Sodium Trimetaphosphate and Hexametaphosphate Impregnated with Silver Nanoparticles: Characteristics and Antimicrobial Efficacy
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Biofouling The Journal of Bioadhesion and Biofilm Research ISSN: 0892-7014 (Print) 1029-2454 (Online) Journal homepage: https://www.tandfonline.com/loi/gbif20 Sodium trimetaphosphate and hexametaphosphate impregnated with silver nanoparticles: characteristics and antimicrobial efficacy Carla Corrêa Mendes-Gouvêa, Jackeline Gallo do Amaral, Renan Aparecido Fernandes, Gabriela Lopes Fernandes, Luiz Fernando Gorup, Emerson Rodrigues Camargo, Alberto Carlos Botazzo Delbem & Debora Barros Barbosa To cite this article: Carla Corrêa Mendes-Gouvêa, Jackeline Gallo do Amaral, Renan Aparecido Fernandes, Gabriela Lopes Fernandes, Luiz Fernando Gorup, Emerson Rodrigues Camargo, Alberto Carlos Botazzo Delbem & Debora Barros Barbosa (2018) Sodium trimetaphosphate and hexametaphosphate impregnated with silver nanoparticles: characteristics and antimicrobial efficacy, Biofouling, 34:3, 299-308, DOI: 10.1080/08927014.2018.1437146 To link to this article: https://doi.org/10.1080/08927014.2018.1437146 Published online: 27 Feb 2018. Submit your article to this journal Article views: 116 View Crossmark data Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=gbif20 BIOFOULING, 2018 VOL. 34, NO. 3, 299–308 https://doi.org/10.1080/08927014.2018.1437146 Sodium trimetaphosphate and hexametaphosphate impregnated with silver nanoparticles: characteristics and antimicrobial efficacy Carla Corrêa Mendes-Gouvêaa, Jackeline Gallo do Amarala, Renan Aparecido Fernandesb, Gabriela Lopes Fernandesb, Luiz Fernando Gorupc, Emerson Rodrigues Camargoc , Alberto Carlos Botazzo Delbema and Debora Barros Barbosab aDepartment of Pediatric Dentistry and Public Health, School of Dentistry Araçatuba, São Paulo State University (UNESP), Araçatuba, Brazil; bDepartment of Dental Materials and Prosthodontics, School of Dentistry Araçatuba, São Paulo State University (UNESP), Araçatuba, Brazil; cDepartment of Chemistry, Federal University of São Carlos (UFSCar), São Carlos, Brazil ABSTRACT ARTICLE HISTORY This study aimed to synthesize and characterize materials containing silver nanoparticles (AgNP) Received 15 August 2017 with polyphosphates (sodium trimetaphosphate (TMP) or sodium hexametaphosphate (HMP), and Accepted 31 January 2018 evaluate their effect against Candida albicans and Streptococcus mutans. The minimum inhibitory KEYWORDS concentration (MIC) was determined, which was followed by the quantification of the biofilm by Polyphosphates; silver counting colony-forming units (CFUs), the amount of metabolic activity, and the total biomass. The nanoparticles; biofilm; MICs revealed greater effectiveness of composites containing 10% Ag (TMP + Ag10% (T10) and Candida albicans; HMP + Ag10% (H10)) against both microorganisms. It was observed that T10 and H10 reduced the Streptococcus mutans formation of biofilms by 56–76% for C. albicans and by 52–94% for S. mutans for total biomass and metabolic activity. These composites promoted significant log reductions in the number of CFUs, between 0.45–1.43 log10 for C. albicans and 2.88–3.71 log10 for S. mutans (p < .001). These composites demonstrated significant antimicrobial activity, especially against S. mutans, and may be considered a potential alternative for new dental materials. Introduction hydroxyl (Souza et al. 2013; Delbem et al. 2014) and amino (Leone et al. 2008; Li and Chang 2008) attachment sites. Streptococcus mutans is the main cariogenic microor- Added to low concentrations of fluoride in an appropriate ganism, and its capacity to produce acid and survive at proportion in oral health products, they provide a greater low pH, as well as its ability to synthesize extracellular protective effect than conventional fluoride products (da polysaccharides, are important virulence factors which Camara et al. 2014; Manarelli et al. 2015; Takeshita et al. contribute to the colonization of the tooth surface, devel- 2015; Conceicao et al. 2015; da Camara et al. 2016). opment of pathogenic biofilms, and enamel demineral- The development of multifunctional biomaterials to be ization (Leone et al. 2008, Li and Chang 2008; Delbem used in the health field is the focus of increasing attention. et al. 2014). Although the development of dental caries Added to that, the exponential growth of nanotechnology, is related to the formation of biofilm, the progression of and the use of silver nanoparticles (AgNP) as an antimi- established lesions may include species of Candida, par- crobial agent allowing for increased surface-to-volume ticularly Candida albicans (Shibata and Morioka 1982; ratios and specific interactions with species of bacteria Monteiro et al. 2015). and fungi (Baker et al. 2005, Morones et al. 2005; Lok Composites containing calcium and phosphate have et al. 2006), new approaches could be useful to prevent shown promising anticariostatic effects (do Amaral et al. and control dental caries. A material containing AgNP as 2013; Takeshita et al. 2015; da Camara et al. 2016). Among an antimicrobial agent, and sources of fluoride and phos- the phosphate salts, sodium trimetaphosphate (TMP) and phate to prevent the demineralization of dental enamel, sodium hexametaphosphate (HMP) have been effective in could achieve promising results. Thus, this study aimed reducing enamel demineralization and improving remin- to synthesize composites containing AgNP, F, and TMP or eralization. They are inorganic cyclophosphates with the HMP, and to characterize and evaluate their effect against capacity to adsorb hydroxyapatite and protein through Streptococcus mutans and Candida albicans. CONTACT Debora Barros Barbosa [email protected] This article was originally published with errors. This version has been corrected. Please see Corrigendum (https://doi.org/10.1080/08927014.2018.1471022) © 2018 Informa UK Limited, trading as Taylor & Francis Group .C. C. MENDES-GOUVÊA ET AL 300 Materials and methods Minimum inhibitory concentration (MIC) Synthesis and characterization of the The microdilution method was performed following the nanocomposites Clinical Laboratory Standards Institute guidelines (CLSI: M27-A2 and M07-A9). The nanocomposites were diluted Nanocomposites were synthesized based on the pro- in deionized water in geometric progression, from 2 to tocol proposed by Miranda et al. (2010), with some 1,024 times. Afterwards, each nanocomposite concen- modifications. The synthesis was carried out in an alco- tration obtained previously was diluted (1:5) in RPMI holic medium (isopropanol) using sodium borohydride 1640 medium (Sigma-Aldrich) for Candida albicans (NaBH , Sigma-Aldrich, St Louis, MO, USA) as a reducing 4 (ATCC 10231), and in brain heart infusion (BHI, Difco, agent. Suspensions containing sodium trimetaphosphate Le Pont de Claix, France) for Streptococcus mutans (ATCC (TMP, Sigma-Aldrich, CAS 7785-84-4) or sodium hex- 25175). Inocula from 24-h cultures were adjusted to the ametaphosphate (HMP, Sigma-Aldrich, CAS 68915-31-1), 0.5 McFarland standard turbidity equivalent in saline fluoride (NaF, Merck, Darmstadt, Germany), and silver solution (0.85% NaCl). Suspensions of each strain were nitrate (AgNO , Merck) were prepared in the presence of a 3 diluted (1:5) in saline and subsequently diluted (1:20) in surfactant (ammonium salt of polymethacrylic acid (NH- RPMI 1640 for C. albicans or BHI broth for S. mutans. PM), Polysciences, Inc., Warrington, PA, USA). AgNO 3 Each microorganism suspension (100 μl) was added in the was employed at 1% or 10% of the phosphate weight. The wells of microtiter plates containing 100 μl of each nano- molar proportions between phosphates and fluoride was composite concentration. Polyphosphate/fluoride, AgNP, 1.2:1 TMP/NaF or 0.62:1 HMP/NaF, and between silver and chlorhexidine gluconate (CHG) (Periogard-Colgate, ions (Ag+) and NaBH it was 1:1.26. Samples were dried 4 New York, NY, USA) were tested as controls. The micr- overnight at 70°C. AgNP with no polyphosphates was also otiter plates were incubated at 37°C, and the MICs were synthesized. determined visually as the lowest concentration of nano- The morphology of the nanocomposites was character- composites with no microorganism growth after 48 h. ized by scanning electron microscopy (SEM) on a Zeiss Supra 35VP microscope (S-360 microscope, Leo, MA, USA) with field emission gun electron effect (FEG-SEM, Biofilm formation and treatment 10 kV). A drop of each sample was placed on a silicon substratum and dried at 40°C for 12 h. Two-dimensional Single species biofilms were grown in 96-well microtiter images were constructed by analyzing the P Kα, Ag Lα1, plates (Costar®, Corning Incorporated, Tewksbury, MA, and O Kα energy emissions using energy dispersive X-ray USA) containing 0.2 ml of C. albicans cell suspension 7 −1 detector (EDX) analysis with mapping. Samples were also (1 × 10 cells ml in artificial saliva) (Silva et al. 2010) or 8 −1 characterized by X-ray diffraction (XRD) (Rigaku dif- 0.2 ml of S. mutans (1 × 10 cells ml in artificial saliva) fractometer [Rigaku Corporation, Tokyo, Japan], DMax (Arias et al. 2016). The plates were incubated at 37°C for 2500PC) with CuK radiation (λ = 1.5406 Å) generated at 24 h. After this period, the biofilm-containing wells were a voltage of 40 kV and a current of 40 mA. The scanning washed with 0.2 ml of phosphate buffered saline (PBS; range (2θ) was from 20° to 110° with a step size of 0.2°, a pH 7, 0.1 M) to remove the non-adherent cells. Next, the 1.0 mm divergent slit, 0.2 mm collection slot, and accu- nanocomposites were