Atividade antimicrobiana de diferentes concentrações da prata coloidal aplicada em implantes dentários com plataforma do tipo hexágono externo: estudo in vitro
Carregando...
Data
2020-02-05
Autores
Pignataro, Rossana Reim Del Gaudio
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
Os implantes dentários osseointegrados apresentam altas taxas de sucesso no tratamento reabilitador a longo prazo. No entanto, complicações biológicas estão relacionadas com a ocorrência de peri-implantite, perda óssea marginal e insucessos clínicos. A busca de uma condição que diminua a colonização microbiana e estabelecimento de biofilmes tem crescido. O tratamento com nanoprata tem sido amplamente estudado como agente antimicrobiano por apresentar seu largo espectro de ação contra bactérias Gram positivas, Gram negativas e fungos. O presente estudo teve por objetivo avaliar, in vitro, a ação antimicrobiana de nanopartículas de prata coloidal, sobre Enterococcus faecalis e a ocorrência de infiltração bacteriana na interface dos implantes de plataforma hexágono externo (HE) e pilares protéticos. Foi determinada a concentração inibitória mínima da solução de nanoprata coloidal. A citotoxicidade foi avaliada sobre células Vero (ECACC 84113001) pela técnica de MTT. A infiltração bacteriana na interface implante/pilar (TiBase®, Conexão Sistemas de Prótese), foi avaliada utilizando 40 conjuntos de implantes e pilares HE (n=10) divididos em 4 grupos de acordo com o tratamento: (1) prata coloidal 29 ppm; (2) prata coloidal 58 ppm; (3) controle negativo (sem tratamento) e (4) controle positivo (digluconato de clorexidina 2%). Inicialmente, da suspensão padronizada 1x106 células/mL de Enterococcus faecalis foi inoculada no interior dos implantes, e, a seguir, os respectivos pilares (TiBase®, Conexão Sistemas de Prótese) foram instalados com torque de 30N de acordo com o fabricante. Os conjuntos foram testados para contaminação externa imediata, suspensos em tubos de ensaio contendo 3 mL de meio de cultura BHI (Brain Heart Infusion), incubados por 24h a 37ᵒC em estufa de CO2. A turvação do caldo indicou infiltração bacteriana. A seguir, os corpos de prova foram incubados em aerobiose a 37ᵒC, por 120 horas. Após incubação, os pilares foram desparafusados e o conteúdo interno dos implantes foi coletado com auxílio de cone de papel estéril e semeado em ágar para a determinação do número de células viáveis. Os dados de log10 UFC/mL dos grupos foram comparados estatisticamente com auxílio do programa GraphPad Prism versão 6.0 (GraphPad Software Inc., San Diego, CA, USA). Para tanto, foi realizado um teste de normalidade, e, a seguir, foram aplicados os testes estatísticos Kruskal-Wallis e Dunn’s post hoc com nível de significância de 5%. Resultados: Nenhum corpo de prova apresentou contaminação externa imediata. O número de células viáveis foi significativamente menor no grupo tratado com a nanoprata coloidal a 58 ppm em relação ao grupo controle negativo. O grupo tratado com digluconato de clorexidina a 2% obteve a contagem significativamente menor em relação aos demais grupos testados. A nanoprata coloidal 58 ppm foi severamente citotóxica. Conclui-se que a nanoprata coloidal na concentração de 58 ppm apresentou atividade antimicrobiana sobre o Enterococcus faecalis, porém com elevada citotoxicidade para células eucarióticas.
Osseointegrated dental implants have high success rates in long-term rehabilitative treatments However; oral conditions could affect peri-implant tissues health and are associated to the occurrence of peri- implantitis, marginal bone loss and clinical failures. The search for a condition that can decrease the microbial colonization and the establishment of biofilms increased. Nanosilver has been widely used as an antimicrobial agent due large spectrum of action against Gram positive, Gram-negative bacteria and fungi. The present study aimed to evaluate, in vitro, the antimicrobial action of colloidal silver nanoparticles on Enterococcus faecalis and on the infiltration at the interface between hexagonal external platform (EH) implants and prosthetic abutments. To obtain the optimal silver concentration, the minimum inhibitory concentration. Cytotoxicity evaluated on Vero cells (ECACC 84113001) by MTT assay. Bacterial infiltration through implant / abutment interface (TiBase®, Conexão Sistemas de Prótese) was evaluated using 40 sets of HE implants and abutments (n = 10) divided into 4 groups according to the treatment: (1) colloidal silver 29 ppm; (2) 58 ppm colloidal silver; (3) negative control (no treatment) and (4) positive control (2% chlorhexidine digluconate). Initially, under sterile conditions, 1 µL of a standardized 1x106 cels / mL suspension of Enterococcus faecalis was inoculated inside the implants, and the respective abutments (TiBase®, Conexão Sistemas de Prótese) were torqued to 30N according to the manufacturer’s instructions. The systems were tested for immediate external contamination, suspended in test tubes containing 3 mL Brain Heart Infusion broth. The tubes were incubated for 24h at 37ᵒC under a CO2 atmosphere. Turbidity indicated external contamination. After the incubation period, the abutments unscrewed, and the internal content of the implants was collected with the aid of a sterile paper point. The bacterial cells were dispersed, and the suspension was plated to quantify the viable cells. Values of log10CFU / mL obtained for the groups were statistically compared using the GraphPad Prism version 6.0 software (GraphPad Software Inc., San Diego, CA, USA). A normality test was performed followed by Kruskal-Wallis and post hoc statistical tests with significance level of 5%. Results: No specimen showed immediate external contamination. Viable cells count was significantly lower in the 58-ppm colloidal nano-silver group when compared to the negative control group. The group treated with 2% chlorhexidine digluconate showed significantly lower counts when to the other groups tested. Colloidal nano silver 58 ppm was severely cytotoxic. It concluded that the 58-ppm colloidal nano silver presented antimicrobial activity on Enterococcus faecalis however with high cytotoxicity to eucariotic cells.
Osseointegrated dental implants have high success rates in long-term rehabilitative treatments However; oral conditions could affect peri-implant tissues health and are associated to the occurrence of peri- implantitis, marginal bone loss and clinical failures. The search for a condition that can decrease the microbial colonization and the establishment of biofilms increased. Nanosilver has been widely used as an antimicrobial agent due large spectrum of action against Gram positive, Gram-negative bacteria and fungi. The present study aimed to evaluate, in vitro, the antimicrobial action of colloidal silver nanoparticles on Enterococcus faecalis and on the infiltration at the interface between hexagonal external platform (EH) implants and prosthetic abutments. To obtain the optimal silver concentration, the minimum inhibitory concentration. Cytotoxicity evaluated on Vero cells (ECACC 84113001) by MTT assay. Bacterial infiltration through implant / abutment interface (TiBase®, Conexão Sistemas de Prótese) was evaluated using 40 sets of HE implants and abutments (n = 10) divided into 4 groups according to the treatment: (1) colloidal silver 29 ppm; (2) 58 ppm colloidal silver; (3) negative control (no treatment) and (4) positive control (2% chlorhexidine digluconate). Initially, under sterile conditions, 1 µL of a standardized 1x106 cels / mL suspension of Enterococcus faecalis was inoculated inside the implants, and the respective abutments (TiBase®, Conexão Sistemas de Prótese) were torqued to 30N according to the manufacturer’s instructions. The systems were tested for immediate external contamination, suspended in test tubes containing 3 mL Brain Heart Infusion broth. The tubes were incubated for 24h at 37ᵒC under a CO2 atmosphere. Turbidity indicated external contamination. After the incubation period, the abutments unscrewed, and the internal content of the implants was collected with the aid of a sterile paper point. The bacterial cells were dispersed, and the suspension was plated to quantify the viable cells. Values of log10CFU / mL obtained for the groups were statistically compared using the GraphPad Prism version 6.0 software (GraphPad Software Inc., San Diego, CA, USA). A normality test was performed followed by Kruskal-Wallis and post hoc statistical tests with significance level of 5%. Results: No specimen showed immediate external contamination. Viable cells count was significantly lower in the 58-ppm colloidal nano-silver group when compared to the negative control group. The group treated with 2% chlorhexidine digluconate showed significantly lower counts when to the other groups tested. Colloidal nano silver 58 ppm was severely cytotoxic. It concluded that the 58-ppm colloidal nano silver presented antimicrobial activity on Enterococcus faecalis however with high cytotoxicity to eucariotic cells.
Descrição
Palavras-chave
Implantes dentários, Microinfiltração, Microrganismos, Nanopartícula prata, Peri-implantite, Dental implants, Microleakage, Microorganisms, Silver nanoparticle, Peri-implantitis