Effects of artificial honey and epigallocatechin-3-gallate on streptococcus pyogenes.

BACKGROUND: Streptococcus pyogenes is an important global human pathogen that causes pharyngitis, and antibacterial therapy has become an important part of the overall therapy for pharyngitis. As natural derivatives, honey and green tea are often recommended for patients with pharyngitis in traditional Chinese medicine without experimental theoretical basis on wether the combined effect of honey and green tea on pharyngitis is better than they alone. The aims of this study were to explore the effects of artificial honey (AH) and epigallocatechin-3-gallate (EGCG) on S. pyogenes and elucidate the possible mechanisms, which were investigated using MIC (the minimum inhibitory concentration), FIC (fractional inhibitory concentration) index, growth pattern, biofilm formation and RT-qPCR. RESULTS: The MIC of AH on S. pyogenes was 12.5% (v/v) and the MIC of EGCG was 1250 µg/ml. The FIC index of AH and EGCG was 0.5. The planktonic cell growth, growth pattern and biofilm formation assays showed that AH and EGCG mixture had stronger inhibitory effect on S. pyogenes than they alone. RT-qPCR confirmed that the expression of hasA and luxS gene were inhibited by AH and EGCG mixture. CONCLUSIONS: AH and EGCG mixture can inhibit the planktonic cell growth, biofilm formation and some virulence genes expression of S. pyogenes, better than they alone. The combination of honey and green tea have the potential to treat pharyngitis as natural derivatives, avoiding drug resistance and double infection.

Evaluating hop extract concentrations found in commercial beer to inhibit Streptococcus mutans biofilm formation.

AIMS: The purpose of this study was to compare the effect of hop extracts with diverse ß-acid concentrations on Streptococcus mutans biofilm formation. METHODS AND RESULTS: Ten different hop extracts, with α-acid concentrations similar to those found in commercial beer products and ß-acid concentrations ranging from 2.6 to 8.1%, were added to distilled water to make standardized concentrations. S. mutans isolates were treated with hop extract dilutions varying from 1:2 to 1:256. The minimum inhibitory, minimum bactericidal and minimum biofilm inhibitory concentrations were determined and the optical density was evaluated. Live/dead staining confirmed the bactericidal effects. Biofilm formation of several strains of S. mutans was significantly inhibited by hop extract dilutions of 1:2, 1:4, 1:8, 1:16 and 1:32. Strong negative correlations were observed between α- and ß-acid concentrations of the hop extracts and S. mutans total growth and biofilm formation. CONCLUSIONS: The use of hop extracts prepared similarly to commercial beer decreased S. mutans biofilm formation. SIGNIFICANCE AND IMPACT OF THE STUDY: The inclusion of hops in the commercial beer products may provide beneficial health effects. Further studies are warranted to determine an effect in vivo on the development of dental caries.

The influence of biofilm maturation on fluoride's anticaries efficacy.

OBJECTIVES: (1) To explore the influence of biofilm maturation and timing of exposure on fluoride anticaries efficacy and (2) to explore biofilm recovery post-treatment. METHODS: Bovine enamel specimens were utilized in a pH cycling model (28 subgroups [n = 18]). Each subgroup received different treatments [exposure]: sodium fluoride [NaF]; stannous fluoride [SnF2]; amine fluoride [AmF]; and de-ionized water [DIW], at a specific period: early: days 1-4; middle: days 3-6; and late: days 7-10. During non-exposure periods, pH cycling included DIW instead of fluorides. Objective 1: part 1 (cycling for 4, 6, or 10 days). Part 2 (cycling for 10 days). Objective 2: early exposure: three sample collection time points (immediate, 3 days, and 6 days post-treatment); middle exposure: two sample collection time points (immediate, 4 days post-treatment). The enamel and biofilm were analyzed ([surface microhardness; mineral loss; lesion depth]; [lactate dehydrogenase enzyme activity; exopolysaccharide amount; viability]). Data were analyzed using ANOVA (p = 0.05). RESULTS: Objective 1: Early exposure to fluorides produced protective effects against lesion progression in surface microhardness and mineral loss, but not for lesion depth. Objective 2: Early exposure slowed the demineralization process. SnF2 and AmF were superior to NaF in reducing LDH and EPS values, regardless of exposure time. They also prevented biofilm recovery. CONCLUSION: Earlier exposure to SnF2 and AmF may result in less tolerant biofilm. Early fluoride treatment may produce a protective effect against demineralization. SnF2 and AmF may be the choice to treat older biofilm and prevent biofilm recovery. CLINICAL RELEVANCE: The study provides an understanding of biofilm-fluoride interaction with mature biofilm (e.g., hard-to-reach areas, orthodontic patients) and fluoride's sustainable effect hours/days after brushing.

Effects of casein phosphopeptide-amorphous calcium phosphate crème on nicotine-induced Streptococcus mutans biofilm in vitro.

OBJECTIVES: The aim of this study was to test the effects of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) crème, or MI Paste™ (MIP), on nicotine-induced Streptococcus mutans biofilm. The experiment utilized S. mutans biofilm assays with varying concentrations of nicotine and MIP aqueous concentrate levels. First hand exposure to nicotine has been demonstrated to significantly increase S. mutans biofilm formation, while the active component, CPP-ACP, in MIP has been shown to reduce S. mutans biofilm formation. MATERIALS AND METHODS: A 24-h culture of S. mutans UA159 in microtiter plates were treated with varying nicotine concentrations (0-32 mg/ml) in Tryptic Soy Broth supplemented with 1% sucrose (TSBS) with or without MIP aqueous concentrate. A spectrophotometer was used to determine total growth absorbance and planktonic growth. The microtiter plate wells were washed, fixed, and stained with crystal violet dye and the absorbance measured to determine biofilm formation. RESULTS: The presence of MIP aqueous concentrate inhibits nicotine-induced S. mutans biofilm formation at different concentrations of nicotine (0-32 mg/ml). CONCLUSION: The results demonstrated nicotine-induced S. mutans biofilm formation is decreased in the presence of MIP. This provides further evidence about the cariostatic properties of CPP-ACP, the active soluble ingredient in the MIP, and reconfirms the harmful effects of nicotine. CLINICAL SIGNIFICANCE: Smokers may gain dual benefits from the use of MIP, as a remineralization agent and as a cariostatic agent, by inhibiting nicotine-induced S. mutans biofilm formation.

Levels of growth factors from platelet-rich fibrin from chronic periodontitis versus periodontally healthy subjects: a pilot study.

OBJECTIVES: This study aimed to (1) compare the amounts of growth factors from platelet-rich fibrin (PRF) between chronic periodontitis and periodontally healthy subjects and (2) evaluate the relationships between the amounts of growth factors from PRF with complete blood counts (white blood cell (WBC) and platelet counts) and the serum concentrations of IL-1ß, IL-6, and tumor necrosis factor-α (TNF-α). MATERIALS AND METHODS: Venous blood was collected from chronic periodontitis (test) and periodontally healthy subjects (control). PRF and serum were collected from the centrifuged blood. Liquid exudates from the compression of PRF were collected. The compressed PRF membranes were incubated in saline, and eluted aliquots were collected at 1, 24, and 72 h, and the membranes were then digested with trypsin. Epidermal growth factor, insulin-like growth factor-1, platelet-derived growth factor-BB, transforming growth factor-ß1, and vascular endothelial growth factor in the exudates and eluents were quantified by ELISA. Serum was used for IL-1ß, IL-6, and TNF-α quantifications. Complete blood counts were measured. RESULTS: There were no significant differences in the amounts of growth factors from PRF exudates and membranes measured between groups (all p > 0.05). The test group had significantly higher WBC (p < 0.05). However, there was no significant correlation between the WBC and the amounts of the growth factors from PRF (all p > 0.05). CONCLUSIONS: PRF can be utilized as an autologous source of growth factors not affected by periodontal condition and WBC level. CLINICAL RELEVANCE: The amounts of growth factors from PRF were not affected by the periodontal condition of the patient.

Impact of caffeine on metabolic activity and biofilm formation of Candida albicans on acrylic denture resin in the presence of nicotine.

STATEMENT OF PROBLEM: Candida albicans has been implicated in denture stomatitis, and this effect is exacerbated by nicotine exposure. However, studies have also suggested that caffeine exposure inhibits the growth of C. albicans. The interaction effects of nicotine and caffeine are not yet clear on the growth of C. albicans. PURPOSE: The purpose of this in vitro study was to determine the effect of caffeine on metabolic activity and biofilm formation of C. albicans growing on acrylic denture resin while simultaneously exposed to nicotine and, if an effect were to be identified, whether this effect would vary depending on the caffeine concentration. MATERIAL AND METHODS: A total of 240 acrylic resin specimens were divided into 2 equal groups (120 each). Specimens in one group were processed to measure C. albicans metabolic activity, and those in the other group were processed to measure C. albicans biofilm attachment. Ten subgroups (n=12) were established within each group with different concentration combinations of nicotine and caffeine to test the interaction effect. The first subgroup was designed as a negative control, containing 0 mg/mL of nicotine and caffeine. The following subgroups all contained 8.00 mg/mL of nicotine, and the caffeine concentrations were prepared at the following 9 levels: 0, 0.25, 0.50, 1.00, 2.00, 4.00, 8.00, 16.00, and 32.00 mg/mL. Metabolic activity was measured by using a 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-carboxanilide (XTT) assay. Biofilm attachment was measured by using spiral plating and calculated in terms of the number of colony-forming units (CFUs)/mL. Descriptive statistics and a 2-way ANOVA were conducted to determine whether the concentrations of nicotine and caffeine used affected the biofilm attachment and metabolic activity of C. albicans (α=.05). RESULTS: The presence of 8 mg/mL of nicotine increased the metabolic activity and biofilm formation of C. albicans. When compared with the 0 mg/mL of caffeine and 8.00 mg/mL of nicotine group, caffeine from 1.00 to 4.00 mg/mL significantly increased C. albicans biofilm metabolic activity. Caffeine at 16.00 and 32.00 mg/mL significantly decreased C. albicans biofilm metabolic activity in the presence of 8 mg/mL of nicotine. Caffeine from 1.00 to 32.00 mg/mL significantly decreased the biofilm formation of C. albicans in the presence of 8 mg/mL of nicotine. CONCLUSIONS: The presence of 8 mg/mL of nicotine alone increased the metabolic activity and biofilm formation of C. albicans. In the presence of 8 mg/mL of nicotine with different caffeine concentrations, the results suggest that, overall, caffeine at higher concentrations (16 and 32 mg/mL) inhibited the metabolic activity and biofilm formation of C. albicans on acrylic denture resin most.

The Impact of Nicotine and Cigarette Smoke Condensate on Metabolic Activity and Biofilm Formation of Candida albicans on Acrylic Denture Material.

PURPOSE: Smokers have increased denture stomatitis caused primarily by Candida albicans. The primary aim of this study was to demonstrate the impact of a wide range of nicotine and cigarette smoke condensate (CSC) concentrations on biofilm formation and metabolic activity of C. albicans on acrylic denture material. MATERIALS AND METHODS: C. albicans (ATCC strain 10231) was used. Standardized denture acrylic (PMMA) specimens (total of 135 specimens) were incubated with C. albicans and exposed to nicotine and CSC at different concentrations (0, 0.25, 0.5, 1, 2, 4, 8, 16, and 32 mg/ml) and (0, 0.25, 0.5, 1, 2, and 4 mg/ml), respectively. For each experiment, 3 samples per nicotine and CSC concentration and a total of 45 specimens (27 specimens for the nicotine and 18 specimens for the CSC-treated samples) were used and were selected randomly for each group. The control group consisted of 0 mg/ml of nicotine or CSC. The viability of C. albicans was measured using spiral plating on blood agar plates. The effect of nicotine and CSC concentrations on planktonic cells was were measured using a microplate reader. Metabolic activity of 24-hour-old established C. albicans biofilm exposed to nicotine and CSC for 24 hours in microtiter plates was determined using a 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-carboxanilide (XTT) reduction assay. RESULTS: The viability of C. albicans increased concomitant with increasing concentrations of CSC and nicotine, particularly at 0.5 and 2 mg/ml, respectively. Concentrations of CSC and nicotine above this resulted in an inhibitory effect on C. albicans viability. CSC and nicotine at 4 and 16 mg/ml, respectively, increased C. albicans biofilm metabolic activity. CONCLUSION: Nicotine and CSC up to certain concentrations caused increases in biofilm formation, metabolic activity, viability, and planktonic cell absorbance of C. albicans. This in vitro study demonstrates the effectiveness of tobacco on promoting the growth of C. albicans and suggests their potential contributing factor in C. albicans biofilm related infections in smokers.

The antimicrobial properties, cytotoxicity, and differentiation potential of double antibiotic intracanal medicaments loaded into hydrogel system.

OBJECTIVE: This study evaluated the antimicrobial properties, cytotoxicity, and mineralization potential of methylcellulose hydrogels loaded with low concentrations of double antibiotic pastes (DAP). MATERIALS AND METHODS: The direct and residual antibacterial effects of 1, 5, and 10 mg/mL of DAP loaded into hydrogels as well as calcium hydroxide (Ca(OH)2) were tested against single-species biofilms of Enterococcus faecalis and dual-species biofilms (Enterococcus faecalis and Prevotella intermedia). The effects of DAP hydrogels on proliferation and mineralization of dental pulp stem cells (DPSC) were tested using MTT assays, alkaline phosphate activity (ALP), and alizarin red staining. Fisher's exact tests, Wilcoxon rank sum tests, and one-way ANOVA were used for statistical analyses (α = 0.05). RESULTS: All tested concentrations of DAP hydrogels as well as Ca(OH)2 demonstrated significant direct antibacterial effects against single- and dual-species biofilms. However, only 5 and 10 mg/mL of DAP hydrogels exhibited significant residual antibacterial effects against both types of tested biofilms. Only 1 mg/mL of DAP hydrogels did not have significant negative effects on DPSC viability, ALP activity, and mineralization nodule formation. However, 5 and 10 mg/mL of DAP hydrogels caused significant negative effects on cytotoxicity and mineralization nodule formation of DPSC. CONCLUSIONS: Hydrogels containing 1 mg/mL DAP offered significant direct antibacterial effects against single- and dual-species biofilms without causing significant negative effects on viability, ALP activity, and mineralization nodule formation of DPSC. CLINICAL RELEVANCE: The methylcellulose-based hydrogel proposed in this study can be used clinically as a biocompatible system to deliver controlled low concentrations of DAP.

Nicotine Upregulates Coaggregation of Candida albicans and Streptococcus mutans.

PURPOSE: Denture stomatitis is a condition of painless inflammation of denture-bearing mucosa. Reports indicate that nicotine, the major psychoactive ingredient in tobacco, increases growth of Streptococcus mutans and Candida albicans in denture biofilm. The purpose of this study was to determine the in vitro effects of nicotine on coaggregation of C. albicans with S. mutans. MATERIAL AND METHODS: C. albicans strain ATCC 10231, S. mutans strain UA159 (ATTC 700610), and nicotine dilutions (ranging from 0 to 32 mg/ml) were used for this study. Both microorganisms were grown for 24 hours in dilutions of nicotine (0 to 32 mg/ml) made in tryptic soy broth (TSB) or TSB supplemented with 1% sucrose (TSBS; S. mutans) or yeast peptone dextrose broth (YPD; C. albicans). Suspensions of the nicotine-treated cells were prepared, mixed together and incubated for up to 24 hours to determine if there was an increase in coaggregation of nicotine-treated cells compared to the no nicotine control cells. Qualitative analysis of coaggregation was performed using a visual aggregation assay and light microscopic observation. A spectrophotometric assay was used to provide a quantitative analysis of the coaggregation. RESULTS: The visual aggregation assay indicated a significant increase in coaggregation between C. albicans and S. mutans with increasing incubation time (0 to 24 hours) and nicotine concentrations (0 to 4 mg/ml). Microbial growth in nicotine at 4 mg/ml demonstrated a significant increase in coaggregation after 24 hours of incubation. The numbers of coaggregated S. mutans/C. albicans cells exhibited a significant increase with incubation time and nicotine concentrations when the samples were examined microscopically. More coaggregation of S. mutans and C. albicans was observed with incubation time and increased nicotine compared to the 0 mg/ml nicotine group. There was a noticeable increase of coaggregation when cells were grown in TSBS compared to TSB. Absorbance of nicotine-treated cells (0.25 to 4 mg/ml) exhibited a decrease in values compared to 0 mg/ml at 0 hours of incubation, confirming increased coaggregation. CONCLUSION: These results demonstrated the effect of nicotine in increasing the coaggregation of S. mutans with C. albicans. Coaggregation increased with incubation time and nicotine concentration. Coaggregation was increased with S. mutans grown in TSBS compared to TSB, suggesting that growth in sucrose media leads to an increase in receptors responsible for coaggregation.

In Vitro Effects of Plantago Major Extract, Aucubin, and Baicalein on Candida albicans Biofilm Formation, Metabolic Activity, and Cell Surface Hydrophobicity.

PURPOSE: To determine the in vitro effectiveness of Plantago major extract, along with two of its active components, aucubin and baicalein, on the inhibition of Candida albicans growth, biofilm formation, metabolic activity, and cell surface hydrophobicity. MATERIALS AND METHODS: Twofold dilutions of P. major, aucubin, and baicalein were used to determine the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), and the minimum biofilm inhibitory concentration (MBIC) of each solution. Separately, twofold dilutions of P. major, aucubin, and baicalein were used to determine the metabolic activity of established C. albicans biofilm using a 2,3-bis (2- methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-carboxanilide reduction assay. Twofold dilutions of P. major, aucubin, and baicalein were used to determine the cell surface hydrophobicity of treated C. albicans biofilm by a two-phase assay using hexadecane. The hydrophobicity percentage of the cell surface was then calculated. A mixed-model ANOVA test was used for intergroup comparisons. RESULTS: The MICs of P. major extract (diluted 1:2 to 1:8), aucubin (61 to 244 µg/ml), and baicalein (0.0063 to 100 µg/ml) on the total growth of C. albicans were noticeable at their highest concentrations, and the inhibition was dose dependent. The MFC was evaluated after 48 hours of incubation, and aucubin (244 µg/ml) exhibited a strong fungicidal activity at its highest concentration against C. albicans growth. The MBIC indicated no growth or reduced growth of C. albicans biofilm at the highest concentrations of aucubin (61 to 244 µg/ml) and baicalein (25 to 100 µg/ml). Similarly, the effects of these reagents on C. albicans biofilm metabolic activity and hydrophobicity demonstrated high effectiveness at their highest concentrations. CONCLUSION: P. major extract, aucubin, and baicalein caused a dose-dependent reduction on the total growth, biofilm formation, metabolic activity, and cell surface hydrophobicity of C. albicans. This demonstrates their effectiveness as antifungals and suggests their promising potential use as solutions for C. albicans biofilm-related infections.

Effects of Cola-Flavored Beverages and Caffeine on Streptococcus mutans Biofilm Formation and Metabolic Activity.

OBJECTIVE: To examine the effects of cola-flavored beverages and caffeine on growth and metabolism of Streptococcus mutans biofilm. This study was designed to determine if carbonated beverages or caffeine can increase S. mutans growth and biofilm formation and metabolic activity in vitro, potentially leading to increased S. mutans-associated cariogenicity in children that consume them. STUDY DESIGN: Six different cola-flavored products, plus pure caffeine, and pure high fructose corn syrup (HFCS), at different concentrations similar to those in the beverages were tested. A 16-hour culture of S. mutans was treated with different dilutions in bacteriological media. To test for the effect on biofilm formation, the biofilm was stained with crystal violet. The absorbance was determined to evaluate biofilm growth. Biofilm metabolic activity was measured based on biofilm having the ability to reduce XTT to a water-soluble orange compound. RESULTS: The inclusion of HFCS in the beverages, as well as pure HFCS, significantly enhanced bacterial biofilm formation and metabolic activity. Pure caffeine and the presence of caffeine in beverages did not significantly increase biofilm formation, but pure caffeine significantly increased metabolism, and Diet Coke had significantly greater metabolic activity than Caffeine-Free Diet Coke. CONCLUSIONS: HFCS increases both the biofilm formation and metabolism of S. mutans, and caffeine in some cases increases metabolism of S. mutans.

Effect of Commonly Prescribed Liquid Medications on Streptococcus mutans Biofilm. An in vitro study.

OBJECTIVE: This study addressed the effect of pediatric liquid antibiotic medications on Streptococcus mutans UA159. These suspensions commonly contain sugars such as sucrose to make them more palatable for children. The study was designed to evaluate the effects of oral liquid antibiotics on Streptococcus mutans growth and biofilm formation. STUDY DESIGN: A 24 hour culture of S. mutans was treated with various concentrations of liquid medications commonly prescribed to children for odontogenic or fungal infections- amoxicillin, penicillin VK, clindamycin, and nystatin. The study was conducted in sterile 96-well flat bottom microtiter plates. The minimum inhibitory and biofilm inhibitory concentrations (MIC/MBIC) of S. mutans were determined for each medication. S. mutans was cultured with and without the test drugs, the amount of total growth measured, the biofilms washed, fixed, and stained with crystal violet. The absorbance was determined to evaluate biofilm formation. RESULTS: Higher concentrations of amoxicillin, penicillin VK and clindamycin had decreased biofilm and overall growth than the control. The MICs were 1:2,560 (1.95 ug/ml), 1:2,560 (1.95 ug/ml) and 1:40 (9.375 ug/ml), while the MBIC were 1:640 (7.8 ug/ml), 1:1,280 (3.9 ug/ml) and 1:20 (18.75 ug/ml), respectively. Lower concentrations provided increased biofilm and overall growth. Nystatin induced significantly more biofilm and overall growth than the control at all concentrations. CONCLUSION: At high concentrations, approximately at the levels expected to be present in the oral cavity of children, amoxicillin, penicillin, and clindamycin inhibited S. mutans biofilm and overall growth due to their antibiotic activity, while at lower concentrations the three antibiotics demonstrated an increase in biofilm and growth. The increase in S. mutans biofilm and overall growth is most likely attributed to the sugar content in the medications. Nystatin provided an increase in biofilm and growth at each concentration tested.

Photo Inactivation of Streptococcus mutans Biofilm by Violet-Blue light.

Among various preventive approaches, non-invasive phototherapy/photodynamic therapy is one of the methods used to control oral biofilm. Studies indicate that light at specific wavelengths has a potent antibacterial effect. The objective of this study was to determine the effectiveness of violet-blue light at 380-440 nm to inhibit biofilm formation of Streptococcus mutans or kill S. mutans. S. mutans UA159 biofilm cells were grown for 12-16 h in 96-well flat-bottom microtiter plates using tryptic soy broth (TSB) or TSB with 1 % sucrose (TSBS). Biofilm was irradiated with violet-blue light for 5 min. After exposure, plates were re-incubated at 37 °C for either 2 or 6 h to allow the bacteria to recover. A crystal violet biofilm assay was used to determine relative densities of the biofilm cells grown in TSB, but not in TSBS, exposed to violet-blue light. The results indicated a statistically significant (P < 0.05) decrease compared to the non-treated groups after the 2 or 6 h recovery period. Growth rates of planktonic and biofilm cells indicated a significant reduction in the growth rate of the violet-blue light-treated groups grown in TSB and TSBS. Biofilm viability assays confirmed a statistically significant difference between violet-blue light-treated and non-treated groups in TSB and TSBS. Visible violet-blue light of the electromagnetic spectrum has the ability to inhibit S. mutans growth and reduce the formation of S. mutans biofilm. This in vitro study demonstrated that violet-blue light has the capacity to inhibit S. mutans biofilm formation. Potential clinical applications of light therapy in the future remain bright in preventing the development and progression of dental caries.

Evaluation of selected properties of a new root repair cement containing surface pre-reacted glass ionomer fillers.

OBJECTIVE: This study evaluated selected properties of a prototype root repair cement containing surface pre-reacted glass ionomer fillers (S-PRG) in comparison to mineral trioxide aggregate (MTA) and intermediate restorative material (IRM). MATERIALS AND METHODS: The antibacterial effect of S-PRG, MTA, and IRM cements was tested against Porphyromonas gingivalis and Enterococcus faecalis after 1 and 3 days of aging of the cements. The set cements were immersed in distilled water for 4 h to 28 days, and ion-releasing ability was evaluated. Initial and final setting times of all cements were evaluated using Gilmore needles. The push-out bond strength between radicular dentin and all cements was tested at different levels of the roots. RESULTS: S-PRG and IRM cements, but not MTA cement, demonstrated significant antibacterial effect against P. gingivalis. All types of cements exhibited significant antibacterial effect against E. faecalis without being able to eliminate the bacterium. S-PRG cement provided continuous release of fluoride, strontium, boron, sodium, aluminum, and zinc throughout all tested time points. Both initial and final setting times were significantly shorter for S-PRG and IRM cements in comparison to MTA. The push-out bond strength was significantly lower for S-PRG cement in comparison to MTA and IRM at coronal and middle levels of the roots. CONCLUSIONS: S-PRG cement demonstrated significant antibacterial effects against endodontic pathogens, multiple ion-releasing ability, relatively short setting time, and low bonding strength. CLINICAL RELEVANCE: S-PRG cement can be used as a one-visit root repair material with promising antibacterial properties and ion-releasing capacity.

Antibacterial TAP-mimic electrospun polymer scaffold: effects on P. gingivalis-infected dentin biofilm.

OBJECTIVES: This study sought to investigate, in vitro, the effects of a recently developed triple antibiotic paste (TAP)-mimic polymer nanofibrous scaffold against Porphyromonas gingivalis-infected dentin biofilm. MATERIALS AND METHODS: Dentin specimens (4 × 4 × 1 mm(3)) were prepared from human canines. The specimens were sterilized, inoculated with P. gingivalis (ATCC 33277), and incubated for 1 week to allow for biofilm formation. Infected dentin specimens were exposed for 3 days to the following treatments: antibiotic-free polydioxanone scaffold (PDS, control), PDS + 25 wt% TAP [25 mg of each antibiotic (metronidazole, ciprofloxacin, and minocycline) per mL of the PDS polymer solution], or a saturated TAP-based solution (50 mg of each antibiotic per mL of saline solution). In order to serve as the negative control, infected dentin specimens were left untreated (bacteria only). To determine the antimicrobial efficacy of the TAP-mimic scaffold, a colony-forming unit (CFU) per milliliter (n = 10/group) measurement was performed. Furthermore, additional specimens (n = 2/group) were prepared to qualitatively study biofilm inhibition via scanning electron microscopy (SEM). Statistics were performed, and significance was set at the 5% level. RESULTS: Both the TAP-mimic scaffold and the positive control (TAP solution) led to complete bacterial elimination, differing statistically (p < 0.05) from the negative control group (bacteria only). No statistical differences were observed for CFU per milliliter data between antibiotic-free scaffolds (2.7 log10 CFU/mL) and the negative control (5.9 log10 CFU/mL). CONCLUSIONS: The obtained data revealed significant antimicrobial properties of the novel PDS-based TAP-mimic scaffold against an established P. gingivalis-infected dentin biofilm. CLINICAL RELEVANCE: Collectively, the data suggest that the proposed nanofibrous scaffold might be used as an alternative to the advocated clinical gold standard (i.e., TAP) for intracanal disinfection prior to regenerative endodontics.

Synthesis and characterization of CaO-loaded electrospun matrices for bone tissue engineering.

OBJECTIVES: This study aims to synthesize and characterize biodegradable polymer-based matrices loaded with CaO nanoparticles for osteomyelitis treatment and bone tissue engineering. MATERIALS AND METHODS: Poly(ε-caprolactone) (PCL) and PCL/gelatin (1:1, w/w) solutions containing CaO nanoparticles were electrospun into fibrous matrices. Scanning (SEM) and transmission (TEM) electron microscopy, Fourier transformed infrared (FTIR), energy dispersive X-ray spectroscopy (EDS), contact angle (CA), tensile testing, and antibacterial activity (agar diffusion assay) against Staphylococcus aureus were performed. Osteoprecursor cell (MC3T3-E1) response (i.e., viability and alkaline phosphatase expression/ALP) and infiltration into the matrices were evaluated. RESULTS: CaO nanoparticles were successfully incorporated into the fibers, with the median fiber diameter decreasing after CaO incorporation. The CA decreased with the addition of CaO, and the presence of gelatin made the matrix very hydrophilic (CA = 0°). Increasing CaO concentrations progressively reduced the mechanical properties (p ≤ 0.030). CaO-loaded matrices did not display consistent antibacterial activity. MC3T3-E1 cell viability demonstrated the highest levels for CaO-loaded matrices containing gelatin after 7 days in culture. An increased ALP expression was consistently seen for PCL/CaO matrices when compared to PCL and gelatin-containing counterparts. CONCLUSIONS: Despite inconsistent antibacterial activity, CaO nanoparticles can be effectively loaded into PCL or PCL/gelatin fibers without negatively affecting the overall performance of the matrices. More importantly, CaO incorporation enhanced cell viability as well as differentiation capacity, as demonstrated by an increased ALP expression. CLINICAL SIGNIFICANCE: CaO-loaded electrospun matrices show potential for applications in bone tissue engineering.

Effect of Infant Formula on Streptococcus Mutans Biofilm Formation.

OBJECTIVE: This study investigated the effect that infant formula had on biofilm growth of Streptococcus mutans. Specifically, it compared biofilm growth in media containing lactose-based and sucrose-based formulas. It also analyzed biofilm formation with formulas of varying iron content. Biofilm growth was tested with the specific infant formula components sucrose, lactose, and ferric chloride. The study was designed to determine if these types of infant formulas and components affected S. mutans biofilm formation differently. STUDY DESIGN: A 24-hour culture of S. mutans was treated with various concentrations of infant formula diluted in bacteriological media. To test for biofilm formation, S. mutans was cultured with and without the infant formula and formula components. The biofilms were washed, fixed, and stained with crystal violet. The absorbance was measured to evaluate biofilm growth and total absorbance. RESULTS: Sucrose-based formulas provided significant increases in biofilm growth when compared to lactose-based formulas at two dilutions (1:5, 1:20). Similac Sensitive RS (sucrose-based) at most dilutions provided the most significant increase in biofilm growth when compared to the control. Sucrose tested as an individual component provided more of a significant increase on biofilm growth than lactose or iron when compared to the control. A low iron formula provided a significant increase in biofilm growth at one dilution (1:5) when compared to formula containing a normal iron content. There was no significant difference in biofilm growth when comparing high iron formula to normal iron formula or low iron formula. There was no significant difference when comparing Similac PM 60/40 (low iron formula) to Similac PM 60/40 with additional ferric chloride. CONCLUSION: The results of this study demonstrated that sucrose-based formula provided more of a significant increase in biofilm growth compared to lactose-based formula. Sucrose alone provided a significant increase of biofilm growth at more dilutions when compared to the control than lactose and iron. The amount of iron in formula had a significant effect on biofilm formation only when comparing low iron formula to normal iron formula at the highest concentration (1:5). There was no significant difference in biofilm growth when iron was added to the low iron formula. The information obtained expands current knowledge regarding the influence of infant formula on the primary dentition and reinforces the importance of oral hygiene habits once the first tooth erupts.

The effect of diluted triple and double antibiotic pastes on dental pulp stem cells and established Enterococcus faecalis biofilm.

OBJECTIVES: To investigate the effect of various dilutions of antibiotic medicaments used in endodontic regeneration on the survival of human dental pulp stem cells (DPSCs) and to determine their antibacterial effect against established Enterococcus faecalis biofilm. MATERIALS AND METHODS: The cytotoxic and antibacterial effects of different triple (TAP) and double antibiotic paste (DAP) dilutions (0.125, 0.25, 0.5, 1, and 10 mg/ml) were tested against Enterococcus faecalis established biofilm and DPSC. Established bacterial biofilm were exposed to antibiotic dilutions for 3 days. Then, biofilms were collected, spiral plated, and the numbers of bacterial colony forming units (CFU/ml) were determined. For the cytotoxic effect, lactate dehydrogenase activity assays (LDH) and cell viability assays (WST-1) were used to measure the percentage of DPSC cytotoxicity after 3-day treatment with the same antibiotic dilutions. A general linear mixed model was used for statistical analyses (α = 0.05). RESULTS: All antibiotic dilutions significantly decreased the bacterial CFU/ml. For WST-1 assays, all antibiotic dilutions except 0.125 mg/ml significantly reduced the viability of DPSC. For LDH assays, the three lowest tested concentrations of DAP (0.5, 0.25, 0.125 mg/ml) and the two lowest concentrations of TAP (0.25 and 0.125 mg/ml) were non-toxic to DPSC. CONCLUSIONS: All tested dilutions had an antibacterial effect against E. faecalis. However, 0.125 mg/ml of DAP and TAP showed a significant antibacterial effect with no cytotoxic effects on DPSCs. CLINICAL RELEVANCE: Using appropriate antibiotic concentrations of intracanal medicament during endodontic regeneration procedures is critical to disinfect root canal and decrease the adverse effects on stem cells.

Biodegradable nanofibrous drug delivery systems: effects of metronidazole and ciprofloxacin on periodontopathogens and commensal oral bacteria.

OBJECTIVES: The purposes of this study were to fabricate biodegradable polydioxanone (PDS II®) electrospun periodontal drug delivery systems (hereafter referred to as matrices) containing either metronidazole (MET) or ciprofloxacin (CIP) and to investigate the effects of antibiotic incorporation on both periodontopathogens and commensal oral bacteria. MATERIALS AND METHODS: Fibrous matrices were processed from PDS polymer solution by electrospinning. Antibiotic-containing PDS solutions were prepared to obtain four distinct groups: 5 wt.% MET, 25 wt.% MET, 5 wt.% CIP, and 25 wt.% CIP. Pure PDS was used as a control. High-performance liquid chromatography (HPLC) was done to evaluate MET and CIP release. Dual-species biofilms formed by Lactobacillus casei (Lc) and Streptococcus salivarius (Ss) were grown on the surface of all electrospun matrices. After 4 days of biofilm growth, the viability of bacteria on biofilms was assessed. Additionally, antimicrobial properties were evaluated against periodontopathogens Fusobacterium nucleatum (Fn) and Aggregatibacter actinomycetemcomitans (Aa) using agar diffusion assay. RESULTS: A three-dimensional interconnected porous network was observed in the different fabricated matrices. Pure PDS showed the highest fiber diameter mean (1,158 ± 402 nm) followed in a descending order by groups 5 wt.% MET (1,108 ± 383 nm), 25 wt.% MET (944 ± 392 nm), 5 wt.% CIP (871 ± 309 nm), and 25 wt.% CIP (765 ± 288 nm). HPLC demonstrated that groups containing higher amounts (25 wt.%) of incorporated drugs released more over time, while those with lower levels (5 wt.%) the least. No inhibitory effect of the tested antibiotics was detected on biofilm formation by the tested commensal oral bacteria. Meanwhile, CIP-containing matrices inhibited growth of Fn and Aa. CONCLUSION: CIP-containing matrices led to a significant inhibition of periodontopathogens without negatively impairing the growth of periodontal beneficial bacteria. CLINICAL RELEVANCE: Based on the proven in vitro inhibition of periodontitis-related bacteria, future in vivo research using relevant animal models is needed to confirm the effectiveness of these drug delivery systems.