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Doctoral Thesis
DOI
https://doi.org/10.11606/T.58.2021.tde-30112022-185449
Document
Author
Full name
Luiz Fernando Ferreira de Oliveira
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2021
Supervisor
Committee
Messora, Michel Reis (President)
Fonseca, Douglas Campideli
Gaio, Eduardo Jose
Salvador, Sergio Luiz de Souza
Title in Portuguese
Efeitos da administração sistêmica de Bifidobacterium animalis subsp. lactis HN019 na periodontite experimental em ratos. Estudo histológico, microtomográfico, imunológico e microbiológico
Keywords in Portuguese
Bifidobacterium
Imunologia
Periodontite
Ratos
Resposta do hospedeiro
Abstract in Portuguese
Este estudo avaliou os efeitos da administração sistêmica de Bifidobacterium animalis subsp. lactis HN019 (HN019) na periodontite experimental em ratos. 32 ratos foram alocados nos grupos CS, DPS, CS-HN019 e DPS-HN019. No dia 0 do experimento, os animais dos grupos CS-HN019 e DPS-HN019 começaram a administração diariamente de B. lactis HN019 na água contendo 1x10^9UFC/mL até o final do experimento (28 dias). No dia 14 os animais dos grupos DPS e DPS-HN019 receberam as ligaduras de seda ao redor dos primeiros molares inferiores (PMIs). Todos os animais foram submetidos à eutanásia no dia 28 do experimento. O tecido gengival, soro sanguíneo, hemi-mandíbulas e biofilme bucal foram coletados para avaliação dos seguintes parâmetros: i) microarquitetura óssea e volume ósseo (microtomografia computadorizada por transmissão de raios X - micro-CT); ii) níveis do osso alveolar na área de furca (análise histomorfométrica); iii) microbiota bucal (checkerboard DNA-DNA hybridization); iv) expressão de citocinas inflamatórias e fatores de transcrição (análise imunoenzimática - Multiplex e reação em cadeia da polimerase por transcriptase reversa em tempo real); iii) padrão de imunomarcação para beta defensinas (BD), receptores do tipo Toll (TLR) e grupamentos de diferenciação (CD) (reações imunohistoquímicas). O grupo DPS-HN019 apresentou menor porosidade óssea, bem como maior volume ósseo quando comparado ao Grupo DPS (p < 0,05). O Grupo DPS-HN019 apresentou menor percentual de espécies semelhantes à F. periodonticum e maior percentual de espécies semelhantes à A. gereneseriae, A.israelli e S. gordonii quando comparado ao Grupo DPS. Nas análises de parâmetros imunoinflamatórios, no tecido gengival os resultados mostram valores aumentados na razão IL-1β/IL-10, IL-1β/IL-4, GROKC/IL-10, RANKL/OPG no Grupo DPS quando comparado ao grupo DPS-HN019 (p <0,05). Nas razões IL-10/TNFα, IL-6/TNFα, MCSF/TNFα, IL-4/ TNFα, TGFβ/TNFα estão reduzidas no grupo DPS quando comparado ao Grupo DPS-HN019 (p <0,05). A razão IL-4/TNFα se mostrou aumentada no CS-HN019 quando comparado ao grupo CS (p <0,05). No soro os resultados mostram valores aumentados na razão IL-1β/IL-4, IL-1β/IL-10 no soro do Grupo DPS quando comparado ao grupo DPS-HN019 (p <0,05). O grupo DPS-HN019 apresentou maior imunomarcação de BDs quando comparado ao grupo DPS (p <0,05). Em relação TLR e CD não houve diferença entre os grupos. Os dados histomorfométricos mostram uma menor área sem osso (ASOL) no grupo DPS-HN019 quando comparado ao grupo DPS (p <0,05). O uso sistêmico de B. lactis HN019 modifica os parâmetros imunoinflamatórios e microbiológicos, promovendo um efeito protetor contra a perda óssea alveolar em ratos com periodontite experimental.
Title in English
Effects of systemic administration of Bifidobacterium animalis subsp. lactis HN019 in experimental periodontitis in rats. Histological, microtomographic, immunological and microbiological study
Keywords in English
Bifidobacterium
Host response
Immunology
Periodontitis
Rats
Abstract in English
This study evaluated the effects of systemic administration of Bifidobacterium animalis subsp. lactis HN019 (HN019) in experimental periodontitis (DP) in rats. 32 rats were allocated to groups CS, CS-HN019, DPS and DPS-HN019. On day 0 of the experiment, the animals in the CS-HN019 and DPS-HN019 groups received daily administration of B. lactis HN019 in water containing 1x10^9 CFU / mL until the end of the experiment (28 days). On day 14, animals in the DPS and DPS-HN019 groups received silk ligatures around the lower first molars (LFM). All animals were euthanized on day 28 of the experiment. Gingival tissue, blood serum, hemi-mandibles and oral biofilm were collected to assess the following parameters: i) bone microarchitecture and bone volume (X-ray transmission computerized microtomography - micro-CT); ii) alveolar bone levels in the furcation area (histomorphometric analysis); iii) oral microbiota (checkerboard DNA-DNA hybridization); iv) expression of inflammatory cytokines and transcription factors (immunoenzymatic analysis - Multiplex and real-time reverse transcriptase polymerase chain reaction); iii) immunostaining pattern for beta defensins (BD), Toll-like receptors (TLR) and differentiation clusters (CD) (immunohistochemical reactions). The DPS-HN019 group had lower bone porosity as well as greater bone volume when compared to the DPS group (p < 0.05). The DPS-HN019 Group had a lower percentage of species similar to F. periodonticum and a higher percentage of species similar to A. gereneseriae, A.israelli and S. gordonii when compared to the DPS Group. In the analysis of immunoinflammatory parameters, in the gingival tissue, the results show increased values in the ratio IL-1β/IL-10, IL-1β/IL-4, GROKC/IL-10, RANKL/OPG in the DPS group when compared to the DPS- group HN019 (p<0.05). In the IL-10/TNFα, IL-6/TNFα, MCSF/TNFα, IL-4/ TNFα, TGFβ/TNFα ratios are reduced in the DPS group when compared to the DPS-HN019 group (p <0.05). The IL-4/TNFα ratio was increased in CS-HN019 when compared to the CS group (p < 0.05). In serum, the results show increased values in the ratio IL-1β/IL-4, IL-1β/IL-10 in the serum of the DPS Group when compared to the DPS-HN019 group (p <0.05). The DPS-HN019 group showed higher immunostaining of BDs when compared to the DPS group (p < 0.05). In relation to TLR and CD there was no difference between groups. Histomorphometric data show a smaller area without bone (ASOL) in the DPS-HN019 group when compared to the DPS group (p < 0.05). The systemic use of B. lactis HN019 modifies immunoinflammatory and microbiological parameters, promoting a protective effect against alveolar bone loss in rats with experimental periodontitis.
 
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Publishing Date
2022-12-01
 
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