Vascular Endothelial Growth Factor Immunostaining Research Articles

Effect of a portable light emitting diode device on wound healing in a rat model.

Light-emitting diode (LED) lights produce a variety of wavelengths that have demonstrable efficacy in therapeutic and aesthetic fields. However, a repetitive treatment regimen is required to produce treatment outcomes, which has created a need for portable LED devices. In this study, we aimed to develop a portable therapeutic LED device and investigate its healing effect on excisional wounds in a rat model. The 35 × 35 mm-sized LED device was used on a total of 30 rats with full-thickness wounds that were divided into two groups depending on radiation intensity (11.1 and 22.2 mW/cm2 group). LED irradiation was performed every 24 h for 30 min, over 14 days, in direct contact with the wound. Percentage wound closure was measured by photographic quantification and was assessed histologically using haematoxylin and eosin (H&E) and Masson's Trichrome staining, and immunohistochemistry for Vascular endothelial growth factor (VEGF) and CD31. Percentage wound closure was significantly higher in 22.2 mW/cm2 irradiated wounds than that in the control wounds on days 7 and 10. The area of collagen deposition was remarkably larger in 22.2 mW/cm2 irradiated wounds than that in the control, with more horizontally organized fibres. CD31 immunostaining confirmed a significant increase in the number of microvessels in 22.2 mW/cm2 irradiated wounds than that in the control wounds, although there was no difference in VEGF immunostaining. Our novel portable LED device accelerates wound healing in a rat model, raising the possibility that portable LED devices can combine convenience with accessibility to play an innovative role in wound dressing.

Immunostaining for VEGF and Decorin Predicts Poor Survival and Recurrence in Canine Soft Tissue Sarcoma

The aim of this study was to investigate whether using immunohistochemistry to detect the angiogenic proteins vascular endothelial growth factor (VEGF) and decorin can help predict the risk of local recurrence of, or death from, canine soft tissue sarcoma (STS). VEGF and decorin were detected using validated immunohistochemical methods on 100 formalin-fixed paraffin-embedded samples of canine STS. The tumours had been resected previously, with clinical outcome determined by questionnaire. Each slide was assessed by light microscopy and the pattern of immunostaining with VEGF and decorin determined. Patterns of immunostaining were then analysed to detect associations with outcome measures of local recurrence and tumour-related death. High VEGF immunostaining was significantly (p < 0.001) associated with both increased local recurrence and reduced survival time. The distribution of decorin immunostaining within the tumour was significantly associated with survival time (p = 0.04) and local tumour recurrence (p = 0.02). When VEGF and decorin scores were combined, STS with both high VEGF and low decorin immunostaining were more likely to recur or cause patient death (p < 0.001). The results of this study suggest that immunostaining of VEGF and decorin may help predict the risk of local recurrence of canine STS.

Metformin suppresses LRG1 and TGFβ1/ALK1-induced angiogenesis and protects against ultrastructural changes in rat diabetic nephropathy

Diabetic nephropathy (DN) has high prevalence and poor prognosis which make it a research priority for scientists. Since metformin, a hypoglycaemic drug, has been found to prolong the survival of mice with DN. This study aims at investigating the molecular mechanisms leading to DN in rats and to explore the role of leucine-rich α-2-glycoprotein-1 (LRG1), activin-like kinase1 (ALK1), and transforming growth factor–β (TGFβ1) in the pathologic alterations seen in DN. The aim was also extended to explore the protective action of metformin against DN in rats and its influence on LRG1and ALK1/TGFβ1 induced renal angiogenesis. 24 male rats were used. Rats were assigned as, the vehicle group, the diabetic control group and diabetic + metformin (100 and 200 mg/kg) groups. Kidney samples were processed for histopathology, immunohistochemistry and biochemical analysis. Bioinformatic analysis of studied proteins was done to determine protein-protein interactions. Metformin reduced serum urea and creatinine significantly, decreased the inflammatory cytokine levels and reduced LRG1, TGFβ1, ALK1 and vascular endothelial growth factor (VEGF) proteins in rat kidneys. Bioinformatic analysis revealed interactions between the studied proteins. Metformin alleviated the histopathological changes observed in the diabetic rats such as the glomerular surface area and increased Bowman’s space diameter. Metformin groups showed decreased VEGF immunostaining compared to diabetic group. Metformin shows promising renoprotective effects in diabetic model that was at least partly mediated by downregulation of LRG1 and TGFβ1/ALK1-induced renal angiogenesis. These results further explain the molecular mechanism of metformin in DN management.

Vascular endothelial growth factor (VEGF) expression in histological endometritis in the bitch: An immunohistochemical study.

This work aimed to study the immunolocalization of vascular endothelial growth factor (VEGF) and its FLT-1 receptor (VEGFR1) in systemically healthy bitches with and without endometritis (END). Thirty-one uterine samples from mixed-breed, systemically healthy diestrous bitches, were included in the study. The bitches were divided into three groups after histological evaluation (normal endometrium [n=8]; acute-END [n=8]; subacute-END [n=5]; chronic-END [n=10]). The immunohistochemical localization of VEGF and VEGFR1 were detected following standard procedures. Data were analysed using SPSS 26.0. Immunoreactions were detected in cells from the luminal epithelium (LE), epithelium of the superficial glands (ESG) and epithelium of the deep glands (EDG), and some cells localized in the stroma. Immunostaining of VEGF in acute-END was higher than chronic-END (p < .05) and was higher in LE than EDG (p .017). Marked-area was higher in END than normal endometrium in VEGFR-1 (p < .004). The current results provide new information on the END in systemically healthy females. Data reported here indicate that VEGF could be involucrated in the pathogenesis of END. Future studies could provide more information, support our results and elucidate the role of VEGF in the pathogenesis of END.

Expression of vascular endothelial growth factor in follicular cell-derived lesions of the thyroid: Is NIFTP benign or precancerous?

Objective: Vascular endothelial growth factor (VEGF) is an angiogenic factor that plays an important role in physiological and pathological angiogenesis of the thyroid. The aim of the current study was to determine the expression characteristics of VEGF in follicular cell-derived lesions of the thyroid and to assess whether a new entity noninvasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP) is precancerous. Material and Methods: Patients diagnosed with 33 follicular adenomas (FA), 41 invasive follicular variant papillary thyroid cancer (IN-FVPTC), and 40 NIFTP in surgical resection materials were evaluated retrospectively. Immunostaining was performed on 5-μm paraffin tissue sections. The percentages of immunostaing for VEGF were evaluated on pathological materials. We used a percentage of labeled thyrocytes score (0, no labeling; 1, <30%; 2, 31-60%; 3, >60%) and an intensity score (0, no staining; 1, weak; 2, intermediate; 3, strong). The sum of two scores were accepted as the total score. Results: Mean ages of the FA, IN-FVPTC, and NIFTP groups were 44.7 ± 11.7 years, 46.9 ± 13.6 years, 43.2 ± 15.4 years, respectively and the mean VEGF immunostaining scores were 44.7 ± 29.3, 50.2 ± 32.54, 4 ± 26.3 respectively. Although there was no statistically significant difference (p= 0.347), the total score of the NIFTPs was higher than the scores of the FA (mean= 3.9 ± 1.8) and IN-FVPTC(mean= 4.3 ± 1.9) groups with a mean value of 4.6 ± 1.7. This result was remarkable. There was no statistically significant difference between tumor diameters and staining percentages (p= 0.750). Conclusion: Even if there were no statistical differences for VEGF immunostaining, it was high in NIFTPs. Since we know the role of VEGF in tumorigenesis, we can hypothesize that NIPTP can be precancerous. Our argue should be corroborated by a large prospective study.

Comparison of vascular endothelialgrowth factor expression between lesional and non-lesional skin in patients with morphea: a dermoscopy-guided immunohistochemical study.

Morphea is an inflammatory fibrosing disease, initiated by vascular injury resulting in increased collagen formation and decreased collagen degradation. This study was designed to evaluate the role of angiogenic vascular endothelial growth factor (VEGF) in the vascular changes which are dermoscopically evident in morphea lesions, compared with that in non-lesional skin, by assessing its expression immunohistochemically on tissue blood vessels. Twenty patients with morphea were subjected to clinical and dermoscopic examinations. Two skin biopsies from lesional and non-lesional skin were obtained and stained with hematoxylin and eosin (H&E) and immunohistochemically with VEGF. Dermoscopic examination showed linear blood vessels in 90% of the lesions. No significant difference in the number of VEGF-stained and unstained blood vessels, was observed between the lesional and non-lesional skin (p = 0.475 and 0.191, respectively). A weak inverse correlation was found between the total number of blood vessels positive for VEGF and the disease duration, (r = - 0.48; p = 0.032). Significant differences were found between different stages of morphea and total number of blood vessels negative for VEGF, (p = 0.017). In conclusion, VEGF immunostaining, which represents the newly formed blood vessels, showed no difference between lesional and non-lesional skin in patients with morphea. Thus, the dermoscopically observable blood vessels in lesions compared with non-lesional skin are not due to angiogenesis, but rather due to the thinning and atrophy of the overlying epidermis in morphea cases, rendering the blood vessels more obvious.

Thyroid Gland Alterations in Old-Aged Wistar Rats: A Comprehensive Stereological, Ultrastructural, Hormonal, and Gene Expression Study.

The aim of the present study was to determine and elaborate on all changes in old-aged (OA) versus young-aged (YA) rat thyroids by using stereological, ultrastructural, hormonal, and gene expression analyses. We used 4- and 24-month-old male Wistar rats in our evaluation, presenting all changes in comparison with YA rats. Results showed that the thyroid parenchyma was characterized by higher absolute volumes of the gland, colloid, epithelium, and interstitium by 135, 135, 140, and 142% (p < 0.05) respectively, while the relative volumes of colloid and glands were unchanged. Ultrastructural analysis revealed less active glands, with smaller amounts of lysosomes, thyroglobulin (Tg) granules, and microvilli in the luminal colloid. Optical density values for thyroid peroxidase (TPO), Tg, and vascular-endothelial growth factor immunostaining remained unchanged; however, TPO and Tg exhibited visually stronger expression in small active follicles. Thyroxine (T4)-Tg, the relative intensity of fluorescence (RIF), serum T4, and the sodium-iodide symporter immunohistochemical and gene expressions decreased by 20, 40, 29, and 31% (p < 0.05), respectively, in OA thyroids. Pituitary thyroid-stimulating hormone (TSH) RIF increased by 44% (p < 0.05), but the TSH serum concentration remained unchanged. In conclusion, the obtained results indicate depression of the thyroid gland synthetic and secretory capacity with advanced age.

Angiotensin-Converting Enzyme Inhibitor, Captopril, Improves Scar Healing in Hypertensive Rats.

Pathological cutaneous scars, with aberrant extracellular matrix accumulation, have multiple origins. Antihypertensive medications, such as calcium channel blockers, have been used to treat pathological scars. However, a relationship between angiotensin-converting enzyme (ACE) inhibitors, pathological scars, and blood pressure (BP) has never been reported. Here, we aimed to compare the differences in scar development and the effects of the administration of systemic ACE inhibitor on scar tissue in a normotensive rat, the Wistar Kyoto rat (WKY), a hypertensive rat, and the spontaneously hypertensive rat (SHR). Using an 8-mm punch, we created two full-thickness skin defects in a total of 32 rats (16 WKY and 16 SHR) to obtain a total of 64 wounds. We established control WKY (n = 16), captopril-treated WKY (n = 16), control SHR (n = 16), and captopril-treated SHR (n = 16) groups and started captopril (100 mg/g per day) treatment on day 21 in the appropriate groups. The BP of all groups was measured at 0, 3, and 5 weeks. The scar area was measured by histopathological examination, and scarring was expressed in terms of scar area and fibroblast and capillary counts. The expression of heat shock protein (HSP) 47, type I and III collagens, alpha-smooth muscle actin (α-SMA), Ki67, and vascular endothelial growth factor (VEGF) was investigated using immunohistochemistry. The scar area and fibroblast count were significantly higher in control SHR than in control WKY. The scar area, fibroblast count, and capillary count were significantly smaller in captopril-treated SHR than in control SHR. Immunostaining for α-SMA, Ki67, and VEGF also showed a noticeable decrease in scarring in the treated SHR compared with that in control SHR. Thus, BP affects scar development in a rat model, and an ACE inhibitor is more effective at reducing scars in hypertensive rats than in normotensive rats.

Phosphodiesterase-5 inhibition improves bone regeneration at the early stages of ischemic osteonecrosis of the femoral head in rats.

Posttraumatic osteonecrosis of the femoral head (ONFH) affects patients at different ages and may lead to functional limitation and joint replacement, with total hip arthroplasty, which is a costly procedure. Proposed methods to optimize ischemic tissue regeneration have been reported. Phosphodiesterase-5 inhibitors act by inhibiting the degradation of guanosine 3',5'-cyclic monophosphate in the nitric oxide pathway, increasing its bioavailability and promoting vascular endothelial growth factor (VEGF)-mediated neovascular recruitment and the induction of tissue regeneration in the traumatized bone. Thirty male Sprague-Dawley rats (6 months old) were subjected to an experimental model of traumatic ONFH divided into two groups, according to the administration of 5 mg/kg sildenafil or water (control group). Rats were then killed at 7, 14, and 21 days. Histological (Goldner's trichrome), histochemical (periodic acid-Schiff [PAS]), and immunohistochemical (VEGF and osteopontin [OPN]) techniques were used to quantify bone and vascular responses. Higher levels of VEGF (p < 0.01) and OPN (p < 0.01) immunostaining in the epiphysis, the greater formation of osteoid tissue (p < 0.01 on Day 7; p < 0.05 on Day 14), and higher levels of PAS staining (p < 0.01 on Day 7) were observed in the sildenafil-treated group. The present study demonstrated that sildenafil optimized bone tissue regeneration by increasing VEGF signaling and OPN expression, with increased bone formation (osteoid and carbohydrate macromolecule deposition) in the early stages following traumatic ischemic insult. Thus, sildenafil treatment may improve the prognosis of patients with osteonecrosis.

Perigestational alcohol consumption induces altered early placentation and organogenic embryo growth restriction by disruption of trophoblast angiogenic factors

Research questionMaternal alcohol consumption produces fetal retardation and malformations, probably associated with placental defects. Does perigestational alcohol consumption up to organogenesis lead to abnormal placentation and embryo growth restriction by disrupting the vascular endothelial growth factor (VEGF) system in embryo–placental development? DesignFemale mice were treated with 10% ethanol in drinking water before and up to day 10 of gestation. Control mice received ethanol-free water. After treatment, the trophoblastic tissue, embryo growth and the angiogenic VEGF pathway were analysed. ResultsFemale mice who had received treatment had resorbed and delayed implantation sites with poor ectoplacental cone development. Reduced trophoblastic area tissue from female mice who had received treatment had abnormal junctional zone and diminished labyrinthine vascularization. After treatment, the labyrinth had increased chorionic trophoblast proliferation, hypoxia inducible factor-1α immunoexpression but reduced apoptosis. The embryo growth was reduced concomitantly with low VEGF immunostaining but high endothelial nitric oxide synthase (eNOS) expression. In junctional and labyrinth of treated female mice, gene and protein immunoexpression of VEGF was reduced and the protein expression of FLT-1 increased compared with controls. Increased activation of kinase insert domain receptor receptor (phosphorylated KDR) and expression of eNOS were observed in placenta of treated female mice. Immunoexpression of metalloproteinase-9, however, was reduced in junctional zone but increased in labyrinth, compared with controls. ConclusionsThese data reveal inadequate expression of VEGF/receptors and angiogenic eNOS and metalloproteinase factors related to abnormal early placentation after perigestational alcohol ingestion, providing insight into aetiological factors underlying early placentopathy associated with intrauterine growth restriction caused by maternal alcohol consumption.

The Effect of EGF on VEGF expression on Submandibular Salivary Gland of Albino Rats Receiving Doxorubicin

Background: Vascular endothelial growth factor (VEGF) has two prominent biological effects of inducing mitogenesis and migration of vascular endothelial cells and to produce permeability on the microvasculature. Epidermal growth factor (EGF), known by its mitogenic activity on different types of cells, could induce the proliferation and differentiation of the epithelial cells and stimulate its regeneration. Doxorubicin (DXR) is a widely used anticancer drug and is one of most commonly used chemotherapeutic drugs, however it may cause some serious complications such as toxicity in various tissues in the body.Aims: The present study aims to investigate the effect of EGF on the VEGF immunoexpression in submandibular salivary gland tissue of rats receiving Doxorubicin (DXR).Methods and Material: 30 male Albino rats, two months old and weighing 200-250 gm body weight, were divided equally into three groups as follows: Group I (control group). Group II: rats received 20 mg/kg body weight DXR as a single intra peritoneal injection. Whereas, group III: rats received the same dose of DXR and on the next day they were injected intraperitoneally with EGF in a daily dose of 10μg/kg body weight for 7 consecutive days.Submandibular salivary glands were dissected out and processed for histological investigation and immunohistochemical localization of VEGF in the glandular tissue.Statistical analysis used: ANOVA was used to compare the mean area percentage of VEGF immunostaining among the specimens of different groups followed by Tukey’s Multiple Comparison Test.Results: VEGF expression in submandibular salivary glands of rats significantly decreased after DXR injection. However, daily intraperitoneal injections of EGF restored normal levels of VEGF expression in the glandular tissue.Conclusions: EGF preserved glandular architecture after DXR injection and consequently the immunohistochemical expression of VEGF in the glands to approximately the normal level.

Thalidomide Reduces Vascular Endothelial Growth Factor Immunostaining in Canine Splenic Hemangiosarcoma.

Hemangiosarcomas (HSA) are common neoplasms of dogs that often metastasize and are typically fatal. Recently it was demonstrated that thalidomide extends the survival time of dogs with HSA, potentially due to thalidomide-induced inhibition of vascular endothelial growth factor (VEGF) production by the neoplastic cells. To investigate this, immunostaining was used to evaluate VEGF within HSA metastases that developed after thalidomide treatment. The immunostaining was then compared to VEGF immunostaining in primary tumors from the same dogs prior to treatment with thalidomide and in metastatic tumors from untreated dogs with splenic HSA. Immunostaining was scored from 1 to 4 for each sample. Immunostaining in the metastatic lesions that had been treated with thalidomide had a mean immunostaining score of 1.4 which was significantly lower than the mean score in the corresponding primary splenic HSA (3.8, p = 0.02) and in metastases from untreated dogs (3.5, p = 0.02). This supports the hypothesis that thalidomide prolongs survival time in dogs with HSA due to inhibition of VEGF production by the neoplastic cells. As VEGF remained visible within HSAs exposed to thalidomide, additional treatments to inhibit VEGF production may further prolong survival times of dogs with these common canine neoplasms.

Comparison of Volume Retention and Biocompatibility of Acellular Dermal Matrix/Hyaluronic Acid Filler to Autologous Fat Grafts in a Mouse Model.

Although fat grafting has become a widely used technique in aesthetic and reconstructive surgeries, resorption is still a challenge. Micronized acellular dermal matrix (ADM) has been considered as a stable, biocompatible soft tissue coverage material that can be used as a volume filler. Here, we compared the bioacceptance and sustainability of ADM hyaluronic acid (HA) filler with human fat graft in a mouse model. Harvested human fat and ADM/HA filler were injected randomly on the dorsal side of mice. Thirty-two mice were analyzed over a 7-week period with respect to volume, weight and microscopic evaluations with hematoxylin and eosin (H&E), epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) staining. The volume and weight were maintained at 80% and 83%, respectively, in the ADM/HA filler group and 56.5% and 49.7%, respectively, in the fat-grafted group by the 7th week. There were no significant microscopic differences in inflammatory changes and granulation via H&E staining. At 1 and 2weeks, EGF and VEGF immunostaining intensity scores were lower in the ADM/HA filler group. At 7weeks, there were no significant differences in immunostaining intensity scores between the two groups. During the 7-week experimental period, the ADM/HA filler showed no foreign body reaction and the proper volume was well maintained. This suggests that the ADM/HA filler can possibly be used in small amounts as an alternative to autologous fat grafts. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authorshttp://www.springer.com/00266.

Immunohistochemical staining of mTOR, vascular endothelial growth factor, CD34 and microvascular density in clear cell renal cell carcinoma and its correlation with clinicopathological profile of the tumour

Background: The study analyses immunoexpression of vascular endothelial growth factor (VEGF) and mammalian target of rapamycin (mTOR) based pathways which are mainstay of drug therapy in advanced and metastatic clear cell renal cell carcinoma (ccRCC) and to delineate their role as prognostic markers in a cohort of primary ccRCC. Methods: Descriptive study wherein immunohistochemical profiling of 100 cases of primary ccRCC was carried out using primary antibodies against mTOR, VEGF and CD34. Mean vascular density (MVD) and immunoreactivity was correlated with grading, staging, risk stratification and survival data. Results: mTOR and VEGF immunostaining was positive in 54% and 70% cases respectively with strong immunoreactivity correlating with high grade, high stage and high risk tumours with low survival. Inverse association was observed with MVD wherein low grade, low stage and low risk ccRCCs were associated with higher MVD. Conclusion: mTOR and VEGF expression in ccRCCs is associated with unfavourable prognosis. High MVD correlates with better outcome though concordance is limited by tumour heterogeneity.

Low intensity pulsed ultrasound increases mandibular height and Col-II and VEGF expression in arthritic mice

ObjectiveRheumatoid arthritis (RA) is a chronic inflammatory disease involving persistent inflammation resulting in cartilage and bone damage. RA can affect the temporomandibular joint (TMJ), and damage to the TMJ condyle can lead to craniofacial developmental disturbances, causing micrognathia, malocclusion, retrognathia, and increased overjet. Current treatments of TMJ arthritis are unsatisfactory. This pilot study aimed to investigate the effect of low intensity pulsed ultrasound (LIPUS) on the mandible and TMJ condyles in an RA mouse model using micro-computed tomography (Micro-CT), histologic, and immunohistochemical analyses. MethodsMRL-lpr/lpr mice received LIPUS application to their TMJs for 20 min/day for 2 and 4 weeks. Micro-CT analysis measured condylar length and width, posterior mandibular height (P.M.H), mandibular ramus length (M.R.L), effective mandibular length (Ef.M.L), angular process length (A.P.L), mandibular plane (M.P), mandibular axis (M.Ax), and lower incisor height (L.I.H). Condylar cartilage thickness was histologically measured, and type II collagen (Col-II), vascular endothelial growth factor (VEGF), nuclear factor kappa-B ligand (RANKL), and osteoprotegerin (OPG) expression was analyzed using immunohistochemistry. ResultsComparing the LIPUS-treated group with the control, P.M.H, M.R.L, and M.P were significantly greater in the LIPUS-treated group. Immunostaining for Col-II and VEGF was stronger in the LIPUS-treated group after 4 weeks. OPG showed slightly more expression in the LIPUS group. ConclusionsLIPUS may enhance mandibular and TMJ condylar bone formation in this RA mouse model by preventing any growth disturbances involved in inflammation. Further studies are recommended to analyze the effect of LIPUS on TMJ of RA in other animal models.

Expression of VEGF in breast lesions: an immunohistochemical study

Background: Breast cancer is the most frequent cancer in India. During the last few years, several investigators have focused on tumor angiogenesis as a critical step in cancer development and progression. Among these, vascular endothelial growth factor (VEGF) is emerging as a prognostic marker in patients with several type of cancer including breast cancer. The aim of the study was to analyse the expression of VEGF in human breast cancer as compared to normal breast tissue and benign breast lesions by immunohistochemistry. Also, to assess the usefulness of VEGF as a predictor of aggressiveness of breast lesions.Methods: Formalin fixed paraffin embedded sections of 10 cases of normal breast tissue, 20 cases of benign breast lesions and 20 cases of malignant breast lesions were taken up for the study and subjected to immunohistochemistry using VEGF.Results: The intensity of VEGF immunostaining in normal breast, benign and malignant breast lesions was evaluated and scoring was graded as 0, 1+, 2+, 3+ and 4+. Statistical analysis was performed with Chi-Square test and significant differences were noted between these 3 groups (p value &lt;0.05).Conclusions:VEGF expression correlated well with the grade and stage of tumor indicating that VEGF positive tumors are biologically aggressive and are associated with poor prognosis but little is known about the implication of genetic alterations of VEGF in benign breast lesions.

Hypoxia-inducible factor 1-alpha and vascular endothelial growth factor in cartilage tumors

ABSTRACTNeoangiogenesis has been demonstrated in chondrosarcoma. Anti-angiogenic therapies are being tested in clinical trials for chondrosarcomas. Studies of the underlying mechanisms have been performed almost exclusively in cell lines. We immunostained 20 samples of chondrosarcoma and 20 samples of enchondromas with antibodies against hypoxia-inducible factor 1-alpha (HIF-1-alpha) and vascular endothelial growth factor (VEGF). The immunohistochemical staining of HIF-1-alpha and VEGF were highly correlated. Enchondromas were HIF-1-alpha and VEGF negative, whereas all chondrosarcoma exhibited HIF-1-alpha and VEGF immunostaining. HIF-1-alpha/VEGF double positive cases were almost exclusively chondrosarcomas with a high tumor grade. We suggest that HIF-1-alpha is a marker of malignancy in chondrosarcomas that correlates with tumor neo-angiogenesis. Our findings also suggest that a HIF-1-alpha/VEGF angiogenic pathway may exist in chondrosarcoma in vivo as in other malignant tumors. The inclusion of novel inhibitors to HIF-1-alpha and other factors may optimize anti-angiogenic interventions in chondrosarcoma.

Euterpe oleracea extract inhibits tumorigenesis effect of the chemical carcinogen DMBA in breast experimental cancer

BackgroundAmong the processes involved in the breast tumor microenvironment, angiogenesis and inflammation play a central role, and the main factors of these processes are the vascular endothelial growth factor (VEGF), cyclooxygenase 2 (COX-2) and macrophages. Recently, the extract of Euterpe oleracea (açaí), a fruit that is widely found in the Amazon region, already showed antitumorigenic effects in vitro in human breast cancer cell lines. The present study aimed to investigate the effect of açaí on breast cancer using a chemically DMBA (7,12-dimethylbenzanthracene) experimental model.MethodsOne day after initiation of treatment with açaí, mammary carcinogenesis was induced in female Wistar rats using a subcutaneous injection of 25 mg/kg of DMBA in the mammary gland. Forty rats were randomized into two groups: treated with 200 mg/kg of either açaí extract or vehicle, via gastric tube for 16 consecutive weeks. After treatment, the tumor was collected for macroscopic, histological and immunohistochemical (VEGF, vascular endothelial growth factor receptor 2 -VEGFR-2, COX-2 and matrix metalloproteinase -MMP-9) analyses; peritoneal fluid was subjected to flow cytometry (F4–80/MAC-2+) and ELISA immunoassay (VEGF, prostaglandin E2 -PGE2 and interleukin-10 -IL-10). Heart, liver and kidney samples were collected for histological analysis.ResultsAfter 16 weeks of induction, the mammary carcinoma was confirmed by macroscopic and histological evaluation. Survival analysis indicates that açaí increased the survival (P = .0002, long-rank test) and reduced the deaths number (P = .0036, Chi-square test). Açaí treatment decreased the number of inflammatory cells and macrophage positive cells (Mac-2 + F4–80+), as well as promoting a reduction in immunostaining of VEGF, VEGFR-2 and COX-2. The açaí group also exhibited lower concentrations of PGE2, VEGF and IL-10 compared to the control. The histopathological results of the liver and kidneys showed protective effect of açaí, since in the control group, there was an increase in fibrosis, atypical cells and hemorrhagic microenvironment.ConclusionThe results of this study demonstrated the antiangiogenic and anti-inflammatory potential of açaí, like due to the decreases of the number of activated macrophages, resulting in the inhibition of DMBA carcinogenicity in breast cancer.

Expression and clinical correlation of NGAL and VEGF in endometrial carcinoma.

To determine the expression of neutrophil gelatinase-associated lipocalin (NGAL) and vascular endothelial growth factor (VEGF) in endometrial carcinoma. The clinical features (FIGO staging, pathological type, differentiation level, myometrial invasion depth and tumor size) and immunostaining analysis of endometrial carcinoma (30 cases), atypical hyperplasia (30 cases) and proliferative endometrial tissues (30 cases) were analyzed. Immunohistochemistry and reverse transcription PCR (RT-PCR) were performed to detect the expression of NGAL and VEGF. The positive tissue immunostaining and mRNA expression of NGAL and VEGF in endometrial carcinoma were significantly higher than in either atypical hyperplasia or proliferative endometrial tissues (p<0.05). The relative expression level of NGAL and VEGF was positively correlated with worse FIGO staging, higher differentiation level and a greater myometrial invasion depth (p<0.05); but not with patient age, pathological type or tumor size (p>0.05). The abnormal high expression of NGAL and VEGF observed in the endometrial carcinoma may be an important biomarker for early tumor diagnosis or as a novel target for therapeutic intervention.

Vitamin E improves testicular damage in streptozocin-induced diabetic rats, via increasing vascular endothelial growth factor and poly(ADP-ribose) polymerase-1.

The precise mechanism by which diabetes impairs spermatogenesis and testicular function is not exactly known. Vascular endothelial growth factor (VEGF) and poly(ADP-ribose) polymerase-1 (PARP-1) are important for germ cell homeostasis and repair of DNA respectively. The aim of this study was to investigate the correlation between diabetes-induced testicular damage and testicular VEGF and PARP-1 expression and the possible protective role of vitamin E supplementation. A total of 45 male Wistar albino rats were randomly divided into three groups: Group I (nondiabetic rats), Group II (streptozocin-induced diabetic rats) and Group III (streptozocin-induced diabetic rats treated orally with 0.4mg/kg vitamin E). Five weeks later, testicular tissue was used for assessment of MDA concentration by colorimetry, histopathological examination and immunostaining for PARP-1 and VEGFIn diabetic rats, testicular weight, seminiferous tubule diameter and germinal epithelial thickness were decreased, basement membrane was thickened and Johnsen score decreased. Reduced VEGF and PARP-1 immunostaining were associated with decreased Johnsen score in diabetic rats. Vitamin E administration was protective against oxidative stress-associated damage evidenced by lower MDA levels, improved testicular weight, spermatogenesis and higher immunostaining for VEGF and PARP-1. Testicular VEGF and PARP-1 might therefore be helpful biomarkers for diabetic testicular damage. Administration of vitamin E may have a protective role against diabetes-induced testicular damage.