Vascular Endothelial Growth Factor Concentrations Research Articles

Abstract 14789: Vascular Growth Factors and Their Association With Arterial Stiffness in the 1986 Jamaica Birth Cohort: A Longitudinal Analysis

Introduction: Vascular growth factors (VasGFs) and increased arterial stiffness provide an early indication of structural and functional blood vessel changes that lead to cardiovascular disease. We aimed to determine if there was an association between serum levels of vascular endothelial growth factor (VEGF), epidermal growth factor (EFG), angiopoietin 1 (Angp1) and angiopoietin 2 (Angp2) with arterial stiffness in 28-30-year-old adults from a birth cohort. Methods: Participants were a subset of members of a longitudinal study of children born in Jamaica between September and October 1986. Sociodemographic and anthropometric measurements were taken at the 18-20 and 28-30 follow-up visits, while serum was collected at the 18-20 year old visit. The pulse wave velocity (Arteriograph TM ) was used to measure arterial stiffness at the 28-30-year-old visit. Analyses were limited to participants who had pulse wave velocity measurements. Multiple imputation using chained equations was done to account for missing data on the explanatory variables. Linear regression models were used to explore bivariate associations. The final multivariable model included VEGF, EGF, Angp1, Angp2, age, sex, BMI, SBP, DBP, smoking and education. Results: Analyses included 387 participants (47% male). At 18-20yrs median serum concentrations of VEGF 75.4 pg/ml; EGF 951.4 pg/ml; Angp1 67863.7 pg/ml; and Angp2 691 pg/ml. At 28-30 there were sex differences in BMI (M:25.2 vs F:28.8kg/m 2 ), SBP (M:114 vs F:108 mmHg), current smokers (M:29% vs F: 5.4%) tertiary education (M:22% vs F:39%) and arterial stiffness (M:6.4 vs F:6.8m/s). In the final multivariable model both VEGF and EGF were associated with arterial stiffness. VEGF coef (95%CI) 0.095 (0.014, 0.176) p= 0.022 and EGF -0.087 (-0.169, -0.004) p =0.039. There was no association between angp1 or angp2 and pulse wave velocity. Conclusion: VEGF concentrations at 18-20 of was positively associated with arterial stiffness at 28-30 years old, while and EGF showed inverse association. VEGF and EGF may be useful biomarkers of vascular function in young adults.

Medical Cannabis Use and Inflammatory Cytokines and Chemokines Among Adult Chronic Pain Patients.

Background: Utilizing cannabis as a therapeutic option for chronic pain (CP) has increased significantly. However, data regarding the potential immunomodulatory effects of cannabis in CP patients remain scarce. We aimed at exploring the relationship between cannabis use and inflammatory cytokines and chemokines among a cohort of CP patients. Methods: Adult patients with a CP diagnosis and medical authorization of cannabis were enrolled. Patients completed validated clinical questionnaires and self-reported the effectiveness of cannabis for symptom management. Patients' blood and cannabis samples were analyzed for the presence of four major cannabinoids, two major cannabinoid metabolites, 29 different cytokines/chemokines, and cortisol. The multivariable linear regression model was used to identify cannabis and patient factors associated with immune markers. Results: Fifty-six patients (48±15 years; 64% females) were included, with dried cannabis (53%) being the most common type of cannabis consumed. Seventy percent of products were considered delta-9-tetrahydrocannabinol (Δ9-THC)-dominant. The majority of patients (96%) self-reported effective pain management, and 76% reported a significant decrease in analgesic medication usage (p≤0.001). Compared with males, female patients had higher plasma levels of cannabidiol (CBD), cannabidiolic acid, Δ9-THC, and 11-hydroxy-Δ9-tetrahydrocannabinol but lower concentrations of delta-9-tetrahydrocannabinolic acid and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH). Females had significantly lower eotaxin levels (p=0.04) in comparison to male patients. The regression analysis indicated that high cannabis doses were related to increased levels of interleukin (IL)-12p40 (p=0.02) and IL-6 (p=0.01), whereas female sex was associated with decreased eotaxin (p≤0.01) concentrations. Blood CBD levels were associated with lower vascular endothelial growth factor (p=0.04) concentrations, and THC-COOH was a factor related to decreased tumor necrosis factor alpha (p=0.02) and IL-12p70 (p=0.03). Conclusion: This study provides further support for the patient-perceived effectiveness of cannabis in managing CP symptoms and reducing analgesic medication consumption. The results suggest a potential sex difference in metabolizing cannabinoids, and the varying immune marker concentrations may support a possible immunomodulatory effect associated with patient sex and cannabis product type. These preliminary findings provide grounds for further validation using larger, well-designed studies with longer follow-up periods.

Peculiarities of serum levels of vascular growth factor and its receptors in dialysis patients

Recent research has shown that VEGF may influence the condition and survival of patients with chronic kidney disease (CKD), including those treated with dialysis procedures. Identifying the value of this factor is important to uncover the mechanisms of CKD progression and improve treatment strategies.
 The present study aimed to determine the mean serum levels of vascular endothelial growth factor (VEGF) and its receptor (VEGF-R) and their correlations with pro-inflammatory cytokines (TNF-α, MCP-1, IL -6, IL -8) in dialysis patients stratified by dialysis modality and Carnivit use.
 Methods. In this prospective cohort study, serum levels of VEGF, VEGF-R, and proinflammatory cytokines were determined in 41 hemodialysis (HD) patients (group 1) and 14 peritoneal dialysis (PD) patients (group 2), 28 of whom were treated with Carnivit. The assay was performed with the enzyme immunoassay "SunRise TouchScreen" using IBL International (Germany) test systems. The limits of normal values (reference range) were determined based on the results of a study with 20 conditionally healthy individuals.
 Results. All included patients had significantly higher serum levels of VEGF compared to healthy donors (p=0.006); a significant increase in this mediator was observed in both HD and PD treated subjects (p=0.002 and p=0.007, respectively). Carnivit treatment resulted in a significant decrease (p=0.023) in this mediator to normal levels in all patients. Subgroup analysis showed a significant decrease in VEGF (p=0.016) and VEGF-R (p=0.005) in HD patients after therapy. Mean concentrations of growth factor in HD patients (p=0.098) and PD patients (p=0.160) did not differ from those of healthy subjects.
 A positive correlation was observed between serum levels of VEGF and its receptors in the studied patients both before and after treatment (p < 0.0001).
 A direct correlation between IL -8 and VEGF and VEGF-R was observed in HD patients and an inverse correlation between MCP-1 and VEGF-R was observed in PD patients. After Carnivit treatment, the correlation between serum levels of IL -8 and VEGF-R (0.782, p=0.01) and between VEGF and VEGF-R (0.770, p=0.01) remained in group 1 and between VEGF and VEGF-R (0.829, p=0.03) in group 2.
 Conclusion. Increased concentrations of VEGF and its receptors were detected in the blood of dialysis patients, with a positive correlation between them and the pro-inflammatory IL -8. Treatment with Carnivit resulted in a significant decrease in cytokines with the maintenance of the balance between vascular growth factor and its receptors, which we consider a positive effect for the inhibition of inflammation with the progression of endothelial dysfunction.

Effect of triamcinolone acetonide on retinal inflammation and angiogenesis induced by pericyte depletion in mouse

Triamcinolone acetonide (TA) has been shown to improve morphological and functional outcome in diabetic macular edema (DME) patients. However, the functional mechanism of TA has not been elucidated yet. In this study we investigated the detailed functional mechanism of TA using culture cells and retinopathy mouse models in which retinal inflammation and abnormal angiogenesis were induced by pericyte depletion. TA significantly prevented retinal hemorrhage, edema and partially improved abnormal angiogenesis. TA decreased retinal vascular endothelial growth factor (VEGF) concentration, presumably by preventing recruitment of macrophages into retina and TA also inhibited expression of inflammatory cytokines in retina. TA inhibited proliferation/migration of vascular endothelial cells and vessel sprouting. No direct inhibition of VEGF receptor 2 (VEGFR2) autophosphorylation was observed by TA. These results suggested that TA improved inflammatory retinal events which were induced in pericyte-deleted mice by mainly decreasing macrophage-derived VEGF and expression of inflammatory cytokines followed by attenuation of vascular permeability and proliferation/migration of endothelial cells. Furthermore, in these processes, translocation of glucocorticoid receptor (GR) was partially involved.

Immunological detection of Platelet-derived growth factor and Vascular endothelial growth factor in Iraqi women with Polycystic Ovarian Syndrome

Background: platelet-derived growth factor (PDGF) signaling controls a wide range of the mesenchymal cell origin lineages throughout the development and in the females with the polycystic ovarian syndrome. Vascular endothelial growth factor (VEGF) has an important impact on vascularization of the tissues generally, and in pathogenesis of a wide range of the illnesses. The PCOS has been found related to high VEGFexpression levels. Aims and Objectives: for the detection and studying of immunological effects of PDGF and VEGF in the Iraqi females with the PCOS. Materials and Methods: 250 Iraqi females, aged between 20 and 50 years have been divided to twogroups, which are: study group (n=125, PCOS) and control group (n=125, which mainly consisted from normal females), the samples of the blood have been obtained in 2ndmenstruate cycle day, VEGF and PDGF concentrations have been estimated for every one of the groups. Results: the PCOS patients have shown a considerable difference in comparison to the other group. The level has been 71.22±65.23Pg/ml and 243± 153.44Pg/ml, for the controls and PCOS patients respectively. In addition to that, the PCOS patients have shown a considerable difference in comparison to the other group. . The level has been 71.22±65.23Pg/ml and 243± 153.44Pg /ml, for the controls and PCOS patients respectively. Conclusions: Increases concentration of PDGF and VEGF in PCOS women related to ovulation lack and that lead to increases the risk of PCOS appearance.

Optimal timing of preoperative intravitreal anti-VEGF injection for proliferative diabetic retinopathy patients.

To analyze concentrations of vascular endothelial growth factor (VEGF) and fibrosis-related factors in vitreous fluid of proliferative diabetic retinopathy (PDR) patients pre-treated with intravitreal anti-VEGF injections (IVI) at different time periods prior to pars plana vitrectomy (PPV), and their correlation with the degree of vitreoretinal fibrosis and explore the optimal timing of preoperative IVI. The prospective case-control study from January 2019 to July 2020 included 31 eyes with PDR-related complications (PDR group) and 21 eyes with non-diabetic ocular disease (control group) requiring PPV. PDR eyes were divided into four groups based on timing of PPV: 3d after IVI (3-day group); 5d after IVI (5-day group); 7 or more days after IVI (≥7-day group); and no IVI. Vitreous fluid samples (0.5-1.0 mL) were collected prior to switching on the infusion before routine 23-G PPV. Concentrations of VEGF, basic fibroblast growth factor (bFGF), periostin (PN), interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α were measured by immunoassay, and concentration differences for each cytokine were compared among the groups. The degree of vitreoretinal fibrosis was graded intraoperatively, and the correlation between the changes in cytokine levels and the severity of vitreoretinal fibrosis was analyzed by univariate ordinal logistic regression analysis. PDR eyes without IVI had significantly higher VEGF, bFGF, PN, and IL-6 concentrations than non-diabetic eyes (all P<0.05), and had a significantly higher concentration of VEGF (P<0.05) and a significantly lower concentration of IL-8 (P<0.05) than PDR eyes with IVI. Statistically significant differences were also observed for concentrations of VEGF, bFGF, PN, IL-6, and IL-8 among 3-day, 5-day, and ≥7-day groups (all P<0.05); meanwhile there was no significant difference in TNF-α among groups (P=0.226). The 5-day group had the lowest concentration of VEGF and the ≥7-day group had the highest concentration of bFGF and PN. The degree of vitreoretinal fibrosis was significantly higher in the ≥7-day group compared to the 3-day (P=0.015) and 5-day group (P=0.039), and vitreoretinal fibrosis correlated significantly with concentrations of bFGF, PN, IL-6, and IL-8 (all P<0.05). Univariate ordinal logistic regression analysis showed that bFGF was an independent risk factor for the severity of vitreoretinal fibrosis in PDR patients pre-treated with IVI. The vitreous concentrations of VEGF, bFGF, PN, IL-6, and IL-8 change after pretreatment with IVI before PPV in PDR patients. The degree of vitreoretinal fibrosis is higher in patients with a longer time between IVI treatment and PPV, which may be related to the angio-fibrosis switch. The results suggest that PPV should be performed 5d after IVI administration in PDR patients.

Estimation of the concentration of some biomarkers in serum of women with rheumatoid arthritis

The current study was conducted on women with rheumatoid arthritis (RA) in Basrah city, Iraq. The study aimed to evaluate the relationship between some biomarkers such as adiponectin hormone, anti-citrullinated protein antibody (ACPA), acute phase C-reactive protein (CRP) and vascular endothelial growth factor (VEGF) in the serum of female patients. In this study, 80 serum samples were collected from women and were distributed as follows: 56 serum samples from women with RA and 24 serum samples from non-infected women who were considered as a healthy group. The patient samples were divided into three categories depending on age, disease severity and duration. The samples were obtained from Basrah General Hospital. The results of the current study showed a significant increase in the concentration of adiponectin, ACPA, CRP and VEGF in RA patients compared to healthy subjects. In addition, there were significant differences in the concentration of these biomarkers between some of the three age groups and between the disease of severity and duration. The study concluded that all the biomarkers under investigation have increased their concentrations in the serum of women suffering from RA.

VEGF Polymorphism rs3025039 and Human T-Cell Leukemia Virus 1 (HTLV-1) Infection among Older Japanese Individuals: A Cross-Sectional Study.

Previous studies have reported a close correlation between vascular endothelial growth factor (VEGF), which plays an important role in angiogenesis, and human T-cell leukemia virus 1 (HTLV-1). However, an association between genetic characteristics related to VEGF and HTLV-1 infection has not yet been reported. Because the VEGF polymorphism rs3025039 is inversely associated with serum concentrations of VEGF, we focus on rs3025039 in the present study. To clarify the association between the VEGF polymorphism rs3025039 and HTLV-1 infection, a cross-sectional study of 1924 Japanese individuals aged 60–79 years who participated in general health check-ups was conducted. Using logistic regression, odds ratios (ORs) and 95% confidence intervals (CIs) for HTLV-1 infection in relation to rs3025039 genotype were calculated with adjustment for known confounders. Compared with rs3025039 CC-homozygotes, (T) allele carriers had a significantly lower OR for HTLV-1 infection. The adjusted OR and 95% CI for HTLV-1 infection was 0.70 (0.54–0.91) (p = 0.009). Genetic characteristics related to lower angiogenesis activity might be associated with a lower chance of establishing HTLV-1 infection. Although further investigation is necessary, angiogenesis might play a crucial role in the establishment of HTLV-1 infection.

Effect of pan-retinal laser photocoagulation on intravitreal vascular endothelial growth factor concentration in proliferative diabetic retinopathy.

Objective: This study aimed to determine the intravitreal concentration of VEGF in eyes with PDR and to evaluate the effects of previous PRP on its level. Methods: It was a cross-sectional study performed at the Clinical Centre University. It included 90 eyes surgically treated with PPV, divided into three groups, group A - patients with PDR with previous PRP, group B - patients with PDR without previous PRP, and group C - PPV performed due to the indication unrelated to diabetes. A vitreous sample was obtained during PPV, and the VEGF concentration was determined using an Enzyme-linked immunosorbent assay test (ELISA). Shapiro-Wilk, nonparametric tests Kruskal-Wallis, Mann-Whithney U test, ANOVA and Spearman's correlation test were used. Results: The highest vitreous VEGF concentration was in group B - 972.96 (743.33-1149.13) and was higher than in group A - 69.22 (37.33-225.15) and in group C - 19.93 (1.15-32.17) (p<0.001). There was a positive correlation between VEGF vitreous concentration and glucose level in group A patients (Rho=0.410; p=0.027). Conclusion: As a treatment before PPV surgery, PRP showed to be effective in the reduction of VEGF levels, which also highlighted a decrease in complications during and postoperatively. Abbreviations: DRS = Diabetic Retinopathy Study, PDR = proliferative diabetic retinopathy, VEGF = vascular endothelial growth factor, PRP = pan-retinal photocoagulation, PPV = pars plana vitrectomy, HbA1c = glycosylated hemoglobin, ETDRS = Early treatment diabetic retinopathy study, ESR = erythrocyte sedimentation rate, BCVA = best corrected visual acuity, OCT = optical coherent tomography, ILM = internal limiting membrane, PHACO = phacoemulsification, IOL = intraocular lens, ELISA = Enzyme-linked immunosorbent assay test, AUC = area under the curve, DME = diabetic macular oedema, TDR = tractional retinal detachment, VMT = vitreomacular traction.

Association of Serum Vascular Endothelial Growth Factor Levels with Nephropathy and Retinopathy in Type 2 Diabetic Patients

Introduction: Vascular endothelial growth factor (VEGF), protein originally referred to as vascular permeability factor, maybe a signal protein produced by cells that stimulate the formation of blood vessels and a sub-family of growth factors. They are important signaling proteins engaged in both vasculogenesis and angiogenesis. It is a part of the system that restores the oxygen supply to tissues when blood circulation is insufficient like in hypoxic conditions. The serum concentration of VEGF is high in asthma and DM. The aim of this study to explore the relevance of VEGF level with the complication of diabetes mellitus (diabetic retinopathy [DR] and diabetic nephropathy (DN)). Methodology: Case–control study carried out in Khartoum–Sudan from August 2018 to August 2020. One hundred and four patients divided into four groups, 3 ml of blood were collected from each subject in sterile plain vacationer tube (3 ml) then was centrifuged, and the serum was stored at −20°C to measure VEGF level by using ELISA kits (E-EL-H0111, 96T). Results: We have confirmed previous observations of increased plasma VEGF Levels in patients with DR and DN. A total of 103 quality controls divided into four groups (diabetics with DR, diabetics with DN diabetics without DR or DN, and health individuals) were observed to determine the relationship between serum of VEGF levels and diabetes complication DR and DN. Conclusions: High level of VEGF plays a major role in the pathogenesis of DR and DN.

Development of surface modified nanoparticles of curcumin for topical treatment of diabetic retinopathy: In vitro, ex vivo and in vivo investigation

Diabetic retinopathy (DR) is the worst microvascular complication of diabetes that results in blindness. Present treatment of DR is invasive and associated with pain and inflammation. There is a need to develop painless treatment. The surface-modified PLGA (50:50) nanoparticles of Curcumin, which is a peroxisome proliferator-activated receptor-gamma agonist, were prepared to target DR by topical administration. Nanoparticles were optimized by a 32 factorial design. The Concentration of PLGA and number of HPH cycles at fixed pressure were independent variables whereas particle size. PDI and % entrapment efficiency were the dependent variables. Batch F6 with particle size, PDI, and % EE 194.7 nm, 0.231, and 90% respectively was selected as the optimized batch. The optimized batch was further subjected to spray drying and evaluated for various parameters. Residual solvent analysis was carried out by Gas chromatography. In vitro drug release study showed a biphasic drug release pattern of 70.17 ± 1.38% cumulative drug release at the end of 10 h. The release profile of curcumin from nanoparticles appeared to fit best with the Higuchi model. XRD study confirmed the molecular dispersion of the drug. The formulation showed satisfactory results in sterility testing, histology and isotonicity testing. In vivo study on rats showed dose dependent reduction in vascular endothelial growth factor (VEGF) concentration in vitreous fluid. Based on available evidences it can be concluded that the prepared formulation possesses great potential to manage diabetic retinopathy.

Tridax procumbens Ameliorates Streptozotocin-Induced Diabetic Neuropathy in Rats via Modulating Angiogenic, Inflammatory, and Oxidative Pathways.

Tridax procumbens (TP) is a traditional Indian therapeutic plant and was evaluated for its blood glucose lowering abilities, as well as for its ability to curb diabetic neuropathy (DN). Administrating 45 mg/kg body weight of streptozotocin (STZ) intraperitoneally for four weeks, DN was induced in Wistar rats. After the rats' tails were clipped, the blood glucose levels were measured. Body weight and urine volume were also assessed. Oxidative stress makers such as superoxide dismutase (SOD), thiobarbituric acid reactive substances (TBARS), catalase (CAT), inflammatory cytokines for instance tumor necrosis factor (TNF)-α, and interleukin (IL)-1β were estimated. Further, protein kinase C (PKC-β) and vascular endothelial growth factor (VEGF) were also estimated as angiogenic markers. Behavioral parameters were also evaluated by using cold allodynia using acetone test, hot allodynia using Eddy's hot plate, grip strength test using Rota rod, and hyperalgesia test using Tail flick technique. The statistical assessment of findings was done employing one-way (ANOVA) analysis of variance, and subsequently Turkey as post hoc with GraphPad Prism software package. The ingestion of TP for 1 month in DN rats stemmed in a substantial decline in blood glucose concentrations matched to nontreated rats with DN. There had been a considerable improvement in DN as evident from the finding from biochemical markers. The serum level of antioxidant defense enzymes was significantly increased, while the activities of TBARS had been substantially reduced in the TP treated rats with DN. TP averted DN-triggered surge levels of TNF-α and IL-6 in the serum. Further, PKC-β and VEGF concentrations had been also reduced by the treatment TP. The findings of this research demonstrated that the restorative impact of TP on DN rats might be linked to the anti-inflammatory and antioxidative antiangiogenic retorts.

Serum Levels of Vascular Endothelial Growth Factor, Platelet Activating Factor and Eosinophil-Derived Neurotoxin in Chronic Spontaneous Urticaria-A Pilot Study in Adult Patients.

Chronic spontaneous urticaria (CSU) is a skin disease characterized by the presence of wheals, angioedema, or both for at least 6 weeks. Although, CSU is often regarded as autoimmune in nature, its etiology is not fully explained and interactions between various small molecules are still taken under account. The aim of this research was to investigate the mean serum concentration of vascular endothelial growth factor (VEGF), platelet activating factor (PAF), and eosinophil-derived neurotoxin (EDN) in relation to the disease activity and pruritus intensity in adult patients with CSU. Fifteen patients with CSU and 15 healthy subjects participated in this pilot study. Blood samples were taken to examine the mean serum levels of VEGF, PAF, and EDN by the enzyme-linked immunosorbent assay test (ELISA). The Urticaria Activity Score (UAS7) and The Visual Analogue Scale (VAS) were used to assess the disease activity and the pruritus intensity, respectively. Obtained results revealed that VEGF, PAF, and EDN concentrations were higher in patients with CSU compared with those of the control group, but only for VEGF it was statistically significant (p = 0.008). However, levels of all investigated cytokines were not significantly correlated neither with the disease activity nor with the pruritus intensity. Our results showed higher serum levels of VEGF, PAF, and EDN among CSU patients which may highlight a functional role of these cytokines in the disease’s pathogenesis. In contrast, VEGF, PAF, or EDN might not be useful to reflect the severity of symptoms.

Cytoplasm or Supernatant: Where Is the Treasury of the Bioactive Antiaging Factor from Mesenchymal Stem Cells?

Cell-free compounds of mesenchymal stem cells (MSCs) could be a safer and cheaper substitution for MSC transplantation and have gained substantial research interest for antiaging skin treatments. However, whether those bioactive components should be obtained from the cytoplasm or supernatant is yet to be determined. In this study, we examined the ingredients of the MSC cytoplasm extract (MSC-ex) and MSC supernatant (MSC-s) and evaluated their effect in a photoaging model. Although MSC-ex has a richer protein composition than MSC-s, the latter has a proteome associated with wound healing and blood vessel development. Over 85% of the proteins in MSC-s were also found in MSC-ex, including extracellular matrix protein and various growth factors. The results of real-time PCR and western blot also demonstrate that both MSC-s and MSC-ex can upregulate collagen, transforming growth factor β (TGF-β), and vascular endothelial growth factor (VEGF) and downregulate IL-1β and matrix metalloproteinase-1 (MMP-1), which were considered critical for antiphotoaging. This supports our observations in the Hematoxylin and Eosin (HE) and Masson staining assay that they have a comparable effect as MSCs in terms of enhancing dermal thickness, and stimulating collagen regeneration. Although MSC-s and MSC-ex showed a weaker immunosuppression effect than MSCs, moisture measurement showed that they repair damage more rapidly than MSCs. Furthermore, the histological results showed that MSC-s maintains a super effect on immunosuppression, epidermal repair, and angiogenesis. That may be associated with the higher content of laminin, TGF-β, and VEGF in MSC-s, as well as its super cytokine transcriptional regulation ability. Thus, both MSC-s and MSC-ex can safely and effectively promote the repair of skin light injury, similar to MSCs. Our findings can broaden the range of active factors available in cell-free treatment, determine the difference between MSC-s and MSC-ex, and provide a reference for the development of similar products in regenerative medicine.

Novel Potency Assay for MSC Secretome-Based Treatment of Idiopathic Male Infertility Employed Leydig Cells and Revealed Vascular Endothelial Growth Factor as a Promising Potency Marker.

Idiopathic male infertility is a highly prevalent diagnosis in developed countries with no specific treatment options. Although empirical medical treatment is widely used to restore male fertility, its efficacy remains limited and inconclusively proven. Therefore, the development of novel therapeutic approaches in this field is a high-priority task. Since the failure of testicular microenvironment components might be involved in the pathogenesis of idiopathic male infertility, application of mesenchymal stromal cells (MSCs) as well as the MSC secretome is worth considering. Previously, we showed that the intratesticular injection of MSCs or the MSC secretome led to the recovery of spermatogenesis at least through replenishing the testicular microenvironment and its maintenance by MSC-secreted paracrine factors. However, the clinical use of such products has been limited to single trials to date. This may be due to the lack of relevant potency tests reflecting mechanisms of action of the MSC secretome in male infertility models. Based on the presumptive MSC secretome mode of action on the testicular microenvironment, we suggest a novel approach to test the potential efficacy of the MSC secretome for idiopathic male infertility treatment. It represents a potency assay based on evaluation of testosterone production by isolated Leydig cells. We demonstrated that the MSC secretome stimulated testosterone secretion by Leydig cells in vitro. We then hypothesized that among the major factors of the MSC secretome, vascular endothelial growth factor (VEGF) could be responsible for the observed effects, which we confirmed by the revealed correlation between the extent of stimulated testosterone production and VEGF concentration in the MSC secretome. The pilot results obtained from the doxorubicin-induced male infertility murine model also indicate the important impact of VEGF in the MSC secretome’s regenerative effects. Utilizing VEGF as a surrogate factor, a novel approach to study the potency of MSC secretome-based products for idiopathic male infertility treatment is suggested. Further validation is required for its implementation into the biopharmaceutical manufacturing process.

Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate.

Successful translation of multipotent mesenchymal stromal cell (MSC)-based therapies into clinical reality relies on adequate cell production procedures. These should be available not only for human MSC, but also for MSC from animal species relevant to preclinical research and veterinary medicine. The cell culture medium supplementation is one of the critical aspects in MSC production. Therefore, we previously established a scalable protocol for the production of buffy-coat based equine platelet lysate (ePL). This ePL proved to be a suitable alternative to fetal bovine serum (FBS) for equine adipose-derived (AD-) MSC culture so far, as it supported AD-MSC proliferation and basic characteristics. The aim of the current study was to further analyze the functional properties of equine AD-MSC cultured with the same ePL, focusing on cell fitness, genetic stability and pro-angiogenic potency. All experiments were performed with AD-MSC from n = 5 horses, which were cultured either in medium supplemented with 10% FBS, 10% ePL or 2.5% ePL. AD-MSC cultured with 2.5% ePL, which previously showed decreased proliferation potential, displayed higher apoptosis but lower senescence levels as compared to 10% ePL medium (p < 0.05). Non-clonal chromosomal aberrations occurred in 8% of equine AD-MSC cultivated with FBS and only in 4.8% of equine AD-MSC cultivated with 10% ePL. Clonal aberrations in the AD-MSC were neither observed in FBS nor in 10% ePL medium. Analysis of AD-MSC and endothelial cells in an indirect co-culture revealed that the ePL supported the pro-angiogenic effects of AD-MSC. In the 10% ePL group, more vascular endothelial growth factor (VEGF-A) was released and highest VEGF-A concentrations were reached in the presence of ePL and co-cultured cells (p < 0.05). Correspondingly, AD-MSC expressed the VEGF receptor-2 at higher levels in the presence of ePL (p < 0.05). Finally, AD-MSC and 10% ePL together promoted the growth of endothelial cells and induced the formation of vessel-like structures in two of the samples. These data further substantiate that buffy-coat-based ePL is a valuable supplement for equine AD-MSC culture media. The ePL does not only support stable equine AD-MSC characteristics as demonstrated before, but it also enhances their functional properties.

Analysis of the Cytokine Expression in the Aqueous Humor of Individuals with BRVO-Associated Macular Edema.

Purpose This study aimed to determine the expression levels of vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in the aqueous humor of patients with macular edema (ME) caused by branch retinal vein occlusion (BRVO), as well as to investigate the relationship between the cytokines as mentioned earlier and best-corrected visual acuity (BCVA), ME, and the degree of ME from the molecular level. Methods In a prospective observational study, fluorescein fundus angiography (FFA) and optical coherence tomography (OCT) were used to classify 58 patients with non-ischemic BRVO-ME into three groups according to the degree of ME: 14-mild, 17-moderate, and 27-severe. The specific concentration of IL-6, VEGF, ICAM-1, and VCAM-1 in the aqueous humor was detected using the BD CSCanto™ II Flow Cytometer (US). Spearman or Pearson correlation analysis was used to test the correlation between the levels of BCVA and severity of ME and the expression levels of IL-6, VEGF, ICAM-1, and VCAM-1 in the aqueous humor. Results According to the obtained data, BCVA did not correlate with the severity of ME, and these four cytokines expression levels in patients' aqueous humor (P > 0.05). Moreover, BCVA did not correlate with mild, moderate, or severe ME as well (P > 0.05). However, the levels of these four cytokines were correlated with the severity of the ME. These underlined cytokines were linked to the mild, moderate, and severe degrees of ME. VEGF was also significantly correlated (r > 0.8, P < 0.0001) with the severity of ME. Conclusions This study suggests that the severity of ME in BRVO-ME patients is significantly correlated with the expression levels of IL-6, VEGF, ICAM-1, and VCAM-1 in the aqueous humor. Lowering the level of disease-associated cytokines may potentially reduce the degree of ME. Therefore, an in-depth study of the levels and the relationship may provide some evidence for the pathogenesis, treatment, and prevention of BRVO-ME.

The Effect and Mechanism of External Use Ulcer Powder on Diabetic Mice.

Objective Through the preparation of the diabetic mice skin ulcer model, we investigated the effect of Mongolian medicine external ulcer powder (WYK) on the treatment of diabetic skin ulcers and the expression of angiogenesis-related factors such as vascular endothelial growth factor (VEGF) and extracellular regulated protein kinases (ERK).Methods Thirty male clean Kunming mice were randomly divided into normal control group (group C), diabetic control group (group HC), and diabetic topical ulcer powder group (group HW). After successful modeling in the HC group and the HW group, the rats in the HW group were given external ulcer powder, which was applied to the back of the mice once a day. In addition, the rats in group C and group HC were treated with gentamicin injection external application once a day. The mice were sacrificed on the 3rd, the sixth, and the ninth day of dosing, and samples were taken. The adopted methods included protein immunoblotting (western blot) and reverse transcription-polymerase chain reaction (RT-PCR). The expression differences of angiogenesis-related factors such as VEGF and ERK in the repair process were detected. SPSS 13 software was used to analyze the results of angiogenesis-related factors VEGF and ERK.ResultsComparison of VEGF and ERK ContentsThe serum VEGF content of mice in the HC group was significantly lower than that in the C group on days 3, 6, and 9 (p <0.05). The VEGF content in the HW group was significantly higher than that in the HC group (p <0.05). The content of ERK in serum was basically consistent with that of VEGF. The results of the western blot assay were consistent with those of the RT-PCR assay.Conclusion WYK can effectively promote the healing of skin ulcer wounds in diabetic mice, accelerate the proliferation of granulation tissue, enrich the contents of capillary blood tubes and collagen fibers, and increase the microvascular content. WYK can improve the expression level of VEGF and ERK in the serum of mice and advance the peak value of protein expression.

An in vitro study on the effect of bevacizumab on endothelial cell proliferation and VEGF concentration level in patients with hereditary hemorrhagic telangiectasia.

Previous studies have demonstrated that vascular endothelial growth factor (VEGF) is upregulated in patients with hereditary hemorrhagic telangiectasia (HHT). The use of Bevacizumab as an anti-angiogenic treatment agent seems promising. The purpose of the present in vitro study was to determine the efficacy and potential toxicity levels of bevacizumab on cell proliferation and VEGF concentrations in endothelial cells of HHT patients. In this in vitro study, endothelial cells from patients with HHT and HUVECs (control) were incubated with different concentration levels of bevacizumab (2, 4, 6, 8 or 10 mg/ml). After 24, 48 or 72 h, the cell proliferation was assessed by Alamar Blue® Assay and the VEGF levels in the cell culture supernatants were measured by VEGF-ELISA. All endothelial cells incubated with bevacizumab showed an initial decrease in cell proliferation. Cell proliferation recovered within 72 h in cell cultures incubated with concentration levels of up to 4 mg/ml bevacizumab, whereas those incubated with higher concentration levels showed a continuous decline in cell proliferation. VEGF expression decreased after 24 h in cell cultures incubated with bevacizumab concentration levels of 2 and 4 mg/ml but increased again after 48 h. Cell cultures incubated with bevacizumab concentration levels of 10 mg/ml showed a constant decline in VEGF expression without any tendency for recovery. Translating these results into daily clinical practice, the present study suggests that the intranasal submucosal injection of bevacizumab in HHT patients should not exceed a concentration level of 4 mg/ml. Overall, higher bevacizumab concentration levels not only reduce VEGF expression but pose a higher risk of toxic effects on endothelial cells as they jeopardize cell proliferation.

Dynamics in Circulating Proinflammatory Biomarkers for Prognostic Assessment of Patients With Advanced HCC – A Substudy From the SORAMIC Trial

IntroductionPrediction of response to treatment in patients with advanced hepatocellular carcinoma (HCC) may assist in the selection of personalized management.ObjectiveThis exploratory analysis of the palliative arm of the SORAMIC trial (ClinicalTrials.gov NCT01126645) evaluated the prognostic potential of basal and dynamic changes in systemic levels of interleukin 6 (IL-6), interleukin 8 (IL-8), systemic vascular endothelial growth factor (VEGF), and lipopolysaccharide (LPS).MethodsWe evaluated the correlations between overall survival (OS) and concentrations of IL-6, IL-8, VEGF, and LPS at follow-up approximately 7-9 weeks after treatment initialization (FU) compared to baseline (BL) in 90 patients treated either with 90Yttrium (90Y) microspheres combined with sorafenib (n = 44) or with sorafenib (n = 46) alone.ResultsChanges in IL-6 concentration during treatment showed correlations with the outcome. An increase in IL-6 concentration of less than 16.8 pg/mL over baseline readings was associated with better survival [median OS 16.3 months compared with 8.9 months (p = 0.0354)]. Correlations with survival were not observed for VEGF or LPS concentrations at baseline, at FU, or changes between these time points.ConclusionsChanges in IL 6 serum levels at 7-9 weeks after treatment initialization but not in IL 8, VEGF, or LPS add important information on the outcome of advanced HCC patients treated palliatively within the SORAMIC trial.