Number Of Endothelial Progenitor Cells Research Articles

FOLATES–RELATED AND BONE MARROW DISORDERS PROMOTE ATRIAL CARDIOMYOPATHY INPATIENTS AFFECTED BY PAROXYSMAL ATRIAL FIBRILLATION VIA BOTH EPCS AND ENDOTHELIAL DYSFUNCTION

Abstract Background Atrial Cardiomyopathy (AC) has been recently postulated to explain those embolic strokes not due to atrial fibrillation (AF) and paradoxical embolism. Pathogenic mechanisms underlying AC are still largely unknown. Folate cycle disorders (FD) are a yet underrated pro–thrombotic dysmetabolism. FD could hinder both endothelial and circulating endothelial progenitor cell (EPCs) functioning, therefore providing one–shot explanation to both atrial fibrosis and endothelial dysfunction (ED). Purpose This study aims to enquire for the hypothesis that: 1) atrial fibrosis (AFib) would relate to FD (intended as both: a)MTHFR C677T inherited mutations and b)bone–marrow function disorders, here referring to erythropoiesis diversions) and 2) AF patients would show dysfunctional EPCs. Methods We studied 59 consecutive patients admitted to the cardiology Unit of the General Hospital“F.Miulli”, with preserved EF, subjected to AF ablation. AFib was quantified in term of percentage of bipolar peak–to–peak voltage points below 0,5mV with respect to the wholeness of picked points. Blood count cell was evaluated at the admission. MTHFR C677T genotypes and serum folate were assessed. EPCs isolation and characterization were performed by cytometry analysis (CD45, CD34, CD133, VEGFR2 and KDR). EPCs functional wound healing assay was performed to determine the percentage of EPCs able to "fix the wound” and compared with matched–controls. Results Baseline characteristics did not differ between Sample and Control groups (Fig. 1 – Left Table). % of Afib significantly differs between C677T MTHFR homozigosis patients (n=15) with respect to non–C677TMTHFR homozygosis patients (n=44) (Fig.1 – Right graph. – p < 0,02). Once univariate analysis was performed, subsequent multivariate analysis highlight highest fit, once merged RBC, RDW–SD and folates values were inputed (Fig.2 – superior graph. – R2=0,39;p=0,0001). Either RBC, RDW–SD and folates coefficient reached significance (p < 0,0001; p < 0,01; p < 0,05 respectively). Number of EPCs signifi cantly differs between AF patients and matched controls (Fig 2 – inferior graph. –p < 0,001). Conclusions Our findings support the hypothesis that genetically determined folates dysmetabolism (MTHFR dysfunction) promotes AFib via a complex cardiac–bone marrow networking involving circulatingEPCs and unraveled by erythropoiesis diversions. Such results suggest a pathogenic role of folate cycle disorders in the AC development.

Patients With Coronary Microvascular Dysfunction Have Less Circulating α-Klotho.

Coronary microvascular dysfunction (CMD) represents an early functional characteristic of coronary vascular aging. Klotho (α-klotho) is a circulating protein inversely linked to physiological aging. We examined low klotho as a potential marker for vascular aging in patients with CMD and no coronary artery disease. Patients undergoing nonurgent angiogram for chest pain who had no coronary artery disease underwent invasive coronary microvascular and endothelial function testing. CMD was defined by ≤50% increase in coronary blood flow (percentage change in coronary blood flow) in response to intracoronary acetylcholine or coronary flow reserve ≤2. Fresh arterial whole blood was used to analyze circulating endothelial progenitor cells with flow cytometry. Stored arterial plasma was used for klotho analysis by ELISA. Participants with CMD (n=62) were compared with those without CMD (n=36). Those with CMD were age 55±10 years (versus 51±11 years; P=0.07) and 73% women (versus 81%; P=0.38). Traditional risk factors for coronary artery disease were similar between groups. Patients with CMD had less klotho (0.88±1.50 versus 1.75±2.38 ng/mL; P=0.03), and the odds of low klotho in CMD were significant in a logistic regression model after adjusting for traditional cardiovascular risk factors (odds ratio [OR], 0.80 [95% CI, 0.636-0.996]; P=0.05). Higher klotho was associated with higher numbers of endothelial progenitor cells with vascular regenerative potential (CD34+ and CD34+CD133+KDR+). Among a subgroup of patients with atherosclerotic cardiovascular disease risk <5% (n=58), CMD remained associated with lower klotho (OR, 0.80 [95% CI, 0.636-0.996]; P=0.047). Klotho may be a biomarker for CMD and may be a therapeutic target for groups of patients without significant traditional cardiovascular risk.

Dulaglutide restores endothelial progenitor cell levels in diabetic mice and mitigates high glucose-induced endothelial injury through SIRT1-mediated mitochondrial fission

Type 2 diabetes mellitus (T2DM) significantly impairs the functionality and number of endothelial progenitor cells (EPCs) and resident endothelial cells, critical for vascular repair and regeneration, exacerbating the risk of vascular complications. GLP-1 receptor agonists, like dulaglutide, have emerged as promising therapeutic agents due to their multifaceted effects, including the enhancement of EPC activity and protection of endothelial cells. This study investigates dulaglutide's effects on peripheral blood levels of CD34+ and CD133+ cells in a mouse model of lower limb ischemia and its protective mechanisms against high-glucose-induced damage in endothelial cells. Results demonstrated that dulaglutide significantly improves blood flow, reduces tissue damage and inflammation in ischemic limbs, and enhances glycemic control. Furthermore, dulaglutide alleviated high-glucose-induced endothelial cell damage, evident from improved tube formation, reduced reactive oxygen species accumulation, and restored endothelial junction integrity. Mechanistically, dulaglutide mitigated mitochondrial fission in endothelial cells under high-glucose conditions, partly through maintaining SIRT1 expression, which is crucial for mitochondrial dynamics. This study reveals the potential of dulaglutide as a therapeutic option for vascular complications in T2DM patients, highlighting its role in improving endothelial function and mitochondrial integrity.

Association between polymorphism in the MTHFR gene and encephaloduroarteriosynangiosis-induced collateral circulation formation.

This study aimed to investigate whether high homocysteine (Hcy) levels associated with the MTHFR gene influence the formation of the collateral vascular network in patients with moyamoya disease (MMD) after encephaloduroarteriosynangiosis (EDAS) by influencing the number of endothelial progenitor cells (EPCs) in peripheral blood. A total of 118 Chinese patients with bilateral primary MMD were prospectively included. Blood samples were collected from the anterior cubital vein before surgery, and MTHFR rs9651118 was genotyped using high-throughput mass spectrometry to determine the genotype of the test specimen. Serum Hcy and EPC levels were measured, the latter with flow cytometry. Digital subtraction angiography was performed 6 months after EDAS, and the formation of collateral circulation was evaluated using the Matsushima grade system. The correlations between MTHFR rs9651118 genotype, Hcy and EPC levels, and Matsushima grade were compared. Among the 118 patients, 53 had the TT genotype (wild type) of MTHFR rs9651118, 33 TC genotype (heterozygous mutation), and 32 CC genotype (homozygous mutation). The mean ± SD Hcy level was 13.4 ± 9.5 μmol/L in TT patients, 9.8 ± 3.2 μmol/L in TC patients, and 8.9 ± 2.9 μmol/L in CC patients (p < 0.001). The level of EPCs in the venous blood of TT patients was 0.039% ± 0.016%, that of TC patients 0.088% ± 0.061%, and that of CC patients 0.103% ± 0.062% (p < 0.001). When the rs9651118 gene locus was mutated, Matsushima grade was better (p < 0.001) but there was no difference between heterozygous and homozygous mutations. The results suggest that the MTHFR rs9651118 polymorphism is a good biomarker for collateral vascular network formation after EDAS in MMD patients.

Transdermal Nicotine Patch Increases the Number and Function of Endothelial Progenitor Cells in Young Healthy Nonsmokers without Adverse Hemodynamic Effects.

Transdermal nicotine patches (TNPs), administering nicotine into the bloodstream through skin, have been widely used as nicotine replacement therapy, and exposure to nicotine can be detected by measurement of plasma cotinine concentration. In animal studies, nicotine treatment could increase the number of endothelial progenitor cells (EPCs), but the effect of TNPs on circulating EPCs and their activity in humans remained unclear. This study aimed to explore the influence of TNPs on circulating EPCs with surface markers of CD34, CD133, and/or KDR, and colony-forming function plus migration activity of early EPCs derived from cultured peripheral blood mononuclear cells before and after TNP treatments in young healthy nonsmokers. In parallel, pulse wave analysis (PWA) was applied to evaluate the vascular effect of TNP treatments. Twenty-one participants (25.8 ± 3.6 years old, 10 males) used TNP (nicotine: 4.2 mg/day) for 7 consecutive days. During the treatment, the CD34+ EPCs progressively increased in number. In addition, the number of EPCs positive for CD34/KDR, CD133, and CD34/CD133 were also increased on day 7 of the treatment. Furthermore, the early EPC colony-forming function and migration activity were increased with the plasma cotinine level positively correlating with change in colony-forming unit number. PWA analyses on day 7, compared with pretreatment, did not show significant change except diastolic pressure time index, which was prolonged and implied potential vascular benefit. In conclusion, 7-day TNP treatments could be a practical strategy to enhance angiogenesis of circulating EPCs to alleviate tissue ischemia without any hemodynamic concern.

Production of angiogenesis mediators and the structure of the vascular wall in the heart in ischemic cardiomyopathy

Background. In the pathogenesis of ischemic cardiomyopathy (ICMP), angiopoiesis remains unexplored.The aim. To describe the vasculature of the heart and the imbalance of angiogenesis mediators in the coronary circulation in association with the number of endothelial progenitor cells (EPC) and desquamated endothelial cells (DEC) in the blood of patients with coronary heart disease (CHD), suffering and not suffering from ICMP.Methods. Fifty-two patients with CHD (30 patients with ICMP, 22 patients without ICMP), 15 healthy donors were examined. The content of EPC (CD14+CD34+VEGFR2+) in the blood from the cubital vein and DEC (CD45–CD146+) in the blood from the coronary sinus and the cubital vein was determined by flow cytometry. The concentrations of VEGF-A (vascular endothelial growth factor A), PDGF (platelet-derived growth factor), and SDF-1 (stromal cell-derived factor 1) in blood plasma were recorded using immunofluorescence assay; the angiopoietin-2, MMP-9 (matrix metallopeptidase 9) were recorded using enzyme immunoassay. In myocardial biopsies the specific area of vessels and the expression of αSMA (smooth muscle alpha-actin) were determined by morphometric and immunohistochemical methods.Results. In the peripheral blood of patients with CHD, regardless of the presence of ICMP, the DEC content exceeded the physiological level, and the VEGF-A, PDGF, angiopoietin-2, and MMP-9 corresponded to the norm. In CHD patients without cardiomyopathy, there was an excess of SDF-1 and EPC in the blood from the cubital vein, and in ICMP, their physiological significance was noted. In the coronary blood flow in patients with CHD without cardiomyopathy, an increase in the concentration of PDGF was found, which was not determined in patients with ICMP, who had an increased content of DEC, angiopoietin-2 and MMP-9. The specific area of the vessels in the patients of the two groups was comparable; the expression of αSMA in ICMP was 6.2 times lower than in patients with CHD without cardiomyopathy.Conclusion. The development of ICMP is accompanied by impaired maturation of vessels in the myocardium, associated with the absence of a compensatory reaction of activation of cellular and humoral factors of angiogenesis.

TLQP-21 facilitates diabetic wound healing by inducing angiogenesis through alleviating high glucose-induced injuries on endothelial progenitor cells.

Diabetes mellitus (DM) is a metabolic disease with multiple complications, including diabetic cutaneous wounds, which lacks effective treating strategies and severely influences the patients' life. Endothelial progenitor cells (EPCs) are reported to participate in maintaining the normal function of blood vessels, which plays a critical role in diabetic wound healing. TLQP-21 is a VGF-derived peptide with promising therapeutic functions on DM. Herein, the protective effects of TLQP-21 on diabetic cutaneous wound and the underlying mechanism will be investigated. Cutaneous wound model was established in T2DM mice, followed by administering 120 nmol/kg and 240 nmol/kg TLQP-21 once a day for 12 days. Decreased wound closure, reduced number of capillaries and EPCs, declined tube formation function of EPCs, and inactivated PI3K/AKT/eNOS signaling in EPCs were observed in T2DM mice, which were sharply alleviated by TLQP-21. Normal EPCs were extracted from mice and stimulated by high glucose (HG), followed by incubated with TLQP-21 in the presence or absence of LY294002, an inhibitor of PI3K. The declined cell viability, increased apoptotic rate, reduced number of migrated cells, declined migration distance, repressed tube formation function, and inactivated PI3K/AKT/eNOS signaling observed in HG-treated EPCs were markedly reversed by TLQP-21, which were dramatically abolished by the co-culture of LY294002. Collectively, TLQP-21 facilitated diabetic wound healing by inducing angiogenesis through alleviating HG-induced injuries on EPCs.

Increasing Physical Tolerance during Cardiac Rehabilitation Helps to Restore Endothelial Function in Patients after Acute Coronary Syndrome

In recent years, many studies have been aimed at exploring the possibilities of cardiac rehabilitation as a tool to improve the prognosis in patients after acute coronary syndrome (ACS). Endothelial dysfunction is one of the initiating mechanisms of cardiovascular diseases, and myocardial infarction in particular, so it is important to assess the dynamics of changes in the number of endothelial progenitor cells (EPCs) in patients during cardiac rehabilitation with the increase in physical activity.&#x0D; The aim. To establish the relationship between the level of recovery of exercise tolerance and the recovery of endothelial function by determining the number of EPCs in patients undergoing cardiac rehabilitation after ACS.&#x0D; Materials and methods. The study included 44 patients with ST-elevation myocardial infarction who underwent urgent stenting of the infarct-related artery, with a mean age of 59 years (Q1-Q3; 51-64). All the study participants underwent laboratory tests (CD45+/CD34+ cell count before and after the exercise test) and instrumental tests (echocardiography, bicycle ergometry, coronary angiography). Statistical processing was carried out using SPSS Statistics 23 (trial version).&#x0D; Results. According to the results of the exercise test at the first examination, the patients were divided into 2 groups: group 1 with low exercise tolerance (≤50 W) and group 2 with high exercise tolerance (&gt;50 W). The data obtained indicate a link between better recovery of exercise tolerance after ACS and recovery of endothelial func-tion in patients with high exercise tolerance during follow-up compared to the patients whose exercise tolerance did not exceed 50 W, as evidenced by a statistically higher number of EPCs after exercise in patients with a favor-able course and high exercise tolerance (3633 vs. 2400 cells/ml) (p=0.006). Patients with low exercise tolerance were more likely to be diagnosed with lesions of left anterior descending coronary artery (96% vs. 70%, p=0.02). More severe coronary vascular lesions with stenosis of 75% of two or more arteries showed lower pre-exercise EPCs, but increased post-exercise EPCs (+228 cells/ml), whereas in the group with stenosis of more than 75% of one vessel, a decrease in post-exercise EPCs (–604 cells/ml) was observed (p=0.004). If patients have more than one stent, there is a 2.5-fold increased risk of decreased exercise tolerance to values of 25-50 W (relative risk = 1.8; 95% confidence interval: 1.3-2.4).&#x0D; Conclusions. The data obtained indicate that there is an association between a better recovery of exercise tolerance after ACS and recovery of endothelial function in patients with a favorable course at repeated examination, compared to patients whose level of exercise tolerance did not exceed 50 W (low exercise tolerance), as evidenced by a statistically greater number of EPCs after exercise test in patients with favorable course and high exercise tolerance compared with patients with unfavorable course and low exercise tolerance.

Endothelial Progenitor Cells: A Brief Update.

An enormous amount of current data has suggested involvement of endothelial progenitor cells (EPCs) in neovasculogenesis in both human and animal models. EPC level is an indicator of possible cardiovascular risk such as Alzheimer disease. EPC therapeutics requires its identification, isolation, differentiation and thus expansion. We approach here the peculiar techniques through current and previous reports available to find the most plausible and fast way of their expansion to be used in therapeutics. We discuss here the techniques for EPCs isolation from different resources like bone marrow and peripheral blood circulation. EPCs have been isolated by methods which used fibronectin plating and addition of various growth factors to culture media. Particularly, the investigations which tried to enhance EPC differentiation while inducing with growth factors and endothelial nitric oxide synthase are shared. We also include the cryopreservation and other storage methods of EPCs for a longer time. Sufficient amount of EPCs are required in transplantation and other therapeutics which signifies their in vitro expansion. We highlight the role of EPCs in transplantation which improved neurogenesis in animal models of ischemic stroke and human with acute cerebral infarct in the brain. Accumulatively, these data suggest the exhilarating route for enhancing EPC number to make their use in the clinic. Finally, we identify the expression of specific biomarkers in EPCs under the influence of growth factors. This review provides a brief overview of factors involved in EPC expansion and transplantation and raises interesting questions at every stage with constructive suggestions.

Regular supervised exercise training improves endothelial function, early atherosclerosis, arterial stiffness and endothelial progenitor cells in PCI patients

Abstract Backgrounds A disrupted balance between endothelial injury and repair induces endothelial dysfunction and endothelial progenitor cells (EPC) augment vascular repair. So, exercise improves endothelial function through increased numbers of EPC. We investigated the effect of training on vascular health by measuring the number of circulating EPC, and assessing flow-mediated dilation (FMD), brachial-ankle pulse wave velocity (baPWV) and carotid intima-media thickness (IMT) in patients with percutaneous coronary intervention (PCI). Methods Fifty-nine patients who were done PCI recently were randomized to full medical treatments or additional 3 months regular supervised exercise program (4 times a week, 1 hr a day). FMD, baPWV, carotid IMT, HOMA, hs-CRP and lipid profiles were measured and circulating CD34/KDR, CD34/CD117 and CD34/CD133 EPC were quantified by flow cytometry at baselines and after 3 months. Results Finally 23 patients joined exercise program(control, N=28). There was no significant difference of baseline characteristics and medication between 2 groups. After 3 months, FMD improved significantly in exercise group (6.62% to 11.48%) compared with control group (7.61% to 8.45%) (P&amp;lt;0.001) (fig 1). Carotid IMT reduced significantly in exercise group (0.759mm to 0.707mm) compared with control group (P=0.015) (fig 2). ba-PWV reduced significantly in exercise group (1529cmm/s to 1455 cmm/s) compared with control group (P=0.016) (fig 3). Numbers of CD34/KDR, CD34/CD117 and CD34/CD133 EPCs increased significantly after exercise compared with control group (P= 0.041, 0.011, 0.022, respectively) (fig 4). We also could see to reduce waist to hip ratio in exercise group compared with control group (P&amp;lt;0.001). Conclusions Three months regular supervised exercise improved FMD, ba-PWV, carotid IMT and waist to hip ratio in PCI patients. and it augmented numbers of circulating EPCs. Our results showed that exercise contributes to additional improvement of vascular health in PCI patients with optimal medical treatments.

Abstract 17537: Myocardial Dysfunction is Associated With Impaired Angiogenesis in Patients With Heart Failure With Preserved Ejection Fraction

Background: Recent evidence suggests that myocardial microvascular damage may represent a key parameter in the pathogenesis of heart failure with preserved ejection fraction (HFpEF). Aim: We sought to investigate a potential correlation between parameters of angiogenesis and myocardial dysfunction in HFpEF patients. Methods: We enrolled 30 consecutive HFpEF patients with NYHA class III, left ventricular ejection fraction (LVEF) &gt;50%, E/e' &gt;15, and NT-proBNP &gt;300 pg/ml. In all patients endothelial progenitor cells (EPCs) were mobilized by granulocyte-colony stimulating factor (G-CSF; 10 mcg/kg, 5 days) and collected via apheresis. Before EPC mobilisation we performed echocardiography and measured plasma levels of biomarkers of angiogenesis. All patients underwent electroanatomical mapping for the measurement of myocardial viability; preserved viability was defined as mean unipolar voltage&gt; 8.3 mV. Results: Myocardial viability was preserved in 11 patients (37%; Group A) and decreased in 19 patients (63%; Group B). The groups did not differ in age (63±10 years in Group A vs. 61±8 years in Group B, P=0.78), gender (male: 73% vs. 79%, P=0.70), serum creatinine (1.07±0.34 mg/dL vs. 1.04±0.40 mg/dL, P=0.43), or LVEF (60±7% vs. 57±4%, P=0.32). In Group B we found higher levels of NT-proBNP (2370±1020 pg/mL vs. 920±455 pg/mL in Group A, P=0.04) and E/e’ (23.2±3.1 vs. 17.4±1.8 pg/ in Group A, P=0.02). After G-CSF stimulation, we found significantly lower numbers of EPCs in Group B (53±42 x10 6 cells/L) than in Group A (102±52 x10 6 cells/L, P=0.001). Similarly, apheresis yielded lower numbers of EPCs in Group B (85±82 x10 6 cells vs. 200±93 x10 6 cells in Group A, P=0.003). When compared to Group A, patients in Group B displayed lower levels of pro-angiogenic biomarkers (agiopoietin-1: 7.4±2.8 ng/mL vs. 12.4±5.1 ng/mL, P=0.02; vascular endothelial growth factor: 27.5±20.1 pg/mL vs. 51.6±24.2 pg/mL, P=0.01), and higher levels of anti-angiogenic biomarkers (endostatin: 56.5±19.1 ng/mL vs. 33.2±14.3 ng/mL, P=0.01; thrombospondin-2: 16.4±4.3 ng/mL vs. 9.2±5.0 ng/mL, P=0.03). Conclusion: In patients with HFpEF, myocardial dysfunction appears to correlate with reduced mobilisation of endothelial progenitor cells and impaired angiogenesis.

Abstract 11795: CXCL5 Inhibition Recovers Neovascularization in Diabetes Mellitus

Introduction: Diabetes mellitus (DM) is a chronic inflammatory disease accompanied by dysfunction of vascular endothelial cells. Vascular complications are the main causes of death in patients with DM. CXCL5 is an inflammatory chemokine that upregulated in patients with type 2 DM. However, its role in diabetic vasculopathy was not clarified. This study aimed to explore the impacts and mechanistic insights of CXCL5 in neovasculogenesis in DM. Methods: Endothelial progenitor cells (EPCs) and human aortic endothelial cells (HAECs) were used in vitro. Streptozotocin-induced diabetic mice and dbdb mice were used as type 1 and type 2 DM models. CXCL5 knockout mice were also used to generate the diabetic animal model. The hindlimb ischemia surgery, matrigel plug assay, and aortic ring assay were conducted. Flow cytometry were used to evaluate the circulating number of EPCs. Results: CXCL5 concentrations were increased in plasma and EPCs culture medium from type 2 DM patients. CXCL5 inhibition by neutralizing antibody upregulated VEGF/SDF-1 expression and improved migration and tube formation abilities in EPCs from type 2 DM patients and high glucose-treated EPCs from non-diabetic subjects as well as HAECs. Intraperitoneal injection of CXCL5 neutralizing antibody increased circulating EPC number, enhanced capillary density, and recovered the blood flow after hindlimb ischemia surgery in both type 1 and type 2 diabetic animals. Protein levels of VEGF and SDF-1 in ischemic muscle were upregulated in the CXCL5 inhibition group. Additionally, diabetic mice injected with CXCL5 neutralizing antibody showed higher hemoglobin contents and neovascular branches in the matrigel plug assay and aortic ring assay, respectively. Importantly, CXCL5 knockout diabetic mice showed improved ischemia-induced angiogenesis and higher expression of angiogenic proteins in ischemic muscle compared with wild-type diabetic mice. Conclusion: CXCL5 inhibition can improve neovasculogenesis via protecting vascular endothelial function and upregulating angiogenic protein expressions in both type 1 and type 2 DM. Further clinical studies are needed to verify that CXCL5 may be a potential therapeutic target for DM vascular complications.

Il ruolo dei progenitori delle cellule endoteliali nel diabete mellito

Diabetes is associated with increased risk of cardiovascular morbidity and mortality. Endothelial progenitor cells (EPCs) are bone marrow-derived cells involved in endothelial health, vasculogenesis and damage repair. Both subjects with type 1 and type 2 diabetes showed a lower number of EPCs and impaired EPC function, suggesting a potential pathogenic role of these cells in both micro and macrovascular complications, as well as in sexual dysfunctions in diabetes. Several glucose-lowering drugs have been reported to potentially improve EPCs levels and bioreactivity. However, larger and longer studies are needed to better explain the mechanisms involved.

Tibial cortex transverse transport potentiates diabetic wound healing via activation of SDF-1/CXCR4 signaling.

The current treatments for diabetic foot ulcers have disadvantages of slow action and numerous complications. Tibial cortex transverse transport (TTT) surgery is an extension of the Ilizarov technique used to treat diabetic foot ulcers, and can shorten the repair time of diabetic foot ulcers. This study assessed the TTT technique for its effectiveness in healing diabetic foot ulcer skin lesions and its related molecular mechanisms. Diabetic rat models were established by injecting healthy Sprague-Dawley rats with streptozotocin (STZ). The effects of TTT surgery on the model rats were assessed by recording changes in body weight, analyzing skin wound pictures, and performing H&E staining to assess the recovery of wounded skin. The numbers of endothelial progenitor cells (EPCs) in peripheral blood were analyzed by flow cytometry, and levels of CXCR4 and SDF-1 expression were qualitatively analyzed by immunofluorescence, immunohistochemistry, qRT-PCR, and western blotting. Both the histological results and foot wound pictures indicated that TTT promoted diabetic wound healing. Flow cytometry results showed that TTT increased the numbers of EPCs in peripheral blood as determined by CD34 and CD133 expression. In addition, activation of the SDF-1/CXCR4 signaling pathway and an accumulation of EPCs were observed in skin ulcers sites after TTT surgery. Finally, the levels of SDF-1 and CXCR4 mRNA and protein expression in the TTT group were higher than those in a blank or fixator group. TTT promoted skin wound healing in diabetic foot ulcers possibly by activating the SDF-1/CXCR4 signaling pathway.

Endothelial Progenitor Cells in Autoimmune Disorders.

Circulating endothelial progenitor cells (EPCs) were first described in 1997 by Asahara et al. as "putative endothelial cells" from human peripheral blood. The study of endothelial progenitors is also intensifying in several pathologies associated with endothelial damage, including diabetes, myocardial infarction, sepsis, pulmonary arterial hypertension, obstructive bronchopneumopathy and transplantation. EPCs have been studied in several autoimmune diseases with endothelial involvement such as systemic lupus erythematosus, thrombotic thrombocytopenic purpura, antineutrophil cytoplasmic antibodies, vasculitis, rheumatoid arthritis, Goujerot-Sjögren and antiphospholipid syndrome. Factors involved in endothelial damage are due to overexpression of pro-inflammatory cytokines and/or autoantibodies. Management of these pathologies, particularly the long-term use of glucocorticoids and methotrexate, promote atherosclerosis. A lack of standardized assessment of the number and function of EPCs represents a serious challenge for the use of EPCs as prognostic markers of cardiovascular diseases (CVD). The objective of this review was to describe EPCs, their properties and their involvement in several autoimmune diseases.

Abstract P1070: In Vivo Autotransplantation Of Spleen-derived Endothelial Progenitor Cells To Promote Angiogenesis In Ischemic Sites

Since mononuclear cells were first illuminated as potential endothelial progenitor cells (EPCs), bone marrow (BM) was extracted and processed as BM-derived EPCs for autotransplantation to the ischemic sites in previous studies. Still, numerous clinical trials based on this strategy showed only marginal effects, and the small number of BM-derived EPCs was suggested as a possible limitation. Meanwhile, spleen contains abundant mononuclear cells which can inherently target the ischemic sites, and these cells can be primed by hypoxia. If only they can be primed in vivo, spleen-derived EPCs can be inherently autotransplanted to ischemic sites with various advantages such as plentiful cell number, non-invasive extraction and delivery of these cells. In this study, splenic mononuclear cells were utilized to promote angiogenesis in ischemic sites. In vivo priming of splenic mononuclear cells, their inherent targeting and therapeutic effect on ischemic sites were sequentially explored towards the 'in vivo autotransplantation'. For hypoxic priming, the effect of hypoxia and hypoxic-mimetic agent (cobalt chloride) on isolated splenic mononuclear cells were comparable in both priming methods, regarding the morphology changes and the increase in expression of EPC markers. In align with these in vitro results, both hypoxic-mimetic agent-loaded nanoparticle delivery to spleen and splenic artery ligation resulted in increased expression of EPC markers in mice spleen. Moreover, the proportion and actual numbers of the cells presenting EPC marker were significantly higher in primed splenic cells than in BM cells. After the priming, these primed cells not only maintained the inherent targeting ability but also gained the ability to promote angiogenesis. Consequently, these primed cells presented therapeutic effects as evidenced by higher expression of angiogenic markers with enhanced blood flow recovery in mouse hindlimb ischemia model, compared with untreated or BM-derived EPCs' autotransplantation for conventional strategy. For EPC-based treatment of ischemia, 'In vivo autotransplantation' is simple and non-invasive strategy which can overcome the previous limitations of insufficient cell numbers.

Re-endothelialization of Decellularized Scaffolds With Endothelial Progenitor Cell Capturing Aptamer: A New Strategy for Tissue-Engineered Heart Valve.

Tissue-engineered heart valve (TEHV) is a promising alternative to current heart valve substitute. Decellularized porcine aortic heart valves (DAVs) are the most common scaffolds of TEHV. Hard to endothelialization is one of the disadvantages of DAVs. Therefore, we aimed to immobilize endothelial progenitor cell (EPC)-aptamer onto DAVs for accelerating endothelialization. In this study, three groups of scaffolds were constructed: DAVs, aptamer-immobilized DAVs (aptamer-DAVs), and glutaraldehyde crosslinked DAVs (GA-DAVs). The results of flow cytometry revealed that EPC-aptamer was specific to EPCs and was immobilized onto DAVs. Cells adhesion experiments demonstrated that EPCs adhered more tightly onto aptamer-DAVs group than other two groups of scaffolds. And cell proliferation assay indicated that EPCs seeded onto aptamer-DAVs group grew faster than DAVs group and GA-DAVs group. Moreover, dynamic capture experiment in flow conditions revealed that the number of EPCs captured by aptamer-DAVs group was more than other two groups. In conclusion, aptamer-DAVs could specifically promote adhesion and proliferation of EPCs and had ability to capture EPCs in simulated flow condition. This could promote re-endothelialization of scaffolds.

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.

LncRNA NEAT-2 regulate the function of endothelial progenitor cells in experimental Sepsis model.

Sepsis is a life-threatening disease with a limited effectiveness and the potential mechanism remains unclear. LncRNA NEAT-2 is reported to be involved in the regulation of cardiovascular disease. This study aimed to investigate the function of NEAT-2 in sepsis. We built sepsis animal model with Male Balb/C mice induced by cecal ligation and puncture (CLP). A total of 54 mice were randomly assigned into eight groups: sham operation group (n = 18), CLP group (n = 18), CLP plus si-control group (n = 3), CLP plus si-NEAT2 group (n = 3), CLP plus mimic control group (n = 3), CLP plus miR-320 group (n = 3), CLP plus normal saline group (n = 3), and normal control group (n = 3). The number of peripheral endothelial progenitor cells (EPCs), the expression level of NEAT-2 and miR-320 were detected during progression of sepsis, as well as the number of peripheral EPCs and level of TNF-α, IL-6, VEGF, ALT, AST and Cr. In addition, the function of EPCs was evaluated after NEAT-2 knockdown and miR-320 overexpression in vitro. The number of circulating EPCs increased significantly in sepsis. NEAT-2 expression was significantly increased in the progress of sepsis, accompanied with miR-320 downregulated. NEAT-2 knockdown and miR-320 overexpression attenuated hepatorenal function and increased cytokines in sepsis. Moreover, NEAT-2 knockdown and miR-320 overexpression decreased the proliferation, migration and angiogenesis of endothelial progenitor cells in vitro. LncRNA-NEAT2 regulated the number and function of endothelial progenitor cells via miR-320 in sepsis, which may contribute to the development of novel potential clinical therapy for sepsis.

Exploration of the influence of early rehabilitation training on circulating endothelial progenitor cell mobilization in patients with acute ischemic stroke and its related mechanism under a lightweight artificial intelligence algorithm.

This work aimed to explore the application of lightweight artificial intelligence algorithms in magnetic resonance imaging (MRI) image processing of patients with acute ischemic stroke (AIS) to clarify the effect and mechanism of early rehabilitation training on the mobilization of circulating endothelial progenitor cells (EPCs) in AIS. A total of 98 AIS patients undergoing MRI examination were selected as the research objects and were randomly divided into a rehabilitation group (early rehabilitation training, 50 cases) and a routine group (conventional treatment, 48 cases) by random number table method and lottery method. In this work, based on the convolutional neural network (CNN) algorithm, a low-rank decomposition algorithm was introduced to optimize it, and a lightweight MRI image computer intelligent segmentation model (LT-RCNN) was established. The LT-RCNN model was used in the MRI image processing of AIS patients, and the role of the model in AIS image segmentation and lesion localization was analyzed. Furthermore, flow cytometry was used to detect the number of peripheral circulating EPCs and CD34+KDR+ cells in the two groups of patients before and after treatment. The serum levels of vascular endothelial growth factor (VEGF), tumor necrosis factor-α (TNF-α), interleukin 10 (IL-10), and stromal cell-derived factor-1α (SDF-1α) content were detected by Enzyme-Linked Immunosorbent Assay (ELISA). In addition, the correlation between each factor and CD34+KDR+ was analyzed by Pearson linear correlation. The diffusion-weighted imaging (DWI) signal of MRI images of AIS patients under the LT-RCNN model was high. The location of the lesion could be accurately detected, and the contour of the lesion could be displayed and segmented, and the segmentation accuracy and sensitivity were significantly better than before optimization. The number of EPCs and CD34+KDR+ cells in the rehabilitation group was increased compared with the control group (p<0.01); the expression levels of VEGF, IL-10, and SDF-1α were higher than those of the control group (p<0.001), and TNF-α content was lower than the control group (p<0.001). The number of CD34+KDR+ cells was positively correlated with VEGF, IL-10, and TNF-α contents (p<0.01). The results showed that the computer-intelligent segmentation model LT-RCNN could accurately locate, and segment AIS lesions and the early rehabilitation training could change the expression level of inflammatory factors and further promote the mobilization of AIS circulation EPCs.