Microcosm Biofilm Research Articles

Combined Effectiveness of β-Cyclodextrin Nanoparticles in Photodynamic Antimicrobial Chemotherapy on In Vitro Oral Biofilms.

Objective: This study aimed to investigate if β-cyclodextrin nanoparticles potentiate the photodynamic antimicrobial chemotherapy (PACT) effects in single and microcosm oral biofilms using methylene blue (MB) and a red laser. Background data: Studies of PACT have demonstrated promising effects; however, the association of nanoparticles with photosensitizers could enhance the antimicrobial result. Materials and methods: Biofilms were grown on enamel blocks either with Streptococcus mutans or in a microcosm model (salivary microorganisms) supplemented with sucrose. PACT using 50 μM MB associated or not with 32 μM encapsulated β-cyclodextrin with MB for 5 min, followed by irradiation with red laser (λ = 660 nm, 320 J/cm2), was conducted and the counts of viable microorganisms in proper selective media were determined. Data were analyzed by one-factor ANOVA, followed by Tukey's test, or Kruskal-Wallis test, followed by Dunn's post hoc test, all with a significance level of 5%. Results: In the single-species biofilm model, a significant reduction in S. mutans counts was found for all groups when light was present. In the microcosm biofilm model, no significant difference was found among the groups for total streptococci, but a significant reduction of S. mutans was observed for the PACT group of encapsulated β-cyclodextrin+MB. However, no statistically significant difference was observed among the PACT groups. Conclusions: PACT with β-cyclodextrin mediated with MB associated with a red laser reduced S. mutans in microcosm biofilms. However, the presence of β-cyclodextrin nanoparticles did not potentiate the PACT effects in single or microcosm oral biofilms.

Effect of commercial herbal toothpastes and mouth rinses on the prevention of enamel demineralization using a microcosm biofilm model

This work evaluated the effects of commercial toothpastes and mouth rinses containing natural/herbal agents on biofilm viability, extracellular polysaccharide (EPS) production and on enamel demineralization in vitro. Microcosm biofilm was produced on bovine enamel for 5 days and treated daily with: Orgânico natural® (toothpaste/mouth rinse), Boni Natural Menta & Malaleuca® (toothpaste/mouth rinse), Propolis & Myrrh® (toothpaste), Colgate Total 12 Clean Mint® (toothpaste, positive control), Malvatricin® Plus (mouth rinse), PerioGard® (mouth rinse, positive control) or PBS (negative control). Tom’s Propolis & Myrrh® and Colgate Total 12® toothpastes and Malvatricin® Plus and PerioGard® mouth rinses significantly reduced biofilm viability (p < 0.05). Only PerioGard® had significant effects on biofilm thickness and EPS. Despite the indication that Tom’s Propolis & Myrrh® significantly reduced lesion depth, only Colgate Total 12® significantly reduced mineral loss. Malvatricin® Plus significantly reduced mineral loss and lesion depth, as did PerioGard®. Some herbal products, Malvatricin® Plus and Tom’s Propolis & Myrrh®, showed anticaries effects.

Self-healing adhesive with antibacterial activity in water-aging for 12 months

ObjectiveSecondary caries and micro-cracks are the main limiting factors for dentin bond durability. The objectives of this study were to develop a self-healing adhesive containing dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP), and investigate the effects of water-aging for 12 months on self-healing, dentin bonding, and antibacterial properties for the first time. MethodsMicrocapsules were synthesized with poly (urea-formaldehyde) (PUF) shells containing triethylene glycol dimethacrylate (TEGDMA) and N,N-dihydroxyethyl-p-toluidine (DHEPT). The adhesive contained 7.5% microcapsules, 10% DMAHDM, and 20% NACP (all mass). Specimens were water-aged at 37 °C for 1 day to 12 months. Dentin bond strength was measured using extracted human teeth. A single-edge-V-notched-beam (SEVNB) method was used to measure fracture toughness KIC and self-healing efficiency. A dental plaque microcosm biofilm model was used with human saliva as inoculum. ResultsThe microcapsules + DMAHDM + NACP group showed no decline in dentin bond strength after water-aging for 12 months, which was significantly higher than that of other groups without DMAHDM (p < 0.05). A self-healing efficiency of 67% recovery in KIC was obtained even after 12 months of water immersion, indicating that the self-healing ability was not lost in water-aging (p > 0.1). The bacteria-killing ability of this adhesive did not decline from 1 day to 12 months (p > 0.1), with biofilm CFU reduction by 3–4 orders of magnitude after the resin was water-aged for 12 months, compared to control resin. SignificanceThis novel adhesive with triple merits of self-healing, antibacterial and remineralization functions showed an excellent long-term durability in water-aging for 12 months. This multifunctional adhesive has the potential for dental applications to heal cracks, inhibit bacteria, provide ions for remineralization, and increase the restoration longevity.

Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans.

N-Acetylglucosamine (GlcNAc) and glucosamine (GlcN) enhance the competitiveness of the laboratory strain DL1 of Streptococcus gordonii against the caries pathogen Streptococcus mutans Here, we examine how amino sugars affect the interaction of five low-passage-number clinical isolates of abundant commensal streptococci with S. mutans by utilizing a dual-species biofilm model. Compared to that for glucose, growth on GlcN or GlcNAc significantly reduced the viability of S. mutans in cocultures with most commensals, shifting the proportions of species. Consistent with these results, production of H2O2 was increased in most commensals when growing on amino sugars, and inhibition of S. mutans by Streptococcus cristatus, Streptococcus oralis, or S. gordonii was enhanced by amino sugars on agar plates. All commensals except S. oralis had higher arginine deiminase activities when grown on GlcN and, in some cases, GlcNAc. In ex vivo biofilms formed using pooled cell-containing saliva (CCS), the proportions of S. mutans were drastically diminished when GlcNAc was the primary carbohydrate. Increased production of H2O2 could account in large part for the inhibitory effects of CCS biofilms. Surprisingly, amino sugars appeared to improve mutacin production by S. mutans on agar plates, suggesting that the commensals have mechanisms to actively subvert antagonism by S. mutans in cocultures. Collectively, these findings demonstrate that amino sugars can enhance the beneficial properties of low-passage-number commensal oral streptococci and highlight their potential for moderating the cariogenicity of oral biofilms.IMPORTANCE Dental caries is driven by dysbiosis of oral biofilms in which dominance by acid-producing and acid-tolerant bacteria results in loss of tooth mineral. Our previous work demonstrated the beneficial effects of amino sugars GlcNAc and GlcN in promoting the antagonistic properties of a health-associated oral bacterium, Streptococcus gordonii, in competition with the major caries pathogen Streptococcus mutans Here, we investigated 5 low-passage-number clinical isolates of the most common streptococcal species to establish how amino sugars may influence the ecology and virulence of oral biofilms. Using multiple in vitro models, including a human saliva-derived microcosm biofilm, experiments showed significant enhancement by at least one amino sugar in the ability of most of these bacteria to suppress the caries pathogen. Therefore, our findings demonstrated the mechanism of action by which amino sugars may affect human oral biofilms to promote health.

Antimicrobial and Anti-Caries Effect of New Glass Ionomer Cement on Enamel Under Microcosm Biofilm Model.

The occurrence of caries lesions adjacent to restorations is a serious problem in Dentistry. Therefore, new antimicrobial restorative materials could help to prevent recurrent carious lesions. This study evaluated the effect of a new glass ionomer cement (Ion Z) on the viability of a microcosm biofilm and on the development of enamel demineralization. Enamel samples were filled with the following materials (n=9): A) Ion-Z (FGM Ltda); B) Maxxion R (FGM Ltda); C) Ketac Fil Plus (3M ESPE) and D) no restoration (control). The samples were then exposed to human saliva mixed with McBain saliva (1:50) containing 0.2% sucrose for 14 days. The live and dead bacteria were quantified by fluorescence using a confocal laser-scanning microscope. The enamel demineralization was analyzed using transverse microradiography (TMR). The data were submitted to ANOVA/Tukey or Kruskal-Wallis/Dunn test (p<0.05). Ion Z induced a higher percentage of dead bacteria (60.96±12.0%) compared to the other groups (Maxxion R: 39.8±6.7%, Ketac Fil Plus: 43.7±9.71% and control 46.3±9.5%). All materials significantly reduced the average mineral loss compared to control (Ion-Z 25.0±4.2%vol, Maxxion R 23.4±8.0%vol, Ketac Fil Plus 30.7±7.7 and control 41.2±6.6%vol). Ion-Z was the only material able to significantly improve the mineral content at the surface layer (Zmax: 63.5±18.2%vol) compared to control (38.9±11.3%vol). Ion-Z shows antimicrobial potential, but its anti-caries effect was similar to the other materials, under this model.

Microcosm biofilms cultured from different oral niches in periodontitis patients

ABSTRACTObjective: Periodontal diseases are triggered by dysbiotic microbial biofilms. Therefore, it is essential to develop appropriate biofilm models. Aim of the present study was to culture microcosm biofilms inoculated from different niches in periodontitis patients and compare their microbial composition to those inoculated from subgingival plaque.Methods: Saliva, subgingival plaque, tongue and tonsils were sampled in five periodontitis patients to serve as inocula for culturing biofilms in vitro in an active attachment model. Biofilms were grown for 14 or 28 d and analyzed for their microbial composition by 16S rDNA sequencing.Results: As classified by HOMD, all biofilms were dominated by periodontitis-associated taxa, irrespective which niche had been used for inoculation. There was a low similarity between 14 d biofilms and their respective inocula (Bray-Curtis similarity 0.26), while biofilms cultured for 14 and 28 d shared high similarity (0.69). Principal components analysis showed much stronger clustering per patient than per niche indicating that the choice of patients may be more crucial than choice of the respective niches in these patients.Conclusion: Saliva, tongue scrapings or tonsil swabs may represent sufficient alternative inocula for growing microcosm biofilms resembling periodontitis-associated microbial communities in cases when sampling subgingival plaque is not possible.

Novel self-etch adhesive with antibacterial and protein-repellent functions to prevent enamel demineralization.

Enamel demineralization is the most undesired side-effect of fixed orthodontic treatments. This study were to develop a novel adhesive that is self-etching instead of using the traditional etching method, and to form a sealant on the enamel to prevent demineralization. 2-methacryloyloxyethyl phosphorylcholine (MPC) and quaternary ammonium dimethylaminohexadecyl methacrylate (DMAHDM) were mixed into a self-etch adhesive (Adper Easy One, 3M, St. Paul, MN, USA; referred to as AEO). Enamel shear bond strength (SBS) was measured. Protein adsorption onto the resins was measured. An oral microcosm biofilm model with saliva was tested. Incorporation of 7.5% MPC and 5% DMAHDM into AEO did not reduce the SBS (p>0.1). AEO with 7.5% MPC+5% DMAHDM had protein adsorption that was only 1/18 that of AEO control. AEO with 7.5% MPC+5% DAMHDM had much stronger antibacterial properties (p<0.05). In conclusion, the new self-etch adhesive with MPC and DAMHDM greatly reduced protein adsorption and inhibited biofilm viability.

Effect of water immersion on a dental self-healing and antibacterial resin composite

This investigation aimed to develop a novel self-healing and antibacterial dental resin composite. The effects of water immersion on its properties were also evaluated. Microcapsules filled with healing agent of triethylene glycol dimethacrylate were synthesized on the basis of previous studies. Antibacterial resin composite contained nano-antibacterial inorganic fillers that were modified by quaternary ammonium salt with long-chain alkyl. Microcapsules were incorporated into antibacterial resin composite at mass fraction of 7.5%. A commercial resin composite named Tetric N-Ceram was used as control. The resin samples were immersed in 37 °C distilled water for different periods. A flexural test was used to measure the mechanical properties of the novel resin composite. A single-edge V-notched beam method was used to measure fracture toughness and self-healing efficiency. A dental plaque microcosm biofilm model with human saliva as inoculum was formed. Colony-forming units (CFU) and lactic acid production of biofilm on the novel resin composite were calculated to test the antibacterial property. Mechanical properties and fracture toughness decreased significantly after the composite was immersed in water for 30 days (P<0.05), and no significant reduction was found from then on (P>0.05). Water immersion did not weaken the self-healing capability of the composite (P>0.05), and self-healing efficiency of 64% could still be obtained even after 270 days. The antibacterial resin composite showed a strong inhibition effect on the biofilm metabolic activity versus water immersion time from 1 day to 270 days. Therefore, the composite could still have a promising antibacterial property even after being immersed in water (P<0.05). Water immersion could weaken the mechanical properties of the novel self-healing and antibacterial resin composite, but it insignificantly affected the self-healing and antibacterial properties of the composite.

Protein-repellent nanocomposite with rechargeable calcium and phosphate for long-term ion release

ObjectiveThere has been no report on the effect of incorporating protein repellent 2-methacryloyloxyethyl phosphorylcholine (MPC) into a composite containing nanoparticles of amorphous calcium phosphate (NACP) on calcium (Ca) and phosphate (P) ion rechargeability. The objectives of this study were to develop a Ca and P ion-rechargeable and protein-repellent composite for the first time, and investigate the effects of MPC and NACP on mechanical properties, protein-repellency, anti-biofilm effects, and Ca and P ion recharge and re-release. MethodsNACP were synthesized using a spray-drying technique. The resin contained ethoxylated bisphenol A dimethacrylate (EBPADMA) and pyromellitic glycerol dimethacrylate (PMGDM). Three NACP composites were made with 0 (control), 1.5%, and 3% of MPC. NACP (20%) and glass particles (50%) were also added into the resin. Protein adsorption was measured using a micro-bicinchoninic acid (BCA) method. A human saliva microcosm biofilm model was used to determine biofilm metabolic activity, lactic acid, and colony-forming units (CFU). Ca and P ion recharge and re-release were measured using a spectrophotometric method. ResultsFlexural strengths and moduli of CaP-rechargeable composites matched those of a commercial composite without CaP rechargeability (p>0.1). Adding 1.5% and 3% MPC reduced protein adsorption to 1/3 and 1/5, respectively, that of commercial composite (p<0.05). Adding 3% MPC suppressed biofilm metabolic activity and lactic acid production, and reduced biofilm CFU by nearly 2 logs. All three NACP composites had excellent ion rechargeability and higher levels of ion re-releases. One recharge yielded continuous ion release for 21 days. The release was maintained at the same level with increasing number of recharge cycles, indicating long-term ion release. Incorporation of MPC did not compromise the CaP ion rechargeability. SignificanceIncorporating 3% MPC into NACP nanocomposite greatly reduced protein adsorption, biofilm growth and lactic acid, decreasing biofilm CFU by nearly 2 logs, without compromising Ca and P recharge. This protein-repellent NACP-MPC rechargeable composite with long-term remineralization is promising for tooth restorations to inhibit secondary caries.

Curcumin-mediated antimicrobial photodynamic therapy reduces the viability and vitality of infected dentin caries microcosms

To our knowledge, there is a lack of evidence on the effect of Antimicrobial Photodynamic Therapy (aPDT) by the application of curcumin against complex biofilms of dental caries lesions. This study aimed to evaluate the viability, vitality, and acid metabolism of infected dentin caries microcosms treated with curcumin-mediated aPDT. After microcosm biofilms growing anaerobically on bovine dentin disks immersed in McBain medium with 1% sucrose at 37 °C for 5 days, the biofilms were treated by the association of DMSO water solution or 600 μmol L-1 curcumin with 0, 37.5 or 75 J cm-2 blue LED (455 nm). Then, the colony-forming units (CFU) counts of total microorganisms, total streptococci, mutans streptococci, and total lactobacilli were determined by plating. The lactic acid concentration was analyzed by enzymatic spectrophotometry method, while the vitality of intact biofilms was evaluated by confocal laser scanning microscope (CLSM). Statistical analysis was performed by Kruskal Wallis and post-hoc Dunn's tests (P < 0.05). Curcumin alone did not affect the viability of microorganisms and the vitality of intact biofilms. However, 75 J cm-2 LED alone decreased the total microorganisms and total lactobacilli counts. The combination of curcumin and LED reduced significantly the counts of all microorganism groups and the vitality of intact biofilms. Differences were not observed between the lactic acid concentrations of distinct groups. Therefore, curcumin-mediated aPDT was effective in reducing the viability and the vitality of infected dentin caries microcosms, without interfering in their acidogenicity.

Developing a New Generation of Therapeutic Dental Polymers to Inhibit Oral Biofilms and Protect Teeth.

Polymeric tooth-colored restorations are increasingly popular in dentistry. However, restoration failures remain a major challenge, and more than 50% of all operative work was devoted to removing and replacing the failed restorations. This is a heavy burden, with the expense for restoring dental cavities in the U.S. exceeding $46 billion annually. In addition, the need is increasing dramatically as the population ages with increasing tooth retention in seniors. Traditional materials for cavity restorations are usually bioinert and replace the decayed tooth volumes. This article reviews cutting-edge research on the synthesis and evaluation of a new generation of bioactive dental polymers that not only restore the decayed tooth structures, but also have therapeutic functions. These materials include polymeric composites and bonding agents for tooth cavity restorations that inhibit saliva-based microcosm biofilms, bioactive resins for tooth root caries treatments, polymers that can suppress periodontal pathogens, and root canal sealers that can kill endodontic biofilms. These novel compositions substantially inhibit biofilm growth, greatly reduce acid production and polysaccharide synthesis of biofilms, and reduce biofilm colony-forming units by three to four orders of magnitude. This new class of bioactive and therapeutic polymeric materials is promising to inhibit tooth decay, suppress recurrent caries, control oral biofilms and acid production, protect the periodontium, and heal endodontic infections.

Laser microtextured titanium implant surfaces reduce in vitro and in situ oral biofilm formation.

IntroductionMicro- or nano-topography can both provide antimicrobial properties and improve osseointegration of dental implant titanium surfaces. Laser treatment is one of the best surface microtexturing techniques. The aim of this study was to evaluate in vitro and in situ biofilm formation on a laser-treated titanium surface, comparing it with two conventional surfaces, machined and grit-blasted.MethodsFor the in vitro experiment, an oral microcosm biofilm model was developed on the surface of titanium disks and reference human enamel using a bioreactor for 48 h. For the in situ experiment, titanium implants with laser-treated, machined and grit-blasted surfaces were mounted on intraoral trays and worn by ten volunteers for 48 h. Biofilm formation was quantitatively evaluated, and surfaces were analyzed using confocal laser scanning microscopy, scanning electron microscopy and energy-dispersive X-ray spectroscopy.Results–in vitro studyBiofilm structures with a prevalence of viable cells covered most of the machined, grit-blasted and human enamel surfaces, whereas less dense biofilm structures with non-confluent microcolonies were observed on the laser-treated titanium. Laser-treated titanium showed the lowest biofilm formation, where microorganisms colonized the edges of the laser-created pits, with very few or no biofilm formation observed inside the pits.Results–in situ studyThe biofilm formation pattern observed was similar to that in the in vitro experiment. Confocal laser scanning microscopy showed complete coverage of the implant threads, with mostly viable cells in grit-blasted and machined specimens. Unexpectedly, laser-treated specimens showed few dead microbial cells colonizing the bottom of the threads, while an intense colonization was found on the threading sides.ConclusionThis data suggests that laser-created microtopography can reduce biofilm formation, with a maximum effect when the surface is blasted orthogonally by the laser beam. In this sense the orientation of the laser beam seems to be relevant for the biological interaction with biofilms.

Protein-repelling adhesive resin containing calcium phosphate nanoparticles with repeated ion-recharge and re-releases

ObjectivesThe objectives were to develop a calcium (Ca) and phosphate (P) ion-rechargeable and protein-repellent adhesive containing nanoparticles of amorphous calcium phosphate (NACP) and 2-methacryloyloxyethyl phosphorylcholine (MPC), and investigate the MPC effects on ion recharge and re-releases for the first time. MethodsPyromellitic glycerol dimethacrylate and ethoxylated bisphenol-A dimethacrylate were used to fabricate adhesive PEHB. Six adhesives were tested: (1) Scotchbond (SBMP); (2) PEHB, (3) PEHB + 20%NACP; (4) PEHB + 30%NACP; (5) PEHB + 20%NACP+3%MPC; (6) PEHB + 30%NACP+3%MPC. Dentin shear bond strength, Ca/P ion release, recharge and re-release, and protein adsorption were measured. A microcosm biofilm model was tested for lactic-acid production and colony-forming units (CFU). ResultsAdding NACP + MPC did not negatively affect dentin bond strength (p > 0.1). With increasing the number of recharge/re-release cycles, the Ca/P ion re-release reached similarly higher levels (p > 0.1), indicating long-term remineralization capability. One recharge enabled the adhesives to have continued re-releases for 21 days. Incorporation of 3% MPC yielded 10-fold decrease in protein adsorption, and 1–2 log decrease in biofilm CFU. ConclusionsThe new rechargeable adhesive with MPC + 30%NACP greatly reduced protein adsorption, biofilm growth and lactic acid. Incorporation of MPC did not compromise the excellent Ca/P ion release, rechargeability, and dentin bond strength. Clinical significanceNovel bioactive adhesive containing MPC + NACP is promising to repel proteins and bacteria, and inhibit secondary caries at the restoration margins. The method of NACP + MPC to combine CaP-recharge and protein-repellency is applicable to the development of a new generation of materials including composites and cements to suppress oral biofilms and plaque formation and protect tooth structures.

Chlorhexidine, a Matrix Metalloproteinase Inhibitor and the Development of Secondary Caries Wall Lesions in a Microcosm Biofilm Model

This study investigated the role of a matrix metalloproteinase (MMP) inhibitor (CHX 2%) in the development of secondary caries wall lesions in different interface conditions with small (run 1) and wider gaps (run 2). Dentin discs were restored and pretreated with or without CHX 2%. In run 1, interfaces were made with gaps of 30, 60, or 90 µm. Interfaces with composite placed directly onto the dentin were either bonded (Adper Single Bond 2) or not bonded. In run 2, interfaces were made with gaps of 100 µm, with or without adhesive on the composite side (CLEARFIL SE Bond). Interfaces were either bonded or not bonded, as in run 1. Microcosm biofilms were grown on dentin-composite samples for 14 days. Caries lesion outcomes were analyzed by transversal wavelength-independent microradiography at 3 locations: the outer surface, and the interface wall at a distance of 200 and 500 µm from the gap entrance. Linear regression analyses showed that pretreatment with MMP inhibitor did not influence progression of the wall lesion at any location (p ≥ 0.218). Interfaces with intentional gaps showed positive and significant effect on the wall lesion progression at 200 µm from the gap entrance (p ≤ 0.005). A small trend of increase in wall lesion development was observed at the 200-µm location when bonding was present on the composite side. In conclusion, the dentin pretreatment with CHX 2% was not able to slow down the development of secondary caries wall lesions in small and wide gaps in this biofilm model.

Antimicrobial effect of bioceramic cements on multispecies microcosm biofilm: a confocal laser microscopy study

To assess the viability of multispecies microcosm biofilm after contact with NeoMTA Plus, Biodentine, and MTA Angelus. Fifty-four human dentin blocks (4 × 5 × 4mm) were allocated to Hawley retainers, worn by six volunteers for 72h. The blocks were then individually incubated in BHI broth for 21days at 37°C. At the end of experimental time for biofilm growth, the samples were randomly divided into four groups (n = 12): NeoMTA Plus, Biodentine, MTA Angelus, and negative control. The materials were placed in contact with the blocks. All samples were placed in cell-culture plate wells and incubated in BHI broth for 7days at 37°C. One sample from each volunteer (n = 6) was analyzed by SEM to describe the biofilm morphology. CLSM was performed to determine the percentage of viable biofilm biovolume. The data were statistically analyzed by one-way ANOVA and Tukey's multiple comparison test (α = 5%). SEM showed biofilm formed by spherical and rod-shaped bacteria surrounded by an extracellular matrix. No material was able to kill all biofilm cells, and all groups had more than 50% of viable bacteria. NeoMTA Plus was significantly different from the negative control group (P < .05). All tested materials were not effective against multispecies microcosm biofilm. NeoMTA Plus, Biodentine, and MTA Angelus were not effective against multispecies microcosm biofilm. It is essential to understand that these bioceramic cements are indicated for infected clinical situations. Thus, complementary disinfection procedures should be conducted prior to filling with these materials.

Effects of water-aging for 6 months on the durability of a novel antimicrobial and protein-repellent dental bonding agent

Biofilms at the tooth-restoration bonded interface can produce acids and cause recurrent caries. Recurrent caries is a primary reason for restoration failures. The objectives of this study were to synthesize a novel bioactive dental bonding agent containing dimethylaminohexadecyl methacrylate (DMAHDM) and 2-methacryloyloxyethyl phosphorylcholine (MPC) to inhibit biofilm formation at the tooth-restoration margin and to investigate the effects of water-aging for 6 months on the dentin bond strength and protein-repellent and antibacterial durability. A protein-repellent agent (MPC) and antibacterial agent (DMAHDM) were added to a Scotchbond multi-purpose (SBMP) primer and adhesive. Specimens were stored in water at 37 °C for 1, 30, 90, or 180 days (d). At the end of each time period, the dentin bond strength and protein-repellent and antibacterial properties were evaluated. Protein attachment onto resin specimens was measured by the micro-bicinchoninic acid approach. A dental plaque microcosm biofilm model was used to test the biofilm response. The SBMP + MPC + DMAHDM group showed no decline in dentin bond strength after water-aging for 6 months, which was significantly higher than that of the control (P < 0.05). The SBMP + MPC + DMAHDM group had protein adhesion that was only 1/20 of that of the SBMP control (P < 0.05). Incorporation of MPC and DMAHDM into SBMP provided a synergistic effect on biofilm reduction. The antibacterial effect and resistance to protein adsorption exhibited no decrease from 1 to 180 d (P > 0.1). In conclusion, a bonding agent with MPC and DMAHDM achieved a durable dentin bond strength and long-term resistance to proteins and oral bacteria. The novel dental bonding agent is promising for applications in preventive and restorative dentistry to reduce biofilm formation at the tooth-restoration margin.

Novel dental adhesive resin with crack self-healing, antimicrobial and remineralization properties

ObjectivesSecondary caries at the tooth-restoration margins is a primary reason for restoration failure. Cracks at the margins lead to leakage which can trap bacteria, producing acids to cause caries. To date, there has been no report on developing an adhesive resin that has self-healing, antibacterial and remineralizing capabilities. The objectives of this study were to: (1) develop the first self-healing adhesive with antimicrobial and remineralizing capabilities, and (2) investigate the effects of incorporating microcapsules, dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP) for the first time. MethodsSelf-healing microcapsules were synthesized with poly(urea-formaldehyde) (PUF) shells containing triethylene glycol dimethacrylate (TEGDMA) as the healing liquid. The new adhesive contained 7.5% microcapsules, 10% DMAHDM and 20% NACP. A single edge V-notched beam (SEVNB) method was used to measure the fracture toughness KIC and the autonomous crack-healing efficiency. An oral plaque microcosm biofilm model was tested. ResultsThe new self-healing, antimicrobial and remineralizing dental adhesive matched the dentin bond strength of a commercial control (p > 0.1). The new adhesive achieved successful crack-healing, with an excellent KIC recovery of 67%. The new adhesive had strong antimicrobial activity, reducing biofilm colony-forming units by four orders of magnitude, and reducing biofilm acid production to 1/100th that of biofilms on the commercial control resin. ConclusionsA self-healing adhesive with antibacterial and remineralizing capabilities was developed for the first time. Excellent dentin bond strength, autonomous crack-healing and KIC recovery, and strong anti-biofilm properties were achieved for the new adhesive resin. Clinical significanceThe novel method of using triple agents (self-healing microcapsules + DMAHDM + NACP) is promising for applications in dental adhesives, cements, sealants and composites to combat the two main challenges: fracture and secondary caries.

A Multi-scale Biophysical Approach to Develop Structure-Property Relationships in Oral Biofilms

Over the last 5–10 years, optical coherence tomography (OCT) and atomic force microscopy (AFM) have been individually applied to monitor the morphological and mechanical properties of various single-species biofilms respectively. This investigation looked to combine OCT and AFM as a multi-scale approach to understand the role sucrose concentration and age play in the morphological and mechanical properties of oral, microcosm biofilms, in-vitro. Biofilms with low (0.1% w/v) and high (5% w/v) sucrose concentrations were grown on hydroxyapatite (HAP) discs from pooled human saliva and incubated for 3 and 5 days. Distinct mesoscale features of biofilms such as regions of low and high extracellular polymeric substances (EPS) were identified through observations made by OCT. Mechanical analysis revealed increasing sucrose concentration decreased Young’s modulus and increased cantilever adhesion (p < 0.0001), relative to the biofilm. Increasing age was found to decrease adhesion only (p < 0.0001). This was due to mechanical interactions between the indenter and the biofilm increasing as a function of increased EPS content, due to increasing sucrose. An expected decrease in EPS cantilever contact decreased adhesion due to bacteria proliferation with biofilm age. The application OCT and AFM revealed new structure-property relationships in oral biofilms, unattainable if the techniques were used independently.

Novel rechargeable calcium phosphate nanocomposite with antibacterial activity to suppress biofilm acids and dental caries

ObjectiveRechargeable calcium phosphate (CaP) composites were developed recently. However, none of the rechargeable CaP composites was antibacterial. The objectives of this study were to develop the first rechargeable CaP composite that was antibacterial, and to investigate the effects of adding dimethylaminohexadecyl methacrylate (DMAHDM) into rechargeable CaP composite on ion rechargeability and re-release as well as biofilm properties. MethodsDMAHDM was synthesized via a Menschutkin reaction. Nanoparticles of amorphous calcium phosphate (NACP) were synthesized using a spray-drying technique. The resin contained ethoxylated bisphenol A dimethacrylate (EBPADMA) and pyromellitic glycerol dimethacrylate (PMGDM). Two composites were fabricated: rechargeable NACP composite, and rechargeable NACP-DMAHDM composite. Mechanical properties and ion release and recharge were measured. A dental plaque microcosm biofilm model using saliva was tested. ResultsFlexural strength and elastic modulus of rechargeable NACP and NACP-DMAHDM composites matched commercial control composite (p > 0.1). NACP-DMAHDM inhibited biofilm metabolic activity and lactic acid, and reduced biofilm colony-forming units (CFU) by 3–4 log. NACP and NACP-DMAHDM showed similar Ca and P ion recharge and re-release (p > 0.1). Therefore, adding DMAHDM did not compromise the ion rechargeability. One recharge yielded continuous release for 42 d. The release was maintained at the same level with increasing number of recharge cycles, indicating long-term ion release and remineralization capability. ConclusionsThe first CaP rechargeable and antibacterial composite was developed. Adding DMAHDM into the rechargeable NACP composite did not adversely affect the Ca and P ion release and recharge, and the composite had much less biofilm growth and lactic acid production, with CFU reduction by 3–4 log. Clinical significanceThis novel CaP rechargeable composite with long-term remineralization and antibacterial properties is promising for tooth restorations to inhibit caries.

Effect of a mouthrinse containing Malva sylvestris on the viability and activity of microcosm biofilm and on enamel demineralization compared to known antimicrobials mouthrinses

The purpose of this study was to evaluate the antimicrobial (anti-biofilm) and anti-caries (enamel demineralization prevention) effects of Malva sylvestris (Malvatricin® Plus) compared with known antimicrobial mouthrinses. Microcosm biofilm was produced on enamel, using inoculum from pooled human saliva mixed with McBain saliva (0.2% sucrose) for 14 days. The biofilm was treated with mouthrinses for 1 min day−1. Oral-B® Complete, Listerine® Zero and Malvatricin® Plus had the greatest effect on the reduction of biofilm viability (p < 0.0001). On the other hand, lactic acid production was reduced significantly with PerioGard®, Noplak® Max and Listerine® Zero compared with the control (p < 0.0001). No significant differences were found among the mouthrinses with respect to the colony-forming unit counting (total microorganisms, total streptococci, mutans streptococci and lactobacilli) and extracellular polysaccharide production. Enamel demineralization was reduced significantly with PerioGard®, Noplak® Max and Malvatricin® Plus compared with the control (p < 0.0001). Malva sylvestris has a comparable anti-caries effect to chlorhexidine mouthrinses.