Sai Ma

Effects of a dental adhesive incorporating antibacterial monomer on the growth, adherence and membrane integrity of Streptococcus mutans.

OBJECTIVESnThis study was attempted to incorporate an antibacterial monomer, methacryloxylethyl cetyl dimethyl ammonium chloride (DMAE-CB), into a commercial dental adhesive and to evaluate the antibacterial activity of the DMAE-CB-incorporated adhesive after being cured against Streptococcus mutans.nnnMETHODSnDMAE-CB was incorporated at 3% (w/v) into a clinically used dental adhesive, Single Bond 2. Single Bond 2 without DMAE-CB was served as a negative control; Clearfil Protect Bond, containing an intensively researched antibacterial monomer MDPB, was enrolled as a positive control. The effects of the cured adhesives and their eluents on the growth of S. mutans were determined by film contact test and absorbance measurement, respectively. The effects of the cured adhesives on the adherence and membrane integrity of S. mutans were investigated using confocal laser scanning microscopy (CLSM) in conjunction with fluorescent indicators.nnnRESULTSnCompared with negative control, the cured DMAE-CB-incorporated dental adhesive and positive control were found to exhibit inhibitory effect on the growth of S. mutans (P<0.05), whereas their eluents did not show detectable antibacterial activity. Moreover, the fluorescence analysis of CLSM images demonstrated that the cured DMAE-CB-incorporated adhesive and positive control could hamper the adherence of S. mutans and exert detrimental effect on bacterial membrane integrity (P<0.05).nnnCONCLUSIONSnThe incorporation of DMAE-CB can render dental adhesive with contact antibacterial activity after polymerization via influencing the growth, adherence and membrane integrity of S. mutans.

Anti-biofilm Effect of Dental Adhesive with Cationic Monomer

The incorporation of polymerizable cationic monomers has been attempted to generate dental resinous materials with antibacterial activity. This study tested the hypothesis that a dental adhesive containing a cationic monomer, methacryloxylethyl cetyl dimethyl ammonium chloride (DMAE-CB), would influence biofilm formation and gtf gene expression of Streptococcus mutans. The effect of the photo-polymerized DMAE-CB-incorporated adhesive on in vitro biofilm accumulation was investigated with spectrophotometry and scanning electron microscopy. The relative level of gtf gene expression by Streptococcus mutans in the biofilm was quantified by real-time reverse-transcription polymerase chain-reaction. The DMAE-CB-incorporated adhesive significantly decreased bio-film accumulation on its surface (P < 0.05), and suppressed the expression of gtfB and gtfC of Streptococcus mutans in the biofilm (P < 0.05). The results suggest that the cured DMAE-CB-incorporated adhesive may hamper biofilm accumulation via selective down-regulation of the expression of gtf genes in Streptococcus mutans.

Therapeutic polymers for dental adhesives: Loading resins with bio-active components

OBJECTIVESnMany recent adhesives on the market exhibit reasonable clinical performance. Future innovations in adhesive materials should therefore seek out novel properties rather than simply modifying existing technologies. It is proposed that adhesive materials that are bio-active could contribute to better prognosis of restorative treatments.nnnMETHODSnThis review examines the recent approaches used to achieve therapeutic polymers for dental adhesives by incorporating bio-active components. A strategy to maintain adhesive restorations is the focus of this paper.nnnRESULTSnMajor trials on therapeutic dental adhesives have looked at adding antibacterial activities or remineralization effects. Applications of antibacterial resin monomers based on quaternary ammonium compounds have received much research attention, and the loading of nano-sized bioactive particles or multiple ion-releasing glass fillers have been perceived as advantageous since they are not expected to influence the mechanical properties of the carrier polymer.nnnSIGNIFICANCEnThe therapeutic polymer approaches described here have the potential to provide clinical benefits. However, not many technological applications in this category have been successfully commercialized. Clinical evidence as well as further advancement of these technologies can be a driving force to make these new types of materials clinically available.

Antibacterial activity and bonding ability of an adhesive incorporating an antibacterial monomer DMAE-CB

This study evaluated the antibacterial effect and microtensile bond strength of a resin-based adhesive containing an antibacterial monomer DMAE-CB (methacryloxylethyl cetyl dimethyl ammonium chloride). Cured specimens of 1, 2, and 3% DMAE-CB-containing Single Bond 2 (crosslinking monomer: Bis-GMA, dimethacrylates; functional monomer: HEMA) were prepared, and their antibacterial effects on Streptococcus mutans ATCC 25175 were investigated. Antibacterial property after 0, 30, 90, and 180 days of aging was also tested. Bonding ability of the experimental adhesive incorporating 3% DMAE-CB was evaluated by microtensile bond strength test. The cured experimental adhesive exhibited an inhibitory effect on S. mutans growth, and the adhesive containing 3% DMAE-CB showed higher antibacterial efficiency compared with those incorporating 1 or 2% anibacterial monomer. Antibacterial activities of the specimens lasted for at least 180 days. Microtensile bond strength test revealed that the bonding ability of the experimental adhesive was not significantly adversely affected by the incorporation of DMAE-CB. Therefore, dental adhesives with strong and long-lasting bacteriostatic property could be achieved by incorporating DMAE-CB without negatively influencing bonding ability.

Antibacterial activity and cytotoxicity of two novel cross-linking antibacterial monomers on oral pathogens

OBJECTIVESnThe antibacterial activity and cytotoxicity of two novel cross-linking antibacterial monomers, 2-methacryloxylethyl dodecyl methyl ammonium bromide (MAE-DB) and 2-methacryloxylethyl hexadecyl methyl ammonium bromide (MAE-HB) were tested in this study.nnnDESIGNnThe minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of unpolymerized MAE-DB and MAE-HB against eight strains of oral bacteria were tested using a broth dilution test. Time-kill determinations were performed to examine the kinetics of unpolymerized MAE-DB and MAE-HB against Streptococcus mutans UA159 and Streptococcus sanguinis ATCC6715. Bacterial morphology was observed using a field emission scanning electron microscope (Fe-SEM). The cytotoxicity of unpolymerized two new monomers and Bis-GMA on the human gingival fibroblast cell line H2620 was assessed using a methyl thiazolyl tetrazolium assay.nnnRESULTSnUnpolymerized MAE-DB and MAE-HB showed strong bactericidal activity against oral bacteria. The MBC value of MAE-DB ranged from 12.2 to 24.4μg/ml and the MBC value of MAE-HB ranged from 6.2 to 48.8μg/ml. Time-kill determinations indicated that unpolymerized MAE-DB and MAE-HB had rapid killing effects against S. mutans UA159 and S. sanguinis ATCC6715 at the concentration of 4× MBC. The Fe-SEM observation showed that MAE-DB and MAE-HB could disturb the integrity of bacteria and cause lysis of bacterial cells. The median lethal concentration values on human gingival fibroblast for both monomers were between 10 and 20μg/ml, and greater than that of Bis-GMA.nnnCONCLUSIONSnUnpolymerized MAE-DB and MAE-HB monomers had strong bactericidal activity against eight strains of oral bacteria. Their cytotoxicities were less than that of Bis-GMA.

Quaternary ammonium-based biomedical materials: State-of-the-art, toxicological aspects and antimicrobial resistance

n Abstractn n Microbial infections affect humans worldwide. Many quaternary ammonium compounds have been synthesized that are not only antibacterial, but also possess antifungal, antiviral and anti-matrix metalloproteinase capabilities. Incorporation of quaternary ammonium moieties into polymers represents one of the most promising strategies for preparation of antimicrobial biomaterials. Various polymerization techniques have been employed to prepare antimicrobial surfaces with quaternary ammonium functionalities; in particular, syntheses involving controlled radical polymerization techniques enable precise control over macromolecular structure, order and functionality. Although recent publications report exciting advances in the biomedical field, some of these technological developments have also been accompanied by potential toxicological and antimicrobial resistance challenges. Recent evidenced-based data on the biomedical applications of antimicrobial quaternary ammonium-containing biomaterials that are based on randomized human clinical trials, the golden standard in contemporary medicinal science, are included in the present review. This should help increase visibility, stimulate debates and spur conversations within a wider scientific community on the implications and plausibility for future developments of quaternary ammonium-based antimicrobial biomaterials.n n

The bonding property and cytotoxicity of a dental adhesive incorporating a new antibacterial monomer

The incorporation of polymerisable cationic monomers has been attempted to generate dental resinous materials with antibacterial activity. This study tested whether the incorporation of a cationic monomer, methacryloxylethyl cetyl dimethyl ammonium chloride (DMAE-CB), into a commercial dental adhesive would influence the bonding property and biocompatibility of the parental adhesive, and whether DMAE-CB could leach out from the cured modified adhesive. Microtensile bond strengths of DMAE-CB-incorporated adhesive and its parental adhesive to dentin were compared. Dentin discs bonded with modified adhesive were incubated in artificial saliva at three different temperatures for 90 days to obtain eluents. The cytotoxicity of DMAE-CB monomer and adhesive eluents were tested with 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cleavage assay (MTT assay). High-performance liquid chromatography (HPLC) was performed with the eluents of the modified adhesive. The results indicated that the incorporation of DMAE-CB into the dental adhesive did not adversely influence its bonding strength to dentin (P > 0·05). Although DMAE-CB monomer exhibited toxicity against L929 cells at the concentration of 2 μg mL(-1) or higher (P < 0·05), the eluents of DMAE-CB-incorporated adhesive did not show significant influence on cell growth (P > 0·05). Moreover, HPLC analysis detected four substances peak baseline separation for the eluents of modified adhesives, which was identical to the eluent of the parental adhesive, indicating no detectable DMAE-CB monomer leaching even after soaking for 90 days. Those results suggest that DMAE-CB could be incorporated into the dental adhesive stably without compromising the bonding efficiency and biocompatibility of its parental adhesive.

Mechanism of detoxification of the cationic antibacterial monomer 12-methacryloyloxydodecylpyridiniumbromide (MDPB) by N-acetyl cysteine

OBJECTIVESnThe protective effects of N-acetyl cysteine (NAC) against cytotoxicity induced by conventional dental resin monomers have been widely documented. However, its effectiveness to detoxify cationic antibacterial monomers has not yet been elucidated. The aim of the present study was to investigate the possible protective effects of NAC against the cytotoxicity of 12-methacryloyloxydodecylpyridiniumbromide (MDPB) and explore the role of adduct formation in NAC-directed detoxification.nnnMETHODSnThe influences of NAC on the cytotoxicity of MDPB were studied in mouse osteoblast-like MC3T3-E1 cells using the MTT assay. Ultra-performance liquid chromatography (UPLC) and liquid chromatography-mass spectrometry (LC-MS) analysis were performed to investigate the possible chemical reaction between NAC and MDPB.nnnRESULTSnWhile only slight reduction in the cytotoxicity of MDPB by NAC was observed immediately after mixing with MDPB, remarkable protection against MDPB-induced cell death was detected when the mixture was tested after 24h of pre-incubation. UPLC and LC-MS analysis revealed that chemical binding of MDPB and NAC occurred under neutral conditions after 24h of pre-incubation.nnnSIGNIFICANCEnOur findings suggest that NAC reduces the toxicity of the cationic antibacterial monomer MDPB, and adduct formation is partially responsible for the detoxification ability of NAC against MDPB-induced cell damage.

N-Acetyl Cysteine (NAC)-Directed Detoxification of Methacryloxylethyl Cetyl Ammonium Chloride (DMAE-CB).

Methacryloxylethyl cetyl ammonium chloride (DMAE-CB) is a polymerizable antibacterial monomer and has been proved as an effective strategy to achieve bioactive bonding with reliable bacterial inhibitory effects. However, the toxicity of DMAE-CB may hamper its wide application in clinical situations. Thus, this study was designed to investigate the toxicity of DMAE-CB and explore the possible protective effects of N-acetyl cysteine (NAC). High performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) analysis showed that chemical binding of NAC and DMAE-CB occurred in a time dependent manner. Pre-incubation of fourty-eight hours is required for adequate reaction between DMAE-CB and NAC. DMAE-CB reduced human dental pulp cells (hDPCs) viability in a dose-dependent manner. The toxic effects of DMAE-CB were accompanied by increased reactive oxygen species (ROS) level and reduced glutathione (GSH) content. NAC alleviated DMAE-CB-induced oxidative stress. Annexin V/ Propidium Iodide (PI) staining and Hoechst 33342 staining indicated that DMAE-CB induced apoptosis. Collapsed mitochondrial membrane potential (MMP) and activation of caspase-3 were also observed after DMAE-CB treatment. NAC rescued hDPCs from DMAE-CB-induced apoptosis, accompanied by lower level of MMP loss and caspase-3 activity. This study assists to elucidate the mechanism underlying the cytotoxic effects of DMAE-CB and provides theoretical supports for the searching of effective strategies to reduce toxicity of quaternary ammonium dental monomers.

The influences of N-acetyl cysteine (NAC) on the cytotoxicity and mechanical properties of Poly-methylmethacrylate (PMMA)-based dental resin

Objectives. This study aimed to investigate the influences of N-acetyl cysteine (NAC) on cytotoxicity and mechanical properties of Poly-methylmethacrylate (PMMA) dental resins. Methods. Experimental PMMA resin was prepared by incorporating various concentrations of NAC (0, 0.15, 0.3, 0.6 and 0.9 wt.%). MTT assay was performed to investigate viability of human dental pulp cells after exposure to extract of PMMA resin with or without NAC. Cell adhesion on resin specimens was examined with scanning electron microscopy. Degree of conversion was studied with Fourier Transform Infrared Spectroscopy (FTIR). Flexural strength, microhardness and surface roughness was evaluated using a universal testing machine, microhardness tester and optical profilometer, respectively. Results. Incorporation of NAC into PMMA resin significantly reduced its cytotoxicity and enhanced cell adhesion on its surface. NAC induced negative influences on the mechanical and physical properties of PMMA resin in a dose-dependent manner. The degree of conversion for all experimental PMMA resins reached as high as 72% after 24 h of polymerization. All the tested properties were maintained when the concentration of incorporated NAC was 0.15 wt.%. Conclusion. The addition of 0.15 wt.% NAC remarkably improved biocompatibility of PMMA resin without exerting significant negative influence on its mechanical and physical properties.