Antibacterial effect of Softsoap used to disinfect dentures.

PURPOSE: To evaluate the in vitro antibacterial effect of Softsoap and Efferdent used as solutions to disinfect Lucitone 199 poly(methyl methacrylate) (PMMA) resin used for dentures. METHODS: S. mutans and plaque bacteria were grown for 24 hours, and suspended to a concentration of 1x106 cells/ml. Bacterial suspensions (0.2 mL) were added to the decontaminated PMMA discs placed in a 48-well culture plate and incubated for 3 days at 37°C. The discs were rinsed to remove the unbound bacterial cells and then incubated for 60 minutes with 5% and 1% dilutions (triplicates) of each of the detergent solutions (0.3 ml). Discs were rinsed and then MTT reagent (0.2 ml) was added and incubated for 2 hours, then overnight with a solubilizing agent. An aliquot from each well (0.1 ml) was transferred to a 96-well flat bottom plate and absorbance was measured to OD @ 595 nm (MTT) of four samples for each data point. Normalized data was compared and statistically analyzed using a three-way ANOVA with Student-Newman-Keuls on Rank data with P< 0.05 for significance. Additionally, data were double-checked with the Holm-Sidak test. RESULTS: There was no statistically significant difference between testing media for C. albicans and mixed plaque (P= 0.078) or testing duration in time at 24 hours and 21 days (P= 0.07). Statistically significant differences were found between all treatment solutions group combinations (P< 0.001) except for 30% Softsoap versus Efferdent (P= 0.056). CLINICAL SIGNIFICANCE: There was no statistically significant difference between testing media for C. albicans and mixed plaque (P= 0.078) or testing duration at 24 hours and 21 days (P= 0.07). Statistically significant differences were noted between all treatment solutions group combinations (P< 0.001), However, there was no difference between 30% Softsoap and Efferdent (P= 0.056).

Doxycycline Release of Dental Implants With Nanotube Surface, Coated With Poly Lactic-Co-Glycolic Acid for Extended pH-controlled Drug Delivery.

When dental implants become infected, the progression of the disease is rapid. Commercially available dental implant surfaces can be easily contaminated, resulting in rapid progression of peri-mucositis and peri-implantitis. The aim of this study was to evaluate, in vitro, the pattern of doxycycline release from by dental implants with titanium nanotube surface (DINS) at different pHs to examine novel drug loading and chemical coating techniques. Nine DINS were loaded with doxycycline and subsequently coated with polylactic-co-glycolic acid (PLGA). High-performance liquid chromatography (HPLC) was used to measure the amounts of released doxycycline in a 30-day period. Cytotoxicity of the DINS was evaluated by an assay using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT). The results showed that the experimental DINS coated with doxycycline and PLGA showed a mean drug release during the experimental period for the groups: pH 7.4 (8.39 µg/mL), pH 6.4 (8.63 µg/mL). The pH 5.4 (15.18 µl/mL) doxycycline release from DINS was faster at pH 5.4 than those at pHs 6.4 and 7.4 (P = .0031 and .0034, respectively). This new surface treatment of dental implants with titanium nanotubes and subsequent drug loading demonstrated biocompatibility and sustained doxycycline release over a 30-day period. Additional studies are needed in order to adopt a stable drug release at neutral pH environment while warranting a constant drug release in an acidic pH environment.

Antibacterial properties of silver-loaded gelatin sponges prepared with silver diamine fluoride.

PURPOSE: To evaluate the antibacterial efficiency of silver-loaded gelatin sponges prepared from gelatin and silver diamine fluoride, Ag(NH3)2F. METHODS: A series of novel silver-loaded gelatin sponges were prepared from gelatin and silver diamine fluoride. They were characterized using Fourier transform infrared spectroscopy (FT-IR), scanning electronic microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). The antibacterial activities against one oral bacteria model S. mutans were evaluated using the agar disk diffusion method and biofilm-grown bacteria assay. For the purpose of comparison, a second series of silver-loaded sponges were prepared using silver nitrate (AgNO3) as the silver source. RESULTS: FT-IR, SEM and EDX results confirmed the successful preparation of silver-loaded gelatin sponges from both silver diamine fluoride and silver nitrate. Agar disk diffusion assays revealed that the antibacterial activity of silver-loaded sponges was strongly correlated with the silver content, and also depending on the silver source used in the preparation of sponges. Sustained inhibition of S. mutans in agar plates was observed for silver-loaded gelatin sponges containing about 5 wt% Ag or more. Biofilm assays showed different viabilities when treated with different formulations, with the viability of 11.4±3.1% for the sponges containing 5.30±1.18 wt% Ag prepared from silver diamine fluoride, and the viability of 15.4±3.1% for the sponges containing 29.13±10.34 wt% Ag prepared from silver nitrate. CLINICAL SIGNIFICANCE: The silver loading contents had a significant effect on the antibacterial activities of silver-loaded gelatin sponges prepared with silver diamine fluoride. In addition, silver diamine fluoride was a superior silver source to prepare antibacterial silver-loaded gelatin sponges when compared with silver nitrate.

Modified Polymeric Nanoparticles Exert In Vitro Antimicrobial Activity Against Oral Bacteria.

Polymeric nanoparticles were modified to exert antimicrobial activity against oral bacteria. Nanoparticles were loaded with calcium, zinc and doxycycline. Ions and doxycycline release were measured by inductively coupled plasma optical emission spectrometer and high performance liquid chromatography. Porphyromonas gingivalis, Lactobacillus lactis, Streptoccocus mutans, gordonii and sobrinus were grown and the number of bacteria was determined by optical density. Nanoparticles were suspended in phosphate-buffered saline (PBS) at 10, 1 and 0.1 mg/mL and incubated with 1.0 mL of each bacterial suspension for 3, 12, and 24 h. The bacterial viability was assessed by determining their ability to cleave the tetrazolium salt to a formazan dye. Data were analyzed by ANOVA and Scheffe’s F (p < 0.05). Doxycycline doping efficacy was 70%. A burst liberation effect was produced during the first 7 days. After 21 days, a sustained release above 6 µg/mL, was observed. Calcium and zinc liberation were about 1 and 0.02 µg/mL respectively. The most effective antibacterial material was found to be the Dox-Nanoparticles (60% to 99% reduction) followed by Ca-Nanoparticles or Zn-Nanoparticles (30% to 70% reduction) and finally the non-doped nanoparticles (7% to 35% reduction). P. gingivalis, S. mutans and L. lactis were the most susceptible bacteria, being S. gordonii and S. sobrinus the most resistant to the tested nanoparticles.

Effects of atmospheric non-thermal argon/oxygen plasma on biofilm viability and hydrophobicity of oral bacteria.

PURPOSE: To evaluate the bactericidal effects of atmospheric non-thermal argon/oxygen plasma on in vitro oral biofilms constructed from S. mutans and/or S. sanguinis, and the influence of the plasma on the virulence properties of A. oris. METHODS: In vitro oral biofilms were constructed in the wells of 48-well plates from S. mutans and/or S. sanguinis. The wells containing constructed biofilms and various amounts of phosphate-buffered saline (PBS) were treated with non-thermal argon/oxygen plasma brush for 2 minutes. The methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and Live/Dead assay were used to evaluate the viability of biofilms in those wells after the plasma treatments. Meanwhile, A. oris suspensions were treated with the plasma and then evaluated for their virulence properties by measuring the hydrophobicity and co-aggregation capability of treated A. oris. RESULTS: The MTT assay showed that exposure to non-thermal plasma for 2 minutes significantly reduced the viability of bacteria in both single-species and two-species biofilms of S. mutans and S. sanguinis with the reductions of up to 99%. Meanwhile, plasma treatment also altered the hydrophobicity of A. oris, and reduced their capability to co-aggregate with S. sanguinis. CLINICAL SIGNIFICANCE: The results from this study demonstrated that atmospheric non-thermal argon/oxygen plasma could effectively deactivate oral bacteria biofilm by decreasing bacterial viability as well as reducing their hydrophobicity and co-aggregation capability.

Effects of trimethylsilane plasma coating on the hydrophobicity of denture base resin and adhesion of Candida albicans on resin surfaces.

STATEMENT OF PROBLEM: Candida-associated denture stomatitis is the most common oral mucosal lesion among denture wearers. Trimethylsilane (TMS) plasma coating may inhibit the growth of Candida albicans on denture surfaces. PURPOSE: The purpose of this in vitro study was to investigate whether TMS plasma coatings can effectively reduce C albicans adhesion on denture base acrylic resin surfaces. MATERIAL AND METHODS: Sixty denture base acrylic resin disks with smooth and rough surfaces were prepared and were either left untreated (control group) or coated with TMS monomer (experimental group) by using plasma. Contact angles were measured immediately after TMS plasma coating. The morphology of C albicans adhesion was observed with scanning electron microscopy (SEM). Energy-dispersive spectroscopy (EDS) was used to characterize the elemental composition of the specimen surface. An adhesion test was performed by incubating the resin disk specimens in C albicans suspensions (1×107 cells/mL) at 37°C for 24 hours and further measuring the optical density of the C albicans by using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay test. One-way ANOVA and 2-way ANOVA were followed by a post hoc test analysis (α=.05). RESULTS: The group with TMS coating exhibited a more hydrophobic surface than the control group. EDS analysis revealed successful TMS plasma coating. The difference in the mean contact angles between the uncoated group and the TMS-coated group was statistically significant (P<.05), 79.0 ±2.9 degrees versus 105.7 ±1.5 degrees for the smooth surface and 90.2 ±7.6 degrees versus 131.5 ±2.1 degrees for the rough surface. In SEM analysis, the C albicans biofilm was found to grow more on the surface of the denture base resin without the TMS coating than on the surfaces of the experimental group. In the adhesion test, the amount of C albicans adhering to the surface of denture base resin with the TMS coating was significantly less than that on the surfaces without TMS coating (P<.05). CONCLUSIONS: TMS coating significantly reduced the adhesion of C albicans to the denture base resin and may reduce denture stomatitis.

Efficiency of Nanotube Surface-Treated Dental Implants Loaded with Doxycycline on Growth Reduction of Porphyromonas gingivalis.

PURPOSE: The prevalence of peri-implant infection in patients with dental implants has been shown to range from 28% to 56%. A nanotube-modified implant surface can deliver antibiotics locally and suppress periodontal pathogenic bacterial growth. The aim of this study was to evaluate the deliverability of antibiotics via a nanotube-modified implant. MATERIALS AND METHODS: Dental implants with a nanotube surface were fabricated and loaded with doxycycline. Afterward, each dental implant with a nanotube surface was placed into 2-mL tubes, removed from solution, and placed in a fresh solution daily for 28 days. Experimental samples from 1, 2, 4, 16, 24, and 28 days were used for this evaluation. The concentration of doxycycline was measured using spectrophotometric analysis at 273-nm absorbance. The antibacterial effect of doxycycline was evaluated by supplementing Porphyromonas gingivalis (P gingivalis) growth media with the solution collected from the dental implants at the aforementioned time intervals for a period of 48 hours under anaerobic conditions. A bacterial viability assay was used to evaluate P gingivalis growth at 550-nm absorbance. RESULTS: Doxycycline concentration varied from 0.33 to 1.22 µg/mL from day 1 to day 28, respectively. A bacterial viability assay showed the highest P gingivalis growth at day 1 (2 nm) and the lowest at day 4 (0.17 nm), with a gradual reduction from day 1 to day 4 of approximately 87.5%. The subsequent growth pattern was maintained and slightly increased from baseline in approximately 48.3% from day 1 to day 24. The final P gingivalis growth measured at day 28 was 29.4% less than the baseline growth. CONCLUSION: P gingivalis growth was suppressed in media supplemented with solution collected from dental implants with a nanotube surface loaded with doxycycline during a 28-day time interval.

Efficacy of 4 Irrigation Protocols in Killing Bacteria Colonized in Dentinal Tubules Examined by a Novel Confocal Laser Scanning Microscope Analysis.

INTRODUCTION: The aim of this study was to determine the efficiency of 4 irrigation systems in eliminating bacteria in root canals, particularly in dentinal tubules. METHODS: Roots of human teeth were prepared to 25/04, autoclaved, and inoculated with Enterococcus faecalis for 3 weeks. Canals were then disinfected by (1) standard needle irrigation, (2) sonically agitating with EndoActivator, (3) XP Endo finisher, or (4) erbium:yttrium aluminum garnet laser (PIPS) (15 roots/group). The bacterial reduction in the canal was determined by MTT assays. For measuring live versus dead bacteria in the dentinal tubules (4 teeth/group), teeth were split open and stained with LIVE/DEAD BackLight. Coronal, middle, and apical thirds of the canal dentin were scanned by using a confocal laser scanning microscope (CLSM) to determine the ratio of dead/total bacteria in the dentinal tubules at various depths. RESULTS: All 4 irrigation protocols significantly eliminated bacteria in the canal, ranging from 89.6% to 98.2% reduction (P < .001). XP Endo had the greatest bacterial reduction compared with other 3 techniques (P < .05). CLSM analysis showed that XP Endo had the highest level of dead bacteria in the coronal, middle, and apical segments at 50-µm depth. On the other hand, PIPS had the greatest bacterial killing efficiency at the 150-µm depth in all 3 root segments. CONCLUSIONS: XP Endo appears to be more efficient than other 3 techniques in disinfecting the main canal space and up to 50 µm deep into the dentinal tubules. PIPS appears to be most effective in killing the bacteria deep in the dentinal tubules.

Assessment of the Effect of CO2 Laser Irradiation on the Reduction of Bacteria Seeded on Commercially Available Sandblasted Acid-Etched Titanium Dental Implants: An In Vitro Study.

PURPOSE: To evaluate the capability of carbon dioxide (CO2) laser in reducing Escherichia coli on sandblasted acid-etched (SAE) titanium dental implants. MATERIALS AND METHODS: SAE dental implants were contaminated with E coli, incubated in a sterile bacterial culture medium for 24 hours, and then exposed to CO2 laser (10,600-nm wavelength) in superpulsed waves (SPW) at 1.5, 1.7, and 2 W at 100-Hz frequency and continuous wave (CW) at 1.5, 2, and 2.5 W. The presence of bacteria trapped in the implant surfaces after contamination and decontamination was verified using spectrophotometry. Scanning electron microscopy (SEM) was used to evaluate the topography of laser irradiation. After implant surface contamination was verified, implants were exposed to CO2 laser irradiation, and bacterial growth was measured with spectrophotometry. RESULTS: The control implants showed the highest bacterial growth (100% growth). Implants exposed to laser showed progressive increase in the percentage of decontamination (DC%) corresponding to the higher wattage in the SPW and CW groups. The DC% were 20.4%, 49.6%, and 51.7% in the SPW group at 100 Hz, at 1.5, 1.7, and 2 W of power, respectively. In the CW group, the DC% were 34.3%, 69.9%, and 85.5% at 1.7, 2, and 2.5 W, respectively. Kruskal-Wallis statistical analysis showed a significant difference between the groups (P < .05). In the pulsed mode (100-Hz) group, statistical analysis showed that the DC% of 1.5 W was significantly lower than the 2 W power. In the CW group, statistical analysis showed that the DC% at 1.7 W was significantly lower (P < .05) than with the other treatments. SEM assessment showed craterlike wear damages and accretions to the implant surfaces that increased progressively as the laser wattage increased. CONCLUSION: CO2 laser irradiation failed to completely decontaminate the implant surfaces. SEM analysis demonstrated damage to the top of the dental implant threads at all settings studied. Thus, CO2 laser irradiation may not be the optimal method to decontaminate implants.

Assessment of the effect of chemical agents used in dentistry on the removal of Porphyromonas gingivalis and Escherichia coli from sandblasted acid-etched titanium dental implants: an in vitro study.

PURPOSE: The aim of this study was to evaluate the capability of chemicals to decontaminate Escherichia coli (E coli) or Porphyromonas gingivalis (P gingivalis) from sandblasted acid-etched (SAE) titanium dental implants. MATERIALS AND METHODS: SAE titanium dental implants were contaminated with E coli or P gingivalis and incubated in a sterile bacterial culture media under aerobic and anaerobic conditions, respectively. The implants were treated with 10 different conditions: calcium hydroxide [Ca(OH)2] paste for 1 minute and saline irrigation for 1 minute; Ca(OH)2 paste for 1 minute and 0.2% chlorhexidine digluconate (CHXD) irrigation for 1 minute; 0.2% CHXD for 1 minute; Dakin's solution for 1 minute; tetracycline hydrochloride (T-HCl) as a 1 g per 20 mL solution for 1, 2, and 3 minutes; and T-HCl paste for 1, 2, and 3 minutes. All implants were irrigated with 1 mL of saline solution and incubated under aerobic and anaerobic conditions for 24 hours or 48 hours for E coli- and P gingivalis-contaminated implants, respectively. The control group was submitted to all procedures except for the chemical treatments. Aliquots were removed, and turbidity was measured by spectrophotometry. The level of bacterial growth in control cultures was considered to have a decontamination percentage (DC%) of 0. RESULTS: Spectrophotometric analysis showed that all chemical treatments resulted in significantly higher DC% compared to controls for SAE implants contaminated with E coli (P < .05) or P gingivalis (P < .05). For the P gingivalis experiments, SAE implants treated with Ca(OH)2 paste and saline solution had a lower DC% (39.3%) than those in the other treatment groups. In the E coli experiments, DC% was significantly lower for SAE implants treated with Ca(OH)2 paste and saline solution (48.7%), Dakin's solution (92.7%), or T-HCl paste for 1 minute (96.6%) than those in the other groups. CONCLUSION: The DC% of SAE implants contaminated with E coli or P gingivalis by means of chemicals commonly used in dentistry is high, with the exception of Ca(OH)2 paste burnished for 1 minute and then irrigated with saline solution for 1 minute.

Inhibitory effects of a cured antibacterial bonding system on viability and metabolic activity of oral bacteria.

OBJECTIVES: To evaluate the antimicrobial efficacy of Clearfil SE Protect (CP) and Clearfil SE Bond (CB) after curing and rinsed against five individual oral microorganisms as well as a mixture of bacterial culture prepared from the selected test organisms. METHODS: Bacterial suspensions were prepared from single species of Streptococcus mutans, Streptococcus sobrinus, Streptococcus gordonii, Actinomyces viscosus and Lactobacillus lactis, as well as mixed bacterial suspensions from these organisms. Dentin bonding system discs (6 mm×2 mm) were prepared, cured, washed and placed on the bacterial suspension of single species or multispecies bacteria for 15, 30 and 60 min. MTT, Live/Dead bacterial viability (antibacterial effect), and XTT (metabolic activity) assays were used to test the two dentin system's antibacterial effect. All assays were done in triplicates and each experiment repeated at least three times. Data were submitted to ANOVA and Scheffe's f-test (5%). RESULTS: Greater than 40% bacteria killing was seen within 15 min, and the killing progressed with increasing time of incubation with CP discs. However, a longer (60 min) period of incubation was required by CP to achieve similar antimicrobial effect against mixed bacterial suspension. CB had no significant effect on the viability or metabolic activity of the test microorganisms when compared to the control bacterial culture. CP was significantly effective in reducing the viability and metabolic activity of the test organisms. SIGNIFICANCE: The results demonstrated the antimicrobial efficacy of CP both on single and multispecies bacterial culture. CP may be beneficial in reducing bacterial infections in cavity preparations in clinical dentistry.

In vitro comparison of commercial oral rinses on bacterial adhesion and their detachment from biofilm formed on hydroxyapatite disks.

PURPOSE: This in vitro study was designed to assess the effectiveness of three oral rinses on bacterial adherence to epithelial cells and hydroxyapatite surfaces. The role of oral rinses on the detachment of bacteria from biofilm was also evaluated. MATERIALS AND METHODS: The efficacy of three oral rinses, Acclean, Noplak and Prevention were tested against a wide range of oral bacteria. Oral rinse antimicrobial activity was determined by an MTT assay for bacterial viability, by live/ dead staining and by measuring the bacterial metabolic activity using an XTT assay. RESULTS: The two oral rinses that contained 0.12% chlorhexidine had the greatest antibacterial activity on both planktonic and bio lm-grown organisms when compared to the Prevention oral rinse. CONCLUSION: Both Acclean and Noplak were extremely effective in lowering the number of bacteria attached to buccal epithelial cells and pelllicles. In addition, these two oral rinses were also effective against the biofilm bacteria.

Inhibition of Streptococcus gordonii metabolic activity in biofilm by cranberry juice high-molecular-weight component.

Previous studies demonstrated that a cranberry high-molecular-mass, nondialyzable material (NDM) can inhibit adhesion of numerous species of bacteria and prevents bacterial coaggregation of bacterial pairs. Bacterial coaggregation leads to plaque formation leading to biofilm development on surfaces of oral cavity. In the present study, we evaluated the effect of low concentrations of NDM on Streptococcus gordonii metabolic activity and biofilm formation on restorative dental surfaces. We found that the NDM selectively inhibited metabolic activity of S. gordonii, without affecting bacterial viability. Inhibiting the metabolic activity of bacteria in biofilm may benefit the health of the oral cavity.

Chitosan coatings deliver antimicrobials from titanium implants: a preliminary study.

OBJECTIVE: Chitosan was investigated as a coating for local delivery of antimicrobials for prevention of acute implant infection. The objectives of this study were to (1) measure the release of 2 antimicrobials from chitosan coatings, (2) determine efficacy of eluted antimicrobials against bacteria, in vitro, and (3) evaluate toxicity of eluted drugs to host cells/tissues. METHODS: Chitosan coatings (80.7% deacetylated, 108 kDa) containing 20% tetracycline or 0.02% chlorhexidine digluconate were bonded to titanium via silane reactions. After elution in culture medium for 7 days, eluates were tested against model pathogens Actinobacillus actinomycetemcomitans and Staphylococcus epidermidis in turbidity tests and in 24-hour cytotoxicity tests using human osteoblasts and fibroblasts. Finally, antibiotic-loaded chitosan-coated titanium pins were implanted for 7 days in muscle of Sprague-Dawley rats to evaluate the initial tissue response. RESULTS: Coatings released 89% of tetracycline in 7 days and 100% chlorhexidine in 2 days. Released tetracycline inhibited growth (95%-99.9%) of pathogens for up to 7 days with no cytotoxicity to human cells. Released chlorhexidine was active against pathogens for 1 to 2 days (56%-99.5% inhibition) but was toxic to cells on the first day of elution. Typical acute inflammatory response was observed to antimicrobial-loaded chitosan coatings similar to unloaded coatings. CONCLUSION: These preliminary data support the hypothesis that chitosan coatings have the potential to locally deliver antimicrobials to inhibit bacteria without being toxic to host cells/tissues and warrant additional studies to evaluate the ability of the coatings to prevent/resist infection and promote osseointegration.