Efeito da administração local ou sistêmica do Lactobacillus reuteri, no tratamento da doença periodontal experimentalmente induzida em ratos, submetidos à quimioterapia com 5-fluorouracil: estudo histopatológico, histométrico e imunoistoquímico
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Data
2018-10-19
Autores
Miessi, Daniela Maria Janjacomo [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Os probióticos (PROs) estão sendo intensamente avaliados na prevenção ou tratamento de doenças da cavidade oral, que estão associadas a uma mudança na composição e atividade microbiana do biofilme e resposta do hospedeiro. O objetivo desse estudo foi avaliar o efeito do Lactobacillus reuteri aplicado localmente ou sistemicamente como coadjuvante ao tratamento periodontal não cirúrgico de raspagem e alisamento radicular (RAR), na periodontite experimentalmente induzida em animais submetidos à quimioterapia com 5-fluorouracil (5-FU). A indução da periodontite experimental (PE) foi realizada com a colocação de um fio de algodão ao redor dos molares inferiores esquerdos de 108 ratos, que permaneceu por 7 dias. Todos os animais receberam o quimioterápico no momento da indução da PE (60mg/kg) e 48 horas após (40mg/kg) e foram separados aleatoriamente em 6 grupos com 18 animais cada um, com os seguintes tratamentos: 5FU: animais tratados com 5-FU; 5FU/RAR: animais tratados com 5-FU, que receberam tratamento com RAR seguido de 1 aplicação de solução fisiológica salina (SS; 0,16mL); Grupo 5FU/SSL: animais tratados com 5-FU, que receberam tratamento com RAR e 4 aplicações locais de SS (0,16mL); 5FU/SSS: animais tratados com 5-FU, que receberam tratamento com RAR e 4 gavagens com SS (0,16mL); 5FU/PL: animais tratados com 5-FU, que receberam tratamento com RAR e 4 aplicações locais de PRO (0,16 mL); 5FU/PS: animais tratados com 5-FU, que receberam tratamento com RAR e 4 gavagens com PRO (0,16mL). 6 animais de cada grupo foram submetidos à eutanásia aos 7, 15 e 30 dias após os tratamentos. A área da furca dos molares foi submetida às análises histopatológica, histométrica e dos padrões de imunomarcação para TRAP, PCNA, RANKL, OPG, OCN e TGF-β1. Os dados obtidos foram submetidos à análise estatística (α=5%). O grupo 5FU/PS apresentou melhora no processo inflamatório em todos os períodos. Houve maior perda óssea alveolar (PO) nos espécimes do grupo 5FU/SSS comparado ao grupo 5FU/SSL (p˂0,01) no período de 7 dias de avaliação. No período de 15 dias os espécimes dos grupos 5FU/SSL e 5FU/SSS demonstraram maior PO comparado aos grupos 5FU (p˂0,05) e 5FU/PS (p˂0,05). Aos 30 dias a PO mostrou-se maior nos espécimes do grupo 5FU/PL comparado aos grupos 5FU/RAR (p˂0,05), 5FU/SSS (p˂0,05) e 5FU/PS (p˂0,05). Na análise intragrupo houve menor PO no grupo 5FU/RAR e 5FU/PS aos 30 dias em relação aos 7 dias (p˂0,05). Na imunomarcação de células TRAP-positivas não foi evidenciada diferença significante entre os grupos e períodos; houve maior imunomarcação de células PCNA-positivas aos 7 dias no grupo 5FU/RAR comparado ao 5FU/SSS. Prevaleceu baixo padrão de imunomarcação de TGF-β1 e OCN nos grupos 5FU, 5FU/RAR, 5FU/SSL e 5FU/SSS em todos os períodos. O grupo 5FU/PS apresentou aos 7 dias um moderado padrão de imunomarcação e aos 15 e 30 dias um alto padrão de imunomarcação de TGF-β1 e OCN; o grupo 5FU/PL apresentou um moderado padrão de imunomarcação de TGF-β1 e OCN aos 7,15 e 30 dias. Na análise de RANKL prevaleceu um alto padrão de imunomarcação nos grupos 5FU, 5FU/RAR, 5FU/SSL, 5FU/SSS em todos os períodos e o grupo 5FU/PS apresentou aos 7 e 15 dias um moderado padrão de imunomarcação e aos 30 dias predominou um padrão de imunomarcação que variou do moderado ao baixo. Prevaleceu um padrão baixo de imunomarcação em todos os grupos experimentais e períodos, com exceção dos grupos 5FU/PL e 5FU/PS aos 30 dias (moderado padrão de imunomarcação) na análise de OPG. Diante dos resultados obtidos e com a metodologia empregada pode ser concluído que o uso sistêmico do Lactobacillus reuteri promoveu redução da inflamação e beneficiou o processo de reparação dos tecidos periodontais, porém não foi capaz de reduzir a PO na região de furca, demonstrando ser uma terapia periodontal coadjuvante promissora em animais submetidos à quimioterapia com 5-FU.
Probiotics (PROs) have been intensively evaluated to prevent or treat oral cavity diseases, associated with a change in the composition and microbial activity of the biofilm and response of the host. The aim of this study was to evaluate the effect of Lactobacillus reuteri applied locally or systemically as a coadjuvant to the non-surgical periodontal treatment of scaling and root planing (SRP) in experimentally induced periodontitis in animals treated with 5-fluorouracil (5-FU). Induction of experimental periodontitis (EP) was performed by placing a cotton thread around the left lower molars of 108 rats, which were kept there for 7 days. All animals underwent chemotherapy twice. They were 60mg/kg at the time they were subjected to ligature induction and again another 40 mg/kg, 48 hours later. They were randomly divided into 6 groups with 18 animals each and they received the following treatments: 5FU: treated animals with 5-FU; 5FU/SRP: 5-FU treated animals receiving SRP treatment followed by physiological saline (SS) solution; 5FU/SSL group: animals treated with 5-FU, who received SRP treatment and local SS applications; 5FU/SSS: animals treated with 5-FU, who received SRP treatment and systemic treatment with SS; 5FU/PL: animals treated with 5-FU, who received SRP treatment and local applications of PRO; 5FU/PS: animals treated with 5-FU, who received SRP treatment and systemic treatment with PRO (0.16ml x 4 days). Six animals from each group were submitted to euthanasia at 7, 15 and 30 days after treatments. The area of the molar furcation was submitted to histopathological, histometric and immuno-labeling analysis for TRAP, PCNA, RANKL, OPG, OCN and TGF-β1. The data were submitted to statistical analysis (α = 5%). There was greater bone loss (BL) in the 5FU/SSS group compared to the 5FU/SSL group (p˂0.01) in the 7-day evaluation period. In the 15-day period, specimens from the 5FU/SSL and 5FU/SSS groups showed a higher BL compared to the 5FU (p˂0.05) and 5FU/PS (p˂0.05) groups. At 30 days the BL was higher in the 5FU/PL group compared to the 5FU/SRP (p˂0.05), 5FU/SSS (p˂0.05) and 5FU/PS groups (p˂0.05). In the intragroup analysis there was a lower BL in the 5FU/SRP group and 5FU/PS at 30 days compared to 7 days (p˂0.05).In the immunostaining of TRAP-positive cells no significant difference between groups and periods was evidenced; there was greater immunolabeling of PCNA-positive cells at 7 days in the 5FU/SRP group compared to 5FU/SSS. Low prevalence of TGF-β1 and OCN immunostaining in the 5FU, 5FU/RAR, 5FU/SSL and 5FU/SSS groups prevailed at all periods. The 5FU/PS group presented at 7 days a moderate pattern of immunostaining and at 15 and 30 days a high standard of immunoblotting of TGF-β1 and OCN; the 5FU/PL group presented a moderate pattern of TGF-β1 and OCN immunoregulation at 7, 15 and 30 days; for RANKL a high standard of immunostaining in the 5FU, 5FU/SRP, 5FU/SSL, 5FU/SSS groups prevailed at all periods, 5FU/PL prevailed at 7 days a high standard of immunostaining and a moderate immunostaining pattern at 15 and 30 days and 5FU/PS showed a moderate immunostaining pattern at 7 and 15 days, and at 30 days a predominance of the immunoblot pattern varied from moderate to low; for OPG a low pattern of immunoblotting prevailed in all experimental groups and periods, except for 5FU/PL and 5FU/PS at 30 days that presented a moderate pattern of immunostaining. Based on the results obtained in this study and the methodology used to work on it, it was concluded that the use of Lactobacillus reuteri promoted reduction of inflamation and benefited the process of repairing periodontal tissues, but it was not able to reduce the BL in the furca region, proving to be a therapy promising adjuvant periodontal in animals submitted to 5-FU chemotherapy.
Probiotics (PROs) have been intensively evaluated to prevent or treat oral cavity diseases, associated with a change in the composition and microbial activity of the biofilm and response of the host. The aim of this study was to evaluate the effect of Lactobacillus reuteri applied locally or systemically as a coadjuvant to the non-surgical periodontal treatment of scaling and root planing (SRP) in experimentally induced periodontitis in animals treated with 5-fluorouracil (5-FU). Induction of experimental periodontitis (EP) was performed by placing a cotton thread around the left lower molars of 108 rats, which were kept there for 7 days. All animals underwent chemotherapy twice. They were 60mg/kg at the time they were subjected to ligature induction and again another 40 mg/kg, 48 hours later. They were randomly divided into 6 groups with 18 animals each and they received the following treatments: 5FU: treated animals with 5-FU; 5FU/SRP: 5-FU treated animals receiving SRP treatment followed by physiological saline (SS) solution; 5FU/SSL group: animals treated with 5-FU, who received SRP treatment and local SS applications; 5FU/SSS: animals treated with 5-FU, who received SRP treatment and systemic treatment with SS; 5FU/PL: animals treated with 5-FU, who received SRP treatment and local applications of PRO; 5FU/PS: animals treated with 5-FU, who received SRP treatment and systemic treatment with PRO (0.16ml x 4 days). Six animals from each group were submitted to euthanasia at 7, 15 and 30 days after treatments. The area of the molar furcation was submitted to histopathological, histometric and immuno-labeling analysis for TRAP, PCNA, RANKL, OPG, OCN and TGF-β1. The data were submitted to statistical analysis (α = 5%). There was greater bone loss (BL) in the 5FU/SSS group compared to the 5FU/SSL group (p˂0.01) in the 7-day evaluation period. In the 15-day period, specimens from the 5FU/SSL and 5FU/SSS groups showed a higher BL compared to the 5FU (p˂0.05) and 5FU/PS (p˂0.05) groups. At 30 days the BL was higher in the 5FU/PL group compared to the 5FU/SRP (p˂0.05), 5FU/SSS (p˂0.05) and 5FU/PS groups (p˂0.05). In the intragroup analysis there was a lower BL in the 5FU/SRP group and 5FU/PS at 30 days compared to 7 days (p˂0.05).In the immunostaining of TRAP-positive cells no significant difference between groups and periods was evidenced; there was greater immunolabeling of PCNA-positive cells at 7 days in the 5FU/SRP group compared to 5FU/SSS. Low prevalence of TGF-β1 and OCN immunostaining in the 5FU, 5FU/RAR, 5FU/SSL and 5FU/SSS groups prevailed at all periods. The 5FU/PS group presented at 7 days a moderate pattern of immunostaining and at 15 and 30 days a high standard of immunoblotting of TGF-β1 and OCN; the 5FU/PL group presented a moderate pattern of TGF-β1 and OCN immunoregulation at 7, 15 and 30 days; for RANKL a high standard of immunostaining in the 5FU, 5FU/SRP, 5FU/SSL, 5FU/SSS groups prevailed at all periods, 5FU/PL prevailed at 7 days a high standard of immunostaining and a moderate immunostaining pattern at 15 and 30 days and 5FU/PS showed a moderate immunostaining pattern at 7 and 15 days, and at 30 days a predominance of the immunoblot pattern varied from moderate to low; for OPG a low pattern of immunoblotting prevailed in all experimental groups and periods, except for 5FU/PL and 5FU/PS at 30 days that presented a moderate pattern of immunostaining. Based on the results obtained in this study and the methodology used to work on it, it was concluded that the use of Lactobacillus reuteri promoted reduction of inflamation and benefited the process of repairing periodontal tissues, but it was not able to reduce the BL in the furca region, proving to be a therapy promising adjuvant periodontal in animals submitted to 5-FU chemotherapy.
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Palavras-chave
Probióticos, Doença periodontal, 5-fluorouracil, Lactobacillus reuteri, Probiotics, Periodontal disease