Arleta Kulwas

Circulating endothelial progenitor cells and angiogenic factors in diabetes complicated diabetic foot and without foot complications

INTRODUCTION Data about angiogenic factors in diabetic foot syndrome (DFS) are insufficient. Therefore, in the present study we focus on circulating endothelial progenitor cells (EPCs) and two major angiogenic factors: vascular endothelial growth factor (VEGF-A) and fibroblast growth factor (FGF-2) in patients with DFS. MATERIALS AND METHODS We included 75 subjects: 45 patients with type 2 diabetes and 30 controls. The study group was divided into 2 subgroups: 23 patients with diabetic foot and 22 patients without diabetic complications. The concentration of VEGF-A, soluble VEGF receptor 2 (sVEGF-R2) and FGF-2 were measured in plasma samples. The number of circulating EPCs was determined in peripheral venous blood. The number of endothelial progenitor cells was measured with FACSCalibur flow cytometer using monoclonal antibodies directed against antigens specific for EPCs. RESULTS In our study we observed significant higher levels of VEGF-A and FGF-2 and lower sVEGF-R2 concentration in patients with T2DM compared to healthy subjects. The conducted analysis showed decreased levels of VEGF-A and elevated levels of FGF-2 in patients with DM complicated DFS compared to diabetic patients without DFS. Increased circulating EPCs number was reported in patients with DFS, and the difference was almost statistically significant. CONCLUSIONS The high concentration of VEGF-A and FGF-2, and a positive correlation between them indicate their participation in the process of angiogenesis in T2DM. Decreased sVEGF-R2 may result from inactivation of VEGF-A during complexes formation.

Adiponectin and endothelial markers in postmenopausal women taking oral or transdermal hormone therapy

To assess the concentration of adiponectin, soluble E‐selectin, soluble thrombomodulin and tissue activator plasminogen antigen in postmenopausal women who received oral or transdermal hormone therapy.

Assessment of ghrelin and leptin receptor levels in postmenopausal women who received oral or transdermal menopausal hormonal therapy

ObjectiveIn postmenopausal women, an increased leptin concentration and reduced levels of ghrelin and adiponectin were observed. The aim of this study was to evaluate the concentrations of the active form of ghrelin, total ghrelin, leptin receptor, lipoprotein(a) (Lp(a)), and plasminogen activator inhibitor type 1 (PAI-1) in postmenopausal women who received oral or transdermal menopausal hormonal therapy (MHT).MethodsThe study involved 76 healthy women: 46 women aged from 44 to 58 years who received oral (26) or transdermal (20) MHT; the control group consisted of 30 women aged from 44 to 54 years who did not receive MHT. The plasma concentrations of total ghrelin, the active form of ghrelin, Lp(a), and PAI-1:Ag were measured by enzyme-linked immunosorbent assay (ELISA). The concentration of the leptin receptor was measured by enzyme immunometric assay (EIA).ResultsWe observed a significantly higher concentration of total ghrelin and the active form of ghrelin in women who received transdermal MHT in comparison with those who took oral MHT. We also found a significantly lower concentration of total ghrelin in women who received oral MHT compared with the control group. A higher concentration of PAI-1:Ag was found in the group of women who took transdermal MHT in comparison with those who took oral MHT and with the control group. The differences were statistically significant. Additionally, we found a significant negative correlation between the concentrations of total ghrelin and PAI-1:Ag and a positive correlation between the concentrations of total ghrelin and leptin receptor in women who received transdermal MHT.ConclusionsThe study showed that women who used transdermal MHT had higher levels of total ghrelin than women who took oral MHT. This indicates a beneficial effect of the transdermal route of MHT. However, transdermal therapy was associated with adverse effects with regard to the observed higher levels of PAI-1:Ag, which in turn, can lead to a reduction in fibrinolytic activity.

VEGF-A and PDGF-BB--angiogenic factors and the stage of diabetic foot syndrome advancement.

INTRODUCTION In patients with diabetic foot syndrome (DFS), an inadequate angiogenic response is observed. The aim of this study was to evaluate the concentrations of VEGF-A, PDGF-BB, sVEGF-R2 and sVEGF-R1 in patients with diabetes-complicated diabetic foot syndrome and analyse them using selected clinical data. MATERIAL AND METHODS Forty seven diabetic patients, 25 women mean age 63 and 20 men mean age 60.5, with diabetic foot syndrome (DFS) were enrolled in the experimental group. To evaluate angiogenesis factors depending on Wagner grade, the subjects were divided into three subgroups: I - patients with 0 Wagner grade (n = 14); II - patients with 1,2,3 Wagner grades (n = 15); and III - patients with 4,5 Wagner grades (n = 18). The control group consisted of 20 healthy volunteers. The material for research was blood. RESULTS Significantly higher levels of VEGF-A and PDGF-BB in the DFS cases compared to controls were observed (VEGF-A p = 0.000001; PDGF-BB p = 0.000051). Analysis of angiogenic parameters according to the stage of diabetic foot syndrome advancement showed higher VEGF-A level (I: p = 0.000867; II: p = 0.001827; III: p = 0.000024) and PDGF-BB (respectively p = 0.004113, p = 0.004224, p = 0.002480) in all the subgroups. Decreased sVEGF-R2 concentrations were observed in the I (p = 0.054) subgroup and the III (p = 0.03524) subgroup. In this study, a strong positive correlation between VEGF-A and PDGF-BB was observed (R = 0.66; p = 0.000001). CONCLUSIONS Our study revealed that proangiogenic factor levels were increased in DFS. This is associated with lower limb ischaemia and hypoxic conditions. The stage of diabetic foot syndrome advancement influenced VEGF-A and PDGF-BB concentrations.

Tissue plasminogen activator (t-PA) and plasminogen activator inhibitor type 1 (PAI-1) in diabetic foot syndrome

PURPOSE The aim was to evaluate tissue plasminogen activator (tPA) and plasminogen activator inhibitor type 1 (PAI-1) concentration using enzyme linked immunosorbent assay method (ELISA) in diabetic foot syndrome (DFS) as compared to a group of healthy people and patients with diabetes mellitus without symptomatic vascular complications (DM2T). MATERIAL/METHODS Venous blood samples were collected from 90 patients with type 2 diabetes mellitus (30 - DM2T; 60 - DFS). Age-matched controls were also included (n=30). tPA and PAI-1 plasma levels were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS We found a significantly lower concentration of tPA:Ag in patients with DFS in comparison to the DM2T group; tPA concentrations were significantly higher in DM2T as compared to the control group. We observed significantly lower concentration of PAI-1:Ag in DF patients treated for hypertension as compared to patients without hypertension. The tPA:Ag and PAI-1:Ag concentration analysis in DFS depending on age, gender and BMI did not show any significant differences. CONCLUSIONS A lower concentration of tPA in patients with DFS may be associated with damage to the endothelial cells, especially in the microvasculature, and the sympathetic nervous system.