Endothelial Growth Factor Gene Expression Research Articles

In vitro effects of photobiomodulation applied to gingival fibroblasts cultured on titanium and zirconia surfaces and exposed to LPS from Escherichia coli.

Photobiomodulation (PBM) therapy is used to stimulate cell proliferation and metabolism, as well as reduce inflammatory cytokine synthesis, which plays a main role in the long-term stability of implants. This study assessed the response of gingival fibroblasts cultured on titanium (Ti) and zirconia (ZrO2), submitted to PBM and exposed to lipopolysaccharide (LPS). Cells seeded on Ti and ZrO2 were irradiated (InGaAsP; 780nm, 25mW) 3 times, using 0.5, 1.5, and 3.0J/cm2 doses, and exposed to Escherichia coli LPS (1μg/mL). After 24h, cell viability (alamarBlue, n = 8), interleukin 6 (IL-6) and 8 (IL-8) synthesis (ELISA, n = 6), and IL-6 and vascular endothelial growth factor (VEGF) gene expression (qPCR, n = 5) were assessed and statistically analyzed (one-way ANOVA, α = 0.05). Cell morphology was evaluated by fluorescence microscopy. Increased cell viability occurred in all groups cultured on Ti compared with that of the control, except for cells exposed to LPS. Fibroblasts cultured on ZrO2 and LPS-exposed exhibited reduced viability. PBM at 3.0J/cm2 and 1.5J/cm2 downregulated the IL-6 synthesis by fibroblasts seeded on Ti and ZrO2, as well as IL-8 synthesis by cells seeded on ZrO2. Fibroblasts seeded on both surfaces and LPS-exposed showed increased IL-6 gene expression; however, this activity was downregulated when fibroblasts were irradiated at 3.0J/cm2. Enhanced VEGF gene expression by cells seeded on Ti and laser-irradiated (3.0J/cm2). Distinct patterns of cytoskeleton occurred in laser-irradiated cells exposed to LPS. Specific parameters of PBM can biomodulate the inflammatory response of fibroblasts seeded on Ti or ZrO2 and exposed to LPS.

Maternal omega-3 fatty acids and vitamin E improve placental angiogenesis in late-onset but not early-onset preeclampsia.

Abnormal placental vasculature is associated with preeclampsia. Preeclampsia is of two types, i.e., early- and late-onset preeclampsia (LOP), both having different etiologies. We have earlier demonstrated low levels of omega-3 fatty acids and vitamin E in women with preeclampsia. The current study examines the effect of maternal omega-3 fatty acids and vitamin E supplementation on angiogenic factors in a rat model of preeclampsia. Pregnant rats were divided into a total of five groups control, early-onset preeclampsia (EOP); LOP; EOP supplemented with omega-3 fatty acid and vitamin E and LOP supplemented with omega-3 fatty acid and vitamin E. Preeclampsia was induced by administering L-nitroarginine methylester (L-NAME) at the dose of 50mg/kg body weight/day. The vascular endothelial growth factor gene expression and protein levels were lower (p < 0.01 for both) in animals from both EOP as well as LOP groups (p < 0.01). In the EOP group, the protein levels of VEGF receptor-1 were also lower (p < 0.01). Supplementation of omega-3 fatty acids and vitamin E to LOP improved the levels of VEGF and VEGF receptor-1 only in the LOP but not in the EOP group. In the EOP group, the gene expression of hypoxia inducible factor 1 alpha (HIF-1α) in the placenta was higher (p < 0.05) and supplementation normalized these levels. Our findings indicate that maternal supplementation of omega-3 fatty acids and vitamin E has differential effect on preeclampsia subtypes.

Caffeine is associated with improved alveolarization and angiogenesis in male mice following hyperoxia induced lung injury

BackgroundCaffeine therapy for apnea of prematurity reduces the incidence of bronchopulmonary dysplasia (BPD) in premature neonates. Several mechanisms, including improvement in pulmonary mechanics underly beneficial effects of caffeine in BPD. As vascular development promotes alveologenesis, we hypothesized that caffeine might enhance angiogenesis in the lung, promoting lung growth, thereby attenuating BPD.MethodsC57Bl/6 mice litters were randomized within 12 h of birth to room air (RA) or 95%O2 to receive caffeine (20 mg/kg/day) or placebo for 4 days and recovered in RA for 12wks. The lung mRNA and protein expression for hypoxia-inducible factors (HIF) and angiogenic genes performed on day 5. Lung morphometry and vascular remodeling assessed on inflation fixed lungs at 12wks.ResultsCaffeine and hyperoxia in itself upregulate HIF-2α and vascular endothelial growth factor gene expression. Protein expression of HIF-2α and VEGFR1 were higher in hyperoxia/caffeine and angiopoietin-1 lower in hyperoxia. An increase in radial alveolar count, secondary septal count, and septal length with a decrease in mean linear intercept indicate an amelioration of hyperoxic lung injury by caffeine. An increase in vessel surface area and a significant reduction in smooth muscle thickness of the pulmonary arterioles may suggest a beneficial effect of caffeine on vascular remodeling in hyperoxia, especially in male mice.ConclusionsPostnatal caffeine by modulating angiogenic gene expression early in lung development may restore the pulmonary microvasculature and alveolarization in adult lung.

Vitamin C and E supplementation effects on secretory and molecular aspects of vascular endothelial growth factor derived from peritoneal fluids of patients with endometriosis

Endometriosis is an extremely heterogeneous disease and affects about ten percent of the female population during their reproductive years. Recent studies showed that endometriosis is an angiogenesis-dependent disease. Peritoneal macrophages are a well-characterised source of vascular endothelial growth factor (VEGF). The aim of this study was to determine the VEGF gene expression and production in peritoneal macrophages of patients with endometriosis under the effects of vitamins C and E in comparison with control. The lab trial study carried out on 50 patients undergoing laparoscopy and peritoneal fluid samples were collected from them. We compared the VEGF gene expression and production in peritoneal macrophages among groups by using real-time polymerase chain reaction and enzyme-linked immunosorbent assay methods, respectively. Our results showed that gene expressions influenced by vitamin C increased in different concentrations and incubation times, except for the incubation time after 48 h. In the case of vitamin E, this was evident with the exception of vitamin E 50 μM after 24 h and vitamin E 100 μM after 48 h. Our findings indicated that vitamin C and E in different concentrations and incubation times altered VEGF gene expression in the peritoneal macrophages but they had not affected on VEGF productions.Impact statementWhat is already known on this subject? Previous studies showed that antioxidants play a key role in the inhibition of oxidative stress-induced damages and the reduction of pelvic pain in patients with endometriosis. Vitamin E and vitamin C are the main components in neutralising free radicals. Also, antioxidant consumption such as vitamin C and vitamin E in women with endometriosis showed an inverse correlation between antioxidant intake and endometriosis pathology.What do the results of this study add? Vitamin C and E in different concentrations and times of incubation altered vascular endothelial growth factor gene expression and production in peritoneal macrophages.What are the implications of these findings for clinical practice and/or further research? Further studies are needed to determine the effects of C and E vitamins in different concentrations on vascular endothelial growth factor gene expression and production in peritoneal macrophages and the possible roles of these vitamins in treating endometriosis.

Effects of islet neogenesis associated protein depend on vascular endothelial growth factor gene expression modulated by hypoxia-inducible factor 1-alpha

BackgroundPharmacology has provided efficient tools to improve insulin effect/secretion but the decrease in β-cell mass remains elusive. INGAP-PP could provide a therapeutic alternative to meet that challenge. AimTo further understand the mechanism that links INGAP-PP effects upon β-cell mass and function with islet angiogenesis. MethodologyNormal male Wistar rats were divided into 2 groups and injected with a single dose of 100 mg/Kg suramin or saline. Both groups were divided into 2 subgroups that received daily doses of 2 mg/kg INGAP-PP or saline for ten days. Plasma glucose, triacylglycerol, TBARS, and insulin levels were measured. Pancreas immunomorphometric analyses were also performed. Pancreatic islets were isolated to measure glucose-stimulated insulin secretion (GSIS). Specific islet mRNA levels were studied by qRT-PCR. Statistical analysis was done using ANOVA. ResultsNo differences were recorded in body weight, food intake, or any other plasma parameter measured in all groups. Islets from INGAP-PP-treated rats significantly increased GSIS, β-cell mass, and mRNA levels of Bcl-2, Ngn-3, VEGF-A, VEGF-R2, CD31, Ang1 and Ang2, Laminin β-1, and Integrin β-1, and decreased mRNA levels of Caspase-8, Bad, and Bax. Islets from suramin-treated rats showed significant opposite effects, but INGAPP-PP administration rescued most of the suramin effects in animals treated with both compounds. ConclusionOur results reinforce the concept that INGAP-PP enhances insulin secretion and β-cell mass, acting through PI3K/Akt/mTOR pathways and simultaneously activating angiogenesis through HIF-1α-mediated VEGF-A secretion. Therefore, INGAP-PP might be a suitable antidiabetic agent able to overcome two major alterations present in T2D.

Analysis of Cell Viability and Gene Expression After Continuous Ultrasound Therapy in L929 Fibroblast Cells.

This study aimed to analyze cell viability and gene expression interleukin 6 and vascular endothelial growth factor after continuous ultrasound therapy of 1 and 3 MHz in L929 fibroblast cells. The L929 cells were cultivated in 12-well plates and divided into the following five groups: Group 1 (G1), nonirradiated; G2, 0.2 W/cm-1 MHz; G3, 0.5 W/cm-1 MHz; G4, 0.2 W/cm-3 MHz; and G5, 0.5 W/cm-3 MHz. The cells were irradiated at 24 and 48 hrs. Cell viability was evaluated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method. The gene expression analysis was assessed using real-time polymerase chain reaction. The G2 and G3 showed a decrease in cell viability when compared with the G1 at 48 hrs (P < 0.01). The G4 and G5 presented an increase in viability (P = 0.01 and P = 0.03, respectively) in 24 to 48 hrs. The cells irradiated at an intensity of 0.5 W/cm-3 MHz at 48 hrs showed a 0.21-fold decrease in interleukin-6 gene transcripts and a 1.67-fold increase in vascular endothelial growth factor gene transcripts. Continuous ultrasound therapy with a frequency of 3 MHz at an intensity of 0.5 W/cm stimulates cell proliferation, decreases interleukin-6 gene expression, and increases vascular endothelial growth factor gene expression in L929 fibroblast cells.

Shenqi Fuzheng Injection (SFI) Enhances IFN-α Inhibitory Effect on Hepatocellular Carcinoma Cells by Reducing VEGF Expression: Validation by Gene Silencing Technique.

Shenqi Fuzheng Injection (SFI) is a traditional Chinese medicine injection with anticancer properties and is mainly composed of ginseng and astragalus. Its efficacy has been confirmed in clinical trials, but the mechanism remains unclear. We investigated the effect of SFI on vascular endothelial growth factor (VEGF) gene expression in hepatocellular carcinoma (HCC) cells and identified its possible mechanism of synergistic effects when combined with the chemotherapeutic drug interferon (IFN-) α. An MTT assay was used to measure the inhibition effects of low-dose IFN-α (6000 IU) with or without SFI (0.5 g/L) on the HCC cell line MHCC97. VEGF-silenced MHCC97L-mir200 cell lines were prepared using lentiviral vectors and evaluated by real-time PCR to determine the inhibition effect. We examined MHCC97L-mir200 and MHCC97L cells by MTT assay, using IFN-α alone or in combination with SFI. The inhibition ratio of IFN-α (6000 IU) was -29.5%, while that for IFN-α (6000 IU) + SFI (0.5 g/L) was 17.0%, which was significantly higher than that for the IFN-α group (P < 0.01). The VEGF gene was silenced successfully in MHCC97-L cells. After interference of VEGF, the inhibition by SFI and IFN-α in MHCC97L-mir200 did not differ from that in MHCC97-L cells (P > 0.05). SFI can reduce the expression of VEGF in HCC, which can increase the efficacy of IFN-α, providing a theoretical basis for clinical application.

Association of vascular endothelial growth factor (VEGF) Gene polymorphisms and expression with the risk of endometriosis: a case-control study.

Endometriosis is a polygenic and multifactorial gynecology situation which might be associated with angiogenesis. In the current study we assess the role of vascular endothelial growth factor (VEGF) - 2578 A/C, and + 936 C/T polymorphisms in susceptibility to endometriosis and checking the expression of VEGF mRNA in eutopic tissue of endometrium with and without endometriosis. The study was comprised of 300 patients who underwent laparascopic or laparotomy surgery with 100 cases who had confirmed histological diagnosis of endometriosis, and 200 controls with no histological diagnosis of disease. The genotyping of VEGF polymorphisms was done by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique and the gene expression in tissue was determined using Real-Time PCR assay. There was no important difference of allele distribution of the - 2578 A/C (P = 0.7) and + 936 C/T (P = 0.5) polymorphisms among endometriosis cases and controls. Study of VEGF expression during the menstrual cycle, showed that endometrial tissue in cases group expressed more VEGF mRNA at the secretory phase compared to the proliferative phase (P = 0.03). Our results suggest that - 2578 A/C and + 936 C/T polymorphisms of VEGF did not seem to have impact on endometriosis predisposition in our study population. Also we did not find any link between VEGF mRNA expression and risk of endometriosis.

The anti-invasive activity of Robinia pseudoacacia L. and Amorpha fruticosa L. on breast cancer MDA-MB-231 cell line

The paper investigates anticancer effects of methanol extracts of invasive plant species Robinia pseudoacacia L. (RpE) and Amorpha fruticosa L. (AfE) on breast cancer MDA-MB-231 and healthy MRC-5 cells. The anticancer activity was evaluated through examination of cytotoxic effects, anti-invasive potential and impact on redox status in comparative analysis using their chemical composition. According to the IC50 values, the investigated plants had no significant cytotoxic effects either on healthy cell line MRC-5 or on MDA-MB-231 cancer cells, but they showed great anti-invasive potential by suppressing all investigated parameters of tumor invasion and metastases (Matrix Metalloproteinases (MMP), protein concentration and MMP-9, C-X-C Motif Chemokine Ligand 12 (CXCL-12), Vascular Endothelial Growth Factor (VEGF-A) and Hypoxia-Inducible Factor (HIF-1α) gene expression) in MDA-MB-231 cells. Based on their remarkable anti-invasive potential, RpE and AfE are suitable for use as potential supplements in anticancer therapy or as nutritional food supplements.

Evaluation of Retinal Pigment Epithelial Cell Cytotoxicity of Recombinant Tissue Plasminogen Activator Using Human-Induced Pluripotent Stem Cells.

Purpose The evaluation of drug-induced cytotoxicity is of great importance for the clinical application of pharmaceutical products, and human-induced pluripotent stem cells (hiPSCs) have received considerable scrutiny as a cell source for in vitro cytotoxicity testing. The aim of this study is to validate the concept of cytotoxicity testing using hiPSC-derived retinal pigment epithelium (hiPSC-RPE) by comparing the responsiveness of human fetal RPE (hfRPE) and human RPE cell line (ARPE19) to recombinant tissue plasminogen activator (rtPA). Methods HfRPE, two types of hiPSC-RPE, and ARPE19 were cultured in media with or without rtPA. A lactate dehydrogenase release assay was performed to investigate the dose- and time-dependent effects of rtPA on cell death. RPE function was evaluated by measuring the secretion of pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor (VEGF) and RPE-specific gene expression. Results Rates of cell damage in hfRPE and both hiPS-RPE were increased by rtPA supplementation (2000 and 4000 μg/ml) for 1 hour, whereas ARPE19 cell damage was increased by supplementation with rtPA at concentrations higher than 50 μg/ml. Although 100 μg/ml rtPA for 24 hours did not affect RPE cell function, sustained rtPA exposure induced prolonged cytotoxic effects in hfRPE and two hiPSC-RPE, but not ARPE19. Conclusion The responsiveness of hiPSC-RPE to rtPA is similar to that of hfRPE in terms of cell death and cell function. Thus, hiPSC-RPE is a valuable cell source for in vitro cytotoxicity testing.

The effect of Hydro alcoholic Olive Leaf Extract on Vascular Endothelial Growth Factor GeneExpression Level in Ischemic Muscle in Hind Limb Ischemic Rat Model

The effect of Hydro alcoholic Olive Leaf Extract on Vascular Endothelial Growth Factor GeneExpression Level in Ischemic Muscle in Hind Limb Ischemic Rat Model

Ultrasound-targeted microbubble destruction improved the antiangiogenic effect of Endostar in triple-negative breast carcinoma xenografts.

Ultrasound-targeted microbubble destruction (UTMD) has been reported to be a meritorious technique for drug targeting delivery. In this study, we aimed to evaluate the synergistic antiangiogenic effect of UTMD combined with Endostar on triple-negative breast carcinoma tumors. The lipid-shelled microbubbles (MBs) conjugated with Endostar were constructed using a biotin-avidin bridging chemistry method, and the morphological characteristics and drug-conjugating content were determined. MBs were administered intravenously to nude mice bearing MDA-MB-231 breast carcinoma xenografts and ultrasound exposure followed. The tumor microcirculation was observed by contrast-enhanced ultrasonography (CEUS) and the Endostar biodistribution was detected by enzyme-linked immunosorbent assay. Twenty-four breast carcinoma-bearing nude mice were divided into four groups. After treatment, every 3days for 15days the in vivo antitumor effects were assessed by calculating the tumor growth inhibition rate (TGIR). The tumor microcirculation was observed by CEUS, the tumor microvessel density (MVD) was calculated by immunohistochemistry under a microscope, and the vascular endothelial growth factor (VEGF) gene expression was detected by real-time quantitative polymerase chain reaction. The prepared Endostar-conjugated MBs were round and well-dispersed with a mean size of 2.8 ± 0.7µm and a drug conjugating content of 800.72 ± 70.53µg/108MBs. UTMD blocked the tumor microcirculation, and improved Endostar release in the targeted tumor tissue with a drug content of 1.12 ± 0.43µg/gram protein, which was about three times higher than that in Endostar group or Endostar conjugated MBs group. Endostar-conjugated MBs combined with UTMD treatment achieved the optimal antitumor effects in vivo with a TGIR of 46.29%, and apparent antiangiogenic effects with minimal tumor blood perfusion, MVD and VEGF gene expression level. UTMD can improve Endostar delivery in the targeting tumor tissue and mediate synergistic antiangiogenetic and antitumor effects, which may be a potential therapeutic strategy for refractory breast cancer.

Angiogenic and Osteogenic Synergy of Human Mesenchymal Stem Cells and Human Umbilical Vein Endothelial Cells Cocultured on a Nanomatrix

To date, bone tissue regeneration strategies lack an approach that effectively provides an osteogenic and angiogenic environment conducive to bone growth. In the current study, we evaluated the osteogenic and angiogenic response of human mesenchymal stem cells (hMSCs) and green fluorescent protein-expressing human umbilical vein endothelial cells (GFP-HUVECs) cocultured on a self-assembled, peptide amphiphile nanomatrix functionalized with the cell adhesive ligand RGDS (PA-RGDS). Analysis of alkaline phosphatase activity, von Kossa staining, Alizarin Red quantification, and osteogenic gene expression, indicates a significant synergistic effect between the PA-RGDS nanomatrix and coculture that promoted hMSC osteogenesis. In addition, coculturing on PA-RGDS resulted in enhanced HUVEC network formation and upregulated vascular endothelial growth factor gene and protein expression. Though PA-RGDS and coculturing hMSCs with HUVECs were each previously reported to individually enhance hMSC osteogenesis, this study is the first to demonstrate a synergistic promotion of HUVEC angiogenesis and hMSC osteogenesis by integrating coculturing with the PA-RGDS nanomatrix. We believe that using the combination of hMSC/HUVEC coculture and PA-RGDS substrate is an efficient method for promoting osteogenesis and angiogenesis, which has immense potential as an efficacious, engineered platform for bone tissue regeneration.

THE PROLIFERATIVE ACTIVITY (KI-67) AND THE LEVEL OF VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) GENE EXPRESSION IN GEMISTOCYTIC ASTROCYTOMAS

Background. Gemistocytic astrocytomas (GA) are a variant of diffuse astrocytomas GII (WHO, 2016), which is characterized by a shorter overall survival of patients than diffuse astrocytoma. The role of vascular endothelial growth factor (VEGF) gene expression and its association with the index of proliferative activity (Ki-67) in GA has not been studied Objective. To estimate the correlation of VEGF gene expression on Ki-67 in GA. Design and methods . A retrospective and prospective analysis of morphological and molecular-genetic features of tumors was performed on 9 patients with the diagnosis of GA. Patients over 18 years of age with supratentorial tumor localization were included in the study. The patients were aged from 27 to 37 years, the median age was 32 years. There were 4 men. Fragments of tumor tissue obtained during surgery were subjected to histological, immunohistochemical (Ki-67 and GFAP) and molecular genetic studies. By real-time PCR method was used to determine the level of the VEGF gene expression and the presence of a mutation in IDH1/IDH2 genes. Histological diagnosis was made in accordance with the classification of tumors of the CNS WHO (2016) and the AFIP atlas (2007). Results. The tumor most often affected several lobes of the brain (44,4%). The first clinical manifestations of the disease in the vast majority of cases were epileptic and hypertensive syndromes (55,6%). For the proliferative activity index Ki-67 >5% (71,4% of cases) the moderate proliferation of vascular endothelium was noted in 85,7% of cases and the middle/high level of VEGF gene expression (except one case). In patients with low Ki-67 (≤5%), the low level of the VEGF gene expression was determined in 100% cases, the proliferation of the vascular endothelium was absent. Conclusion. High Ki-67 (>5%), the proliferation of the vascular endothelium, middle/high level of VEGF gene expression are typical attributes of anaplastic astrocytomas. That’s why some GA have features of astrocytomas GIII.

Inhibition of VEGF gene expression in osteoblast cells by different NSAIDs

ObjectiveTo determine the effect of different nonsteroidal anti-inflammatory drugs (NSAIDs) on vascular endothelial growth factor (VEGF) gene expression in two osteoblast cell populations. DesignOsteoblasts obtained by primary culture (HOp) and human osteosarcoma cell line MG63 (MG-63), which were treated with 10 μM doses of acetaminophen, indomethacin, ketoprofen, diclofenac, ibuprofen, ketorolac, naproxen or piroxicam. At 24 h of treatment, their gene expression of VEGF was evaluated by real-time polymerase chain reaction (RT-PCR) and compared with the expression in untreated cells (control group). ResultsThe treatment with the different NSAIDs significantly reduced VEGF expression regardless of the cell line and NSAID studied. ConclusionThe results of this study suggest that these drugs may have undesirable effects on the osteoblast and its bone-forming capacity, given the effect of this growth factor on these cells. Further studies are warranted to determine their repercussions on bone tissue and to elucidate the cell signaling mechanism/s involved.

Determining the Significance of a Novel Disease Inducing Rodent Diet on Adenocarcinoma Growth In Vivo

Diet and diet‐induced morbidities (such as obesity) are leading risk factors for the development of various forms of cancer worldwide. However, the physiological mechanisms responsible for the interaction of diet and cancer growth are still unclear. Mechanistic studies in animals support the interaction of diet with cancer development and progression, however the animal diets used do not adequately model the true nutritional quality of many Americans. We have developed a novel disease inducing rodent diet referred to as the Americanized diet (AD) that embodies the complex nutritional profile of Americans while also accounting for the differences in human and rodent nutritional requirements. We set out to determine how consumption of the AD would influence the growth of the MC‐38 murine adenocarcinoma cells in vivo. All animal experiments were performed following animal protocols approved by the Liberty University IACUC and conform to the FASEB Statement of Principles for the use of animals in research and education. Weanling C57Bl/6 male mice were given ad libitum access to the AD (n=5) or standard rodent chow (n=5) and ddH2O. Food intake and body weights were calculated weekly and circulating total cholesterol (CHOL), HDL‐cholesterol (HDL), triglycerides, and glucose were quantified at baseline and again at the end of the study. After 6 weeks, mice were injected with 0.5×106 MC‐38 adenocarcinoma cells in the rear flank and injection site was monitored for the presence of a palpable tumor. Tumor volume was calculated using calipers and, after 3 weeks, tumor blood flow was estimated using contrast‐enhanced ultrasonography. Animals were then euthanized and tumor tissues collected for assessment of leukocyte content by flow cytometry and quantification of interleukin‐10 (IL‐10), major histocompatibility complex H2K, and vascular endothelial growth factor (VEGF) gene expression by RT‐PCR. Body weights and tumor volumes were analyzed using generalized linear models and the remaining parameters were analyzed using GLM multivariate analyses in SPSS. Mice assigned to ad libitum feeding of the AD consumed more calories and gained more weight (P=0.01) as compared to mice consuming standard chow. After 6 weeks of consuming the diet, mice fed the AD had greater HDL cholesterol (P=0.03); however this difference was lost after MC‐38 innoculation (P=0.2). MC‐38 growth was significantly reduced (P=0.004 and 0.07 at weeks 1 and 2, respectively) in mice consuming the AD as compared to mice consuming chow. Interestingly, no palpable tumor was observed in 2 of the 5 mice consuming the AD while all 5 mice consuming chow developed tumors. There was a greater relative tissue perfusion (P=0.02) in the tumor‐bearing mice fed the AD as compared to the mice fed chow. No difference was observed in the proportion of tumor leukocytes amongst the diets. Similarly, RT‐PCR revealed no difference in IL‐10, H2K or VEGF gene expression between treatment groups (P= 0.25, 0.98, 0.62 respectively). Based on this study, mice fed the AD had reduced growth of the MC‐38 murine adenocarcinoma cell line in vivo despite having greater body weight and caloric intake. While we were unable to detect a significant difference in tumor leukocyte content and/or the expression of IL10, H2K, and VEGF at the end of the study, it is possible these outcomes differed during earlier time points (weeks 1 and 2) and contributed to the reduced tumor growth observed in mice fed the AD.Support or Funding InformationThis work was supported by Liberty University, College of Osteopathic Medicine intramural grant #2016‐07.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Anti-Inflammatory and Tissue Regenerative Effects of Topical Treatment with Ozonated Olive Oil/Vitamin E Acetate in Balanitis Xerotica Obliterans.

Balanitis xerotica obliterans (BXO) is a chronic inflammatory skin disorder, considered the male genital variant of lichen sclerosus. Anti-inflammatory drugs are commonly used in BXO. We evaluated the effects of an innovative formulation of ozonated olive oil with vitamin E acetate (OZOILE®) on the inflammatory status and tissue remodeling in male children with BXO. The mRNA transcripts of proteins involved either in inflammation or in dynamics of tissue regeneration were analyzed by quantitative real-time PCR, in foreskins affected by BXO removed from patients untreated or treated with OZOILE® cream for 7 days before circumcision. We found a significant reduction in mRNA levels of IL-1β, TNF-α, INF–γ, transglutaminase 2 and NOS2 in foreskins treated with OZOILE® in comparison to untreated ones (p < 0.001). No significant differences were observed in NF-κB activation in the specimens obtained from treated and untreated patients. Hence, OZOILE® treatment up-regulated hypoxia-inducible factor (HIF)-1alpha, vascular endothelial growth factor (VEGF) and E-cadherin gene expression (p < 0.001). The treatment with OZOILE® showed effective results in children affected by BXO by reducing the inflammatory process and stimulating mechanisms for tissue regeneration of the foreskin. A randomized clinical trial on a large number of children affected by BXO might be useful to verify the efficacy of topical treatment with OZOILE®.

Evaluation of the effect of hesperidin on vascular endothelial growth factor gene expression in rat skin animal models following cobalt-60 gamma irradiation.

Skin is highly prone to radiation damage. Radiation burn is defined as damage to the skin or other biological tissues induced by radiofrequency or ionizing radiation. Vascular endothelial growth factor (VEGF) is a heparin-binded pro-angiogenic factor. Flavonoids belong to a family of polyphenol chemical compounds that are frequently present in fruits and vegetables. Hesperidin is an agent belonging to the flavonoid family. The aim of this study is to investigate whether hesperidin can affect the VEGF gene expression in rat skin following gamma irradiation or not. A total number of 36 male Sprague-Dawley rats were divided into three groups. First group: radiation group (n = 12), second group: radiation + hesperidin-treated group (n = 12), and third group: untreated control group (n = 12). The hesperidin administration dose was 100 mg/kg body weight. The rats received a 22 Gy single dose at a dose rate of about 0.3 Gy/min using a cobalt-60 external beam radiotherapy unit. The animals were euthanized 24 h postirradiation. VEGF gene expression data were analyzed using the equation 2-ΔΔCT, where ΔΔCT = (Threshold cycle [CT], of target gene - CT of housekeeping gene)treated group- (CT of target gene - CT of housekeeping gene)untreated control group. Glyceraldehyde-3-phosphate dehydrogenase gene was used as a housekeeping gene. VEGF gene in the radiation + hesperidin group overexpressed 25-fold relative to the control group. In addition, VEGF gene in the radiation group underexpressed 0.15-fold relative to the control group. When the three groups were compared relative to each other using the Kruskal-Wallis test, P < 0.001 was obtained. Based on the Mann-Whitney U-test, when all groups were compared to each in a binary model, P = 0.001 was achieved. These tests all showed statistically significant changes in VEGF gene expression. We can conclude that hesperidin is a potent angiogenic factor. Hesperidin as a radioprotector can initiate angiogenesis by VEGF gene induction. It may stimulate epithelialization, collagen deposition, and enhanced cellular proliferation. These changes can together accelerate wound healing, in particular, radiation-induced skin damage.

Tumoral Necrosis Factor (TNF) mRNA gene expression in relation to Vascular Endothelial Growth Factor (VEGF) and chemokines mRNA gene expression in tumoral microenvironment of endometrial cancer versus normal endometrium: Preliminary results

Tumoral Necrosis Factor (TNF) mRNA gene expression in relation to Vascular Endothelial Growth Factor (VEGF) and chemokines mRNA gene expression in tumoral microenvironment of endometrial cancer versus normal endometrium: Preliminary results

Cell and gene therapy with reporter gene imaging in myocardial ischemia.

Reporter gene/probe systems have proved to be reliable for monitoring gene/cell therapy. We sought to evaluate whether a reporter gene/probe system, namely the human estrogen receptor ligand binding domain (hERL)/16α-18F fluoro-17β-estradiol (18F-FES), could be used for monitoring vascular endothelial growth factor (VEGF) gene expression and response to bone marrow mesenchymal stem cell (MSCs) therapy in ischemic heart disease. Reporter gene hERL and therapeutic gene VEGF165 were linked through internal ribosome entry site (IRES), and then the recombinant adenovirus vector Adenovirus 5-hERL-IRES-VEGF (Ad5-EIV) was constructed and transfected into MSCs, and named Ad5-EIV-MSCs. Rat myocardial infarction was induced by coronary arterial branch ligature, and Ad5-EIV-MSCs were transplanted by injection into the peripheral myocardium, while non-transfected MSCs transplantation used as controls. Fluorine-18-FDG micro-PET imaging was performed to confirm myocardial infarction 1 day after surgery. Fluorine-18-FES micro-PET/CT images were acquired 2 days after Ad5-EIV-MSCs transplantation. Myocardial specimens were obtained and stained with hematoxylin-eosin (H&E) staining to verify the myocardial infarction. The expression of estrogen receptor (ER) and VEGF was detected using immunohistochemistry (IHC). Rat myocardial infarction models were successfully produced and confirmed by H&E staining. Images of 18F-FDG PET showed obvious reduced or absent uptake of 18F-FDG on the infarct myocardium, while uniform and well-distribution on the normal myocardium. 18F-FES micro-PET/CT showed the tracer notable accumulated in the apical region where Ad5-EIV-MSCs were injected with an uptake value of 0.38±0.09%ID/g, which was much higher than that of surrounding normal myocardium with nearly no uptake of 18F-FES (0.10±0.03%ID/g, n=5, P<0.05). In the group of non-transfected MSCs, the apical uptake was similar to that of normal myocardium. Immunohistochemistry studies demonstrated positive expression of both ER and VEGF in the involved region accompanied by active angiogenesis. This study confirmed that hERL/18F-FES could be used as a reporter gene/probe system for monitoring gene and cell therapy in the ischemic heart disease.