Number Of Endothelial Progenitor Cells Research Articles

Effects of metoprolol, methyldopa, and nifedipine on endothelial progenitor cells in patients with gestational hypertension and preeclampsia

Endothelial progenitor cells (EPCs) are critical for vascular regeneration and function, but are reduced in hypertensive disorders of pregnancy. We aimed to determine the possible effects of antihypertensive drugs, such as metoprolol, methyldopa, and nifedipine, on EPC number and functions in patients with gestational hypertension and preeclampsia. We collected blood samples from 30 normal pregnant women, 67 patients with gestational hypertension and 48 patients with preeclampsia. The patients received no drug or an antihypertensive drug, such as metoprolol, methyldopa, or nifedipine, between 20 and 24weeks of gestation. The number of EPCs and circulating endothelial cells (CECs) in the blood was measured by flow cytometry. Moreover, colony formation and migration assays were performed on the isolated EPCs. Both the systolic and diastolic blood pressure (BP) increased, while the percentage of flow-mediated vasodilatation (FMD) decreased in patients with gestational hypertension and preeclampsia, compared to the healthy controls at 20weeks of gestation. CEC number increased in the patients, whereas EPC counts decreased. Furthermore, EPC colony formation and migration abilities were also impaired in the patients. However, administration of metoprolol, methyldopa, or nifedipine effectively restored the systolic and diastolic BP, FMD%, EPCs, and CEC numbers, as well as EPC migration capacity. Endothelial progenitor cells colony formation ability selectively improved with methyldopa and nifedipine. In patients receiving no drugs, most of these indexes worsened within 4weeks (study duration). This study revealed a new pharmacological action of these antihypertensive drugs against gestational hypertension and preeclampsia, thus supporting their clinical use.

Serum miR-122 levels correlate with diabetic retinopathy.

Diabetic retinopathy is the most severe ocular complication of diabetes and may lead to visual disability and blindness. Proliferative diabetic retinopathy (PDR) is characterized by ischemia-induced neovascularization with associated complications. An association was established between the presence of PDR, cardiovascular disease, and mortality among patients with type 1 diabetes mellitus and type 2 diabetes mellitus in epidemiological studies. However, the mechanism underlying increased cardiovascular risk in patients with PDR is still unknown. In recent years, a group of miRNAs has been linked to the pathology of diabetes mellitus. Besides, miRNAs in biofluids such as serum have been suggested as potential minimally invasive biomarkers of diabetes and vascular complications. This was a prospective study that recruited 40 human subjects: 10 healthy subjects, 10 with diabetes but without retinopathy (NDR), 10 with diabetic non-proliferative retinopathy (NPDR), and 10 with proliferative diabetic retinopathy (PDR). To examine whether serum miRNAs show altered levels at different stages of diabetic retinopathy, seven specific miRNA candidates (miR-126-3p, miR-130a-3p, miR-21-1, let-7f-5p, miR-122, miR-30c and miR-451a) were measured by qRT-PCR in RNA isolated from sera of all subjects. miR-122 levels increased in parallel with retinopathy severity: from healthy controls to NDR and from NDR to NPDR. However, when the disease progressed to PDR a marked decrease in miR-122 level was noted. This decrease was significant both compared to NPDR samples (p = 0.016) and to all non-PDR samples (p = 0.0002). Additionally, a positive trend was observed comparing miR-122 levels and the number of endothelial progenitor cells in the sera of all subjects. A significant increase in miR-122 was found in patients with diabetic retinopathy that may be related to its role in preventing angiogenesis and proliferation. The dramatic decline in patients with PDR may represent an inhibition or exhaustion of the anti-angiogenic anti-proliferative defense system. Further studies are needed to understand whether miRNA-122 has a role in the pathogenesis of diabetic retinopathy.

Evaluation the effect of testosterone on the number of endothelial progenitor cells and amount of SDF-1α, PDGF, bFGF, and NO.

Background:Recent therapeutic advances in cardiovascular disease, thanks to the discovery of endothelial progenitor cells (EPCs). Stromal cell-derived factor-1α (SDF-1α), platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), and nitric oxide (NO) play a role in migration, homing, and differentiation of EPCs into mature endothelial cells. The incidence of cardiovascular disease is higher in men than in women. This fact suggests the influence of sex hormones on incidence of cardiovascular disease.Methods:Twenty-four female wistar rats weighing 160–180 g were randomly divided into four groups (N = 6): 1. sham-treated by sesame oil, 2. ovariectomized (OVX)-treated by sesame oil, 3. OVX-treated by 10 μg/kg/day testosterone, and 4. OVX-treated by 100 μg/kg/day testosterone. After 21 days, the animals were euthanized and blood samples were saved for determination of EPC count and serum levels of SDF-1α, PDGF, bFGF, and NO production.Results:High-dose testosterone induced significant increase in EPC count in OVX rats (P < 0.05). Also 100 μg/kg/day testosterone increased serum level of SDF-1α more than OVX-treated by 10 μg/kg/day testosterone (P < 0.05). But 10 μg/kg/day testosterone increased significantly the serum level of PDGF >100 μg/kg/day testosterone-treated group (P < 0.05). The serum level of bFGF in sham-treated by sesame oil was equal with its concentration in OVX-treated by 100 μg/kg/day testosterone. And the serum concentration of NO production in testosterone-treated groups were significantly less than other groups (P < 0.05).Conclusions:This study suggests that testosterone might be effective on cardiovascular disease in females by increasing EPC count through SDF-1α and PDGF mechanisms which are some of the vascular healing factors.

Positive Correlation between Very Small Embryonic Stem Cell, Hematopoietic Stem Cell, and Endothelial Progenitor Cell in Umbilical Cord Blood Unit

BACKGROUND: Umbilical Cord Blood (UCB) has been widely use as regenerative medicine due to the content of undifferentiated cell which have capability to do self-renewal and differentiation into various type of cell called stem cells. Recent studies show that UCB contains not only hematopoietic stem cell (HSC) but also others stem cell and progenitor cell such as endothelial progenitor cell (EPC) and very small embryonic-like stem cell (VSEL). It is beliefs that HSC and EPC shared the same progenitor. In this study, correlation between the cell number of HSC, EPC and VSEL is analyzed in umbilical cord blood as the source of stem cell for clinical application.METHODS: The cell number of HSC, EPC and VSEL is counted from cryopreserved UCB collected from 22 women delivered via cesarean section which already stored for more than 2 years in this study. Sample were incubated with antibodies such as cluster of differentiation (CD)34-phycoerythrin (PE)/CD45-fluorescein isothiocyanate (FITC), CD133/1 (anti-CD (AC)133)-antigen-presenting cell (APC) and, CD184 (C-X-C chemokine receptor (CXCR)4)-PE.Vio770 to detect the present of HSC, EPC and VSEL in UCB. Sample were analyze using flowcytometer BD FACS Canto II.RESULTS: The cell population of HSC and late-EPC is 0.009% and 0.01% of total cell in UCB. VSEL only represented 0.001% from total cell in UCB, showing the lowest number of cell population in UCB. The correlation between the cell number of HSC and EPC is r=0,483*, p=0.023) and between HSC and VSEL is r=0.510*, p=0.015.CONCLUSION: In this study, both EPC and VSEL have a significant positive correlation with HSC.KEYWORDS: stem cell, umbilical cord blood, endothelial progenitor, flowcytometry

Reduced mRNA and Protein Expression Levels of Tet Methylcytosine Dioxygenase 3 in Endothelial Progenitor Cells of Patients of Type 2 Diabetes With Peripheral Artery Disease

Endothelial progenitor cells (EPCs) with immunological properties repair microvasculature to prevent the complications in patients with diabetes. Epigenetic changes such as DNA methylation alter the functions of cells. Tet methylcytosine dioxygenases (TETs) are enzymes responsible for the demethylation of cytosine on genomic DNA in cells. We hypothesized that EPCs of diabetic patients with peripheral artery disease (D-PAD) might have altered expression levels of TETs. Subjects who were non-diabetic (ND, n = 22), with diabetes only (D, n = 29) and with D-PAD (n = 22) were recruited for the collection of EPCs, which were isolated and subjected to analysis. The mRNA and protein expression levels of TET1, TET2, and TET3 were determined using real-time PCR and immunoblot, respectively. The TET1 mRNA expression level in ND group was lower than that in the D and D-PAD groups. The TET3 mRNA level in the ND group was higher than that in the D group, which was higher than that in the D-PAD group. The TET1 protein level in the D-PAD group, but not the D group, was higher than that in the ND group. The TET2 protein level in the D-PAD group, but not the D group, was lower than that in the ND group. The TET3 protein level in the ND group was higher than that in the D group, which was higher than that in the D-PAD group, which is the lowest among the three groups. The changes of TETs protein levels were due to the alterations of their transcripts. These probably lead to epigenetic changes, which may be responsible for the reductions of EPCs numbers and functions in patients with the D-PAD. The expression pattern of TET3 mRNA and TET3 protein in EPCs may be a biomarker of angiopathy in diabetic patients.

Involvement of endothelial nitric oxide synthase pathway in IGF‑1 protects endothelial progenitor cells against injury from oxidized LDLs.

A high level of oxidized low‑density lipoproteins (oxLDLs) is an independent risk factor for cardiovascular disease. The aim of the present study was to investigate whether insulin‑like growth factor‑1 (IGF‑1) protected endothelial progenitor cells (EPCs) from injury caused by ox‑LDLs, and whether the endothelial nitric oxide synthase (eNOS)/nitric oxide (NO) pathway was involved in this process. EPCs were isolated from human peripheral blood and characterized. In order to evaluate the effect of IGF‑1 on EPCs, cells were incubated with ox‑LDLs (100mg/ml) for 24h to induce a model of EPC dysfunction invitro, which demonstrated a decrease in the number of EPCs, concomitant with increased apoptosis and decreased proliferation rates. IGF‑1 dose‑dependently increased the number of EPCs. Concurrently, IGF‑1 decreased the levels of apoptosis of EPCs and improved EPCs proliferation following ox‑LDLs challenge. In addition, IGF‑1 significantly increased NO levels in ox‑LDLs‑treated EPCs, accompanied by an upregulation in eNOS expression. The protective effects of IGF‑1 on EPCs and NO production were abolished by L‑NAME, a specific eNOS inhibitor. These results suggested that IGF‑1 protects EPCs from dysfunction induced by oxLDLs through a mechanism involving the eNOS/NO pathway.

Improved angiogenic activity of endothelial progenitor cell in diabetic patients treated with insulin plus metformin.

Type 2 diabetes (T2DM) is associated with an increased vascular disease. Moreover, endothelial progenitor cell (EPC) function is impaired in diabetic patients. Decreased EPC number plays a critical role in reduced endothelial repair and development of the vascular disorder. To determine the effect of metformin and insulin plus metformin on functional activity of EPCs, 130 participants were divided into three groups (group 1: healthy control; group 2: metformin; group 3: insulin plus metformin). The concentration of EPCs in the circulation was first quantified. Thereafter, circulating EPCs (cEPCs) were harvested and the biological features of these cells including proliferative, clonogenicity, tubulogenic, and migratory properties were analyzed after expansion. The serum protein levels of some proangiogenic factors were also measured. Our results showed greater numbers of cEPCs in control and in diabetic patients treated with insulin plus metformin than in metformin-treated patients. Insulin plus metformin therapy was associated with augmented proliferative, clonogenicity, migratory, and tubulogenic activity of cEPCs in patients with T2DM. Increased serum concentrations of angiogenic factors were also observed in patients treated with insulin plus metformin. Western blot analysis showed increased protein levels of pTie-2/Tie2 and Pakt/AKT in cEPCs harvested from T2DM, treated with insulin metformin plus. This study showed that treatment with insulin plus metformin in diabetic patients is associated with increased mobilization of EPCs into the circulation, with potential beneficial effect in vascular protection in diabetic patients.

Dysfunctional high density lipoprotein failed to rescue the function of oxidized low density lipoprotein-treated endothelial progenitor cells: a novel index for the prediction of HDL functionality

Lipid metabolic disorders play critical roles in atherogenesis. Traditionally, it has been suggested that reduced high density lipoprotein (HDL) levels might be an important morbidity indicator for cardiovascular diseases. Therefore, it has been argued that therapeutically raising HDL levels may reduce atherogenesis in patients with dyslipidemia. However, recent clinical trials to elevate serum HDL levels by pharmacologic approaches failed to demonstrate clinical efficacy. Thus, to investigate the functionality of HDL and to explore the possible clinical relevance as well as to define an effective indicator that can represent HDL function may provide another key and reference to disclose the clinical treatment of dyslipidemia. We analyzed the association between the data of dichlorofluorescein assay (assay the functionality of HDL), the effect of HDL on oxidized low density lipoprotein (oxLDL)-stimulated endothelial progenitor cells (EPCs) in vitro, levels of circulating EPCs, and ex vitro EPC colony forming units of each case, we defined the indicator (relative HDL index (RHDL index) = dichlorofluorescein assay result of each subject/dichlorofluorescein assay reading of our young healthy controls) that may represent functionality of HDL. HDL from healthy adults protected oxLDL-treated EPCs by modulating p38 mitogen-activated protein kinase and Rho activation and by promoting nitric oxide production. HDL from subject with RHDL index ≧2 also failed to restore the functionality of oxLDL-treated EPCs via cell-signaling pathways in vitro. The RHDL index significantly correlated with patients' circulating EPC number or EPC colony forming units ex vivo. In conclusions, we explored the RHDL index as a score to predict a patient's EPC functions in vivo and ex vitro.

17β-estradiol promotes recovery after myocardial infarction by enhancing homing and angiogenic capacity of bone marrow-derived endothelial progenitor cells through ERα-SDF-1/CXCR4 crosstalking.

17β-estradiol (E2) has been shown to mediate endothelial progenitor cells (EPCs) to repair infarcted myocardium. Both estrogen receptor α (ERα) and stromal derived factor-1 (SDF-1)/CXCR4 signaling pathways may play a critical role in regulating homing and angiogenesis of EPCs in this process. However, the interaction between ERα and SDF-1/CXCR4 signaling pathways remains unclear. In response to E2, the expression of SDF-1 and CXCR4 in EPCs from ovariectomized BALB/C mice was obviously up-regulated, in addition, the migration and tube formation of EPCs in vitro were also significantly enhanced. However, ERα antagonist (MMP) and CXCR4 inhibitor (AMD3100) significantly decreased the migration and tube length of EPCs, even if mediated by E2. The combined treatment of MMP and AMD3100 exerted more inhibitory effects on migration and tube formation of EPCs induced by E2. In in vivo studies, ovariectomized mice were induced acute myocardial infarction (AMI), and divided into four groups (n = 6): non-preconditioned EPCs (3 × 106) group, E2-preconditioned EPCs group, MMP + AMD3100 preconditioned EPCs group, and EPCs pretreated with E2 + MMP + AMD3100 group. E2 group displayed a greater number of homing EPCs, increased capillary density in infarcted myocardium, decreased left ventricular (LV) fibrosis. Nevertheless, these effects of E2 were almost completely blocked by the combined treatment of MMP and AMD3100. E2 can produce cardiovascular protective effects in AMI setting by enhancing homing and angiogenic capacity of EPCs through ERα and CXCR4 signaling pathways, which means that ERα and CXCR4 pathways are effective targets for the development of treatment strategies for AMI.

Effectiveness of endothelial progenitor cell culture under microgravity for improved angiogenic potential

Endothelial progenitor cell (EPC) transplantation is beneficial for ischemic diseases such as critical limb ischemia and ischemic heart disease. The scarcity of functional EPCs in adults is a limiting factor for EPC transplantation therapy. The quality and quantity culture (QQc) system is an effective ex vivo method for enhancing the number and angiogenic potential of EPCs. Further, microgravity environments have been shown to enhance the functional potential of stem cells. We therefore hypothesized that cells cultured with QQc under microgravity may have enhanced functionality. We cultured human peripheral blood mononuclear cells using QQc under normal (E), microgravity (MG), or microgravity followed by normal (ME) conditions and found that ME resulted in the most significant increase in CD34+ and double positive Dil-Ac-LDL-FITC-Ulex-Lectin cells, both EPC markers. Furthermore, angiogenic potential was determined by an EPC-colony forming assay. While numbers of primitive EPC-colony forming units (pEPC-CFU) did not change, numbers of definitive EPC-CFU colonies increased most under ME conditions. Gene-expression profiling also identified increases in angiogenic factors, including vascular endothelial growth factor, under MG and ME conditions. Thus, QQc along with ME conditions could be an efficient system for significantly enhancing the number and angiogenic potential of EPCs.

Probucol protects circulating endothelial progenitor cells from ambient PM2.5 damage via inhibition of reactive oxygen species and inflammatory cytokine production in vivo.

Bone marrow-derived circulating endothelial progenitor cells (EPCs) contribute to angiogenesis and vascular repair. The number and function of EPCs are significantly decreased following exposure to ambient fine particulate matter of ≤2.5 µm in diameter (PM2.5) through reactive oxygen species (ROS) generation and inflammatory cytokine secretion. The anti-oxidant drug probucol reduces ROS and inflammatory cytokine production. The present study was designed to determine the protective effects of probucol on EPCs from PM2.5-associated impairment in vivo and to explore the potential underlying mechanisms. Male C57BL/6 mice were exposed to ambient air containing PM2.5 for one month with or without probucol treatment. Mice that breathed filtered air were used as a control group. Serum and blood cells were collected for analysis. The results indicated that PM2.5 exposure induced increases in blood intracellular ROS, serum inflammatory cytokine levels and the blood cell apoptotic rate, while it decreased the number and proliferation rate of circulating EPCs in the mice with PM2.5 exposure. These effects were significantly reduced/abrogated by probucol treatment. The present in vivo study suggested that probucol protects EPCs from damage through PM2.5 exposure by inhibiting ROS generation and inflammatory cytokine production.

17. Aberrant corpus luteum number as a contributor to altered maternal vascular health in early pregnancy and preeclampsia risk after assisted reproduction

Introduction Pregnancies conceived after utilization of assisted reproductive technologies (ART) are associated with an increased risk for preeclampsia. The pathophysiologic reasons are mainly unknown. Objective We sought to determine if a non-physiologic hormonal milieu influenced by the number of corpora lutea (CL) and the mode of conception affects maternal vascular health in early pregnancy. Methods Blood pressure, endothelial function, circulating endothelial progenitor cell numbers (CPCs), lipid levels, and CL hormones were compared in a prospective cohort of women at 11 to 14 weeks’ gestation by number of CL and mode of conception (N = 57): 0 CL (programmed frozen embryo transfer (FET), N = 18); 1 CL (spontaneous pregnancy after infertility [N = 16] and modified natural cycle FET [N = 12]); or >3 CL associated with in vitro fertilization [N = 11]. Results Women with 0 or >3 CL lacked the drop in mean arterial blood pressure compared to women with 1 CL (P = 0.05; P = 0.05). Reactive Hyperemia Index (RHI) was impaired in patients lacking a CL compared to women with 1 CL (P = 0.04). Baseline pulse wave amplitude (BPWA) was higher in subjects with >3 CL compared to 1 CL (P = 0.01) or zero (P = 0.01). Suppression of CL development in FET cycles was associated with a lower RHI compared with FETs in a natural cycle (P = 0.03). The number of angiogenic and non-angiogenic CPCs was lower in the absence of a CL in FETs (P = 0.02 and P = 0.04). Relaxin levels correlated with the number of angiogenic CPCs (r = 0.31; P = 0.03). Discussion Maternal vascular health in early pregnancy is altered in women with aberrant numbers of CL (0 or >3), and might represent insufficient cardiovascular adaption leading to an increased risk of preeclampsia.

No Differences in Levels of Circulating Progenitor Endothelial Cells or Circulating Endothelial Cells Among Patients Treated With Ticagrelor Compared With Clopidogrel During Non- ST -Segment-Elevation Myocardial Infarction.

BackgroundTicagrelor use during acute coronary syndromes demonstrated a decrease in all‐cause mortality in the PLATO (Platelet Inhibition and Patient Outcomes) trial. This effect has been attributed to a non–platelet‐derived improvement in endothelial function. The aim of this study was to determine differences in the number of endothelial progenitor cells and/or circulating endothelial cells found in peripheral blood in patients treated with either ticagrelor or clopidogrel during non–ST‐segment–elevation myocardial infarction.Methods and ResultsIn this multicenter, randomized study (NCT02244710), patients were considered for inclusion after non–ST‐segment–elevation myocardial infarction whenever they were P2Y12‐inhibitor naïve. Ticagrelor and clopidogrel were allocated at a 1:1 ratio. Blood samples for determining endothelial progenitor cells and circulating endothelial cells were extracted before the antiplatelet loading dose, 48 hours after presentation of index symptoms, and 1 month after the event. A multichannel cytometer was used for optimal cell characterization. A total of 96 patients fulfilled the inclusion criteria. Circulating endothelial cell levels corrected by white blood cells were as follows at baseline, 48 hours, and 1 month: 44 (28–64), 50 (33–63), and 38 (23–62) cells/mL, respectively, for clopidogrel and 38 (29–60), 45 (32–85), and 35 (24–71) cells/mL, respectively, for ticagrelor (P=0.6). Endothelial progenitor cell levels were 29 (15–47), 27 (15–33), and 18 (10–25) cells/mL, respectively, for clopidogrel and 20 (11–33), 22 (12–32), and 18 (11–29) cells/mL, respectively, for ticagrelor (P=0.9). No differences in intraindividual changes were found.ConclusionsPatients treated with ticagrelor during non–ST‐segment–elevation myocardial infarction, in comparison to clopidogrel, showed similar levels of endothelial progenitor cells and circulating endothelial cells. These data suggest that the endothelial protective effect mediated by ticagrelor is not related to bone marrow physiology modulation.Clinical Trial RegistrationURL: https://www.clinicaltrials.gov. Unique identifier: NCT02244710.

Deregulation of Notch1 pathway and circulating endothelial progenitor cell (EPC) number in patients with bicuspid aortic valve with and without ascending aorta aneurysm

Bicuspid aortic valve (BAV) is frequently associated with the development of ascending aortic aneurysm, even if the underlying mechanisms remain to be clarified. Here, we investigated if a deregulation of Notch1 signaling pathway and endothelial progenitor cells (EPCs) number is associated with BAV disease and an early ascending aortic aneurysm (AAA) onset. For this purpose, 70 subjects with BAV (M/F 50/20; mean age: 58.8 ± 14.8 years) and 70 subjects with tricuspid aortic valve (TAV) (M/F 35/35; mean age: 69.1 ± 12.8 years) and AAA complicated or not, were included. Interestingly, patients with AAA showed a significant increase in circulating Notch1 levels and EPC number than subjects without AAA. However, circulating Notch1 levels and EPC number were significantly lower in BAV subjects than TAV patients either in the presence or absence of AAA. Finally, Notch pathway was activated to a greater extent in aortic aneurysmatic portions with respect to healthy aortic fragments in both BAV and TAV patients. However, the expression of genes encoding components and ligands of Notch pathway in aortic tissues was significantly lower in BAV than TAV subjects. Our study demonstrates that BAV subjects are characterized by a significant decrease in both tissue and circulating levels of Notch pathway, and in blood EPC number than TAV patients, either in presence or absence of AAA disease.

FEATURES OF THE CONTENT OF ENDOTHELIAL PROGENITOR CELLS IN UMBILICAL CORD BLOOD OF PRE-TERM AND FULL-TERM NEWBORNS

The purpose of the study : determination of content of circulating endothelial cellsin umbilical cord blood in orderto assessthe regenerative potential of the endothelium in newborns. 29 newborn babies were examined: 18 pre-term infants (gestational age 30–35 weeks) and 11 full-term infants (gestational age 38–41 weeks). Determination of the number of circulating endothelial cells was determined in mixed umbilical cord blood of all children their phenotyping was carried out by cytofluorometry method using fluorochromes-labeled monoclonal antibodies. Circulating endothelial cells were defined as CD45–, CD133+, VEGFR2, CD34–, endothelial progenitor cells were defined as CD45–, CD133+, VEGFR2+, CD34+. It was revealed that in umbilical cord blood of pre-term infants was decreased both the total number of circulating endothelial cells and endothelial progenitor cells compared to full-term infants. Moreover, in preterm infants was observed higher percentage of endothelial progenitor cells, rather than mature endothelial cells. In blood of full-term infants was observed higher percentage of mature endothelial cells compared to endothelial progenitor cells and their number increases with increasing birth weight of the newborn. The decrease in the number of endothelial progenitor cells in pre-term infants is associated with impaired endothelial function. CD45–CD133+VEGFR2+CD34+ cells are involved in reparation of damaged endothelium and reduction in their number in the blood reflects the reduction of its regenerative potential.

Increased circulating endothelial progenitor cells and improved short-term outcomes in acute non-cardioembolic stroke after hyperbaric oxygen therapy

BackgroundAcute ischemic stroke is a leading cause of mortality and long-term disability, and profiles of endothelial progenitor cells (EPCs) reflect the degree of endothelial impairment. This study tested the hypothesis that hyperbaric oxygen therapy (HBOT) both improves the clinical short-term outcomes and increases the number of circulating EPCs and antioxidant capacity.MethodsThe numbers of circulating EPCs [CD133+/CD34+ (%), KDR+/CD34+ (%)], biomarkers for oxidative stress (thiols and thiobarbituric acid-reactive substances), and clinical scores (National Institutes of Health Stroke Scale [NIHSS], Barthel index [BI], and modified Rankin Scale [MRS]) were prospectively evaluated in 25 patients with acute non-cardioembolic stroke under HBOT at two time points (pre- and post-HBOT). The biomarkers and clinical scores were compared with those of 25 age- and sex-matched disease controls.ResultsThe numbers of KDR+/CD34+ (%) in the HBOT group following HBOT increased significantly, whereas the numbers of CD133+/CD34+ (%) also showed a tendency to increase without statistical significance. The mean high-sensitivity C-reactive protein levels showed significant decrease post-HBOT follow-up in the HBOT group. The changes in KDR+/CD34+EPC (%) numbers were positively correlated with changes in clinical outcomes scores (BI, NIHSS, and MRS) in the HBOT group.ConclusionsBased on the results of our study, HBOT can both improve short-term clinical outcomes and increase the number of circulating EPCs in patients with acute non-cardioembolic stroke.

Expression of endothelial progenitor cells and vascular endothelial growth factor according to genomic instability in colorectal liver metastases

Introduction: Increased numbers of endothelial progenitor cells and high level of vascular endothelial growth factor may indicate on higher tumor propensity to angiogenesis. The study aimed to evaluate the possible role of genomic instability in stimulation of angiogenesis in liver metastases of colorectal carcinoma. Methods: Among 80 randomly selected patients liver metastases, 70% and 30% were recognized as microsatellite stable and unstable, respectively. Samples of 2 ml of fresh blood were collected preoperatively. The cell fraction positive to CD309 (EPCS) was enumerated by flow cytometer and expressed as the percentage of WBC. Serum samples for evaluation of the vascular endothelial growth factor (VEGF) concentration were collected simultaneously for ELISA. A P-value < 0.05 was adopted as statistically significant in multiparametric analysis to reveal the association of genomic instability and of angiogenesis. A control group of 36 volunteers (18 F, 18 M, ma 38) was also measured. Results: The expression of EPCs and VEGF were significantly higher in MSI-H than MSS patients (0.01% versus 0.004% for EPCs, P< 0.008; 420.9 versus 300.2 pg/ml for VEGF). Multiparametric analysis showed angiogenesis nine times more active in MSI-H patients (OR=9.1, P< 0.01). They were characterized by more frequent metastases in both liver lobes (OR=32.8 P< 0.001) and metastases outside the liver (OR=8.3 P< 0.01). Conclusions: Patients with CRC metastases with MSI-H have high circulating numbers of EPCs and serum levels of VEGF. This may facilitate they regrowth. The data does point to the potential utility of anti-angiogenesis therapy in MSI-H metastatic disease.

Endothelial progenitor cells in cardiovascular diseases.

Cardiovascular diseases (CVDs) are the leading cause of death in both developed and developing countries. Endothelial progenitor cells (EPCs) are derived from hematopoietic stem cells with powerful function of angiogenesis. There are many studies on the relation between coronary heart disease and circulating EPCs. In this review, we discuss biological characteristics of endothelial progenitor cells, some influencing factors of the number and function of EPCs, and the role of EPCs in the treatment of cardiovascular disease. At last, we bring some perspectives on the future of endothelial progenitor cell therapy.

Microparticles harbouring Sonic hedgehog morphogen improve the vasculogenesis capacity of endothelial progenitor cells derived from myocardial infarction patients.

Endothelial progenitor cells (EPC) play a role in endothelium integrity maintenance and regeneration. Decreased numbers of EPC or their impaired function correlates with an increase in cardiovascular events. Thus, EPC are important predictors of cardiovascular mortality and morbidity. Microparticles carrying Sonic hedgehog (Shh) morphogen (MPShh+) trigger pro-angiogenic responses, both in endothelial cells and in ischaemic rodent models. Here, we propose that MPShh+ regulates EPC function, thus enhancing vasculogenesis, and correcting the defects in dysfunctional EPC obtained from acute myocardial infarction (AMI) patients. The mechanisms underlying Shh pathway function and nitric oxide (NO) production in EPC were evaluated. MPShh+ increased both the in vitro and in vivo vasculogenic capacity of EPC isolated from adult human peripheral blood samples. MPShh+ treatment significantly increased the expression of Shh signalling pathway genes (PTCH1, SMO, and GLI1) and masters of pro-angiogenic genes (NOS3, VEGFA, KDR, and KLF2) in EPC. Moreover, MPShh+ increased both the protein expression and activity of eNOS, resulting in increased NO production. Most importantly, MPShh+ improved the vasculogenic capacity of EPC from AMI patients to levels similar to that of EPC from healthy patients. All these effects were due to the activation of Shh pathway. MPShh+ increase both the vasculogenesis of EPC and their capacity to produce NO, including EPC from patients who have recently suffered an AMI. This study emphasizes MPShh+ and EPC as potential therapeutic tools for improving vascular regeneration as a treatment for cardiovascular ischaemic disease.

Association between Type I interferon and depletion and dysfunction of endothelial progenitor cells in C57BL/6 mice deficient in both apolipoprotein E and Fas ligand

Patients with systemic lupus erythematosus (SLE) have a tremendously increased risk for cardiovascular disease (CVD), which could not be accounted in entirety by traditional Framingham risk factors. To study whether the accelerated atherosclerosis in SLE patients is mediated by type I interferon (IFN-I) through the regulation of endothelial progenitor cells (EPCs), we created a line of C57BL/6 mice with deficiency in both apolipoprotein E (ApoE−/−) and fas ligand (FasL−/−, gld.). As expected, the resultant gld. ApoE−/− mice exhibited both aggravated lupus-like disease and atherosclerosis under normal diet. Increased expression of IFN-I-stimulated genes (ISGs) was closely associated with depletion and dysfunction of EPCs, as well as with accelerated atherosclerotic lesion in gld. ApoE−/− mice. While only IFN-α instead of other interventions, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IRS423 and IRS661, impaired EPC function in vitro. Mechanistically, activation or inhibition of the TLR7/9 signaling could modulate EPC number and function in vivo. Decreased proliferation rate and increased apoptotic rate of EPCs induced by IFN-α might contribute to the results to a certain extent. Thus, our data suggest that excessive expression of IFN-I through the activation of TLR7/9 signaling may induce accelerated atherosclerosis in lupus through the depletion or dysfunction of EPCs, suggesting that targeting IFN-I might have potential therapeutic effects on both lupus disease and premature atherosclerosis in SLE patients.