Resposta imunoinflamatória na periodontite experimental induzida em ratos submetidos à quimioterapia: estudo histomorfométrico e imunoistoquímico
Carregando...
Arquivos
Data
2017-02-21
Autores
Gusman, David Jonathan Rodrigues [UNESP]
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
O presente estudo tem por objetivo avaliar o efeito dos quimioterápicos 5-Fluorouracil (5-FU) e da Cisplatina (CIS) sob os tecidos periodontais saudáveis e durante os estágios inicias da periodontite experimental (PE) induzida em ratos. No presente estudo, foram utilizados 156 ratos machos, distribuídos aleatoriamente em 6 grupos: SPE-SS (n= 26): sem indução da PE e que receberam administração de solução salina (grupo Sham); SPE-CIS (n= 21) – sem indução da PE e que receberam administração de CIS; SPE-5FU (n= 27) – sem indução da PE e que receberam administração de 5-FU; PE-SS (n= 27): indução da PE e que receberam administração de solução salina; PE-CIS (n= 28) - indução da PE e que receberam administração de CIS; PE-5FU (n= 27) - indução da PE e que receberam administração de 5-FU. Os animais receberam 2 injeções para administração dos quimioterápicos com intervalo de 48 horas entre elas (períodos: baseline e 02 dias), aplicando-se as doses de 5 mg/kg e 2,5mg/kg (CIS) e 60 mg/kg e 40 mg/kg (5-FU). Logo após a primeira injeção, foi realizado a indução da PE adaptado-se um fio de algodão número 24 ao redor do primeiro molar inferior esquerdo. Nos períodos de 03, 05 e 07 dias após indução da PE, os animais foram eutanasiados pela administração de dose letal de thiopental (150mg/kg). Para análise hematológica de leucócitos totais (LT), foram obtidas amostras sanguíneas no baseline e no último período de eutanásia e para fosfatase alcalina (FA) no baseline e em todos os períodos experimentais. As mandíbulas obtidas foram fixadas em formaldeído tamponado 4%, descalcificadas em EDTA e incluídas em parafina para confecção de cortes histológicos seriados de 4 μm de espessura no sentido mésio-distal para coloração com Hematoxilina e Eosina e realização de reações imunoistoquímicas. Foi realizado análise histológica (escores), histométrica de porcentagem de osso na furca (POF) e padrões de imunomarcação para TNFα, IL-1β e RANKL (escores). Os dados obtidos foram submetidos à análise estatística (p<0,05) em programa computacional especializado. Observou-se uma diminuição nos níveis de LT nos grupos SPE-5FU e PE-5FU aos 7 dias. A FA sérica diminuiu aos 3, 5 e 7 dias nos grupos SPE-CIS, SPE-5FU, PE-SS, PE-CIS e PE-5FU. Histologicamente, SPE-CIS e SPE-5FU obtiveram uma exacerbação do processo inflamatório comparado ao SPE-SS, isso foi condizente com os resultados imunoistoquímicos, com maior padrão de imunomarcação para TNFα e IL-1β. Para PE-CIS e PE-5FU houve maior exacerbação do processo inflamatório comparados com os demais grupos, isso foi condizente com os resultados imunoistoquímicos, com maior padrão de imunomarcação para TNFα e IL-1β aos 3 e 7 dias e RANKL aos 7 dias. Deste modo, houve menor POF nos grupos PE-CIS e PE-5FU comparado ao PE-SS em todos períodos experimentais, além disso, PE-5FU apresentou menor POF até mesmo comparado ao PE-CIS aos 5 e 7 dias. Diante dos limites do presente estudo, pode-se concluir que os quimioterápicos promoveram uma exacerbação do processo inflamatório nos tecidos periodontais saudáveis e agravaram a progressão da PE em seu estágio inicial.
The aim of the present study was to evaluate the effect of the chemotherapics 5-Fluorouracil (5-FU) and Cisplatin (CIS) on healthy periodontal tissues and during the early stages of induced experimental periodontitis (EP) in rats. 156 male rats were randomly set in 6 groups: NP-SS (n = 26): without EP induction and received saline solution (Sham group); NP-CIS (n = 21) - without EP induction and received CIS; NP-5FU (n = 27) - without EP induction and received 5-FU; EP-SS (n = 27): induction of EP and received saline solution; EP-CIS (n = 28) - induction of EP and received CIS; EP-5FU (n = 27) - induction of PE and received 5-FU. The animals received 2 injections of chemotherapics with 48-hour interval between them, with doses of 5 mg/kg and 2.5 mg/kg (CIS) and 60 mg/kg and 40 mg/kg (5-FU). After the first injection, the induction of EP was performed by the placement of a number 24 cotton thread around the lower left first molar. At 03, 05 and 07 days after EP induction the animals were euthanized by administration of thiopental (150mg/kg). For hematological analysis of total leukocytes (TL), blood samples were obtained at baseline and in the last euthanasia period, and for Alkaline Phosphatase (AP), at baseline and in all experimental periods. The collected mandibles were fixed in buffered 4% formaldehyde, demineralized in EDTA and embedded in paraffin for preparation of serial histological sections of 4 μm thickness in mesiodistal direction for Hematoxylin and Eosin staining and immunohistochemical reactions. Histotologic (scores), histometry of the percentage of bone in the furcation (PBF) and immunolabelling standards for TNFα, IL-1β and RANKL (scores) analyzes were performed. The data were submitted to statistical analysis (p <0.05) in a specialized computer program. A decrease of the TL levels in NP-5FU and EP-5FU groups was observed at 7 days. Serum AP decreased at 3, 5 and 7 days in NP-CIS, NP-5FU, EP-SS, EP-CIS and EP-5FU. Histologically, NP-CIS and NP-5FU presented an exacerbation of the inflammatory process compared to the NP-SS, this was consistent with the immunohistochemical results, with higher immunolabelling pattern of TNFα and IL-1β. In EP-CIS and EP-5FU was observed an increased exacerbation of the inflammatory process compared to the other groups, this was consistent with the immunohistochemical results, with a higher immunolabelling pattern for TNFα and IL-1β at 3 and 7 days, and RANKL at 7 days. Thus, there was a smaller POF in EP-CIS and EP-5FU compared to the EP-SS in all experimental periods, moreover, EP-5FU presented smaller POF even compared to EP-CIS at 5 and 7 days. Given the limitations of the present study, it can be concluded that chemotherapy exacerbated the inflammatory process in healthy periodontal tissues and aggravated the progression of EP in its initial stage.
The aim of the present study was to evaluate the effect of the chemotherapics 5-Fluorouracil (5-FU) and Cisplatin (CIS) on healthy periodontal tissues and during the early stages of induced experimental periodontitis (EP) in rats. 156 male rats were randomly set in 6 groups: NP-SS (n = 26): without EP induction and received saline solution (Sham group); NP-CIS (n = 21) - without EP induction and received CIS; NP-5FU (n = 27) - without EP induction and received 5-FU; EP-SS (n = 27): induction of EP and received saline solution; EP-CIS (n = 28) - induction of EP and received CIS; EP-5FU (n = 27) - induction of PE and received 5-FU. The animals received 2 injections of chemotherapics with 48-hour interval between them, with doses of 5 mg/kg and 2.5 mg/kg (CIS) and 60 mg/kg and 40 mg/kg (5-FU). After the first injection, the induction of EP was performed by the placement of a number 24 cotton thread around the lower left first molar. At 03, 05 and 07 days after EP induction the animals were euthanized by administration of thiopental (150mg/kg). For hematological analysis of total leukocytes (TL), blood samples were obtained at baseline and in the last euthanasia period, and for Alkaline Phosphatase (AP), at baseline and in all experimental periods. The collected mandibles were fixed in buffered 4% formaldehyde, demineralized in EDTA and embedded in paraffin for preparation of serial histological sections of 4 μm thickness in mesiodistal direction for Hematoxylin and Eosin staining and immunohistochemical reactions. Histotologic (scores), histometry of the percentage of bone in the furcation (PBF) and immunolabelling standards for TNFα, IL-1β and RANKL (scores) analyzes were performed. The data were submitted to statistical analysis (p <0.05) in a specialized computer program. A decrease of the TL levels in NP-5FU and EP-5FU groups was observed at 7 days. Serum AP decreased at 3, 5 and 7 days in NP-CIS, NP-5FU, EP-SS, EP-CIS and EP-5FU. Histologically, NP-CIS and NP-5FU presented an exacerbation of the inflammatory process compared to the NP-SS, this was consistent with the immunohistochemical results, with higher immunolabelling pattern of TNFα and IL-1β. In EP-CIS and EP-5FU was observed an increased exacerbation of the inflammatory process compared to the other groups, this was consistent with the immunohistochemical results, with a higher immunolabelling pattern for TNFα and IL-1β at 3 and 7 days, and RANKL at 7 days. Thus, there was a smaller POF in EP-CIS and EP-5FU compared to the EP-SS in all experimental periods, moreover, EP-5FU presented smaller POF even compared to EP-CIS at 5 and 7 days. Given the limitations of the present study, it can be concluded that chemotherapy exacerbated the inflammatory process in healthy periodontal tissues and aggravated the progression of EP in its initial stage.
Descrição
Palavras-chave
Perda óssea alveolar, Doenças periodontais, Quimioterapia, Defeitos de furca, Modelos animais