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Objective: The present study aimed to establish an optimized protocol for biofilm removal from titanium (Ti) surfaces using citric acid (CA) solutions., Background: Biofilm accumulation is the main factor to trigger peri-implant infections and to increase the risk of treatment failures. Although CA has been suggested as the anti-infective agent with highest potential for biofilm removal on Ti, there is no consensus that CA could improve the anti-infective treatment and its effect., Methods: Physical and chemical alterations, electrochemical behavior, cytotoxicity, and antimicrobial effect of CA on Ti discs were evaluated using four concentrations (1, 10, 20, and 40%) and two application methods (immersion and rubbing). Negative control using 0.9% NaCl was used in all experiments. To evaluate whether different application times can have similar response, polymicrobial biofilm (microcosm model) was formed on Ti and treated with CA for 1, 2, 4, and 8 min. An in situ study was conducted to verify whether the established protocol is equally effective in biofilms formed on machined and sandblasted, large-grit, and acid-etched (SLA) Ti surfaces., Results: CA 40% induced significantly higher surface alterations observed by confocal images and profilometry. In general, rubbing protocol decreased the surface roughness and increased the wettability (p < 0.05), exhibiting better surface cleaning by biofilm removal. CA 10% presented no indirect cytotoxicity and, when applied by rubbing for 8 min, presented proper in vitro antibacterial action and potential corrosion inhibition. When CA 10% was rubbed on Ti surfaces for 4 min, it displayed optimum cleaning ability as 8 min, working equally to remove in situ biofilm on machined and SLA surfaces., Conclusions: The application of CA 10% by rubbing for at least 4 min demonstrated to be a promising protocol to eliminate biofilms formed in smooth and rougher surfaces, which could improve implant-related infection therapies., (© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: To evaluate the effect of titanium (Ti) particles and ions on oral biofilm growth and composition., Background: Particles and ions of Ti released from dental implants can trigger unfavorable biological responses in human cells. However, their effect on oral biofilms composition has not been tested., Methods: In this blind in situ study, volunteers wore a palatal appliance containing Ti disks for 7 days to allow biofilm formation. Disks were then collected and biofilms were treated, in vitro, with Ti particles (0.75% and 1%), ions (10 and 20 ppm), or a combination of both (1% particles + 20 ppm ions). Biofilms exposed only to medium was used as control group. After 24 hours, biofilms were collected and analyzed by checkerboard DNA-DNA hybridization. Direct effects of Ti particles and ions on biofilm/cellular morphology were evaluated by transmission electron microscopy (TEM)., Results: Ti particles affected biofilm composition, increasing population of four bacterial species (P < .05), while Ti ions showed higher levels of putative pathogens from the orange complex with reduction in species from the yellow complex (P < .05), compared with control. The combination of particles + ions increased green complex and reduced yellow complex proportions (P < .05). TEM showed clusters of particles agglomerated in extracellular environment, while Ti ions were precipitated in both extracellular and intracellular sites., Conclusions: Ti products, especially Ti ions, have the potential to change the microbiological composition of biofilms formed on Ti surfaces. Therefore, the presence of Ti products around dental implants may contribute to microbial dysbiosis and peri-implantitis., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)