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.

Effect of LLLT on endothelial cells culture.

Growth factors as vascular endothelial growth factor (VEGF), produced by the endothelial cells, take an essential part in pathological and physiological angiogenesis. The possibility of angiogenesis modulation by application of laser radiation may contribute to the improvement of its use in this process. Thus, the aim of the study was to investigate the influence of low-level laser therapy (LLLT) on the proliferation of endothelial cells, secretion of VEGF-A and presence of soluble VEGF receptors (sVEGFR-1 and sVEGFR-2) in the medium after in vitro culture. Isolated human umbilical vein endothelial cells (HUVECs) were irradiated using a diode laser at a wavelength of 635 nm and power density of 1,875 mW/cm(2). Depending on radiation energy density, the experiment was conducted in four groups: I 0 J/cm(2) (control group), II 2 J/cm(2), III 4 J/cm(2), and IV 8 J/cm(2). The use of laser radiation wavelength of 635 nm, was associated with a statistically significant increase in proliferation of endothelial cells (p = 0.0041). Moreover, at 635-nm wavelength, all doses of radiation significantly reduced the concentration of sVEGFR-1 (p = 0.0197).

The number of circulating endothelial progenitor cells in healthy individuals--effect of some anthropometric and environmental factors (a pilot study).

PURPOSE: The aim of the study was the evaluation of the number of circulating endothelial progenitor cells (CEPCs) in healthy people and the assessment of the variability of quantitative of CEPCs after 6 weeks. MATERIAL AND METHODS: The study involved 48 healthy individuals; the group consisted of 24 men and 24 women; the mean age of 34. The criterion for the patients' eligibility for the study was the absence of diabetes, thrombosis and cardiovascular diseases such as atherosclerosis, hypertension, and heart failure. Neither did the respondents take any medication that could clearly affect the value of the results. In the whole blood samples the number of circulating endothelial progenitor cells was determined using flow cytometry. During the analysis the fluorescence of 100,000 cells was measured. CEPCs were identified with immunophenotype CD45-, CD31+, CD34+, CD133+. RESULTS: In the study, the median of the number of circulating endothelial progenitor cells in the whole group was 0.41/µL. There was also recorded an increased number of CEPCs after 6 weeks, as compared to the baseline; the difference was significant. There were no differences in the number of CEPCs between the women and the men. There was found no effect on the number of CEPCs factors such as: smoking, physical activity and alcohol consumption. CONCLUSIONS: The study showed that in healthy individuals the gender had no essential effect on the number of endothelial progenitor cells. Based on the demographic and lifestyle data acquired, it is difficult to explain the increase number of CEPCs after 6 weeks.

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.

PAI-1 and α2-AP in patients with morbid obesity.

BACKGROUND: Obesity is a multifactorial, progressing and life-long illness that consists in an exaggerated collection of fatty tissue. In 1997 WHO acknowledged that overweight and obesity had the character of an epidemic in developed countries. Studies show that in Poland morbid obesity was diagnosed in 2.2% of women and in 0.6% of men. Thromboembolic incidents occur very often in people with obesity, especially with morbid obesity. In hypercoagulability, fibrinolysis process decides about the scale of clinical symptoms of disorders of the hemostasis. OBJECTIVES: The aim of this study was to assess the chosen parameters of the fibrinolysis process in patients with BMI crossing 40, classified for surgical treatment of obesity. MATERIAL AND METHODS: The study was conducted in 50 patients with BMI > 40, including 30 women and 20 men. The mean age of the patients was 38.5 years. The control group was made up of 20 healthy volunteers, with a mean age of 38 years. In the blood of both groups the following examinations were performed: concentration of tissue plasminogen activator antigen (t-PA:Ag), antigen of the plasminogen activator inhibitor type-1 (PAI-1:Ag), D-dimers, fibrinogen and plasminogen, activity of the α2-antiplasmin (α2-AP). RESULTS: The conducted study showed that in patients with morbid obesity there was a higher concentration of tPA:Ag, PAI-1:Ag, D-dimers and a higher activity of α2-AP. CONCLUSIONS: The conducted study demonstrates that the activation of the fibrinolysis process appeared after the coagulation process, indicated by an increase in the t-PA:Ag concentration and D-dimers concentration in patients with morbid obesity. The essential growth of PAI-1:Ag level and α2-AP level shows strong inhibition of fibrinolysis in patients.

Endothelial progenitor cells in diabetic foot syndrome.

In the late 20th century endothelial progenitor cells (EPCs) were discovered and identified as cells capable of differentiating into endothelial cells. Antigens characteristic of endothelial cells and hematopoietic cells are located on their surface. EPCs can proliferate, adhere, migrate and have the specific ability to form vascular structure, and they have a wide range of roles: They participate in maintaining hemostasis, and play an important part in the processes of vasculogenesis and angiogenesis. They are sources of angiogenic factors, especially vascular endothelial growth factor (VEGF). EPCs exist in bone marrow, from which they are recruited into circulation in response to specific stimuli. Tissue ischemia is thought to be the strongest inductor of EPC mobilization. Local ischemia accompanies many pathological states, including diabetic foot syndrome (DFS). Impaired angiogenesis--in which EPCs participate--is typical of DFS. An analysis of the available literature indicates that in diabetic patients the number of EPCs declines and their functioning is impaired. Endothelial progenitor cells are crucial to vasculogenesis and angiogenesis during ischemic neovascularization. The pathomechanisms underlying impaired angiogenesis in patients with DFS is complicated, but the discovery of EPCs has shed new light on the pathogenesis of many diseases, including diabetes foot syndrome.