RESUMO
AIM: Angiogenesis contributes to the development of apical periodontitis, periodontitis, and other oral pathologies; however, it remains unclear how this process is triggered. The aim was to evaluate whether lipopolysaccharide (LPS) from Porphyromonas endodontalis and Porphyromonas gingivalis induced angiogenesis-related effects in vitro via TLR2 and TLR4. METHODOLOGY: Porphyromonas endodontalis LPS (ATCC 35406 and clinical isolate) was purified with TRIzol, whereas P. gingivalis LPS was obtained commercially. The effects of the different LPS (24 h) in endothelial cell migration were analysed by Transwell assays, following quantification in an optical microscope (40×). The effects of LPS on FAK Y397 phosphorylation were assessed by Western blotting. Angiogenesis in vitro was determined in an endothelial tube formation assay (14 h) in Matrigel in the absence or presence of either LPS. IL-6 and VEGF-A levels were determined in cell supernatants, following 24 h treatment with LPS, and measured in multiplex bead immunoassay. The involvement of TLR2 and TLR4 was assessed with blocking antibodies. The statistical analysis was performed using STATA 12® (StataCorp LP). RESULTS: The results revealed that P. endodontalis LPS, but not P. gingivalis LPS, stimulated endothelial cell migration. Pre-treatment with anti-TLR2 and anti-TLR4 antibodies prevented P. endodontalis LPS-induced cell migration. P. endodontalis LPS promoted FAK phosphorylation on Y397, as observed by an increased p-FAK/FAK ratio. Both P. gingivalis and P. endodontalis LPS (ATCC 35406) induced endothelial tube formation in a TLR-2 and -4-dependent manner, as shown by using blocking antibodies, however, only TLR2 blocking decreased tube formation induced by P. endodontalis (clinical isolate). Moreover, all LPS induced IL-6 and VEGF-A synthesis in endothelial cells. TLR2 and TLR4 were required for IL-6 induction by P. endodontalis LPS (ATCC 35406), while only TLR4 was involved in IL-6 secretion by the other LPS. Finally, VEGF-A synthesis did not require TLR signalling. CONCLUSION: Porphyromonas endodontalis and P. gingivalis LPS induced angiogenesis via TLR2 and TLR4. Collectively, these data contribute to understanding the role of LPS from Porphyromonas spp. in angiogenesis and TLR involvement.
Assuntos
Lipopolissacarídeos , Receptor 2 Toll-Like , Lipopolissacarídeos/farmacologia , Receptor 2 Toll-Like/metabolismo , Porphyromonas gingivalis/metabolismo , Porphyromonas endodontalis/metabolismo , Fator A de Crescimento do Endotélio Vascular , Células Endoteliais/metabolismo , Anticorpos Bloqueadores , Interleucina-6 , Receptor 4 Toll-Like/metabolismoRESUMO
ABSTRACT Purpose: To evaluate whether using platelet-rich plasma (PRP) in the graft recipient bed after the resection of a neoplasia can influence its recurrence because this product stimulates angiogenesis, mitogenesis and chemotaxis. Methods: A study with 30 rats Wistar (Rattus norvegicus albinus), which were separated into group A (induction of carcinogenesis, PRP in the postoperative period) and group B (induction of carcinogenesis, absence of PRP in the postoperative period), with 15 animals in each. Carcinogenesis was induced on the skin of the animals' chest by the topical application of 0.5% dimethylbenzantracene (DMBA) diluted in acetone. After surgical resection of the induced neoplasia, PRP was used to stimulate angiogenesis before surgical wound synthesis. Data on the control and experimental groups and macroscopic and microscopic variables were evaluated using analysis of variance and the Tukey's test (5%). Results: It was possible to determine that the use of PRP is good in reconstructive surgeries, but it is contraindicated in patients during tumor resection, as it can cause changes in the surgical bed, in addition to stimulating recurrences and metastases. Conclusions: PRP may interact with tumour cells that were in the recipient site of the surgical wound during the resection of a neoplasia, and a local recurrence process can be triggered by applying this product.
Assuntos
Animais , Ratos , Neoplasias Cutâneas/cirurgia , Neoplasias Cutâneas/induzido quimicamente , Plasma Rico em Plaquetas , Cicatrização , Transplante de Pele , Ratos WistarRESUMO
Abstract Objective: To analyze angiogenesis in the post-extracted tooth of Wistar rats after application of okra (Abelmoschus esculentus) extract. Material and Methods: A total of 18 rats were divided into two groups (control and treatment). Okra extract with a concentration of 30% in gel form was applied on the post-extraction socket of the treatment group. The rats were sacrificed on day-3, day-5, and day-7 after tooth extraction. The newly-formed blood vessels were counted and statistically analyzed by means of One Way ANOVA and Tukey HSD with a significance level set at 5%. Results: The newly-formed capillaries of the control group (4.67 ± 1.53) on day-3 were lower than the treatment group (9.00 ± 1.00). The newly-formed capillaries recorded from the control group, both in day-5 (9.33 ± 1.53) and day-7 (8.67 ± 1.53) were lower than the treatment group, which started to decreased from day-5 (13.67 ± 1.53) to day-7 (12.33 ± 0.58). Significant differences were found in treatment group, on day-3 compared to day-5 (p=0.005), and on day-3 to day-7 (p=0.024). Conclusion: Okra extract in gel form at 30% concentration can increase the angiogenesis during the wound healing process of the extracted tooth on Wistar rats.
Assuntos
Animais , Ratos , Extração Dentária , Cicatrização , Ferimentos e Lesões , Ratos Wistar , Abelmoschus , Análise de Variância , Estatísticas não Paramétricas , IndonésiaRESUMO
Abstract Objective: This study evaluated the angiogenesis-enhancing potential of a tricalcium silicate-based mineral trioxide aggregate (ProRoot MTA), Biodentine, and a novel bioceramic root canal sealer (Well-Root ST) in human dental pulp stem cells (hDPSCs), human periodontal ligament stem cells (hPLSCs), and human tooth germ stem cells (hTGSCs). Methodology: Dulbecco's modified Eagle's medium was conditioned for 24 h by exposure to ProRoot MTA, Biodentine, or Well-Root ST specimens (prepared according to the manufacturers' instructions). The cells were cultured in these conditioned media and their viability was assessed with 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium (MTS) on days 1, 3, 7, 10, and 14. Angiogenic growth factors [platelet-derived growth factor (PDGF), basic fibroblast growth factor (FGF-2), and vascular endothelial growth factor (VEGF)] were assayed by sandwich enzyme-linked immunosorbent assay (ELISA) on days 1, 7, and 14. Human umbilical vein endothelial cell (HUVEC) migration assays were used to evaluate the vascular effects of the tested materials at 6-8 h. Statistical analyses included Kruskal-Wallis, Mann-Whitney U, and Friedman and Wilcoxon signed rank tests. Results: None of tricalcium silicate-based materials were cytotoxic and all induced a similar release of angiogenic growth factors (PDGF, FGF-2, and VEGF) (p>0.05). The best cell viability was observed for hDPSCs (p<0.05) with all tricalcium silicate-based materials at day 14. Tube formation by HUVECs showed a significant increase with all tested materials (p<0.05). Conclusion: The tricalcium silicate-based materials showed potential for angiogenic stimulation of all stem cell types and significantly enhanced tube formation by HUVECs.
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Humanos , Materiais Restauradores do Canal Radicular/farmacologia , Células-Tronco/efeitos dos fármacos , Cerâmica/farmacologia , Silicatos/farmacologia , Compostos de Cálcio/farmacologia , Indutores da Angiogênese/farmacologia , Ligamento Periodontal/citologia , Ligamento Periodontal/efeitos dos fármacos , Germe de Dente/citologia , Germe de Dente/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Teste de Materiais , Fator de Crescimento Derivado de Plaquetas/análise , Fator de Crescimento Derivado de Plaquetas/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Sobrevivência Celular/efeitos dos fármacos , Reprodutibilidade dos Testes , Fator 2 de Crescimento de Fibroblastos/análise , Fator 2 de Crescimento de Fibroblastos/efeitos dos fármacos , Estatísticas não Paramétricas , Neovascularização Fisiológica/efeitos dos fármacos , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Citometria de FluxoRESUMO
Abstract Background: Diabetes mellitus (DM) is one of the major risk factors for cardiovascular disease, leading to endothelial dysfunction and angiogenesis impairment . MiR-126 and miR-210 support angiogenic response in endothelial cells. Objective: The present study sought to explore the effect of garlic and voluntary exercise, alone or together, on miR-126 and miR-210 expressions and cardiac angiogenesis in rats with type 1 diabetes. Methods: Male Wistar rats were divided into five groups (n = 7): Control, Diabetes, Diabetes+Garlic, Diabetes+Exercise, and Diabetes+Garlic+Exercise. Diabetes was induced in the animals by streptozotocin (ip, 50 mg/kg). The rats were then fed raw fresh garlic homogenate (250 mg/kg) or were subjected to voluntary exercise, or to combined garlic and voluntary exercise for 6 weeks. MiR-126 and miR-210 expressions in the myocardium were determined by real time PCR, and the serum lipid profile was measured by enzymatic kits. Angiogenesis was evaluated by immunostaining for PECAM-1/ CD31 in the myocardium. Results: Diabetes reduced both cardiac miR-126 expression and angiogenesis (p < 0.05). On the other hand, there was a miR-210 expression increase in the myocardium of diabetic animals (p < 0.001). However, those effects reversed either with garlic or voluntary exercise (p < 0.01). Moreover, treating diabetic rats with garlic and voluntary exercise combined had an additional effect on the expressions of miR-126 and miR-210 (p < 0.001). Furthermore, both voluntary exercise and garlic significantly improved serum lipid profiles (p < 0.001). Conclusion: The induction of diabetes decreased angiogenesis in the myocardium, whereas our treatment using long-term voluntary exercise and garlic improved myocardial angiogenesis. These changes were possibly owing to the enhancement of myocardial miR-126 and miR-210 expressions.
Resumo Fundamento: O diabetes mellitus (DM) é um dos principais fatores de risco para doenças cardiovasculares, levando à disfunção endotelial e inibição da angiogênese. O miRNA-126 e o miRNA-210 promovem a resposta angiogênica em células endoteliais. Objetivo: O presente estudo buscou explorar o efeito do alho e de exercícios físicos voluntários, isoladamente ou em conjunto, nas expressões do miRNA-126 e do miR-210 e na angiogênese cardíaca em ratos com diabetes tipo 1. Métodos: Ratos Wistar machos foram divididos em cinco grupos (n = 7): Controle, Diabetes, Diabetes+Alho, Diabetes+Exercícios e Diabetes+Alho+Exercícios. Introduziu-se diabetes nos animais por estreptozotocina (ip, 50 mg/kg). Os ratos foram então alimentados com homogenato de alho fresco cru (250 mg/kg), ou foram submetidos a exercícios voluntários, ou a uma combinação de alho e exercícios voluntários, durante 6 semanas. As expressões do miRNA-126 e do miRNA-210 no miocárdio foram determinadas por PCR em tempo real, e o perfil lipídico sérico foi medido por kits enzimáticos. A angiogênese foi avaliada por imunocoloração por PECAM-1/CD31 no miocárdio Resultados: O diabetes reduziu a expressão do miRNA-126 cardíaco e da angiogênese (p < 0,05). Por outro lado, houve um aumento da expressão do miRNA-210 no miocárdio dos animais diabéticos (p < 0,001). No entanto, tais efeitos foram revertidos com alho ou exercícios voluntários (p < 0,01). Além disso, o tratamento de ratos diabéticos conjuntamente com alho e exercícios voluntários teve um efeito adicional sobre as expressões do miRNA-126 e do miRNA-210 (p < 0,001). Além disso, tanto os exercícios voluntários quanto o alho melhoraram significativamente os perfis lipídicos séricos (p < 0,001). Conclusões: A indução de diabetes diminuiu a angiogênese no miocárdio, enquanto nosso tratamento com exercícios voluntários de longa duração e alho melhorou a angiogênese miocárdica. Estas alterações devem-se, possivelmente, ao aumento das expressões do miRNA-126 e do miRNA no miocárdio.
Assuntos
Animais , Masculino , Condicionamento Físico Animal/fisiologia , Neovascularização Fisiológica/fisiologia , Vasos Coronários/fisiopatologia , MicroRNAs/análise , Diabetes Mellitus Tipo 1/fisiopatologia , Alho/química , Triglicerídeos/sangue , Imuno-Histoquímica , Distribuição Aleatória , Colesterol/sangue , Reprodutibilidade dos Testes , Resultado do Tratamento , Ratos Wistar , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análise , MicroRNAs/fisiologia , Diabetes Mellitus Experimental/fisiopatologia , Diabetes Mellitus Tipo 1/terapia , Reação em Cadeia da Polimerase em Tempo Real , Coração/fisiopatologiaRESUMO
Purpose To explore the potential role and unclear molecular mechanisms of vaccarin in wound healing. Methods Rats skin excision model to study the effects of vaccarin on wound healing in vivo . Hematoxylin and eosin staining was performed to evaluate Histopathologic characteristics. Immunohistochemistry was employed to assess the effects of vaccarin in accelerating angiogenesis. Western blot was used to evaluate relative protein expressed levels. Results Vaccarin could significantly promote wound healing and endothelial cells and fibroblasts proliferation in the wound site. Immunohistochemistry and Western blot studies showed that the nodal proteins and receptor (bFGFR) related to angiogenesis signaling pathway were activated, and the microvascular density in the wound site was markedly higher than that in the control group. Conclusions The present study was the first to demonstrate that vaccarin is able to induce angiogenesis and accelerate wound healing in vivo by increasing expressions of p-Akt, p-Erk and p-bFGFR. This process is mediated by MAPK/ERK and PI3K/AKT signaling pathways.(AU)
Assuntos
Animais , Masculino , Ratos , Vaccaria , Cicatrização , Indutores da Angiogênese/uso terapêutico , Modelos AnimaisRESUMO
Abstract Purpose To explore the potential role and unclear molecular mechanisms of vaccarin in wound healing. Methods Rats' skin excision model to study the effects of vaccarin on wound healing in vivo . Hematoxylin and eosin staining was performed to evaluate Histopathologic characteristics. Immunohistochemistry was employed to assess the effects of vaccarin in accelerating angiogenesis. Western blot was used to evaluate relative protein expressed levels. Results Vaccarin could significantly promote wound healing and endothelial cells and fibroblasts proliferation in the wound site. Immunohistochemistry and Western blot studies showed that the nodal proteins and receptor (bFGFR) related to angiogenesis signaling pathway were activated, and the microvascular density in the wound site was markedly higher than that in the control group. Conclusions The present study was the first to demonstrate that vaccarin is able to induce angiogenesis and accelerate wound healing in vivo by increasing expressions of p-Akt, p-Erk and p-bFGFR. This process is mediated by MAPK/ERK and PI3K/AKT signaling pathways.
Assuntos
Animais , Masculino , Cicatrização/efeitos dos fármacos , Extratos Vegetais/farmacologia , Fosfatidilinositol 3-Quinases/efeitos dos fármacos , Quinases de Proteína Quinase Ativadas por Mitógeno/efeitos dos fármacos , Caryophyllaceae/química , Indutores da Angiogênese/farmacologia , Fatores de Tempo , Imuno-Histoquímica , Extratos Vegetais/química , Transdução de Sinais , Western Blotting , Reprodutibilidade dos Testes , Ratos Sprague-Dawley , Fosfatidilinositol 3-Quinases/análise , Quinases de Proteína Quinase Ativadas por Mitógeno/análise , Células Endoteliais/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/análise , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/efeitos dos fármacos , Fibroblastos/efeitos dos fármacosRESUMO
This study aimed to investigate the effects of gallium-aluminum-arsenium (GaAlAs) (670 nm) laser therapy on neoangiogenesis and fibroplasia during tissue remodelling. Forty male Wistar rats underwent cutaneous surgery and were divided into 2 experimental groups: the Control and Laser group (9 mW, 670 nm, 0.031 W/cm2 , 4 J/cm2 ). After 14, 21, 28, and 35 days, the animals were euthanised. Descriptive and quantitative analyses were performed in sections stained with haematoxylin-eosin and Sirius Red, respectively. The amounts of VEGF+ and CD31+ cells were evaluated by immunohistochemistry and histomorphometric analysis, respectively. Statistical analysis was performed using the Mann-Whitney, Friedman, and Spearman correlation test, P < 0.05. The collagen expression was significantly higher in the laser group compared with the control group on days 14 and 21 after the creation of the skin wound (P = 0.008; P = 0.016) and in the control group between 14 and 28 and 14 and 35 days (P = 0.001; P = 0.007). There were more blood vessels in three periods of the study only in the (Laser) treated group, with statistical significance at day 14 (P = 0.016). There was no statistically significant difference in VEGF+ cell count in the different experimental groups throughout the study, although a positive correlation was shown with the area of collagen on days 14 and 28 (P = 0.037). Laser treatment had a positive effect in the late course of healing, particularly with regards to collagen expression and the number of newly formed vessels. VEGF+ cells were present in both experimental groups, and VEGF appeared to influence fibroplasia in the treated group.
Assuntos
Colágeno/efeitos da radiação , Lasers Semicondutores/uso terapêutico , Terapia com Luz de Baixa Intensidade/métodos , Pele/efeitos da radiação , Cicatrização/fisiologia , Ferimentos e Lesões/tratamento farmacológico , Ferimentos e Lesões/radioterapia , Alumínio/uso terapêutico , Animais , Colágeno/efeitos dos fármacos , Gálio/uso terapêutico , Masculino , Modelos Animais , Ratos , Ratos WistarRESUMO
ABSTRACT INTRODUCTION: Bismuth subgallate is a salt derived from heavy metal. The aim of this study was to evaluate the effect of this salt on some phases of healing. OBJECTIVES: To assess the effect of subgallate on mucosa and to evaluate the association between the use of bismuth subgallate and neogenesis of vessels in oral mucosal wounds. METHODS: This was a prospective and experimental study. This study used sixty rats, which were divided into control and experimental groups. The animals were submitted to a surgical procedure, which caused oral mucosal injury. A saline solution was applied on the wound of the control group, and in the experimental group, a solution of bismuth subgallate was administrated. RESULTS: The experimental group showed greater inflammatory reaction with increasing monomorphic proliferation. There was increased vessel proliferation in the control group. CONCLUSION: Bismuth subgallate had a negative influence on the healing process, delaying the rate of new vessel formation and optimal wound healing.
RESUMO INTRODUÇÃO: O subgalato de bismuto é um sal derivado de metal pesado. A ideia desta pesquisa é avaliar sua interferência em alguma das fases da cicatrização. OBJETIVO: Delinear a ação do subgalato em mucosas. Avaliar a relação entre a utilização do subgalato de bismuto e a neoformação de vasos nas feridas em mucosa oral, para evidenciar o possível benefício resultante do seu uso. MÉTODO: Estudo experimental, prospectivo. Utilizou-se sessenta ratos, que foram divididos igualmente em grupo controle e experimento. Foram submetidos a um procedimento cirúrgico onde foi feito uma lesão na mucosa oral dos animais, após, uma solução de soro fisiológico foi aplicada sobre a lesão do grupo controle e sobre a ferida do grupo experimento foi aplicada uma solução de subgalato de bismuto. RESULTADOS: o grupo experimento apresentou maior reação inflamatória com crescente proliferação monomórfica. Vasos: houve maior proliferação no grupo controle. CONCLUSÕES: concluiu-se que o subgalato de bismuto teve uma ação negativa no processo de cicatrização, atrasando a velocidade de formação dos neovasos e a cicatrização ideal da ferida operatória.
Assuntos
Animais , Humanos , Masculino , Ratos , Indutores da Angiogênese/uso terapêutico , Ácido Gálico/análogos & derivados , Compostos Organometálicos/uso terapêutico , Cicatrização/efeitos dos fármacos , Modelos Animais de Doenças , Ácido Gálico/uso terapêutico , Mucosa Bucal/irrigação sanguínea , Mucosa Bucal/cirurgia , Estudos Prospectivos , TonsilectomiaRESUMO
INTRODUCTION: Bismuth subgallate is a salt derived from heavy metal. The aim of this study was to evaluate the effect of this salt on some phases of healing. OBJECTIVES: To assess the effect of subgallate on mucosa and to evaluate the association between the use of bismuth subgallate and neogenesis of vessels in oral mucosal wounds. METHODS: This was a prospective and experimental study. This study used sixty rats, which were divided into control and experimental groups. The animals were submitted to a surgical procedure, which caused oral mucosal injury. A saline solution was applied on the wound of the control group, and in the experimental group, a solution of bismuth subgallate was administrated. RESULTS: The experimental group showed greater inflammatory reaction with increasing monomorphic proliferation. There was increased vessel proliferation in the control group. CONCLUSION: Bismuth subgallate had a negative influence on the healing process, delaying the rate of new vessel formation and optimal wound healing.
Assuntos
Indutores da Angiogênese/uso terapêutico , Ácido Gálico/análogos & derivados , Compostos Organometálicos/uso terapêutico , Cicatrização/efeitos dos fármacos , Animais , Modelos Animais de Doenças , Ácido Gálico/uso terapêutico , Humanos , Masculino , Mucosa Bucal/irrigação sanguínea , Mucosa Bucal/cirurgia , Estudos Prospectivos , Ratos , TonsilectomiaRESUMO
Objective Evaluate the effects of VEGF165 gene transfer in the process of remodeling of the extracellular matrix after an acute myocardial infarct. Methods Wistar rats were submitted to myocardial infarction, after the ligation of the left descending artery, and the left ventricle ejection fraction was used to classify the infarcts into large and small. The animals were divided into groups of ten, according to the size of infarcted area (large or small), and received or not VEGF165 treatment. Evaluation of different markers was performed using immunohistochemistry and digital quantification. The primary antibodies used in the analysis were anti-fibronectin, anti-vimentin, anti-CD44, anti-E-cadherin, anti-CD24, anti-alpha-1-actin, and anti-PCNA. The results were expressed as mean and standard error, and analyzed by ANOVA, considering statistically significant if p≤0.05. Results There was a significant increase in the expression of undifferentiated cell markers, such as fibronectin (protein present in the extracellular matrix) and CD44 (glycoprotein present in the endothelial cells). However, there was decreased expression of vimentin and PCNA, indicating a possible decrease in the process of cell proliferation after treatment with VEGF165. Markers of differentiated cells, E-cadherin (adhesion protein between myocardial cells), CD24 (protein present in the blood vessels), and alpha-1-actin (specific myocyte marker), showed higher expression in the groups submitted to gene therapy, compared to non-treated group. The value obtained by the relation between alpha-1-actin and vimentin was approximately three times higher in the groups treated with VEGF165, suggesting greater tissue differentiation. Conclusion The results demonstrated the important role of myocytes in the process of tissue remodeling, confirming that VEGF165 seems to provide a protective effect in the treatment of acute myocardial infarct. .
Objetivo Avaliar os efeitos da transferência gênica do VEGF165 no processo de remodelamento da matriz extracelular após infarto agudo do miocárdio. Métodos Ratos Wistar foram submetidos ao infarto do miocárdio por ligação da artéria coronária descendente esquerda, e a fração de ejeção de ventrículo esquerdo foi utilizada para classificar os infartos em grandes e pequenos. Os animais foram divididos em grupos de dez animais, de acordo com o tamanho do infarto (grande ou pequeno), e receberam ou não tratamento com o VEGF165. A avaliação dos diferentes marcadores foi realizada por imuno-histoquímica e quantificação digital. Os anticorpos primários utilizados foram antifibronectina, antivimentina, anti- CD44, anti-E-caderina, anti-CD24, anti-alfa-1-actina e anti-PCNA. Os resultados foram representados como média e erro padrão, e analisados por ANOVA, sendo considerado estatisticamente significativo se p≤0,05. Resultados Houve aumento significativo da expressão de marcadores de células indiferenciadas, como fibronectina (proteína presente na matriz extracelular) e CD44 (glicoproteína presente nas células endoteliais). Entretanto, houve diminuição da expressão de vimentina e PCNA, indicando possível diminuição do processo de proliferação celular após o tratamento com VEGF165. Os marcadores de células diferenciadas, E-caderina (proteína de adesão entre as células do miocárdio), CD24 (proteína presente nos vasos sanguíneos) e alfa-1-actina (marcador especifico de miócitos) também apresentaram maior expressão nos grupos submetidos à terapia gênica, comparativamente com o grupo não tratado. O valor obtido pela relação entre alfa-1-actina e vimentina foi aproximadamente três vezes maior nos grupos tratados com VEGF165, indicando maior diferenciação tecidual. Conclusão O papel dos miócitos se mostrou importante no processo de remodelamento tecidual, confirmando que o VEGF165 parece conferir um efeito protetor no tratamento do infarto agudo do miocárdio. .
Assuntos
Animais , Feminino , Matriz Extracelular/fisiologia , Técnicas de Transferência de Genes , Infarto do Miocárdio/terapia , Fator A de Crescimento do Endotélio Vascular/uso terapêutico , Actinas/análise , /análise , /análise , Caderinas/análise , Proliferação de Células/fisiologia , Modelos Animais de Doenças , Fibronectinas/análise , Terapia Genética/métodos , Imuno-Histoquímica , Miócitos Cardíacos/fisiologia , Ratos Wistar , Reprodutibilidade dos Testes , Resultado do Tratamento , Fator A de Crescimento do Endotélio Vascular/genética , Vimentina/análiseRESUMO
OBJECTIVE: The aim of this work was to demonstrate a possible relationship between anti-latency-associated peptide human latent transforming growth factor beta 1 (latent TGF-ß1) expression in megakaryocytes and microvascular density in bone marrow biopsies from patients with essential thrombocythemia and primary myelofibrosis. METHODS: Microvascular density was evaluated by immunohistochemical analysis and the expression of latent TGF-ß1 in samples (100 megakaryocytes per bone marrow sample) from 18 essential thrombocythemia and 38 primary myelofibrosis (19 prefibrotic and 19 fibrotic) patients. Six bone marrow donor biopsies were used as controls. Fibrosis in the bone marrow biopsies was evaluated according to the European Consensus. RESULTS: The average fibrosis grade differed between essential thrombocythemia and primary myelofibrosis groups when compared to the control group. Latent TGF-ß1 expression differed significantly between the fibrotic primary myelofibrosis (PMF) group and the control group (p-value<0.01). A high degree of neo-angiogenesis (demonstrated by analysis of CD34 expression) was detected in patients with myelofibrosis. There were correlations between latent TGF-ß1 expression and microvascular density (r=0.45; p-value<0.0009) and between degree of microvascular density and fibrosis grade (r=0.80; p-value<0.0001). Remarkable differences for neo-angiogenesis were not observed between patients with essential thrombocythemia and controls. CONCLUSION: Angiogenesis participates in the pathogenesis of primary myelofibrosis, in both the prefibrotic and fibrotic stages, while latent TGF-ß is differentially expressed only in the prefibrotic stage.
RESUMO
Objective: The aim of this work was to demonstrate a possible relationship between anti-latency-associated peptide human latent transforming growth factor beta 1 (latent TGF-β1) expression in megakaryocytes and microvascular density in bone marrow biopsies from patients with essential thrombocythemia and primary myelofibrosis. Methods: Microvascular density was evaluated by immunohistochemical analysis and the expression of latent TGF-β1 in samples (100 megakaryocytes per bone marrow sample) from 18 essential thrombocythemia and 38 primary myelofibrosis (19 prefibrotic and 19 fibrotic) patients. Six bone marrow donor biopsies were used as controls. Fibrosis in the bone marrow biopsies was evaluated according to the European Consensus. Results: The average fibrosis grade differed between essential thrombocythemia and primary myelofibrosis groups when compared to the control group. Latent TGF-β1 expression differed significantly between the fibrotic primary myelofibrosis (PMF) group and the control group (p-value < 0.01). A high degree of neo-angiogenesis (demonstrated by analysis of CD34 expression) was detected in patients with myelofibrosis. There were correlations between latent TGF-β1 expression and microvascular density (r = 0.45; p-value < 0.0009) and between degree of microvascular density and fibrosis grade (r = 0.80; p-value < 0.0001). Remarkable differences for neo-angiogenesis were not observed between patients with essential thrombocythemia and controls. Conclusion: Angiogenesis participates in the pathogenesis of primary myelofibrosis, in both the prefibrotic and fibrotic stages, while latent TGF-β is differentially expressed only in the prefibrotic stage...
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Humanos , Indutores da Angiogênese , Fibrose , Mielofibrose Primária , Fator de Crescimento Transformador beta1RESUMO
Angiogenic therapies for critical limb ischemia were tested in a mouse model. The mice were anesthetized and their femoral arteries were ligated. The animals were treated with bone marrow mononuclear cells (BMMCs) alone, BMMCs combined with plasmid vector encoding granulocyte macrophage colony-stimulating factor (GM-CSF), received no treatment, or no intervention (controls). The degree of ischemia was monitored for 4 weeks using a visual scale. Muscle atrophy and strength were assessed at 4 weeks postoperatively; the mice were then killed. In treated animals, total necrosis of the limb was not found, the weight of the gastrocnemius and quadriceps muscles was significantly higher, functional ability and tissue regeneration were significantly increased, and muscle impairment and adipocyte presence were significantly reduced compared with untreated animals. At inducing angiogenesis, the BMMCs alone was more effective than BMMCs combined with plasmid vector encoding GM-CSF. Treated animals showed increased angiogenesis compared with ischemic untreated ones.
Assuntos
Transplante de Medula Óssea , Terapia Genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos/biossíntese , Isquemia/terapia , Neovascularização Fisiológica , Músculo Quadríceps/irrigação sanguínea , Animais , Células Cultivadas , Estado Terminal , Modelos Animais de Doenças , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Membro Posterior , Isquemia/genética , Isquemia/metabolismo , Isquemia/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Força Muscular , Atrofia Muscular/patologia , Atrofia Muscular/fisiopatologia , Atrofia Muscular/terapia , Necrose , Músculo Quadríceps/patologia , Músculo Quadríceps/fisiopatologia , Fatores de Tempo , TransfecçãoRESUMO
INTRODUCTION: Different low-level laser (LLL) irradiation protocols have been tested to accelerate orthodontic tooth movement (OTM). Nevertheless, divergent results have been obtained. It was suggested that the stimulatory action of low level laser irradiation occurs during the proliferation and differentiation stages of bone cellular precursors, but not during later stages. OBJECTIVE: The purpose of this study was to determine the effect of two protocols of LLL irradiation on experimental tooth movement: One with daily irradiations and another with irradiations during the early stages. METHODS: Thirty-six rats were divided into control groups (CG1, CG2, CG3) and irradiated groups (IrG1, IrG2, IrG3) according to the presence of: experimental tooth movement, laser irradiation, type of laser irradiation protocol and date of euthanasia (3th or 8th day of experiment). At the end of experimental periods, a quantitative evaluation of the amount of OTM was made and the reactions of the periodontium were analyzed by describing cellular and tissue reactions and by counting blood vessels. RESULTS: The amount of OTM revealed no significant differences between groups in the same experimental period (p < 0.05). Qualitative analysis revealed the strongest resorption activity in irradiated groups after seven days, especially when using the daily irradiation protocol. There was a higher number of blood vessels in irradiated animals than in animals without orthodontic devices and without laser irradiation (p < 0.05). CONCLUSION: Moreover, angiogenesis was verified in some of the irradiated groups. The irradiation protocols tested were not able to accelerate OTM and root resorption was observed while they were applied.
Assuntos
Terapia com Luz de Baixa Intensidade/métodos , Neovascularização Fisiológica/efeitos da radiação , Ligamento Periodontal/efeitos da radiação , Técnicas de Movimentação Dentária/métodos , Análise de Variância , Animais , Ligamento Periodontal/irrigação sanguínea , Ligamento Periodontal/citologia , Ratos , Reabsorção da RaizRESUMO
PURPOSE: To evaluate the effects of aroeira (Schinus terebinthifolius) ointment on skin wound healing in rats. METHODS: Adult male rats (n=20) were divided into four groups of five animals each, as follows: G4, G7, G14 and G21, which corresponds to 4th, 7th, 14th and 21th days postoperatively. Each animal were made two incisions on the skin, including the subcutaneous tissue, in the right and left sides of thoracic region, separated by a distance of two inches. The right lesion was treated with base ointment (vaseline, lanolin); the left one was treated with base ointment containing 5% of aroeira oil. At the end of each experimental period the lesions were evaluated for the contraction degree. Then held the collection of fragments that were fixed in 10% formalin and processed for paraffin embedding. In the histological sections (5μm) was evaluated the morphology and quantified the collagen and blood vessels. The data obtained were submitted to ANOVA test complemented by Tukey-Kramer test (p<0.05). RESULTS: The contraction of the lesions was higher in wounds treated with aroeira oil than in controls at 7th and 14th days (p<0.01), whereas in the 21st day all lesions were already completely healed. The morphology showed granulation tissue more developed, with fibroblasts more bulky and collagen fibers more arranged in the experimental group at 4th, 7th and 14th days. The morphometry showed a significant increase in the quantification of collagen fibers in the experimental group at 7th and 14th days (p<0.05). CONCLUSION: The aroeira oil accelerates the healing process of wounds as a macroscopic, morphological and morphometrical analysis.
Assuntos
Animais , Masculino , Ratos , Anacardiaceae , Extratos Vegetais/uso terapêutico , Óleos de Plantas/uso terapêutico , Tela Subcutânea/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Indutores da Angiogênese/uso terapêutico , Colágeno/análise , Imuno-Histoquímica , Fitoterapia , Período Pós-Operatório , Tela Subcutânea/patologia , Fatores de Tempo , Resultado do TratamentoRESUMO
INTRODUCTION: Different low-level laser (LLL) irradiation protocols have been tested to accelerate orthodontic tooth movement (OTM). Nevertheless, divergent results have been obtained. It was suggested that the stimulatory action of low level laser irradiation occurs during the proliferation and differentiation stages of bone cellular precursors, but not during later stages. OBJECTIVE: The purpose of this study was to determine the effect of two protocols of LLL irradiation on experimental tooth movement: One with daily irradiations and another with irradiations during the early stages. METHODS: Thirty-six rats were divided into control groups (CG1, CG2, CG3) and irradiated groups (IrG1, IrG2, IrG3) according to the presence of: experimental tooth movement, laser irradiation, type of laser irradiation protocol and date of euthanasia (3th or 8th day of experiment). At the end of experimental periods, a quantitative evaluation of the amount of OTM was made and the reactions of the periodontium were analyzed by describing cellular and tissue reactions and by counting blood vessels. RESULTS: The amount of OTM revealed no significant differences between groups in the same experimental period (p < 0.05). Qualitative analysis revealed the strongest resorption activity in irradiated groups after seven days, especially when using the daily irradiation protocol. There was a higher number of blood vessels in irradiated animals than in animals without orthodontic devices and without laser irradiation (p < 0.05). CONCLUSION: Moreover, angiogenesis was verified in some of the irradiated groups. The irradiation protocols tested were not able to accelerate OTM and root resorption was observed while they were applied.
INTRODUÇÃO: diferentes protocolos de irradiação por laser de baixa potência (LBP) têm sido testados para potencializar o movimento ortodôntico; entretanto, há resultados divergentes. Foi sugerido que seu efeito bioestimulador ocorre nas fases de proliferação e diferenciação celular, não agindo em estágios tardios. OBJETIVO: avaliar o efeito de dois protocolos de irradiação do LBP na movimentação ortodôntica: um com irradiações diárias e outro em que irradiações foram realizadas apenas nos períodos iniciais. MÉTODOS: trinta e seis ratos Wistar foram divididos em grupos controles (GC1, GC2 e GC3) e irradiados (GIr1, GIr2 e GIr3), de acordo com a presença de dispositivo ortodôntico, a presença de irradiação, o tipo de protocolo de irradiação e a data de eutanásia (3º ou 8º dia de experimento). Ao final dos períodos experimentais, foram realizadas mensurações da movimentação dentária, análise qualitativa das reações celulares e teciduais do periodonto e contagem de vasos sanguíneos no ligamento periodontal. RESULTADOS: a quantidade de movimentação não diferiu entre os grupos num mesmo tempo experimental (p < 0,05). A análise qualitativa revelou maior atividade absortiva nos grupos irradiados ao final de 7 dias, especialmente quando as irradiações foram diárias. Nos grupos irradiados diariamente, a contagem de vasos foi aumentada em relação aos animais isentos de dispositivo ortodôntico e de aplicações de LBP (p < 0,05). CONCLUSÃO: apesar de verificada angiogênese em certos grupos irradiados, os protocolos de irradiação testados não foram capazes de acelerar a movimentação dentária, e foi possível verificarem-se absorções radiculares.
Assuntos
Animais , Ratos , Terapia com Luz de Baixa Intensidade/métodos , Neovascularização Fisiológica/efeitos da radiação , Ligamento Periodontal/efeitos da radiação , Técnicas de Movimentação Dentária/métodos , Análise de Variância , Ligamento Periodontal/irrigação sanguínea , Ligamento Periodontal/citologia , Reabsorção da RaizRESUMO
PURPOSE: To evaluate the effects of aroeira (Schinus terebinthifolius) ointment on skin wound healing in rats. METHODS: Adult male rats (n=20) were divided into four groups of five animals each, as follows: G4, G7, G14 and G21, which corresponds to 4th, 7th, 14th and 21th days postoperatively. Each animal were made two incisions on the skin, including the subcutaneous tissue, in the right and left sides of thoracic region, separated by a distance of two inches. The right lesion was treated with base ointment (vaseline, lanolin); the left one was treated with base ointment containing 5% of aroeira oil. At the end of each experimental period the lesions were evaluated for the contraction degree. Then held the collection of fragments that were fixed in 10% formalin and processed for paraffin embedding. In the histological sections (5μm) was evaluated the morphology and quantified the collagen and blood vessels. The data obtained were submitted to ANOVA test complemented by Tukey-Kramer test (p<0.05). RESULTS: The contraction of the lesions was higher in wounds treated with aroeira oil than in controls at 7th and 14th days (p<0.01), whereas in the 21st day all lesions were already completely healed. The morphology showed granulation tissue more developed, with fibroblasts more bulky and collagen fibers more arranged in the experimental group at 4th, 7th and 14th days. The morphometry showed a significant increase in the quantification of collagen fibers in the experimental group at 7th and 14th days (p<0.05). CONCLUSION: The aroeira oil accelerates the healing process of wounds as a macroscopic, morphological and morphometrical analysis.(AU)
Assuntos
Animais , Ratos , Tórax/anatomia & histologia , Ferimentos e Lesões , Pele/anatomia & histologia , Fitoterapia/instrumentação , Anacardiaceae , Cicatrização/fisiologiaRESUMO
PURPOSE: To study the effects of the angico extract (Anadenanthera colubrina var. cebil) on the healing of rat skin. METHODS: Twenty adult rats were divided into four groups of five animals each, the G4, G7, G14 and G21, which corresponds to the respective postoperative days. Each group received two incisions on skin and subcutaneous tissue in the right and left antimere of the thoracic region, separated by a distance of 2 cm. The right lesion was treated daily with saline and the left with the angico alcoholic extract (5%). At the end of each experimental period, animals were euthanized and fragments of the wound area, together with the edges were removed, fixed in 10% formaldehyde solution and processed for paraffin embedding. In the histological sections with 5 µm of thickness, were carried out immunohistochemical methods for detection of blood vessels (VEGF) and stained with hematoxylin and eosin for morphological analysis. Statistical analysis was done by ANOVA and Tukey test (p<0.05). RESULTS: Morphological analysis showed larger fibroblasts and a higher concentration of collagen fibers in days 7 and 14 in wounds treated with the angico extract. Morphometric analysis demonstrated a significant increase in the number of blood vessels in both the seventh and 14th days (p<0.01) in wounds treated with the angico extract. CONCLUSION: The angico alcoholic extract (Anadenanthera colubrina var. cebil) induces the acceleration of wound healing in skin wounds of rats.(AU)
OBJETIVO: Avaliar os efeitos do extrato de angico (Anadenanthera colubrina var. cebil) na cicatrização em pele de ratos. MÉTODOS: Ratos machos adultos (n=20) foram distribuídos em quatro grupos de cinco animais cada, a saber: G4, G7, G14 e G21, o que corresponde a quatro, sete, 14 e 21 dias de pós-operatório. Cada grupo recebeu duas incisões na pele compreendendo o tecido subcutâneo, nos antímeros direito e esquerdo da região torácica, separadas por uma distância de dois cm. A lesão esquerda com extrato alcoólico de angico (5%), iniciando-se logo após a cirurgia por 21 dias consecutivos. Ao final de cada período (4, 7, 14 e 21 de pós-operatório) experimental foram coletados fragmentos da área da ferida, fixada em formol a 10% e processadas para inclusão em parafina. Nos cortes histológicos com 5 µm de espessura, foram realizados métodos imunoistoquímicos para detecção dos vasos sanguíneos (VEGF) e coloração pela hematoxilina para análise morfológica. Os dados obtidos foram submetidos à análise estatística ANOVA complementada pelo teste de Tukey-Kramer (p<0,05). RESULTADOS: A análise morfológica mostrou fibroblastos mais volumosos e alta concentração de fibras colágenas no 7º e 14º dias nas feridas tratadas com extrato de angico. A análise morfométrica demonstrou aumento significativo no número de vasos sanguíneos no sétimo e 14º dias (p<0,01) de pós-operatório em feridas tratadas com extrato de angico. CONCLUSÃO: O extrato hidroalcoólico a 5% da casca e entrecasca do angico (Anadenanthera colubrina var. cebil) acelera a neoangiogênese em feridas cutâneas de ratos.(AU)