Fabrication of pH-Responsive Zn2+-Releasing Glass Particles for Smart Antibacterial Restoratives.

The on-demand release of antibacterial components due to pH variations caused by acidogenic/cariogenic bacteria is a possible design for smart antibacterial restorative materials. This study aimed to fabricate pH-responsive Zn2+-releasing glass particles and evaluate their solubilities, ion-releasing characteristics, and antibacterial properties in vitro. Three kinds of silicate-based glass particles containing different molar ratios of Zn (PG-1: 25.3; PG-2: 34.6; PG-3: 42.7 mol%) were fabricated. Each particle was immersed in a pH-adjusted medium, and the solubility and concentration of the released ions were determined. To evaluate the antibacterial effect, Streptococcus mutans was cultured in the pH-adjusted medium in the presence of each particle, and the bacterial number was counted. The solubility and concentration of Zn2+ released in the medium increased with a decrease in medium pH. PG-3 with a greater content of Zn demonstrated higher concentrations of released Zn2+ compared with PG-1 and PG-2. PG-2 exhibited bactericidal effects at pH 5.1, whereas PG-3 demonstrated bactericidal effects at pH values of 5.1 and 6.1, indicating that PG-3 was effective at inhibiting S. mutans even under slightly acidic conditions. The glass particle with 42.7 mol% Zn may be useful for developing smart antibacterial restoratives that contribute to the prevention of diseases such as caries on root surfaces with lower acid resistance.

Incorporation of chlorhexidine in self-adhesive resin cements.

The aim of this study was to evaluate the maximum amount of chlorhexidine (CHX) that could be incorporated to self-adhesive resin cements to add antibacterial effect without affecting the physical properties. The CHX was incorporated into a commercial self-adhesive resin cement at mass fractions of 0.5-15 wt%, and the CHX-release profile, antibacterial effect, flexural and bond strengths of experimental cements were evaluated. Increasing the CHX content from 5 to 15 wt% resulted in a higher released concentration of CHX. In agar diffusion tests, experimental cements containing 5, 10, and 15 wt% CHX produced inhibition zones against oral bacteria. In flexural strength and shear bond strength to dentin, no significant reduction was observed with the incorporation of 5 wt% CHX. This in vitro study suggests that the addition of 5 wt% CHX yielded an antibacterial self-adhesive cement and had no adverse effect on the flexural and shear bond strengths.

Antibacterial activities and mineral induction abilities of proprietary MTA cements.

Mineral trioxide aggregate (MTA) cements are used in direct pulp capping and many other applications, and several types of these products have been commercialized. The aim of this study was to examine the antibacterial effects and mineral induction abilities of three conventional MTA cements and one resin-modified MTA cement. Agar diffusion tests revealed that, after setting, all four cements exhibited little antibacterial effects against Enterococcus faecalis and Streptococcus mutans, with no significant differences among the materials. After 24 h, E. faecalis and S. mutans suspensions incubated in the presence of each cement did not exhibit reduced numbers of viable bacteria, compared with those same bacterial suspensions incubated without any cement; this indicated that none of the cements inhibited bacterial growth. Furthermore, the resin-modified MTA cement exhibited lower mineral induction ability, compared with that of the three conventional MTA cements.

Development of endodontic sealers containing antimicrobial-loaded polymer particles with long-term antibacterial effects.

OBJECTIVE: The objective of this study is to prepare new dental resins with a long-lasting antimicrobial activity. Specifically, this study evaluates an approach for controlling infection in root canals using sealers containing polyhydroxyethyl methacrylate trimethylolpropane trimethacrylate (polyHEMA/TMPT) particles loaded with cetylpyridinium chloride (CPC). In addition, the physical properties of sealers containing CPC-loaded polyHEMA/TMPT particles (CLP) are determined. METHODS: PolyHEMA/TMPT particles with 10 (10%-CLP) and 25wt.% CPC (25%-CLP) with different particle sizes were fabricated and incorporated in HEMA-based sealers. CPC-release profiles were evaluated over 14 days of immersion in water, followed by 14 days of storage and 14 days of water immersion. The antibacterial activity of these sealers against Enterococcus faecalis in dentinal tubules was assessed using a root-canal-infection model. Their sealing abilities were evaluated by fluid filtration and physical properties were tested according to the ISO 6876 standard. The long-term antibacterial activity of the cured sealer containing 25%-CLP (∼21µm particle diameter) was re-assessed after 1 year of storage. RESULTS: After 28 days of immersion, 25%-CLP exhibited a higher and sustained CPC release unlike 10%-CLP. Residual bacteria in root dentinal tubules were eradicated by obturation with 25%-CLP-containing sealers. The incorporation of 25%-CLP (∼21µm) had no adverse effects on the sealing ability and physical properties of the sealer and resulted in long-term antibacterial activity. SIGNIFICANCE: The incorporation of CPC-loaded particles in HEMA resins yielded endodontic sealers with long-term bactericidal activity against E. faecalis in root canals. These sealers can potentially be used to prevent recurrent apical periodontitis.

Development of novel surface coating composed of MDPB and MPC with dual functionality of antibacterial activity and protein repellency.

Resin-based reconstructive/restorative materials with antibacterial effects are potentially useful for preventing dental and oral diseases. To this end, the immobilization of an antibacterial component on the surface of a resin by incorporating polymerizable bactericide such as a quaternary ammonium compound-monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) is an effective technique. However, the effectiveness of immobilized bactericide is reduced by salivary protein coverage. We address this issue by utilizing 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, which exhibits protein repellency, with MDPB to fabricate a novel copolymer, which served as a surface coating on a methacrylate-based resin. This coating provided a more hydrophilic surface than that provided by MDPB coating and reduced the adsorption of bovine serum albumin and salivary protein. To evaluate bacterial growth on the contact surface, Streptococcus mutans suspension was placed on the coated specimen. After 24-h incubation, MDPB/MPC copolymer exhibited killing effects against S. mutans. Moreover, confocal laser scanning microscopy and scanning electron microscopy were used to evaluate biofilm formation after 48-h incubation in S. mutans suspension, which revealed sparse biofilm and dead bacteria in biofilm on the surface coated with MDPB/MPC. Overall, the proposed surface coating on dental resins exhibited protein-repellent ability and inhibitory effects against bacteria and oral biofilms.

Antibacterial activity and dentin bonding ability of combined use of Clearfil SE Protect and sodium hypochlorite.

The aim of this study was to evaluate the antibacterial activity and dentin bonding ability of a commercial self-etch adhesive Clearfil SE Protect (Kuraray Noritake Dental, Tokyo, Japan) in combination with sodium hypochlorite (NaOCl). Agar disc diffusion tests and measurement of minimum inhibitory/bactericidal concentrations (MIC/MBC) against Streptococcus mutans were performed to evaluate antibacterial effects. The mixture solution of 5.25% NaOCl and the primer of Clearfil SE Protect demonstrated less antibacterial activity than primer only. In microtensile bond strength tests using non-carious human molars, pretreatment with 5.25% NaOCl aqueous solution had no influence on the bond strength of Clearfil SE Protect. These results indicate that pretreatment with NaOCl does not influence the bonding ability of Clearfil SE Protect, while their combined use does not enhance cavity disinfecting effects.

Non-biodegradable polymer particles for drug delivery: A new technology for "bio-active" restorative materials.

To develop dental restorative materials with "bio-active" functions, addition of the capability to release active agents is an effective approach. However, such functionality needs to be attained without compromising the basic properties of the restorative materials. We have developed novel non-biodegradable polymer particles for drug delivery, aimed for application in dental resins. The particles are made using 2-hydroxyethyl methacrylate (HEMA) and a cross-linking monomer trimethylolpropane trimethacrylate (TMPT), with a hydrophilic nature to adsorb proteins or water-soluble antimicrobials. The polyHEMA/TMPT particles work as a reservoir to release fibroblast growth factor-2 (FGF-2) or cetylpyridinium chloride (CPC) in an effective manner. Application of the polyHEMA/TMPT particles loaded with FGF-2 to adhesives, or those loaded with CPC to resin-based endodontic sealers or denture bases/crowns is a promising approach to increase the success of the treatments by conferring "bio-active" properties to these materials to induce tissue regeneration or to inhibit bacterial infection.

SiO2-nanocomposite film coating of CAD/CAM composite resin blocks improves surface hardness and reduces susceptibility to bacterial adhesion.

Composite resin blocks for computer-aided design/computer-aided manufacturing (CAD/CAM) applications have recently become available. However, CAD/CAM composite resins have lower wear resistance and accumulate more plaque than CAD/CAM ceramic materials. We assessed the effects of SiO2-nanocomposite film coating of four types of CAD/CAM composite resin blocks: Cerasmart, Katana Avencia block, Lava Ultimate, and Block HC on surface hardness and bacterial attachment. All composite blocks with coating demonstrated significantly greater Vickers hardness, reduced surface roughness, and greater hydrophobicity than those without coating. Adhesion of Streptococcus mutans to the coated specimens was significantly less than those for the uncoated specimens. These reduced levels of bacterial adherence on the coated surface were still evident after treatment with saliva. Surface modification by SiO2-nanocomposite film coating has potential to improve wear resistance and susceptibility to plaque accumulation of CAD/CAM composite resin restorations.

Antibacterial activity of resin composites containing surface pre-reacted glass-ionomer (S-PRG) filler.

OBJECTIVE: A surface pre-reacted glass-ionomer (S-PRG) filler is a technology of interest for providing bio-functions to restorative materials. Resin composites containing S-PRG filler have been reported to show less plaque accumulation and reduced bacterial attachment. In this study, experimental resin composites containing S-PRG filler at various concentrations were fabricated, and the inhibitory effects on bacterial growth on their surface and the association of ions released from S-PRG filler with antibacterial activity were evaluated. METHODS: Five kinds of experimental resin composites containing S-PRG filler at 0, 13.9, 27.3, 41.8, or 55.9 (vol.%) were fabricated. Streptococcus mutans was cultured on the cured discs for 18h to examine the growth of bacteria in contact with the surface of the experimental resins. The concentrations of Al(3+), BO3(3-), F(-), Na(+), SiO3(2-), or Sr(2+) released from each experimental resin into water were measured. The standardized solutions of each ion were prepared at the concentrations determined to be released from the experimental resin, and their inhibitory effects of single ion species on S. mutans growth were evaluated by using each standardized solution. RESULTS: Resin composites containing S-PRG filler at 13.9 (vol.%) or greater inhibited S. mutans growth on their surface. When S. mutans was incubated in the presence of six kinds of ions at the concentrations released from the resin composite containing S-PRG filler at 55.9 (vol.%), a significant reduction in bacterial number was observed for BO3(3-), F(-), Al(3+), and SiO3(2-). Among these four ions, BO3(3-) and F(-) demonstrated the strongest inhibitory effect on S. mutans growth. SIGNIFICANCE: Our findings suggest that resin composites containing S-PRG filler inhibit the growth of S. mutans on their surface. BO3(3-), F(-), Al(3+) and SiO3(2-) released from S-PRG filler have the ability to inhibit S. mutans growth, and the inhibitory effects are mainly attributed to release of BO3(3-) and F(-).

Evolution of resistance to cationic biocides in Streptococcus mutans and Enterococcus faecalis.

OBJECTIVES: The aim of this study was to investigate whether Streptococcus mutans and Enterococcus faecalis develop resistance to the cationic biocides chlorhexidine (CHX), cetylpyridinium chloride (CPC), and 12-methacryloyloxydodecylpyridinium bromide (MDPB). METHODS: The minimum inhibitory concentrations (MICs) of CHX, CPC, and MDPB were assessed after repeated exposure of S. mutans and E. faecalis to these biocides. Cell-surface hydrophobicity and protein expression profiles of bacterial cells were examined to elucidate possible resistance mechanisms. RESULTS: The MIC of CHX against E. faecalis showed constant increases up to 10 passages. No changes in the MICs of CPC and MDPB against E. faecalis were observed. The MICs of CHX, CPC, and MDPB against S. mutans did not increase. The surface hydrophobicity of E. faecalis significantly increased with increasing exposure to CHX and CPC. However, changes in protein expression profiles were only found in CHX-adapted E. faecalis, as evidenced by the emergence of a novel, approximately 19-kDa band following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. CONCLUSIONS: While E. faecalis and S. mutans did not exhibit increased resistance to CPC or MDPB, repeated exposure of E. faecalis to CHX led to resistance. It is likely that the acquisition of resistance is related to an altered protein composition. CLINICAL SIGNIFICANCE: Alkyl pyridinium compounds, such as CPC and MDPB, could have a lower risk to cause adaptation of E. faecalis, which is advantageous compared with CHX.

Development of sustained antimicrobial-release systems using poly(2-hydroxyethyl methacrylate)/trimethylolpropane trimethacrylate hydrogels.

Reconstructive materials with sustained antimicrobial effects could be useful for preventing infectious diseases in an environment containing indigenous bacteria or fungi such as the oral cavity. With the objective of applying a non-biodegradable hydrogel to resin-based materials as a reservoir for water-soluble antimicrobials, novel hydrogels consisting of 2-hydroxyethyl methacrylate (HEMA) and trimethylolpropane trimethacrylate (TMPT) were fabricated. Cetylpyridinium chloride (CPC) was loaded into five hydrogels comprising different ratios of HEMA/TMPT, and their ability to release as well as to be recharged with CPC was examined in vitro. A polyHEMA/TMPT hydrogel comprising 50% HEMA/50% TMPT could be effectively loaded and recharged with CPC by immersion into a CPC solution, demonstrating the longest release of CPC, above the concentration required to inhibit bacteria and fungi. The binding of CPC to the hydrogels was mainly through hydrophobic interaction. Loading of CPC into a hydrogel by mixing CPC powder with the HEMA/TMPT monomer before polymerization resulted in marked extension of the initial CPC-release period. The CPC-pre-mixed hydrogel was confirmed to exhibit antibacterial activity by agar diffusion tests. It is possible to achieve a sustained release system for antimicrobials by pre-mix loading and recharging CPC into a 50% HEMA/50% TMPT hydrogel.