Pomegranate extract in polyphosphate-fluoride mouthwash reduces enamel demineralization.

OBJECTIVES: To evaluate the anti-demineralizing effect of a mouthwash comprising pomegranate peel extract (PPE 3%), sodium trimetaphosphate (TMP 0.3%), and fluoride (F 225 ppm) in an in situ study, and to assess its irritation potential in an ex vivo study. METHODS: This double-blind crossover study was conducted in four phases with 7 days each. Twelve volunteers used palatal appliances containing enamel blocks, which were subjected to cariogenic challenges. The ETF formulation (PPE + TMP + F, pH 7.0), TF formulation (TMP + F, pH 7.0), deionized water (W, pH 7.0), and essential oil commercial mouthwash (CM, 220 ppm F, pH 4.3) were dropped onto the enamel twice daily. The percentage of surface hardness loss, integrated loss of subsurface hardness, calcium, phosphorus, and fluoride in enamel and biofilms were determined. In addition, alkali-soluble extracellular polysaccharide concentrations were analyzed in the biofilms. The irritation potential was evaluated using the hen's egg chorioallantoic membrane test through the vascular effect produced during 300-s of exposure. RESULTS: ETF was the most efficacious in preventing demineralization. It also showed the highest concentrations of calcium and phosphorus in the enamel and in the biofilm, as well as the lowest amount of extracellular polysaccharides in the biofilm. In the eggs, ETF produced light reddening, whereas CM led to hyperemia and hemorrhage. CONCLUSIONS: The addition of PPE to formulations containing TMP and F increased its anti-demineralizing property, and this formulation presented a lower irritation potential than the CM. CLINICAL RELEVANCE: ETF can be a promising alternative alcohol-free mouthwash in patients at high risk of caries.

Pomegranate Extract Potentiates the Anti-Demineralizing, Anti-Biofilm, and Anti-Inflammatory Actions of Non-Alcoholic Mouthwash When Associated with Sodium-Fluoride Trimetaphosphate.

This study investigated the anti-caries and anti-inflammatory effects of mouthwash formulations containing Punica granatum (pomegranate) peel extract (PPE), sodium-trimetaphosphate, and low concentrations of fluoride. PPE was characterized using high-performance liquid chromatography (ellagic acid and punicalagin). Total phenolics were quantified among formulations, and their stability was analyzed for 28 days. The formulation effects were evaluated as follows: (1) inorganic component concentration and reduced demineralization on bovine enamel blocks subjected to pH cycling; (2) anti-biofilm effect on dual-biofilms of Streptococcus mutans ATCC 25175 and Candida albicans ATCC 10231 treated for 1 and 10 min, respectively; and (3) cytotoxicity and production of inflammatory mediators (interleukin-6 and tumor necrosis factor-alpha). The formulation containing 3% PPE, 0.3% sodium-trimetaphosphate, and 225 ppm of fluoride resulted in a 34.5% surface hardness loss; a 13% (treated for 1 min) and 36% (treated for 10 min) biofilm reduction in S. mutans; a 26% (1 min) and 36% (10 min) biofilm reduction in C. albicans; absence of cytotoxicity; and anti-inflammatory activity confirmed by decreased interleukin-6 production in mouse macrophages. Thus, our results provide a promising prospect for the development of an alcohol-free commercial dental product with the health benefits of P. granatum that have been recognized for a millennium.

Silver nanoparticles associated with a polyphosphate and fluoride enhance the prevention of enamel demineralization and impact on dual-biofilm adhesion.

OBJECTIVES: The aim of this study were to produce a multifunctional nanocomposite combining silver nanoaparticles (Ag), sodium trimetaphosphate (TMP) and fluoride (F), to investigate its effect on dental enamel demineralization and on biofilms of Streptococcus mutans and Candida albicans. METHODS: Bovine enamel blocks were submitted to five pH cycles and treated 2x/day with 100 ppm F, 225 ppm F, 100 ppm F + 0.2%TMP or 100 ppm F + 0.2%TMP+10% Ag (100F/TMP/Ag). Next, surface hardness loss (%SH), integrated loss of subsurface hardness (ΔKHN), enamel fluoride (F) and calcium (Ca) concentration were determined. Biofilms from single and dual species of S. mutans and C. albicans were treated with 100F/TMP/Ag, Ag or chlorhexidine gluconate for 24 h. The antibiofilm effect was evaluated by colony-forming unit counting and Scanning Electron Microscopy. RESULTS: The nanocomposite reduced 43.0% of %SH and was similar with samples treated with 225F, 100F/TMP and 100/TMP/Ag. The attribute of F and/or TMP in reducing ΔKHN in 5-20 µm was not affected by the addiction of Ag (110F = 225F = 100F/TMP = 100F/TMP/Ag > Negative Control). Further, 100F/TMP/Ag strongly reduced viable cells of S. mutans in dual biofilms (∼5 log10cm2) and structurally affected the biofilms. CONCLUSION: The 100F/TMP/F promoted a protective effect against enamel demineralization and was able to significantly inhibit the growth of biofilms of S. mutans and C. albicans. CLINICAL SIGNIFICANCE: The focus on prevention and non-invasive dental treatment is the most effective and least costly way to improve the population's oral health conditions. We present a nanocomposite for a multiple approach in prevention of caries.

Antibiofilm effect of chlorhexidine-carrier nanosystem based on iron oxide magnetic nanoparticles and chitosan.

This study synthesized and characterized a chlorhexidine (CHX)-carrier nanosystem based on iron oxide magnetic nanoparticles (IONPs) and chitosan (CS), and evaluated its antimicrobial effect on mono- and dual-species biofilms of Candida albicans and Streptococcus mutans. CHX was directly solubilized in CS-coated IONPs and maintained under magnetic stirring for obtaining the IONPs-CS-CHX nanosystem. Antimicrobial susceptibility testing for planktonic cells was performed by determining the minimum inhibitory concentration (MIC) of the nanosystem and controls. The effects of the IONPs-CS-CHX nanosystem on the formation of mono- and dual-species biofilms, as well as on pre-formed biofilms were assessed by quantification of total biomass, metabolic activity and colony-forming units. Data were analyzed by the Kruskal-Wallis' test or one-way analysis of variance, followed by the Student-Newman-Keuls' or Holm-Sidak's tests (α = 0.05), respectively. Physico-chemical results confirmed the formation of a nanosystem with a size smaller than 40 nm. The IONPs-CS-CHX nanosystem and free CHX showed similar MIC values for both species analyzed. In general, biofilm quantification assays revealed that the CHX nanosystem at 78 µg/mL promoted similar or superior antibiofilm effects compared to its counterpart at 39 µg/mL and free CHX at 78 µg/mL. These findings highlight the potential of CS-coated IONPs as preventive or therapeutic agents carrying CHX to fight biofilm-associated oral diseases.

Iron Oxide Nanoparticles for Biomedical Applications: A Perspective on Synthesis, Drugs, Antimicrobial Activity, and Toxicity.

Medical applications and biotechnological advances, including magnetic resonance imaging, cell separation and detection, tissue repair, magnetic hyperthermia and drug delivery, have strongly benefited from employing iron oxide nanoparticles (IONPs) due to their remarkable properties, such as superparamagnetism, size and possibility of receiving a biocompatible coating. Ongoing research efforts focus on reducing drug concentration, toxicity, and other side effects, while increasing efficacy of IONPs-based treatments. This review highlights the methods of synthesis and presents the most recent reports in the literature regarding advances in drug delivery using IONPs-based systems, as well as their antimicrobial activity against different microorganisms. Furthermore, the toxicity of IONPs alone and constituting nanosystems is also addressed.

Síntese e avaliação do efeito antibiofilme de um novo nanosistema composto por nanopartículas magnéticas de óxido de ferro, quitosana e clorexidina / Synthesis and evaluation of the antibiofilm effect of a new nanosystem composed by iron oxide magnetic nanoparticles, chitosan and chlorhexidine

Objetivo: Este estudo sintetizou um novo nanosistema carreador de clorexidina (CLX) baseado em nanopartículas magnéticas de óxido de ferro (NMOFs) e quitosana (QS), e avaliou seu efeito antimicrobiano sobre biofilmes de Candida albicans e Streptococcus mutans: Metodologia: O nanosistema NMOFs-QS-CLX foi preparado pela interação de CLX sobre NMOFs revestidas com QS, e caracterizado por difração de raios X, espectroscopia de absorção na região do infravermelho médio com transformada de Fourier, microscopia eletrônica de transmissão e dispersão dinâmica de luz. A concentração inibitória mínima (CIM) do nanosistema NMOFs-QS-CLX capaz de inibir as cepas no estado planctônico foi determinada de acordo com o método de microdiluição em caldo. Na sequência, biofilmes simples e mistos de C. albicans e S. mutans foram formados durante 24 horas em poços de placas de 96 poços na presença do nanosistema contendo CLX a 39 (NMOFs-QSCLX39) ou 78 µg/mL (NMOFs-QS-CLX78). Ainda, biofilmes pré-formados (24 horas) foram tratados durante 24 horas com o nanosistema nas mesmas concentrações. O efeito antibiofilme foi avaliado através da contagem do número de células cultiváveis, quantificação da biomassa total e avaliação da atividade metabólica. Controles apropriados foram incluídos em todas as análises. Os dados foram analisados pelo teste de Kruskal-Wallis e por ANOVA a um critério, seguidos dos testes StudentNewman-Keuls e Holm-Sidak, respectivamente (α = 0,05). Resultados: Os resultados de caracterização confirmaram a formação do nanosistema NMOFs-QSCLX, sem alteração das propriedades cristalinas das NMOFs. Além disso, as bandas de absorção características de cada composto foram identificadas no espectro do infravermelho, e o diâmetro médio do nanosistema foi menor que 40 nm. Os resultados de CIM mostraram que o nanosistema foi ligeiramente mais efetivo do que a CLX na inibição do crescimento dos microrganismos testados. Biofilmes formados na presença do nanosistema NMOFs-QS-CLX39 atingiram patamares quantitativos similares àqueles observados para CLX a 78 µg/mL. Ainda, para os biofilmes simples e mistos, o nanosistema NMOFs-QS-CLX78 mostrou efeitos redutores superiores ou similares àqueles encontrados para CLX a 78 µg/mL e NMOFs-QS-CLX39. Conclusão: O nanosistema NMOFs-QS-CLX foi efetivo tanto na inibição da formação de biofilmes como sobre biofilmes pré-formados de C. albicans e S. mutans em culturas simples ou mistas. O desenvolvimento desse nanosistema instaura inúmeras possibilidades para exploração de terapias baseadas em nanopartículas magnéticas como carreadoras de drogas usadas na área médico-odontológica(AU)

Aim: This study synthesized a new chlorhexidine(CHX)-carrier nanosystem based on iron oxide magnetic nanoparticles (IONPs) and chitosan (CS), as well as evaluated its antimicrobial effect on biofilms of Candida albicans and Streptococcus mutans. Method: The IONPs-CS-CHX nanosystem was prepared by CHX interaction to CScoated IONPs and characterized by X-ray powder diffraction, Fourier transform infrared spectroscopy, transmission electron microscopy and dynamic light scattering. Minimum inhibitory concentration (MIC) of the IONPs-CS-CHX nanosystem was determined according to the broth microdilution assay. After, mono-and dual-species biofilms of C. albicans and S. mutans were formed for 24 hours into wells of 96-well plates in the presence of the nanosystem containing CHX at 39 (IONPs-CS-CHX39) or 78 µg/mL (IONPs-CS-CHX78). Moreover, pre-formed biofilms (24 h) were treated for 24 h with the nanosystem at the same concentrations. The antibiofilm effect was determined by quantification of cultivable cells, total biomass and metabolic activity. Appropriate controls were included in all analyzes. Data were analyzed by Kruskal-Wallis' test and one-way ANOVA, followed by Student-Newman-Keuls and Holm-Sidak tests (α = 0.05). Results: Characterization results confirmed the nanosystem formation without altering the crystalline properties of the IONPs. In addition, the characteristic absorption bands of each compound were identified in the infrared spectrum, and the mean diameter of the nanosystem was lower than 40 nm. MIC results showed that the nanosystem was slightly more effective than CLX in inhibiting the growth of the microorganisms tested. Biofilms formed in the presence of IONPs-CS-CHX39 attained similar quantitative levels to those observed for CHX at 78 µg/mL. Further, for mono- and dual-species biofilms, IONPs-CS-CHX78 showed similar or superior antibiofilm effects when compared with IONPs-CS-CHX39 and free CHX at 78 µg/mL. Conclusion: The IONPs-CS-CHX nanosystem was able to reduce biofilm formation and pre-formed biofilms of C. albicans and S. mutans in single or mixed cultures. The development of this nanosystem establishes several possibilities for exploration of magnetic nanoparticle-based therapy as drug carrier used in health area(AU)