Efecto antiangiogénico de la triamcinolona asociada a lidocaína con epinefrina de uso odontológico en la membrana alantocoriónica de pollo / Antiangiogenic effect of triamcinolone associated with lidocaine with epinephrine for dental use in chicken chorioallantoic membrane

RESUMEN: El proceso angiogénico se define como el proceso en el que los vasos sanguíneos generan brotes dando como resultado neovascularidad. Un desbalance en el proceso angiogénico contribuye a numerosos desórdenes inflamatorios, infecciosos, isquémicos, inmunológicos y malignos. En el territorio maxilofacial se pueden encontrar patologías neoplásicas benignas de desarrollo local con un marcado componente angiogénico que determinan su crecimiento y agresividad. Sin embargo, existe escasa evidencia de cómo tratarlas en base al control de la angiogénesis. Terry & Jacoway (1994) desarrollaron un protocolo de tratamiento para lesiones neoplásicas benignas con un importante componente vascular que se utiliza actualmente. Este protocolo consiste en la infiltración intralesional de una suspensión de triamcinolona 10 mg/ml más una solución de anestésico local de uso odontológico como la lidocaína al 2 % asociada a epinefrina en una concentración de 1:200.000. Sin embargo, el uso de epinefrina podría disminuir la acción antiangiogénica de la triamcinolona al ser un vasoconstrictor. El objetivo de este estudio es comparar el efecto antiangiogénico, en la membrana alantocoriónica de pollo (MAC), de esta suspensión versus el efecto de la triamcinolona sin asociar a anestésicos locales. Los resultados del efecto antiangiogénico en la MAC de pollo, obtenidos en la investigación concluyeron que la suspensión de triamcinolona asociada a lidocaína con epinefrina es similar al de la suspensión de triamcinolona sin asociar a anestésicos locales. Además, se logró determinar que las suspensiones de triamcinolona sin asociar a anestésicos locales y las asociadas a anestésicos locales con o sin vasoconstrictor poseen un marcado efecto antiangiogénico, en la MAC de pollo, en comparación al grupo control.

SUMMARY: Angiogenesis is defined as the process through which new blood vessels form from previously existing vessels. Several inflammatory, infectious, ischemic, immunological and malignant disorders are caused by the lack of adequate angiogenesis balance. In the maxillofacial area, there are invasive benign neoplastic pathologies with a strong angiogenic component, which determines aggressive behavior and growth. Studies in the literature are scarce regarding treatment of these conditions based on angiogenesis control. Currently, the protocol used to treat these maxillofacial benign neoplastic lesions, was developed in 1994 by Terry & Jacoway and has a strong angiogenic component. Consequently lesions are treated via intra-lesion administration of triamcinolone 10 mg / mL, a solution used in dental local anesthetic, such as lidocaine 2 %, in conjunction with epinephrine at a concentration of 1:200,000. The objective of this study was to compare the antiangiogenic effect of this protocol in chicken chorioallantoic membrane (CAM) without the use of local anesthetic. The results of the antiangiogenic effect in the CAM obtained in this study concluded that the effect of the suspension of triamcinolone associated to lidocaine with epinephrine, is similar to the suspension of triamcinolone without associating local anesthetics. Furthermore, it was determined that suspensions of triamcinolone without local anesthetic, and those associated to local anesthetic with, and without vasoconstrictor have a strong antiangiogenic effect in CAM compared to the control group.

Efecto del aliskireno sobre la angiogénesis en modelo de membrana alantocoriónica (MAC) de pollo / Effect of aliskiren on angiogenesis in chick embryo chorioallantoic membrane (CAM) assay

La acción antiangiogénica de los inhibidores del receptor de angiotensina II (ARA II), ha sido documentada previamente, sin embargo, no ha sido descrita la relación entre angiogénesis e inhibidores directos de la renina (DRIs), los cuales participan regulando el sistema renina-angiotensina-aldosterona (SRAA). El objetivo fue demostrar el efecto antiangiogénico de aliskireno, un DRI, en membranas alantocoriónicas (MAC) de pollo, para lo cual fueron instilados aliskireno y enalapril sobre MAC en distintas concentraciones para realizar su comparación posterior. En secciones histológicas seriadas se registró el número de vasos sanguíneos presentes en 9000 µm2 bajo microscopio de luz a máximo aumento, y se realizó análisis estadístico utilizando ANOVA y el test de Tukey para demostrar posibles diferencias. Los receptores tratados con aliskireno, en ambas concentraciones utilizadas, presentaron menor densidad vascular, en comparación con los controles, siendo ésta estadísticamente significativa a mayor concentración. Aliskireno en concentraciones altas tiene un efecto antiangiogénico en un modelo experimental de MAC. Este hallazgo plantea la necesidad de estudios posteriores, dada la proyección que podría tener el uso inhibidores directos de la renina. A partir de estos resultados, se podría pensar en la factibilidad del uso de aliskireno para la modulación de la angiogénesis en diversas enfermedades crónicas no transmisibles.

Angiogenesis is the formation of new blood vessels from pre-existing ones. Antiangiogenic effect of angiotensin receptor blockers has been reported, however, the relationship between direct renin inhibitors and angiogenesis has not been well described. To assess the antiangiogenic effect of aliskiren, a direct renin inhibitor, on chick embryo chorioallantoic membrane (CAM) assay. Aliskiren and enalapril were instilled in different concentrations and compared. In serial histological sections, the number of blood vessels per 9000 µm2 area under observation through optical microscope using maximum zoom, was registered. Statistical analysis using Anova and Tukey test in order to show possible differences, was performed. Receptors treated with aliskiren presented lower vascular density, which was statistically significant when a higher concentration was administered. High concentrations of aliskiren have an antiangiogenic effect on CAM assay. This finding means further studies are needed, because of the usefulness direct renin inhibitors could have. These results, also, might enhance the possibility of using aliskiren for regulating angiogenesis in the context of non-transmissible chronic diseases.

In ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membrane

BACKGROUND: Leptin, the cytokine produced by white adipose tissue is known to regulate food energy homeostasis through its hypothalamic receptor. In vitro studies have demonstrated that leptin plays a major role in angiogenesis through binding to the receptor Ob-R present on ECs by stimulating and initiating new capillary like structures from ECs. Various in vivo studies indicate that leptin has diverse effect on angiogenesis. A few reports have showed that leptin exerts pro angiogenic effects while some suggested that it has antiangiogenic potential. It is theoretically highly important to understand the effect of leptin on angiogenesis to use as a therapeutic molecule in various angiogenesis related pathological conditions. Chicken chorio allantoic membrane (CAM) on 9th day of incubation was incubated with 1, 3 and 5 µg concentration of HRL for 72 h using gelatin sponge. Images where taken after every 24 h of incubation and analysed with Angioguant software. The treated area was observed under microscope and histological evaluation was performed for the same. Tissue thickness was calculated morphometrically from haematoxylin and eosin stained cross sections. Reverse transcriptase PCR and immunohistochemistry were also performed to study the gene and protein level expression of angiogenic molecules. RESULTS: HRL has the ability to induce new vessel formation at the treated area and growth of the newly formed vessels and cellular morphological changes occur in a dose dependent manner. Increase in the tissue thickness at the treated area is suggestive of initiation of new capillary like structures. Elevated mRNA and protein level expression of VEGF165 and MMP2 along with the activation of ECs as demonstrated by the presence of CD34 expression supports the neovascularization potential of HRL. CONCLUSION: Angiogenic potential of HRL depends on the concentration and time of incubation and is involved in the activation of ECs along with the major interaction of VEGF 165 and MMP2. It is also observed that 3 µg of HRL exhibits maximum angiogenic potential at 72 h of incubation. Thus our data suggest that dose dependent angiogenic potential HRL could provide a novel role in angiogenic dependent therapeutics such as ischemia and wound healing conditions.

Angiogenic response of advanced glycation end products (AGEs) involves PPAR

Diabetes is associated with increased formation of advanced glycation end products (AGEs), which have been implicated in micro and macrovascular complications of diabetes. Our earlier reports showed proangiogenic effect of AGE-bovine serum albumin (BSA). In order to understand the mechanism of AGE-mediated angiogenesis, the possibility of involvement of peroxisome prolifeator activated receptor (PPAR) , a ligand activated transcription factor was examined. The angiogenic effect was studied in chick chorio allantoic membrane (CAM) and by analyzing angiogenic markers in human umbilical vein endothelial cells (HUVECs) in culture. The involvement of PPAR was investigated using synthetic PPAR agonist GW 1929 and antagonist GW 9662 and by RT-PCR. In CAM assay, PPAR antagonist GW 9662 reversed the AGE-induced effect on vascularity. In HUVECs in culture, GW 9662 reversed the effect of AGE-BSA and decreased the expression of CD 31, E-Selectin and VEGF. RT-PCR analysis showed that treatment with AGE-BSA caused upregulation of PPAR mRNA levels. The reversal of the effect of AGE on angiogenesis by treatment with PPAR antagonists and up-regulation of PPAR gene in HUVECs treated with AGE-BSA suggested the possible involvement of PPAR -dependent downstream pathway in mediating the angiogenic effect of AGE.

[ effects of different doses of atorvastatin on angiogenesis of chorioallantoic membrane of chick embryo]

Statins promote the proliferation, migration and survival of endothelial cells and bone marrow -derived endothelial progenitor cells [Angioblasts]. Angiogenesis, the formation of new blood vessels, is a dynamic and complex activity which is needed for embryogenesis and other physiological processes. However, in many pathological conditions such as solid tumor progression, the disease appears to be associated with persistent upregulated angiogenesis. In this research we used atorvastatin [0.1microm] and [10 microm] on angiogenesis of chick embryo. In this experimental study 42 Ross fertilized eggs were randomly divided into 3groups as follows: 1] control group, 2] group treated with atorvastatin [0.1microm], 3] group treated with atorvastatin [10microm]. In day 2 a window was opened on eggs in sterile condition and In day 8 gelatin sponge was placed on chorioallantic membrane [CAM] and was soaked with 10 microliters atorvastatin [0.1microm] and [10 microm] in group 2 and group 3. In day12 CAMs were examined and photographed by research photo-stereomicroscope in all cases. Data were analyzed statistically by ANOVA and Tukey tests. The average number and length of vessels in control and the group treated with atorvastatin [0.1microm] showed a significant increase [P<0.05] and the average number and length of vessels in the group treated with atorvastatin [10 microm] showed a significant decrease compare to control group [P<0.05]. The results of this study showed that atorvastatin has a stimulatory effect on angiogenesis in CAM and atorvastatin [10 microm] has an inhibitory effect on angiogenesis in CAM. It is suggested that astatine can be used as a new medicine for angiogenesis balancing in treat ment of diseases related to angiogenesis

Angiogenic activity of Calendula officinalis flowers L. in rats / Atividade angiogênica das flores da Calendula officinalis L. em ratos

Purpose: In this work, angiogenic activity of Calendula officinalis L. (Asteraceae) ethanolic extract and dichloromethane and hexanic fractions were evaluated, considering medicinal properties, especially healing activity, are attributed to this plant. Methods: Models using 36 rats and 90 embryonated eggs were used to evaluate healing and angiogenic activities of extracts and fractions of the plant, through the induction of skin wounds and the chorioallantoic membrane, respectively. The effect of vascular proliferation was also tested from the study to verify the intensity of expression of vascular endothelial growth factor (VEGF) in cutaneous wounds in rats. Results: The angiogenic activity of the extract and the fractions was evidenced in both experimental models. It was verified that this effect is not directly related to the expression of VEGF and it could be associated to other pro-angiogenic factors. Conclusion: The healing activity referred to C. officinalis is related, among other factors, to its positive effect on angiogenesis, characterized by the induction of neovascularization.

Objetivo: Neste trabalho a atividade sobre a angiogênese do extrato etanólico (EEC) e das frações diclorometano e hexânica das flores de Calendula officinalis L. (Asteraceae) cultivada no Brasil foram avaliados, visto que propriedades medicinais têm sido atribuídas às flores da planta, destacando-se a atividade cicatrizante. Métodos: Modelos utilizando 36 ratos e 90 ovos embrionados foram usados para avaliar as atividades cicatrizante e angiogênica dos extratos e frações da planta, por meio da indução de feridas cutâneas e da membrana corioalantóide, respectivamente. O efeito proliferativo vascular foi também testado a partir do estudo imunoistoquímico, realizado para verificar a intensidade da expressão do fator de crescimento endotelial vascular (VEGF) na derme de ratos. Resultados: A atividade angiogênica do extrato e das frações foi evidenciada nos dois modelos experimentais empregados. Foi evidenciado que este efeito não estava diretamente relacionado à expressão do VEGF, podendo estar associado a outros fatores pró-angiogênicos. Conclusão: A atividade cicatrizante referida a C. officinalis está relacionada ao seu efeito positivo sobre a angiogênese, e este foi caracterizado pela indução de neovascularização.

In vivo anti-angiogenic effects further support the promise of the antineoplasic activity of methyl jasmonate / Efeitos antiangiogênicos in vivo convalidam a atividade antineoplásica potencial do metiljasmonato

Molecular plant components have long been aimed at the angiogenesis and anti-angiogenesis pathways, and have been tested as sources for antineoplasic drugs with promising success. The present work deals with the anti-angiogenic effects of Methyl Jasmonate. Jasmonate derivatives were demonstrated to selectively damage the mitochondria of cancer cells. In vitro, 1-10 mM Methyl Jasmonate induced the cell death of the human umbilical vein endothelial cells (HUVEC) and the Murine melanoma cells (B16F10), while micromolar concentrations were ineffective. In vivo, comparable concentrations were toxic and reduced the vessel density of the Chorioallantoic Membrane of the Chicken Embryo (CAM). However, 1-10 µM concentrations produced a complex effect. There was increased capillary budding, but the new vessels were leakier and less organised than corresponding controls. It is suggested that not only direct toxicity, but also the drug effects upon angiogenesis are relevant to the antineoplasic effects of Methyl Jasmonate.

Moléculas de origem vegetal são, há muito, conhecidas como substâncias ativas sobre as vias de angiogênese e antiangiogênese e foram testadas como fonte de drogas antineoplásicas com sucesso promissor. Este trabalho trata dos efeitos antiangiogênicos do Metiljasmonato, um protótipo da família dos derivados do ácido jasmônico, que danificam seletivamente a mitocôndria de células neoplásicas. In vitro, metiljasmonato 1-10 mM promoveu a morte celular de células endoteliais humanas de cordão umbilical (HUVEC) e de melanoma murino (B16F10); concentrações micromolares foram inócuas. In vivo, concentrações equivalentes foram tóxicas e reduziram a densidade de vasos em membranas corioalantoicas de embrião de galinha (CAM). Entretanto, concentrações entre 1-10 µM produziram um efeito complexo. Ocorreu aumento no brotamento capilar, mas os novos vasos apresentaram-se frágeis e menos organizados que os controles correspondentes. Sugere-se que, além da toxicidade direta contra as células tumorais, a ação do metiljasmonato sobre a angiogênese seja relevante para seu efeito antineoplásico.

Angiogenic activity of Synadenium umbellatum Pax latex / Atividade angiogênica do látex de Synadenium umbellatum Pax

Synadenium umbellatum Pax, popularly known as "cola-nota", is a medicinal plant that grows in tropical regions. Latex of this plant is used to treat various diseases such as diabetes mellitus, Hansen´s disease, tripanosomiases, leukemia and several malignant tumors. In the present study, the angiogenic activity of S. umbellatum latex was evaluated using the chick embryo chorioallantoic membrane (CAM) assay. Results showed significant increase of the vascular net (p < 0.05) compared to the negative control (H2O). The histological analysis was in accordance with the results obtained. In conclusion, our data indicate that S. umbellatum latex, under the conditions of this research, presented angiogenic effect.

Synadenium umbellatum Pax, popularmente conhecida como "cola-nota", é uma planta medicinal que cresce em regiões tropicais. O látex desta planta tem sido utilizado no tratamento de várias doenças, como diabetes mellitus, hanseníase, tripanossomíases, leucemia e vários tumores malignos. No presente estudo, a atividade angiogênica do látex de S. umbellatum foi avaliada pelo ensaio da membrana corio-alantóide (MCA) de ovo embrionado de galinha. Os resultados mostraram aumento significativo da rede vascular (p < 0.05) em relação ao controle negativo (H2O). A análise histológica está em concordância com os resultados obtidos. Em conclusão, os dados indicaram que, nas condições experimentais deste estudo, o látex de S. umbellatum exibiu efeito angiogênico.