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      In vitro and in vivo evaluations of glass-ionomer cement containing chlorhexidine for Atraumatic Restorative Treatment

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          Abstract

          Objectives:

          Addition of chlorhexidine has enhanced the antimicrobial effect of glass ionomer cement (GIC) indicated to Atraumatic Restorative Treatment (ART); however, the impact of this mixture on the properties of these materials and on the longevity of restorations must be investigated. The aim of this study was to evaluate the effects of incorporating chlorhexidine (CHX) in the in vitro biological and chemical-mechanical properties of GIC and in vivo clinical/ microbiological follow-up of the ART with GIC containing or not CHX.

          Material and Methods:

          For in vitro studies, groups were divided into GIC, GIC with 1.25% CHX, and GIC with 2.5% CHX. Antimicrobial activity of GIC was analyzed using agar diffusion and anti-biofilm assays. Cytotoxic effects, compressive tensile strength, microhardness and fluoride (F) release were also evaluated. A randomized controlled trial was conducted on 36 children that received ART either with GIC or GIC with CHX. Saliva and biofilm were collected for mutans streptococci (MS) counts and the survival rate of restorations was checked after 7 days, 3 months and one year after ART. ANOVA/Tukey or Kruskal-Wallis/ Mann-Whitney tests were performed for in vitro tests and in vivo microbiological analysis. The Kaplan-Meier method and Log rank tests were applied to estimate survival percentages of restorations (p<0.05).

          Results:

          Incorporation of 1.25% and 2.5% CHX improved the antimicrobial/anti-biofilm activity of GIC, without affecting F release and mechanical characteristics, but 2.5% CHX was cytotoxic. Survival rate of restorations using GIC with 1.25% CHX was similar to GIC. A significant reduction of MS levels was observed for KM+CHX group in children saliva and biofilm 7 days after treatment.

          Conclusions:

          The incorporation of 1.25% CHX increased the in vitro antimicrobial activity, without changing chemical-mechanical properties of GIC and odontoblast-like cell viability. This combination improved the in vivo short-term microbiological effect without affecting clinical performance of ART restorations.

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          Most cited references31

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          Presence of mutans streptococci and Candida spp. in dental plaque/dentine of carious teeth and early childhood caries.

          This study determined the presence of mutans streptococci and Candida spp. in supragingival dental plaque and infected dentine of caries-free children, with early childhood caries and caries. Pooled samples of dental plaque and infected dentine were collected from 56 children aged 1-5 years, which were divided into 3 groups: early childhood caries (ECC); caries and caries-free. Infected dentine was collected in ECC and caries groups to compare the frequency of these microorganisms in the collected sites. The samples were inoculated in SB20 and SA medium, for mutans streptococci and Candida spp., respectively, and incubated at 37 degrees C for 48 h. Colony growth was verified and the identification was performed by biochemical tests and CHROMagar Candida. Fisher's test or chi-square (chi(2)) were applied (p=0.05). The more prevalent species were S. mutans and Candida albicans in ECC (85.4% and 60.4%, respectively), independently of the sample site. S. mutans only was significantly associated with carious teeth, whether in early childhood caries or not. However, the frequency of C. albicans in ECC was higher when compared to caries and caries-free groups. There is a significant association between the presence of C. albicans and early childhood caries.
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            Antimicrobial actions of fluoride for oral bacteria.

            Fluoride is widely used as a highly effective anticaries agent. Although it is felt that its anticaries action is related mainly to effects on mineral phases of teeth and on the process of remineralization, fluoride also has important effects on the bacteria of dental plaque, which are responsible for the acidification of plaque that results in demineralization. The results of recent studies have shown that fluoride can affect bacterial metabolism through a set of actions with fundamentally different mechanisms. It can act directly as an enzyme inhibitor, for example for the glycolytic enzyme enolase, which is inhibited in a quasi-irreversible manner. Direct action seems also to occur in inhibition of heme-based peroxidases with binding of fluoride to heme. The flavin-based peroxidases of many oral bacteria are insensitive to fluoride. Another mode of action involves formation of metal-fluoride complexes, most commonly AlF4-. These complexes are responsible for fluoride inhibition of proton-translocating F-ATPases and are thought to act by mimicking phosphate to form complexes with ADP at reaction centers of the enzymes. However, the actions of fluoride that are most pertinent to reducing the cariogenicity of dental plaque are those related to its weak-acid character. Fluoride acts to enhance membrane permeabilities to protons and compromises the functioning of F-ATPases in exporting protons, thereby inducing cytoplasmic acidification and acid inhibition of glycolytic enzymes. Basically, fluoride acts to reduce the acid tolerance of the bacteria. It is most effective at acid pH values. In the acidic conditions of cariogenic plaque, fluoride at levels as low as 0.1 mM can cause complete arrest of glycolysis by intact cells of Streptococcus mutans. Overall, the anticaries actions of fluoride appear to be complex, involving effects both on bacteria and on mineral phases. The antibacterial actions of fluoride appear themselves to be complex but to be dominated by weak-acid effects.
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              Antibacterial effects and physical properties of glass-ionomer cements containing chlorhexidine for the ART approach.

              Since atraumatic restorative treatment (ART) involves removal of carious lesions with manual instruments, improvement of filling materials to guarantee greater success should be considered. This study aimed to evaluate antibacterial, physical, and bonding properties of glass-ionomer cements (GIC) containing chlorhexidine (CHX), and to determine optimal concentrations for incorporation of agents to obtain antibacterial GICs for use with the ART approach. CHX diacetate combined with CHX dihydrochloride was added to control GIC powder to obtain concentration ratios of 1/0, 2/0, 3/0, 1/1, or 2/2% w/w. Antibacterial activity of each cement against Streptococcus mutans, Lactobacillus casei or Actinomyces naeslundii was examined using agar-diffusion methods, and release of CHX was analyzed by HPLC. Compressive strength, bond strength to dentin, and setting time were measured, and compared with those of control samples. All experimental GICs exhibited inhibition of three bacteria, but sizes of inhibition zones and concentrations of CHX released were not dependent upon CHX content. Incorporation of CHX diacetate at 2% or greater, significantly decreased compressive strength, and bond strength to dentin was adversely affected by addition of CHX diacetate at 2% or more (p<0.05, ANOVA, Fisher's PLSD test), although setting time was extended a little by addition of any concentrations of CHX. The present results demonstrate that experimental GICs containing CHX are effective in inhibiting bacteria associated with caries, and incorporation of 1% CHX diacetate is optimal to give appropriate physical and bonding properties.
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                Author and article information

                Journal
                J Appl Oral Sci
                J Appl Oral Sci
                jaos
                Journal of Applied Oral Science
                Faculdade De Odontologia De Bauru - USP
                1678-7757
                1678-7765
                2017
                2017
                : 25
                : 5
                : 541-550
                Affiliations
                [1 ]Univ. Estadual Paulista (UNESP), Faculdade de Odontologia de Araçatuba, Departamento de Odontologia Infantil e Social, Araçatuba, SP, Brasil
                [2 ]Universidade Federal Fluminense (UFF), Instituto de Saúde de Nova Friburgo, Departamento de Odontologia, Nova Friburgo, RJ, Brasil
                [3 ]Universidade Federal do Rio Grande do Sul (UFRGS), Faculdade de Odontologia, Departamento de Odontologia Conservadora, Porto Alegre, RS, Brasil
                [4 ]Universidade Estadual de Campinas (UNICAMP), Faculdade de Odontologia de Piracicaba, Departamento de Odontologia Infantil, Piracicaba, SP, Brasil
                [5 ]Univ. Estadual Paulista (UNESP), Faculdade de Odontologia de Araraquara, Departamento de Fisiologia e Patologia, Araraquara, SP, Brasil
                Author notes
                Corresponding address: Cristiane Duque, Departmento de Odontologia Infantil e Social. Faculdade de Odontologia de Araçatuba. Universidade Estadual Paulista (UNESP). R. José Bonifácio, 1193 - 16015-050 Araçatuba - SP - Brazil e-mail: cristianeduque@ 123456yahoo.com.br ; cduque@ 123456foa.unesp.br

                Conflicts of interest

                The authors of this manuscript declare that there are no conflicts of interest.

                Article
                1678-7757-2016-0195
                10.1590/1678-7757-2016-0195
                5804391
                29069152
                4cfb602d-05a1-4dd5-8ec8-b1d6ae31369f

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 03 June 2016
                : 20 January 2017
                : 29 January 2017
                Page count
                Figures: 4, Tables: 4, Equations: 0, References: 30, Pages: 10
                Categories
                Original Article

                dental atraumatic restorative treatment,chlorhexidine,glass ionomer cements

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