Increase In Endothelial Progenitor Cells Research Articles

The bone marrow endothelial progenitor cell response to septic infection.

Early increase in the level of endothelial progenitor cells (EPCs) in the systemic circulation occurs in patients with septic infection/sepsis. The significance and underlying mechanisms of this response remain unclear. This study investigated the bone marrow EPC response in adult mice with septic infection induced by intravenous injection (i.v.) of Escherichia coli. For in vitro experiments, sorted marrow stem/progenitor cells (SPCs) including lineage(lin)-stem cell factor receptor (c-kit)+stem cell antigen-1 (Sca-1)-, lin-c-kit+, and lin- cells were cultured with or without lipopolysaccharides (LPSs) and recombinant murine vascular endothelial growth factor (VEGF) in the absence and presence of anti-Sca-1 crosslinking antibodies. In a separate set of experiments, marrow lin-c-kit+ cells from green fluorescence protein (GFP)+ mice, i.v. challenged with heat-inactivated E. coli or saline for 24h, were subcutaneously implanted in Matrigel plugs for 5 weeks. Marrow lin-c-kit+ cells from Sca-1 knockout (KO) mice challenged with heat-inactivated E. coli for 24h were cultured in the Matrigel medium for 8 weeks. The marrow pool of EPCs bearing the lin-c-kit+Sca-1+VEGF receptor 2 (VEGFR2)+ (LKS VEGFR2+) and LKS CD133+VEGFR2+ surface markers expanded rapidly following septic infection, which was supported by both proliferative activation and phenotypic conversion of marrow stem/progenitor cells. Increase in marrow EPCs and their reprogramming for enhancing angiogenic activity correlated with cell-marked upregulation of Sca-1 expression. Sca-1 was coupled with Ras-related C3 botulinum toxin substrate 2 (Rac2) in signaling the marrow EPC response. Septic infection caused a substantial increase in plasma levels of IFN-γ, VEGF, G-CSF, and SDF-1. The early increase in circulating EPCs was accompanied by their active homing and incorporation into pulmonary microvasculature. These results demonstrate that the marrow EPC response is a critical component of the host defense system. Sca-1 signaling plays a pivotal role in the regulation of EPC response in mice with septic infection.

Корекція функції ендотелію - нова ціль профілактики і лікування артеріальної гіпертензії в пацієнтів із серцево-судинною патологією

Purpose - to determine the dynamics of endothelial progenitor cells (EPCs) and asymmetric dimethylarginine (ADMA) in pregnant women with hypertension as markers of the effectiveness of endothelial dysfunction correction on the background of L-arginine therapy (Tivortin, manufactured by Yuria-Pharm). Materials and methods. The study pregnant women were divided into two groups: women with hypertension at 12-15 weeks of gestation (n=29) and women with congenital heart disease at 12-14 weeks of gestation (n=21). The control group consisted of 40 healthy pregnant women. The examination was conducted before the start of oral L-arginine (Tivortin aspartate) and 7-8 weeks after the start of therapy. Tivortin aspartate was administered orally 1 measuring spoon 5 times a day for 6 weeks. The study of the level of EPCs (CD45+/CD34+ phenotype of peripheral blood) was performed by flow cytometry using reagents for the determination of CD34, CD45 differentiation clusters produced by Beckman Coulter Inc. The results are presented in % of ERC from the total number of leukocytes, as well as in absolute values - the number of cells per 1 ml of blood in absolute values. The level of ADMA in plasma was determined by an indirect enzyme-linked immunosorbent assay using the ADMA ELISA test system (manufactured by Immunodiagnostik AG, Germany). The reference values for pregnant women are 0.26-0.60 μmol/l. Results. After treatment with L-arginine (Tivortin), the ADMA index decreased compared to baseline data in the studied groups of pregnant women. Thus, in pregnant women with hypertension, ADMA decreased by almost 15%, which is a marker of improved endothelial function. The same dynamics was observed in the group of women with congenital heart disease. In both study groups of pregnant women, the number of EPCs increased by almost 10% after the addition of L-arginine (Tivortin) to the treatment complex. There was a positive correlation between a decrease in mean systolic blood pressure, an increase in EPCs and a decrease in ADMA after complex treatment. Studies have shown a direct correlation (r=0.75) between blood pressure and the number of ЕРСs and an inverse correlation (r=-0.68) between blood pressure and ADMA. The reduction in blood pressure during treatment was accompanied by an improvement in endothelial function, namely, an increase in the number of ЕРСs, mainly due to the vasodilation potential of the endothelium. There was also a decrease in ADMA against the background of a decrease in systolic blood pressure during treatment. Conclusions. The clinical relevance of asymmetric dimethylarginine as a specific marker in patients with hypertension has been established: a significant increase in its level is observed in patients with cardiovascular disease, in particular, with hypertension, which confirms the role of endothelial dysfunction as a leading pathogenetic factor in the development of hypertension in the absence of anatomical and pathological changes in the vessels. The concept of maintenance therapy in patients with cardiovascular disease aims to restore adequate NO bioavailability by administering exogenous L-arginine. Based on the results of the study of the effect of L-arginine (Tivortin) on the index of ЕРСs, which are actively involved in the repair of damaged endothelium, it can be concluded that this drug has a positive effect on the prognosis of life in patients with hypertension, restoring endothelial function. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of all participating institutions. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

Reduced Endothelial Progenitor Cells: A Possible Biomarker for Idiopathic Fetal Growth Restriction in Human Pregnancies.

Circulating endothelial progenitor cells (EPCs) may be necessary throughout pregnancy by ensuring proper placentation and embryonic growth. The lack of standardized EPC quantification techniques has prevented conclusive proof of an increase in EPC during pregnancy. The purpose of this study was to determine whether EPC levels change for healthy and idiopathic fetal growth restriction (FGR) pregnancies. The study population consisted of 48 healthy pregnant females with no previous history of IUGR (10 in the first trimester, 15 in the second, and 23 in the third), 48 women with pregnancy complicated by idiopathic FGR, and 15 non-pregnant women. By using flow cytometry, EPCs in maternal blood were recognized as CD45dim/CD34/KDR cells. ELISA was used to measure plasmatic cytokines. We ascertained a progressive rise in EPCs in healthy pregnancies that was apparent in the first but more pronounced in the third trimester. At comparable gestational ages, FGR-complicated pregnancies had impaired EPC growth. Placental growth factor and stromal-derived factor-1 levels in the blood were significantly lower in FGR than in healthy pregnancies, which may have contributed to the degradation of the EPCs. The count in EPCs might hold considerable promise toward developing a peculiar authentication marker for observing pregnancies, and could be the focus of cutting-edge tactics for the prognosis and treatment of FGR pregnancies.

Glycaemic Control Achieves Sustained Increases of Circulating Endothelial Progenitor Cells in Patients Hospitalized for Decompensated Diabetes: An Observational Study.

Background and AimDiabetes reduces the levels of circulating endothelial progenitor cells (EPCs), which contribute to vascular homeostasis. In turn, low EPCs levels predict progression of chronic complications. Several studies have shown that hyperglycaemia exerts detrimental effects on EPCs. Improvement in glucose control with glucose-lowering medications is associated with an increase of EPCs, but only after a long time of good glycaemic control. In the present study, we examined the effect of a rapid glycaemic amelioration on EPC levels in subjects hospitalized for decompensated diabetes.MethodsWe used flow cytometry to quantify EPCs (CD34+/CD133+KDR+) in patients hospitalized for/with decompensated diabetes at admission, at discharge, and 2 months after the discharge. During hospitalization, all patients received intensive insulin therapy.ResultsThirty-nine patients with type 1 or type 2 diabetes were enrolled. Average (± SEM) fasting glucose decreased from 409.2 ± 25.9 mg/dl at admission to 190.4 ± 12.0 mg/dl at discharge and to 169.0 ± 10.3 at 2 months (both p < 0.001). EPCs (per million blood cells) significantly increased from hospital admission (13.1 ± 1.4) to discharge (16.4 ± 1.1; p = 0.022) and remained stable after 2 months (15.5 ± 1.7; p = 0.023 versus baseline). EPCs increased significantly more in participants with newly-diagnosed diabetes than in those with pre-existing diabetes. The increase in EPCs was significant in type 1 but not in type 2 diabetes and in those without chronic complications.ConclusionIn individuals hospitalized for decompensated diabetes, insulin therapy rapidly increases EPC levels for up to 2 months. EPC defect, reflecting impaired vascular repair capacity, may be reversible in the early diabetes stages.

Abstract 13035: A Potent Protective Vascular Endothelium Property of Garcinia mangostana L. Extract in High-Risk Framingham Score Patients With Type 2 Diabetes: A Prospective Randomized Control Trial

Background: Endothelial dysfunction is the hallmark of vascular complications in cardiovascular disease with type 2 diabetes as the comorbid. Garcinia mangostana L., or mangosteen, has been known as the queen of fruits from Southeast Asia that owns anti-inflammatory and anti-oxidant properties for years. Objective: This study was aimed to evaluate the role of G. mangostana extract as a potent protective vascular endothelium in patients with type 2 diabetes and high-risk Framingham score. Method: This is a prospective, randomized, single blind, placebo-controlled trial in subjects with type 2 DM and high-risk Framingham score. Subjects were randomly divided into two groups: the treatment group of G. mangostana extract, administered 2,175 mg/day in divided dosage for 90 days, and control group receiving placebo. The primary endpoint was the changes in endothelial progenitor cells (EPCs) and circulating endothelial cells (CECs) levels. We also evaluated the levels of NO, IL-1, IL-6, TNF-α, fasting blood glucose and HbA1c. Study had been approved by local ethics committee. Statistical analysis was done and p&lt;0.05 was considered significant. Results: From 90 subjects with high-risk Framingham score, there were 49 (54.4%) subjects with type 2 diabetes, which randomly divided into two groups. The treatment group (n=23) received G. mangostana extract and control group (n=26) received placebo. Statistical analysis showed that increase of EPC and decrease of CEC levels were significant in treatment group, compared to control (p=0.000 and p=0.002, respectively). The levels of NO increased significantly (p=0.027) and levels of IL-1, IL-6, TNF-α decreased significantly in comparison to placebo (p=0.003, p=0.002, p=0.000, respectively). Analysis also showed that HbA1c decreased significantly (p=0.002) compared to placebo; however, it was insignificant for fasting blood glucose reduction (p=0.336). Conclusion: Garcinia mangostana L. extract, as an adjuvant therapy, plays a role in improving endothelial dysfunction by significant increase in EPCs and decrease in CECs, in patients with type 2 diabetes and high-risk Framingham score.

Effects of structured home-based exercise training on circulating endothelial progenitor cells and endothelial function in patients with intermittent claudication.

Exercise training improves walking capacity in patients with intermittent claudication (IC). Endothelial progenitor cells (EPCs), endothelial microparticles (EMPs), and endothelial dysfunction could play a role in this process. We measured EPCs and EMPs in a group of 60 patients with IC, and in a control group of 20 individuals without IC, before a treadmill test and 2, 24, and 48 hours after the test. Thirty patients with IC were randomly assigned to perform a 12-week home-based exercise training program. The EPC count, flow-mediated dilation (FMD) of the brachial artery, pain-free walking time (PFWT), and maximum walking time (MWT) were measured at the baseline and after the exercise training program. In patients with IC, EMPs significantly increased 2 hours after the treadmill test, whereas EPCs significantly increased after 24 hours. Among the subjects assigned to complete the training program, we observed a significant increase in the number of EPCs after 12 weeks, as well as an improvement in FMD, PFWT, and MWT. A significant correlation between the variation of EPCs, FMD, and MWT was found. The increase of EPCs and FMD were independent determinants of the walking capacity improvement, without significant interaction. Our results suggest that EPCs mobilization contributes to the improvement of walking capacity in patients with IC undergoing structured physical training. A number of different, partly independent, mechanisms are involved in this process, and our results highlight the potential role of EMPs release and endothelial function improvement. ClinicalTrials.gov Identifier: NCT04302571.

The acute and long-term effects of a cardiac rehabilitation program on endothelial progenitor cells in chronic heart failure patients: Comparing two different exercise training protocols.

BackgroundVascular endothelial dysfunction is an underlying pathophysiological feature of chronic heart failure (CHF). Endothelial progenitor cells (EPCs) are also impaired. The purpose of the study was to assess the effect of a cardiac rehabilitation (CR) program on the increase of EPCs at rest and on the acute response after maximal exercise in patients with CHF and investigate whether there were differences between two exercise training protocols and patients of NYHA II and III classes.MethodsForty-four patients with stable CHF enrolled in a 36-session CR program and were randomized in one training protocol; either high-intensity interval training (HIIT) or HIIT combined with muscle strength (COM). All patients underwent maximum cardiopulmonary exercise testing (CPET) before and after the CR program and venous blood was drawn before and after each CPET. Five endothelial cellular populations, expressed as cells/106 enucleated cells, were quantified by flow cytometry.ResultsAn increase in all endothelial cellular populations at rest was observed after the CR program (p < 0.01). The acute response after maximum exercise increased in 4 out of 5 endothelial cellular populations after rehabilitation. Although there was increase in EPCs at rest and the acute response after rehabilitation in each exercise training group and each NYHA class, there were no differences between HIIT and COM groups or NYHA II and NYHA III classes (p > 0.05).ConclusionsA 36-session CR program increases the acute response after maximum CPET and stimulates the long-term mobilization of EPCs at rest in patients with CHF. These benefits seem to be similar between HIIT and COM exercise training protocols and between patients of different functional classes.

Endothelial Progenitor Cells as a Potential Biomarker in Interstitial Lung Disease Associated with Rheumatoid Arthritis.

Interstitial lung disease (ILD) increases morbidity and mortality in patients with rheumatoid arthritis (RA). Although the pathogenesis of ILD associated with RA (RA-ILD+) remains poorly defined, vascular tissue is crucial in lung physiology. In this context, endothelial progenitor cells (EPC) are involved in endothelial tissue repair. However, little is known about their implication in RA-ILD+. Accordingly, we aimed to investigate the potential role of EPC related to endothelial damage in RA-ILD+. EPC quantification in peripheral blood from 80 individuals (20 RA-ILD+ patients, 25 RA-ILD− patients, 21 idiopathic pulmonary fibrosis (IPF) patients, and 14 healthy controls) was performed by flow cytometry. EPC were considered as CD34+, CD45low, CD309+ and CD133+. A significant increase in EPC frequency in RA-ILD+ patients, as well as in RA-ILD− and IPF patients, was found when compared with controls (p < 0.001, p = 0.02 and p < 0.001, respectively). RA-ILD+ patients exhibited a higher EPC frequency than the RA-ILD− ones (p = 0.003), but lower than IPF patients (p < 0.001). Our results suggest that EPC increase may represent a reparative compensatory mechanism in patients with RA-ILD+. The degree of EPC frequency may help to identify the presence of ILD in RA patients and to discriminate RA-ILD+ from IPF.

Preconception preparation for pregnancy

Background. Experience in high-risk obstetrics shows that a significant part of the determining factors affects the woman and the fetus before the first visit to the doctor. Improving a woman’s health before conception can improve her reproductive performance and reduce financial costs spent on obstetric medical aid.&#x0D; Objective. Describe the key concepts of preconception training.&#x0D; Materials and methods. Analysis of literature sources on this topic; own study involving 42 pregnant women (22 women with systemic lupus erythematosus (SLE), 20 women with congenital heart disease (CHD) and hypertensive complications of previous pregnancies), who were divided into two groups. Group 1 received routine drugs and L-arginine (Tivortin, “Yuria-Pharm”) in doses recommended for cardiac patients, and group 2 – only routine drugs.&#x0D; Results and discussion. The components of preconception programs include the individual responsibility of women throughout life, awareness of women, preventive visits, interventions on identified risks, pre-pregnancy examinations and pregnancy supervision, health insurance for low-income women, health programs and strategies, medical research, and the improvement of monitoring. Target groups of preconception include women with unfavorable obstetric history (premature birth, cessation of fetal development, cesarean section, stillbirth, multiple miscarriages, birth of children with birth defects, hypertensive complications of pregnancy) and chronic diseases (type 1 diabetes mellitus, SLE, antiphospholipid syndrome, severe cardiovascular pathology, arterial hypertension, pulmonary hypertension). An important role in the management of pregnant women belongs to the prevention of preeclampsia (PE). Administration of acetylsalicylic acid reduces the likelihood of PE by 10 % in low-risk pregnant women and by 25 % in high-risk pregnant women, but the drug should be started during gestation. The use of calcium (1000 mg) reduces the risk of PE in high-risk pregnant women by 37 %. Preconception preparation is especially important, because some processes, such as the implantation of placental structures, begin and end before a woman learns she is pregnant. Remodeling of the spiral arteries also begins in the early stages of pregnancy, so its correction after the confirmation of pregnancy is less effective than prevention before it occurs. E.E. Camarena Pulido et al. (2016) studied the role of L-arginine (5 tablets of 600 mg per day from the 20th week of gestation before delivery) in the prevention of PE in high-risk women. In the L-arginine group significantly fewer cases of PE (3/49 vs. 11/47 in the placebo group; p=0.01) and a lower incidence of preterm birth were detected. In the another study, pregnant women with chronic hypertension received oral L-arginine or placebo. In the arginine group, there was a lower need for antihypertensive drugs, as well as lower frequency of births before 34 weeks, PE and neonatal complications (Neri I. et al., 2010). In the own study, favorable obstetric results of Tivortin were found. The percentage of births at &gt;37 weeks in the Tivortin group among women with SLE was 90.9 %, and in the group of standard therapy – 50 % (p&lt;0.01), among women with CHD – 90 % and 75 % respectively; p&lt;0.05). No newborns with an Apgar score of &lt;7 were found in the Tivortin group. In the comparison group their number was 27.7 %. Laboratory studies have shown that there is a significant increase in endothelial progenitor cells in the Tivortin group.&#x0D; Conclusions. 1. Preconception preparation of pregnant women is a multifaceted set of measures. 2. A significant number of physiological and pathological processes occur in the early stages of pregnancy, so they can be influenced only in the preconception period. 3. The use of L-arginine during gestation reduces the risk of PE, premature birth and other complications in all women and especially in women with hypertension.

OPN binds alpha V integrin to promote endothelial progenitor cell incorporation into vasculature.

Angiogenesis is fundamental to the expansion of the placental vasculature during pregnancy. Integrins are associated with vascular formation; and osteopontin is a candidate ligand for integrins to promote angiogenesis. Endothelial progenitor cells (EPCs) are released from bone marrow into the blood and incorporate into newly vascularized tissue where they differentiate into mature endothelium. Results of studies in women suggest that EPCs may play an important role in maintaining placental vascular integrity during pregnancy, although little is known about how EPCs are recruited to these tissues. Our goal was to determine the αv integrin mediated effects of osteopontin on EPC adhesion and incorporation into angiogenic vascular networks. EPCs were isolated from 6 h old piglets. RT-PCR revealed that EPCs initially had a monocyte-like phenotype in culture that became more endothelial-like with cell passage. Immunofluorescence microscopy confirmed that the EPCs express platelet endothelial cell adhesion molecule, vascular endothelial cadherin, and von Willebrand factor. When EPCs were cultured on OPN-coated slides, the αv integrin subunit was observed in focal adhesions at the basal surface of EPCs. Silencing of αv integrin reduced EPC binding to OPN and focal adhesion assembly. In vitro siRNA knockdown in EPCs,demonstrated that OPN stimulates EPC incorporation into human umbilical vein endothelial cell (HUVEC) networks via αv-containing integrins. Finally, in situ hybridization and immunohistochemistry localized osteopontin near placental blood vessels. In summary, OPN binds the αv integrin subunit on EPCs to support EPC adhesion and increase EPC incorporation into angiogenic vascular networks.

Clinical significance of endothelial progenitor cells in patients with liver cirrhosis with or without hepatocellular carcinoma.

The role of endothelial progenitor cells in patients with cirrhosis has seldom been investigated. This study was conducted to assess the clinical significance of circulating endothelial progenitor cells in patients with liver cirrhosis with or without hepatocellular carcinoma. A blood sample was collected once from patients with cirrhosis alone (n = 34) or cirrhosis and hepatocellular carcinoma (n = 46) and healthy controls (n = 27) for assessing levels of endothelial progenitor cells and vascular endothelial growth factor. Blood cells staining positive for CD34/CD133/KDR using flow cytometry were characterized as endothelial progenitor cells. Plasma vascular endothelial growth factor was quantified by ELISA. The levels of CD34/KDR-positive endothelial progenitor cells, CD133/KDR-positive endothelial progenitor cells, and vascular endothelial growth factor were higher in patients with cirrhosis ± hepatocellular carcinoma than in healthy controls (P = 0.017, P < 0.001 and P < 0.001, respectively). The levels of endothelial progenitor cells and vascular endothelial growth factor did not show statistical difference according to Child-Turcotte-Pugh class. There was a moderately significant correlation between vascular endothelial growth factor levels and hepatocellular carcinoma stage (ρ = 0.464, P = 0.001). Smoking, ascites, and portal vein thrombosis were independently related to lower levels of circulating CD34/KDR-positive endothelial progenitor cells, higher levels of CD133/KDR-positive endothelial progenitor cells, and higher levels of vascular endothelial growth factor, respectively (P = 0.041, P = 0.023, and P < 0.001, respectively). Circulating endothelial progenitor cells and plasma vascular endothelial growth factor levels were higher in patients with liver cirrhosis ± hepatocellular carcinoma compared to healthy controls. The increase in endothelial progenitor cells and vascular endothelial growth factor may have a possible role in the development of complications, especially ascites and portal vein thrombosis, or in progression of hepatocellular carcinoma.

Sweeteners modulate bioactivity of endothelial progenitor cells but not induce detrimental effects both on inflammation and behavioural changes

This study sought to determine the possible detrimental effects of several low- or non-caloric sweeteners on endothelial progenitor cells (EPCs), inflammation and behavioural changes in mice. C57BL/6 male mice received low and high dose of natural and artificial sweeteners for 4 weeks. EPCs, physical and biochemical variables, inflammation and behavioural changes were evaluated. A significant reduction of about 25% of EPCs was found when mice received a moderate amount of all sweeteners (p < .05). This reduction was more strongly significant when a double dose of glucose, aspartame, rebaudioside A and cyclamate (p < .005) in comparison to fructose and sucrose (p < .05) was administered. During inflammation carrageenan-induced, all sweeteners produced a significant increase of EPCs compared to the control group (p < .05). Consumption of glucose and sugar substitutes affect mouse EPC number according to the absence or presence of an inflammatory status, but does not induce detrimental effects on inflammation and behavioural changes.

High levels of endothelial progenitor cells and circulating endothelial cells in patients with Behcet’s disease and their relationship to disease activity

AbstractBackground: Behcet’s disease is a multisystemic vasculitis, associated with vascular endothelial dysfunction. Currently, the prognosis is unpredictable, because there is still no valid laboratory marker indicating the disease activity in Behcet’s disease. Endothelial progenitor cells and circulating endothelial cells are newly introduced hematological markers which are presumed to take part in the pathogenesis of vasculitis.Objectives: To evaluate the levels of endothelial progenitor cells and subtypes and circulating endothelial cells in patients with Behcet’s disease and to describe their relationship with the disease activity.Methods: A total of 45 patients with Behcet’s disease and 28 healthy controls were included in the study. Endothelial progenitor cells (CD34+CD133+KDR+ as early endothelial progenitor cells and CD34+KDR+ as late endothelial progenitor cells), and circulating endothelial cells (CD34+CD133+) were measured by flow cytometry.Results: The mean plasma level of endothelial progenitor cells and circulating endothelial cells, vascular endothelial growth factor, matrix metalloproteinase-9, C-reactive protein, and erythrocyte sedimentation rate were significantly higher in patients with Behcet’s disease. All of these parameters except circulating endothelial cells were also found to be higher in patients with active disease than in patients with inactive disease. Early endothelial progenitor cells showed significant correlations with C-reactive protein and circulating endothelial cells.Study limitations: The cross-sectional nature of the study and patient characteristics such as being under treatment, which can affect endothelial progenitor cells numbers.Conclusion: The increase in endothelial progenitor cells may play an essential role in the repair of endothelial injury in Behcet’s disease, especially in the active period of the disease. Thus, endothelial progenitor cells can indicate the disease activity. In addition, endothelial progenitor cells and circulating endothelial cells can be used as endothelial repair and injury markers for Behcet’s disease, respectively.

Consumption of aged white wine modulates cardiovascular risk factors via circulating endothelial progenitor cells and inflammatory biomarkers

There is compelling evidence showing that moderate alcohol consumption reduces cardiovascular risk factors related to atherosclerosis. The aim of this study was to evaluate the effects of aged white wine (AWW) and gin on circulating endothelial progenitor cells (EPC) and the expression of cell adhesion molecules, inflammatory cytokines and chemokines related to atherosclerosis in high cardiovascular risk subjects. This was an open, randomized, controlled, crossover study in 38 high-risk male volunteers between 55 and 80 years of age randomized to receive 30g of ethanol/day as AWW or gin during 3 weeks. We used the paired two-tailed t-test to compare differences in outcome variables in response to each intervention. Carryover effects for the two periods were evaluated comparing the outcome variables before the AWW and gin interventions. Compared to gin, AWW intake was associated with a significant 39.6% increase in EPCs. Expression of CD31 and CD40 in T-lymphocytes and of CCR2 and CD36 in monocytes also decreased significantly after AWW intake. In addition, compared to gin, AWW was associated with a significant decrease of plasma pro-inflammatory biomarkers interleukin-8 and interleukin-18 and vascular and intercellular adhesion molecules-1. Lfa-1, Mac-1, VLA4, CD40 and CD31 expression in monocytes and interferon gamma (IFN-γ) concentrations significantly decreased after intake of both alcoholic beverages. AWW shows a greater ability to repair and maintain endothelial integrity compared to gin. This effect is probably due to grape-derived minor components in AWW, which are absent in gin.

Alogliptin and Gliclazide Similarly Increase Circulating Endothelial Progenitor Cells in Type 2 Diabetes Patients.

We investigated the effect of alogliptin and gliclazide on endothelial progenitor cells (EPCs) in type 2 diabetes. Eighty patients with type 2 diabetes and HbA1c between 7.5% and 8.5% were randomized to receive either alogliptin (25 mg/daily) or gliclazide extended-release (30 mg/daily for HbA1c 7.5-8.0% and 60 mg/daily for HbA1c 8.0-8.5%) in combination with metformin for 4 months. At baseline and 4 months, clinical and laboratory parameters of EPCs were determined. After 4 months of treatment, alogliptin and gliclazide resulted in a similar significant reduction in HbA1c (%) (8.0±0.3 vs. 7.1±0.2, and 8.0±0.3 vs. 7.0±0.2, respectively; P<0.05) and a similar and significant increase in EPC count (cells/106 WBC) (CD45-CD133+KDR+ : 2.2±1.2 vs. 3.7±1.6, CD45-CD34+KDR+: 3.3±1.8 vs. 4.9±1.8; P<0.05 for alogliptin; CD45-CD133+KDR+: 2.3±1.3 vs. 3.6±1.5, CD45-CD34+KDR+: 3.1±1.3 vs. 4.6±1.7; P<0.05 for gliclazide). Both alogliptin and gliclazide demonstrated a beneficial effect in increasing EPCs in poorly controlled type 2 diabetes. As alogliptin and gliclazide exhibit different mechanisms of action, the observed increase in EPCs seems to be due to their glucose-lowering effect.

Abstract 27: Loss of GATA4 in Kit Lineage-derived Endothelium Leads to Vascular Permeability and Increased Heterotypic Fusion

The treatment of cardiac ischemic injury using heart-derived c-Kit stem cells showed promising results in clinical trials with scar reduction and improved ejection fraction following coronary infusion. We previously developed a c-Kit lineage tracing mouse model showing that these cells very rarely convert into de novo cardiomyocytes and most lineage-derived cardiomyocytes are due to fusion. To potentially reduce this de novo rate to zero, and unequivocally evaluate the cellular fusion process in vivo , we deleted the essential cardiomyogenic transcription factor Gata4 in c-Kit cells using a Cre-loxP approach (Kit-Gata4 KO). Mice with the tamoxifen inducible Kit-MerCreMer allele crossed into a Gata4 homozygous LoxP targeted background with the Rosa26-eGFP reporter were fed tamoxifen at weaning to delete Gata4 in all c-Kit expressing cells and show them and their progeny as eGFP + . Unexpectedly, we observed a greater than 10-fold increase in Kit lineage-traced cardiomyocytes in Kit-Gata4 KO mice compared to Kit only controls with up to 4 months of tamoxifen treatment (90% v. 81% fusion in Kit-Gata4 KO v. controls). A 6-fold increase in leukocyte (CD45 + ) infiltration was observed suggesting the possibility for greater rates of fusion in the heart. However, bone marrow transplant studies revealed that Kit-Gata4 KO bone marrow does not exhibit increased cardiac infiltration or heterotypic fusion with cardiomyocytes. The reciprocal experiment, donating control bone marrow to recipient mice with Kit-Gata4 KO hearts, showed a 4-fold increase in fusion of control bone marrow showing that increases in fusion was intrinsic to the cells within the heart. Analysis of cardiac endothelial cells in Kit-Gata4 KO mice revealed that total CD31 + cells were increased by 30% with 2 months of tamoxifen treatment with a parallel increase in endothelial progenitor cells (CD133 + ) in the bone marrow. This endothelial phenotype was corroborated using endothelial-specific deletion of Gata4 in a Tie2-CreER mouse which showed a 5-fold increase in both CD31 + cells and vascular permeability after 1 month of tamoxifen. Currently, studies are underway to assess the mechanism of altered endothelial integrity and expansion in the absence of Gata4 in c-kit hemoangioblast progenitors.

Active Stromal Cell-Derived Factor 1α and Endothelial Progenitor Cells are Equally Increased by Alogliptin in Good and Poor Diabetes Control.

Background:It is postulated that the ability of dipeptidyl peptidase-4 inhibitors (DPP-4-i) to increase circulating endothelial progenitor cells (EPCs) may be at least partly mediated by active stromal cell–derived factor 1α (SDF-1α) (a pivotal mediator of stem cell mobilization from the bone marrow). As other DPP-4-i were demonstrated to increase EPC concentrations, in this study, we sought to investigate the ability of the DPP-4-i alogliptin in modifying EPCs and SDF-1α, in patients with good and poor diabetes control.Methods:Two groups of diabetic patients on metformin were divided by hemoglobin A1c (HbA1c): Group A—those with HbA1c ≤6.5% (28 patients) and Group B—those with HbA1c 7.5% to 8.5% (31 patients). Both groups received alogliptin 25 mg/daily for 4 months. At baseline and 4 months later, clinical, laboratory parameters, EPCs, and active SDF-1α were determined.Results:After 4-month treatment with alogliptin, either Group A or Group B showed reduced HbA1c levels and concomitant similar increase in EPCs and active SDF-1α.Conclusions:Alogliptin showed significant benefits in increasing EPCs and active SDF-1α either in good or poor diabetes control. The study demonstrated that similar to other DPP-4-i, also alogliptin is able to increase EPC concentrations, suggesting the existence of a class effect mediated by SDF-1α. The extent of increase in EPCs is independent from baseline diabetes control.

Electronic cigarettes increase endothelial progenitor cells in the blood of healthy volunteers

Background and aimsThe use of electronic cigarettes is increasing dramatically on a global scale and its effects on human health remain uncertain. In the present study, we measured endothelial progenitor cells (EPCs) and microvesicles (MVs) in healthy young volunteers following short-term exposure to inhalation of e-cigarette vapor (ECV) to determine vascular changes. MethodsSixteen healthy seldom smokers were randomized into two groups either exposed or not exposed to 10 puffs of ECV for 10 min, in a crossover design. Blood samples were obtained at baseline and 1, 4 and 24 h following exposure. EPCs (CD34 + CD309) and MVs were analyzed by flow cytometry. MVs were phenotyped according to origin (platelet (CD41), endothelial (CD144), leukocytes (CD45), monocytes (CD14)) and nuclear content (SYTO 13 dye). In addition, expression of inflammation markers such P-selectin (CD62P), E-selectin (CD62E), CD40-ligand (CD154) and HMGB1 was investigated. Fractional exhaled nitric oxide (FeNO) was also measured at baseline and after 24 h. ResultsEPC levels in blood were significantly increased 1 h following exposure to ECV and returned to baseline values after 24 h. Only E-selectin positive MVs (endothelial origin) were slightly elevated (p < 0.038). FeNO was unaffected by exposure to ECV. ConclusionsIn healthy volunteers, ten puffs of e-cigarette vapor inhalation caused an increase in EPCs. This increase was of the same magnitude as following smoking of one traditional cigarette, as we previously demonstrated. Taken together, these results may represent signs of possible vascular changes after short e-cigarette inhalation. Further studies analyzing potential cardiovascular health effects are critical as the e-cigarette market continues to burgeon.

Increase of endothelial progenitor cells in acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation for acute myeloid leukaemia

IntroductionCirculating endothelial progenitor cells (EPCs; CD31+ CD34brightCD133+ CD45dim cells) are novel markers of endothelial dysfunction and related to inflammatory processes such as acute graft-versus-host disease (aGvHD). Patients and methods47 patients with acute myeloid leukaemia (AML) who were in complete remission as they underwent allogeneic hematopoietic stem cell transplantation with myeloablative conditioning with PBSC as stem cell source were enrolled in the study. Blood samples for the quantitative analysis of circulating EPC levels were drawn at different time points in patients with and without aGvHD. CD34+ VEGFR2/KDR+ CD133+ triple-positive cells identified among CD34+ cells by FACS. EPC were quantified and data are presented as cells/ml whole blood. ResultsCirculating EPC levels were not significantly different in patients with and without aGvHD prior to conditioning (baseline) and at the time of engraftment. However, at diagnosis of aGvHD≥grade 2, EPC levels increased whereas in patients without aGvHD the EPC levels remained significantly lower (3021±278 versus 2322±195 cells/ml; p<0.001). Patients with steroid-refractory aGvHD had high levels of EPC throughout. EPC levels fell in responding patients. ConclusionOur results demonstrate that the number of circulating EPCs is increased in patients with aGvHD compared to patients without aGvHD.

Biphasic response in number of stem cells and endothelial progenitor cells after left ventricular assist device implantation: A 6 month follow-up

BackgroundContinuous blood flow could have deleterious effects on endothelium and vascular health. This could have serious consequences in patients with heart failure treated with continuous flow left ventricular assist devices (LVAD). Therefore, we studied effect of LVAD on three circulating vascular biomarkers: stem cells (SC), endothelial progenitor cells (EPC) and microparticles (MP). MethodsIn 23 patients (5 women) with end-stage heart failure, SC, EPC and MP were measured before, and 3 and 6months after implantation of LVAD (HeartMate II). SC were defined using determination of surface antigen expression as mononuclear CD34+/CD45low+ cells and EPC as mononuclear CD34+/CD45low+/KDR+ cells. MP concentrations were determined by ELISA method. ResultsThree months after LVAD implantation numbers of SC and EPC significantly decreased (p=0.01 and p=0.001, respectively). On the contrary, between 3rd and 6th month after implantation they significantly increased (p=0.006 and p=0.003, respectively).MP did not change significantly during the study despite exerting similar trend as SC and EPC. ConclusionsObserved biphasic changes of SC and EPC might reflect two processes. First, shortly after LVAD implantation, improved tissue perfusion could lead to decrease in ischemic stimuli and ensuing decrease of SC and EPC. Second, continuous flow between 3rd and 6th month produced by LVAD could lead to increase of SC and EPC through activation of endothelium. This explanation could be supported also by similar trend in the changes of concentrations of MP.