Influência do estresse crônico na resposta tecidual de cimentos endodônticos
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Data
2023-02-27
Autores
Justo, Mariana Pagliusi [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Objetivo: Este estudo buscou correlacionar o estresse crônico com a resposta tecidual de diferentes tipos de cimentos endodônticos empregando o modelo ISO 10993-6:2016 para testes de implantação em tecido subcutâneo. Métodos: Foram utilizados 28 ratos Wistar divididos em 2 grupos (n=14): ratos controle (rC) e ratos estressados (rE). Em cada rato foram implantados 4 tubos de polietileno, sendo 1 tubo vazio para Controle, e os demais contendo um dos tipos de cimentos endodônticos selecionados para o estudo (Sealapex®, Endofill®, AH Plus®). Os períodos de avaliação foram de 7 e 30 dias (n=7/tempo). Após a eutanásia foi coletado o sangue para dosagem de corticosterona pelo método ELISA e, em seguida, os tubos e o tecido circundante foram removidos e processados para análise histológica e imunoistoquímica. A inflamação foi avaliada em cortes teciduais corados em H.E e pela marcação imunoistoquímica para IL-6, IL-17,TNF-α, CD8 e CD20; o colágeno, por meio da coloração de Picrosírios red (PSR); a biomineralização, por meio da coloração de von Kossa (vK) e em cortes sem coloração pela luz polarizada (POL), assim como também pela análise da osteocalcina (OCN). Para todos os testes empregados foi considerado o nível de significância de 5% (p<0,05). Resultados: Observou-se aumento de corticosterona em rE comparado a rC, apenas no período de 7 dias (p < 0,05). Animais do grupo rE apresentaram menor ganho de peso corporal comparado aos animais em rC (p < 0,05). Na análise histológica em rC aos 7 dias observou-se que o grupo Endofill apresentou infiltrado inflamatório mais intenso comparado ao Controle (p = 0,031). Em rE não houve diferença entre os grupos em nenhum período (p > 0,05). Comparando a resposta de cada grupo em função da presença ou não do estresse, observou-se que apenas aos 7 dias o grupo controle em rE apresentou infiltrado inflamatório mais intenso que o grupo Controle em rC (p < 0,05). Não houve diferença entre os grupos para IL-6 em nenhuma das comparações e períodos (p > 0,05). Para IL-17 em rC, aos 7 dias, o grupo Endofill apresentou infiltrado inflamatório mais intenso comparado ao Controle (p < 0,05). Para TNF-α, observou-se aos 7 dias que o grupo controle em rE apresentou maior imunomarcação comparado ao grupo Controle em rC (p < 0,05). Para CD8, no período 30 dias observou-se que o grupo AH Plus em rC apresentou maior quantidade de células CD8+ comparado ao AH Plus em rE (p < 0,05). Para CD20 em rC, aos 7 dias, foi observado que o grupo Endofill apresentou maior quantidade de células CD20+ quando comparado aos grupos Controle e Sealapex (p < 0,05). Em rE aos 7 dias, foi observado que o grupo Endofill apresentou maior quantidade de células CD20+ quando comparado ao grupo Controle (p < 0,05). Aos 7 dias o grupo Controle em rC apresentou maior quantidade de células CD20+ comparado ao grupo Controle em rE (p < 0,05). Para PSR, na comparação entre os grupos em rC, aos 7 dias, os grupos Sealapex e Endofill apresentaram maior percentual de fibras imaturas e menor percentual de fibras maduras quando comparados aos grupos Controle e AH Plus (p < 0,05). O grupo Controle, Sealapex e Endofill em rE apresentaram maior percentual de fibras imaturas e menor percentual de fibras maduras comparado aos grupos Controle, Sealapex e Endofill em rC (p < 0,05). O grupo AH Plus em rE, tanto aos 7 dias quando aos 30 dias apresentou maior percentual de fibras imaturas e menor percentual de fibras maduras comparado ao grupo AH Plus em rC p < 0,05. Apenas no grupo Sealapex, tanto em rC quando em rE, foram observadas estruturas coradas em negro na coloração von Kossa, cristais birrefringentes sob luz polarizada e imunomarcação positiva para OCN. Além disso, em rC houve maior imunomarcação para OCN comparado a rE (p < 0,05). Conclusão: O estresse crônico influenciou a resposta tecidual de cimentos endodônticos, aumentando o infiltrado inflamatório e retardando o reparo tecidual. Ainda, o estresse crônico reduziu a capacidade de biomineralização do material que possui esta habilidade.
Objective: This study aimed to correlate chronic stress with the tissue response of different types of endodontic cements using the ISO 10993-6:2016 model for subcutaneous tissue implantation tests. Methods: 28 Wistar rats were used, divided into two groups (n=14): control rats (rC) and stressed rats (rE). Four polyethylene tubes were implanted in each rat, 1 empty for control, and the others containing one of the endodontic cements selected for the study (Sealapex®, Endofill® and AH Plus®). The evaluation periods were 7 and 30 days (n=7/time). After euthanasia, blood was collected for corticosterone dosage by the ELISA method, and then the tubes and surrounding tissue were removed and processed for histological and immunohistochemical analysis. Inflammation was assessed in tissue sections stained in H.E. and by immunohistochemical staining for IL-6, IL-17, TNF-α, CD8 and CD20; collagen, by Picrosirius red staining (PSR); biomineralization, by von Kossa staining (vK) and in sections without staining by polarized light (POL), as well as by osteocalcin (OCN) analysis. For all tests employed, a significance level of 5% (p<0.05) was considered. Results: An increase in corticosterone was observed in rE compared to rC, only in the 7-day period (p < 0.05). Animals in the rE group less body weight gain compared to animals in rC (p < 0.05). Histological analysis in rC at 7 days showed that the Endofill group showed more intense inflammatory infiltrate compared to Control (p = 0.031). In rE there was no difference between the groups in any period (p > 0.05). Comparing the response of each group according to the presence or absence of stress, it was observed that only at 7 days the control group in rE presented a more intense inflammatory infiltrate than the Control group in rC (p < 0.005). There was no difference between groups for IL-6 in any of the comparisons and periods (p > 0.05). For IL-17 in rC, at 7 days, the Endofill group showed more intense inflammatory infiltrate compared to the Control (p < 0.05). For TNF-α, it was observed at 7 days that the control group in rE showed greater immunolabeling compared to the Control group in rC (p < 0.05). For CD8, at 30 days, it was observed that the AH Plus group in rC showed a greater amount of CD8+ cells compared to AH Plus in rE (p < 0.05). For CD20 in rC, at 7 days, the Endofill group had more CD20+ cells when compared to the Control and Sealapex groups (p < 0.05). In rE, at 7 days, it was observed that the Endofill group presented a greater amount of CD20+ cells when compared to the Control group (p < 0.05). At 7 days, the Control group in rC presented a greater amount of CD20+ cells compared to the Control group in rE (p < 0.05). For PSR, when comparing the groups in rC, at 7 days, the Sealapex and Endofill groups showed a higher percentage of immature fibers and a lower percentage of mature fibers when compared to the Control and AH Plus groups (p < 0.05). The Control, Sealapex and Endofill groups in rE showed a higher percentage of immature fibers and a lower percentage of mature fibers compared to the Control, Sealapex and Endofill groups in rC (p < 0.05). The AH Plus group in rE, both at 7 days and 30 days showed a higher percentage of immature fibers and a lower percentage of mature fibers compared to the AH Plus group in rC (p < 0.05). Only in the Sealapex group, both in rC and rE, structures stained black in von Kossa staining, birefringent crystals under polarized light and positive immunostaining for OCN were observed. Furthermore, in rC there was greater immunostaining for OCN compared to rE (p < 0.05). Conclusion: Chronic stress influenced the tissue response of endodontic sealers, increasing the inflammatory infiltrate and delaying tissue repair. Still, chronic stress reduced the biomineralization capacity of the material that has this ability.
Objective: This study aimed to correlate chronic stress with the tissue response of different types of endodontic cements using the ISO 10993-6:2016 model for subcutaneous tissue implantation tests. Methods: 28 Wistar rats were used, divided into two groups (n=14): control rats (rC) and stressed rats (rE). Four polyethylene tubes were implanted in each rat, 1 empty for control, and the others containing one of the endodontic cements selected for the study (Sealapex®, Endofill® and AH Plus®). The evaluation periods were 7 and 30 days (n=7/time). After euthanasia, blood was collected for corticosterone dosage by the ELISA method, and then the tubes and surrounding tissue were removed and processed for histological and immunohistochemical analysis. Inflammation was assessed in tissue sections stained in H.E. and by immunohistochemical staining for IL-6, IL-17, TNF-α, CD8 and CD20; collagen, by Picrosirius red staining (PSR); biomineralization, by von Kossa staining (vK) and in sections without staining by polarized light (POL), as well as by osteocalcin (OCN) analysis. For all tests employed, a significance level of 5% (p<0.05) was considered. Results: An increase in corticosterone was observed in rE compared to rC, only in the 7-day period (p < 0.05). Animals in the rE group less body weight gain compared to animals in rC (p < 0.05). Histological analysis in rC at 7 days showed that the Endofill group showed more intense inflammatory infiltrate compared to Control (p = 0.031). In rE there was no difference between the groups in any period (p > 0.05). Comparing the response of each group according to the presence or absence of stress, it was observed that only at 7 days the control group in rE presented a more intense inflammatory infiltrate than the Control group in rC (p < 0.005). There was no difference between groups for IL-6 in any of the comparisons and periods (p > 0.05). For IL-17 in rC, at 7 days, the Endofill group showed more intense inflammatory infiltrate compared to the Control (p < 0.05). For TNF-α, it was observed at 7 days that the control group in rE showed greater immunolabeling compared to the Control group in rC (p < 0.05). For CD8, at 30 days, it was observed that the AH Plus group in rC showed a greater amount of CD8+ cells compared to AH Plus in rE (p < 0.05). For CD20 in rC, at 7 days, the Endofill group had more CD20+ cells when compared to the Control and Sealapex groups (p < 0.05). In rE, at 7 days, it was observed that the Endofill group presented a greater amount of CD20+ cells when compared to the Control group (p < 0.05). At 7 days, the Control group in rC presented a greater amount of CD20+ cells compared to the Control group in rE (p < 0.05). For PSR, when comparing the groups in rC, at 7 days, the Sealapex and Endofill groups showed a higher percentage of immature fibers and a lower percentage of mature fibers when compared to the Control and AH Plus groups (p < 0.05). The Control, Sealapex and Endofill groups in rE showed a higher percentage of immature fibers and a lower percentage of mature fibers compared to the Control, Sealapex and Endofill groups in rC (p < 0.05). The AH Plus group in rE, both at 7 days and 30 days showed a higher percentage of immature fibers and a lower percentage of mature fibers compared to the AH Plus group in rC (p < 0.05). Only in the Sealapex group, both in rC and rE, structures stained black in von Kossa staining, birefringent crystals under polarized light and positive immunostaining for OCN were observed. Furthermore, in rC there was greater immunostaining for OCN compared to rE (p < 0.05). Conclusion: Chronic stress influenced the tissue response of endodontic sealers, increasing the inflammatory infiltrate and delaying tissue repair. Still, chronic stress reduced the biomineralization capacity of the material that has this ability.
Descrição
Palavras-chave
Materiais dentários, Inflamação, Biomineralização, Corticosterona, Corticosterone