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Profile of host cell responses to exposure to stressed bacteria in planktonic; dislodged, and intact biofilm mode
Zancan, Rafaela Fernandes; Ponce, José Burgos; Dionisio, Thiago José; Oliveira, Rodrigo Cardoso de; Silva, Rafaela Alves da; Duque, Jussaro Alves; Duarte, Marco Antonio Hungaro.
  • Zancan, Rafaela Fernandes; University of São Paulo. Bauru Dental School. Department of Restorative Dentistry, Dental Materials and Endodontics. Bauru. BR
  • Ponce, José Burgos; University of São Paulo. Bauru Dental School. Department of Surgery, Stomatology, Pathology and Radiology. BR
  • Dionisio, Thiago José; University of São Paulo. Bauru Dental School. Department of Biological Sciences. BR
  • Oliveira, Rodrigo Cardoso de; University of São Paulo. Bauru Dental School. Department of Pediatric Dentistry, Orthodontics and Public Health. Bauru. BR
  • Silva, Rafaela Alves da; University of São Paulo. Bauru Dental School. Department of Pediatric Dentistry, Orthodontics and Public Health. Bauru. BR
  • Duque, Jussaro Alves; University of São Paulo. Bauru Dental School. Department of Restorative Dentistry, Dental Materials and Endodontics. Bauru. BR
  • Duarte, Marco Antonio Hungaro; University of São Paulo. Bauru Dental School. Department of Restorative Dentistry, Dental Materials and Endodontics. Bauru. BR
Braz. dent. j ; 32(3): 10-20, May-June 2021. tab, graf
Article in English | LILACS, BBO | ID: biblio-1345504
Responsible library: BR1.1
ABSTRACT
Abstract The host defense response to microbial challenge emerging from the root canal system leads to apical periodontitis. The aim of this study was to evaluate the expression of inflammatory cytokines and Nitric Oxide (NO) by macrophages after interaction with Enterococcus faecalis in the plankton and dislodged biofilm mode; intact biofilm mode stimulated by calcium hydroxide (CH), CH and chlorhexidine (CHX) or Triple Antibiotic Paste (TAP). For this purpose, culture of macrophages from monocytes in human peripheral blood (N=8) were exposed to the different modes of bacteria for 24 hours. Subsequently, the cytokines, such as, Tumor Necrotic Factor- alfa (TNF-α), interleukin (IL)-1β, IL-6, IL-10; and NO were quantified by Luminex xMAP and Greiss reaction, respectively. In addition to the potential therapeutic effects of the intracanal medication, their antimicrobial activity against Enterococcus faecalis biofilm were also tested in vitro by confocal microscopy. The experiments` data were analyzed by the Kruskal-Wallis test with the Dunn post hoc test (α < 0.05). Bacteria in dislodged biofilm mode were shown to be more aggressive to the immune system than bacteria in plankton mode and negative control, inducing greater expression of NO and TNF-α. Relative to bacteria in intact biofilm mode, the weakest antimicrobial activity occurred in Group CH. In Groups CH/CHX and TAP the percentage of dead bacteria was significantly increased to the same extent. Interestingly, the biofilm itself did not induce the release of pro-inflammatory cytokines - except for NO - while the biofilm treated with TAP and CH based pastes enhanced the levels of IL-6 and TNF-α; and IL-1 β, respectively. In contrast, the levels of a potent anti-inflammatory (IL-10) were increased in Group TAP.
RESUMO
Resumo A resposta de defesa do hospedeiro ao desafio microbiano que emerge do sistema de canais radiculares leva à periodontite apical. Os objetivos deste estudo foram avaliar a expressão de citocinas pró e anti-inflamatórias e Óxido Nítrico (NO) por macrófagos após interação com Enterococcus faecalis no modo planctônio e de biofilme desalojado; biofilme intacto estimulado por hidróxido de cálcio (CH), CH e clorexidina ou Pasta Tri Antibiótica (TAP). Para isto, a cultura de macrófagos originados de monócitos do sangue periférico de humanos (N=8) foi exposta aos diferentes tipos de bactéria por 24 horas. Então, a quantificação da produção de of Fator de Necrose Tumoral- alfa (TNF-α), interleucina (IL)-1β, IL-6, IL-10 e NO por macrófagos se deu por meio do Luminex xMAP e reação de Greiss, respectivamente. Além dos potenciais efeitos terapêuticos desses compostos, sua atividade antimicrobiana contra E. faecalis também foi testada através microscopia confocal. Os dados dos experimentos foram analisados através do teste de Kruskal-Wallis com Dunn`s post hoc (α < 0.05). Bactéria em modo de biofilme desalojado se mostrou mais agressivo ao sistema imune que as bactérias planctônicas e controle negativo induzindo a maior excreção de NO e TNF-α. Em relação ao biofilme intacto, a atividade antimicrobiana mais fraca ocorreu no grupo de CH. Os grupos CHX e TAP aumentaram significativamente a porcentagem de bactérias mortas na mesma extensão. Interessantemente, o biofilme por ele mesmo não induziu a liberação de citocinas pro-inflamatórias - exceto por NO - enquanto que o biofilme tratado com TAP ou pastas a base de CH aumentaram os níveis de IL-6; e TNF-α e IL-1 β respectivamente. Em contraste, os níveis da potente citocina anti-inflamatória (IL-10) foram aumentados pelo grupo TAP.
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Full text: Available Index: LILACS (Americas) Main subject: Plankton / Biofilms Limits: Humans Language: English Journal: Braz. dent. j Journal subject: Dentistry Year: 2021 Type: Article Affiliation country: Brazil Institution/Affiliation country: University of São Paulo/BR

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Full text: Available Index: LILACS (Americas) Main subject: Plankton / Biofilms Limits: Humans Language: English Journal: Braz. dent. j Journal subject: Dentistry Year: 2021 Type: Article Affiliation country: Brazil Institution/Affiliation country: University of São Paulo/BR