Impacto de andrógenos sobre o processo de reparação tecidual e o mecanismo de reabsorção óssea
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Data
2018-03-27
Autores
Gonçalves, Vinícius de Paiva [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O propósito do trabalho foi avaliar o impacto da depleção de testosterona e sua administração exógena, bem como de drogas bloqueadoras dos receptores de andrógenos (flutamida) e estrógeno (fulvestranto), e inibidoras da conversão de testosterona em estradiol (anastrosol), sobre a reparação tecidual cutânea e reabsorção óssea inflamatória. Foram utilizados 80 ratos, divididos em 8 grupos (n=10/grupo): G1-Sem tratamento, G2-Durateston, G3-Orquiectomia (OQX), G4-OQX+Durateston; G5-Anastrosol, G6-Fulvestranto, G7-Flutamida, G8-Durateston+Anastrosol. Os animais foram submetidos aos diferentes tratamentos por um período de 15 dias e, em seguida, foi realizada a indução de ferida cirúrgica cutânea e reabsorção óssea em calvária durante mais 7 dias. Através dos cortes histológicos foi avaliada a contração das feridas, análise estereométrica e qualitativa do tecido de granulação e mensuração da formação de colágeno; e, ainda, foi realizada reação de imunohistoquímica para os anticorpos PCNA e KGF, e análise da resposta imunoinflamatória por ensaio multiplex. As calvárias foram submetidas a avaliação da porcentagem de tecido ósseo (% BV/TV) através da análise de micromotografia computadorizada (μCT), quantificação do número de osteoclastos e pontos de reabsorção (lacunas de Howship) (H&E), marcação de osteoclastos TRAP+ (imunohistoquímica), e avaliação da resposta imunoinflamatória por ensaio multiplex. Foram utilizados ainda, 60 ratos divididos em 6 grupos (n=10/grupo): 1-Sham, 2-Sham+Ligadura, 3-Orquiectomia (OQX), 4-OQX+Ligadura, 5-OQX+Testosterona, 6-OQX+Testosterona+Ligadura. Os animais foram submetidos aos diferentes tratamentos de acordo com cada grupo e, em seguida, receberam ligadura ao redor dos primeiros molares inferiores, pelo período de 15 dias. Foram analisadas, a expressão de citocinas inflamatórias gengivais através de ensaio ELISA, análise estereométrica do processo inflamatório gengival e quantificação da reabsorção óssea inflamatória por μCT. No modelo de reparo, o tratamento com durateston aumentou a porcentagem de matriz extracelular e vasos sanguíneos, a imunomarcação para KGF e PCNA, estimulou uma maior contração das feridas, e elevou a produção de TNF-α e TGF-β1 (p<0.05). A flutamida aumentou a proliferação de fibroblastos (imunomarcados por PCNA) e imunomarcação para KGF, estimulou a contração das feridas (p<0.05) e, ainda, elevou a produção de TNF-α, TGF-β1 e VEGF, e reduziu IL-6 e PDGF (p<0.05). No modelo de reabsorção óssea em calvária, a OQX aumentou o número de pontos de reabsorção e de osteoclastos, assim como a atividade dos osteoclastos (p<0.05), e o tratamento com durateston reduziu esses eventos e preservou o percentual de volume ósseo (p<0.05). O número de pontos de reabsorção foi elevado pela flutamida, e a droga, assim como o fulvestranto, aumentaram a atividade de osteoclastos e elevaram a expressão de RANKL (p<0.05). No modelo de periodontite experimental, a reabsorção óssea foi significativamente atenuada pela orquiectomia (p<0.05). A periodontite experimental induziu um aumento significativo da interleucina IL-1β, que foi reduzido pela orquiectomia. Os níveis de IL-6 não foram afetados pela OQX, mas foram significativamente reduzidos em animais orquiectomizados e tratados com testosterona. Os achados sugerem que níveis sub ou suprafisiológicos de testosterona circulante, em resposta ao bloqueio ou não dos receptores desses hormônios, alteram parte ou partes dos complexos eventos associados a formação do tecido de granulação e reabsorção óssea inflamatória.
The aim of the study was to evaluate the impact of testosterone depletion and its exogenous administration, as well as androgen receptor blockers (flutamide) and estrogen (fulvestrant), and inhibitors of the conversion of testosterone to estradiol (anastrosol) on the cutaneous tissue repair and inflammatory bone resorption. Eighty rats were divided into eight groups (n=10/group): G1-No treatment, G2-Durateston, G3-Orchiectomy (OQX), G4-OQX+Durateston; G5-Anastrozol, G6-Fulvestrant, G7-Flutamide, G8-Durateston+Anastrozol. The animals were submitted to the different treatments for a period of 15 days and then the induction of cutaneous surgical wound and bone resorption in calvaria was performed for another 7 days. Through the histological sections the wound contraction, stereometric and qualitative analysis of the granulation tissue and measurement of collagen formation were evaluated; and also immunohistochemical reaction was performed for PCNA and KGF antibodies and analysis of the immunoinflammatory response by multiplex assay. The calvaria were submitted to evaluation of the percentage of bone tissue (% BV / TV) through computerized micromotography (μCT) analysis, quantification of the number of osteoclasts and reabsorption points (Howship lacunes) (H&E), labeling of TRAP+ osteoclasts (immunohistochemistry), and evaluation of the immunoinflammatory response by multiplex assay. In addition, 60 rats were divided into 6 groups (n = 10 / group): 1-Sham, 2-Sham+Ligature, 3-Orchiectomy (OQX), 4-OQX+Ligation, 5-OQX+Testosterone, 6-OQX+Testosterone + Ligature. The animals were submitted to different treatments according to each group and then received ligature around the lower first molars, for a period of 15 days. It was performed the expression of gingival inflammatory cytokines by ELISA, stereometric analysis of the inflammatory gingival process and quantification of inflammatory bone resorption by μCT. In the repair model, the durateston treatment increased the extracellular matrix and blood vessels percentage, the immunostaining for KGF and PCNA, stimulated a greater contraction of the wounds, and increased the production of TNF-α and TGF-β1 (p <0.05). Flutamide increased the fibroblasts proliferation (immunolabelled by PCNA) and immunolabelling to KGF, stimulated wound contraction (p <0.05), and increased TNF-α, TGF-β1 and VEGF production and reduced IL-6 and PDGF (p <0.05). In the calvariae bone resorption model, OQX increased the number of resorption points and osteoclasts, as well as the activity of osteoclasts (p <0.05), and the treatment with durateston reduced these events and preserved the percentage of bone volume (p <0.05). The reabsorption points number was elevated by flutamide, and the drug, as well as fulvestrant, increased osteoclast activity and elevated RANKL expression (p <0.05). In the experimental periodontitis model, bone resorption was significantly attenuated by orchiectomy (p <0.05). Experimental periodontitis induced a significant increase in IL-1β interleukin, which was reduced by orchiectomy. IL-6 levels were not affected by OQX, but were significantly reduced in orchiectomized and testosterone-treated animals. The findings suggest that sub- or supraphysiological levels of circulating testosterone, in response to or not blocking the receptors of these hormones, alter part or all of the complex events associated with granulation tissue formation and inflammatory bone resorption.
The aim of the study was to evaluate the impact of testosterone depletion and its exogenous administration, as well as androgen receptor blockers (flutamide) and estrogen (fulvestrant), and inhibitors of the conversion of testosterone to estradiol (anastrosol) on the cutaneous tissue repair and inflammatory bone resorption. Eighty rats were divided into eight groups (n=10/group): G1-No treatment, G2-Durateston, G3-Orchiectomy (OQX), G4-OQX+Durateston; G5-Anastrozol, G6-Fulvestrant, G7-Flutamide, G8-Durateston+Anastrozol. The animals were submitted to the different treatments for a period of 15 days and then the induction of cutaneous surgical wound and bone resorption in calvaria was performed for another 7 days. Through the histological sections the wound contraction, stereometric and qualitative analysis of the granulation tissue and measurement of collagen formation were evaluated; and also immunohistochemical reaction was performed for PCNA and KGF antibodies and analysis of the immunoinflammatory response by multiplex assay. The calvaria were submitted to evaluation of the percentage of bone tissue (% BV / TV) through computerized micromotography (μCT) analysis, quantification of the number of osteoclasts and reabsorption points (Howship lacunes) (H&E), labeling of TRAP+ osteoclasts (immunohistochemistry), and evaluation of the immunoinflammatory response by multiplex assay. In addition, 60 rats were divided into 6 groups (n = 10 / group): 1-Sham, 2-Sham+Ligature, 3-Orchiectomy (OQX), 4-OQX+Ligation, 5-OQX+Testosterone, 6-OQX+Testosterone + Ligature. The animals were submitted to different treatments according to each group and then received ligature around the lower first molars, for a period of 15 days. It was performed the expression of gingival inflammatory cytokines by ELISA, stereometric analysis of the inflammatory gingival process and quantification of inflammatory bone resorption by μCT. In the repair model, the durateston treatment increased the extracellular matrix and blood vessels percentage, the immunostaining for KGF and PCNA, stimulated a greater contraction of the wounds, and increased the production of TNF-α and TGF-β1 (p <0.05). Flutamide increased the fibroblasts proliferation (immunolabelled by PCNA) and immunolabelling to KGF, stimulated wound contraction (p <0.05), and increased TNF-α, TGF-β1 and VEGF production and reduced IL-6 and PDGF (p <0.05). In the calvariae bone resorption model, OQX increased the number of resorption points and osteoclasts, as well as the activity of osteoclasts (p <0.05), and the treatment with durateston reduced these events and preserved the percentage of bone volume (p <0.05). The reabsorption points number was elevated by flutamide, and the drug, as well as fulvestrant, increased osteoclast activity and elevated RANKL expression (p <0.05). In the experimental periodontitis model, bone resorption was significantly attenuated by orchiectomy (p <0.05). Experimental periodontitis induced a significant increase in IL-1β interleukin, which was reduced by orchiectomy. IL-6 levels were not affected by OQX, but were significantly reduced in orchiectomized and testosterone-treated animals. The findings suggest that sub- or supraphysiological levels of circulating testosterone, in response to or not blocking the receptors of these hormones, alter part or all of the complex events associated with granulation tissue formation and inflammatory bone resorption.
Descrição
Palavras-chave
Tecido de granulação. Reabsorção óssea. Periodontite. Inflamação. Androgênios. Ratos.