinical atherosclerosis including carotid intima-media thickness (cIMT), flow-mediated dilatation (FMD), endothelial progenitor cell (EPC) count and cytokine levels were determined at baseline. All measurements were repeated at 6-month follow-up in vitamin D-deficient subjects after vitamin D replacement. Results: Vitamin D deficient premenopausal women had lower FMD (9.9 ± 1.3 vs. 13.8 ± 1.7%, p < 0.001) and EPC counts at baseline. This population also had lower IL-10 and higher IL-17 levels. A 6-month vitamin D replacement therapy resulted in a significant increase in FMD (9.9 ± 1.3 vs. 11.4 ± 1.4%, p < 0.001) and EPC counts. Furthermore, cytokine profile shifted toward a more anti-inflammatory phenotype including elevated IL-10 and decreased IL-17 levels. cIMT was not different between patient and control groups and did not change following vitamin D replacement. Change in 25(OH)D and IL-17 levels were independent predictors of the change in FMD measurements following vitamin D replacement. Conclusion: This study demonstrates that endothelial function is impaired in otherwise healthy vitamin D deficient young premenopausal women and improves with 6-month replacement therapy. Immune-modulatory effects of vitamin D may, at least partly, be responsible for its beneficial effects on vascular health.