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SILVA, P.H.F. Topical use of predatory bacteria reduces periodontal tissue destruction in rats with experimental periodontitis: histological, microtomographic, immunological and microbiological study. 2018. 172f. Dissertação (Mestrado) – Faculdade de Odontologia de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, 2018.SILVA, P.H.F. Topical use of predatory bacteria reduces periodontal tissue destruction in rats with experimental periodontitis: histological, microtomographic, immunological and microbiological study. 2018. 172f. Dissertação (Mestrado) – Faculdade de Odontologia de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, 2018. This study evaluated the effects of topical administration of Bdellovibrio bacteriovorus HD100 on experimental periodontitis in rats. Thirty-two rats were allocated in the CT, DPT, CT-HD100 and DPT-HD100 groups. At day 0 of the experiment, animals of the DPT and DPT-HD100 groups received silk ligatures around the mandibular first molars (MFMs). In the CT-HD100 and DPT-HD100 groups, 1 ml suspensions containing B. bacteriovorus HD100 were topically administered in the subgingival region of MFMs on days 0, 3 and 7. In the CT and DPT groups, topical administrations were performed with a suspension without B. bacteriovorus HD100. All animals were submitted to euthanasia on day 14 of the experiment. The gingival tissue, hemi-mandibles and oral biofilm were collected to evaluate the following parameters: i) bone microarchiteture, bone volume and alveolar bone level (X-ray microcomputed tomography micro-CT); ii) levels of conjunctive insertion (histomorphometric analysis); iii) microbiological profile (checkerboard DNA-DNA hybridization); iv) expression of inflammatory cytokines and transcription factors (immunoenzymatic analysis Multiplex and Real-time reverse transcription polymerase chain reaction); iii) immunostaining pattern for Beta defensins (BD), Toll-like receptors (TLR) and Cluster of differentiation (CD) (immunohistochemical reactions). In vitro tests were also performed to evaluate the antimicrobial potential of B. bacteriovorus HD100 against periodontopathogens. The data were analyzed statistically (p <0.05). The DPT-HD100 Group presented lower bone porosity, trabecular separation, alveolar bone level and connective tissue attachment level, as well as higher bone volume and number of bone trabeculae when compared to the DPT Group (p <0.05). The DPT-HD100 Group presented higher proportions of Actinomyces and Streptococcus-like species and smaller proportions of Prevotella intermedia, Peptostreptococcus micros, Fusobacterium nucleatum, Fusobacterium polymorphum, Eikenella corrodens, Eubacterium nodatum, Campylobacter gracilis, Capnocytophaga sputigena and Veillonella parvula-like species when compared to the DPT Group. In the analysis of immunoinflammatory parameters, the DPT-HD100 Group presented higher levels of Monocyte-1 chemoattractant protein (MCP-1), Regulated on activation, normal T cell expressed and secreted (RANTES), Osteoprotegerin (OPG), Transforming Growth Factor (TGF)-β and Interleukin (IL) -10 and lower levels of Tumor Necrosis Factor (TNF)-α, as well as a higher immunolabeling pattern for BD-1, BD-2 and BD-3 when compared to the DPT Group (p <0.05). For levels of IL-1β, IL-6, Macrophage Colony Stimulating Factor (M-CSF), Receptor activator of nuclear factor kappa-B ligand (RANK-L) and immunolabeling patterns for TLR-2 TLR4, CD-4, CD-8 and CD-57, no differences were observed between the DPT and DPT-HD100 groups. In the analysis of gene expression, the DPT-HD100 group presented higher expression of IL-17, IL-10 and Forkhead box P3 (FOXP3) when compared to the DPT Group (p <0.05). In the in vitro tests, co-cultures of periodontopathogens (F. nucleatum, P. intermedia and A. ctinomycetemcomitans) and B. bacteriovorus HD100 showed lower optical density than isolated cultures of periodontopathogens at 48 hours. Topical use of B. bacteriovorus HD100 modifies immunoinflammatory and microbiological parameters, promoting a protective effect against alveolar bone loss and loss of connective tissue attachment in rats with experimental periodontitis.

Uso tópico de bactérias predadoras reduz a destruição tecidual periodontal em ratos com periodontite experimental: estudo histológico, microtomográfico, imunológico e microbiológico