Yuxing Bai

Effect of quaternary ammonium and silver nanoparticle-containing adhesives on dentin bond strength and dental plaque microcosm biofilms

OBJECTIVE Antibacterial bonding agents are promising to hinder the residual and invading bacteria at the tooth-restoration interfaces. The objectives of this study were to develop an antibacterial bonding agent by incorporation of quaternary ammonium dimethacrylate (QADM) and nanoparticles of silver (NAg), and to investigate the effect of QADM-NAg adhesive and primer on dentin bond strength and plaque microcosm biofilm response for the first time. METHODS Scotchbond Multi-Purpose adhesive and primer were used as control. Experimental adhesive and primer were made by adding QADM and NAg into control adhesive and primer. Human dentin shear bond strengths were measured (n = 10). A dental plaque microcosm biofilm model with human saliva as inoculum was used to investigate biofilm metabolic activity, colony-forming unit (CFU) counts, lactic acid production, and live/dead staining assay (n = 6). RESULTS Adding QADM and NAg into adhesive and primer did not compromise the dentin shear bond strength which ranged from 30 to 35MPa (p>0.1). Scanning electron microscopy (SEM) examinations revealed numerous resin tags, which were similar for the control and the QADM and NAg groups. Adding QADM or NAg markedly reduced the biofilm viability, compared to adhesive control. QADM and NAg together in the adhesive had a much stronger antibacterial effect than using each agent alone (p<0.05). Adding QADM and NAg in both adhesive and primer had the strongest antibacterial activity, reducing metabolic activity, CFU, and lactic acid by an order of magnitude, compared to control. SIGNIFICANCE Without compromising dentin bond strength and resin tag formation, the QADM and NAg containing adhesive and primer achieved strong antibacterial effects against microcosm biofilms for the first time. QADM-NAg adhesive and primer are promising to combat residual bacteria in tooth cavity and invading bacteria at the margins, thereby to inhibit secondary caries. QADM and NAg incorporation may have a wide applicability to other dental bonding systems.

Effect of water-ageing on dentine bond strength and anti-biofilm activity of bonding agent containing new monomer dimethylaminododecyl methacrylate

OBJECTIVES The objectives of this study were to develop bonding agent containing a new antibacterial monomer dimethylaminododecyl methacrylate (DMADDM) as well as nanoparticles of silver (NAg) and nanoparticles of amorphous calcium phosphate (NACP), and to investigate the effects of water-ageing for 6 months on dentine bond strength and anti-biofilm properties for the first time. METHODS Four bonding agents were tested: Scotchbond Multi-Purpose (SBMP) Primer and Adhesive control; SBMP+5% DMADDM; SBMP+5% DMADDM+0.1% NAg; and SBMP+5% DMADDM+0.1% NAg with 20% NACP in adhesive. Specimens were water-aged for 1d and 6 months at 37°C. Then the dentine shear bond strengths were measured. A dental plaque microcosm biofilm model was used to inoculate bacteria on water-aged specimens and to measure metabolic activity, colony-forming units (CFUs), and lactic acid production. RESULTS Dentine bond strength showed a 35% loss in 6 months of water-ageing for SBMP control (mean±sd; n=10); in contrast, the new antibacterial bonding agents showed no strength loss. The DMADDM-NAg-NACP containing bonding agent imparted a strong antibacterial effect by greatly reducing biofilm viability, metabolic activity and acid production. The biofilm CFU was reduced by more than two orders of magnitude, compared to SBMP control. Furthermore, the DMADDM-NAg-NACP bonding agent exhibited a long-term antibacterial performance, with no significant difference between 1d and 6 months (p>0.1). CONCLUSIONS Incorporating DMADDM-NAg-NACP in bonding agent yielded potent and long-lasting antibacterial properties, and much stronger bond strength after 6 months of water-ageing than a commercial control. The new antibacterial bonding agent is promising to inhibit biofilms and caries at the margins. The method of DMADDM-NAg-NACP incorporation may have a wide applicability to other adhesives, cements and composites.

Casein phosphopeptide–amorphous calcium phosphate remineralization of primary teeth early enamel lesions

UNLABELLED Early childhood caries (ECC) is a serious problem that progresses rapidly and often goes untreated. Current traumatic treatments may be replaced by safe and effective remineralization at very early stages. OBJECTIVE The aim of this in vitro study was to evaluate the remineralization effects of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) paste on enamel lesions by assessing ultrastructure, nanomechanical properties, and compound and elemental analysis. METHODS Enamel specimens from 6-year-old children were divided into groups: (1) native enamel; (2) water as negative control; (3) 500ppm NaF as positive control; and (4-7) CPP-ACP paste for 4, 8, 12, and 24h, as test groups. Ultrastructure and roughness were observed by atomic force microscopy (AFM); nanohardness and elastic modulus were measured by nanoindentation; compound and crystal size of enamel surface patterns were investigated by X-ray diffractometer (XRD). An electron microprobe (EPMA) was used for element analysis. Data were analyzed using one-way ANOVA. RESULTS The CPP-ACP paste repaired the microstructure of enamel, including prism and interprism, through significantly increased hydroxyapatite crystal size (12.06±0.21nm) and Ca/P molar ratios (1.637±0.096) as compared with NaF (8.56±0.13nm crystal size and 1.397±0.086 Ca/P, p<0.01). Both CPP-ACP and NaF decrease roughness, and increase the nanohardness and elastic modulus, with no significant differences between the materials. CONCLUSIONS The CPP-ACP paste is more suitable for children than NaF, due to advantages for remineralization. The AFM, nanoindentation, EPMA, and XRD are very helpful methods for further understanding of microscale and nanoscale remineralization mechanisms.

Development of novel dental adhesive with double benefits of protein-repellent and antibacterial capabilities

OBJECTIVES Secondary caries at the tooth-restoration margins remains a main reason for restoration failure. The objectives of this study were to: (1) combine protein-repellent 2-methacryloyloxyethyl phosphorylcholine (MPC) with quaternary ammonium dimethylaminohexadecyl methacrylate (DMAHDM) to develop a new dental adhesive with double benefits of protein-repellent and antibacterial capabilities for the first time; and (2) investigate the effects on protein adsorption, anti-biofilm activity, and dentin bond strength. METHODS MPC and DMAHDM were incorporated into Scotchbond Multi-Purpose (SBMP) primer and adhesive. Dentin shear bond strengths were measured using extracted human molars. Protein adsorption onto the adhesive resin surfaces was determined by the micro bicinchoninic acid (BCA) method. A dental plaque microcosm biofilm model with human saliva as inoculum was used to investigate biofilm metabolic activity, colony-forming unit (CFU) counts, lactic acid production and live/dead staining of biofilms on resins. RESULTS Incorporation of 7.5% MPC and 5% DMAHDM into primer and adhesive did not adversely affect the dentin shear bond strength (p>0.1). The resin with 7.5% MPC+5% DMAHDM had protein adsorption that was nearly 20-fold less than SBMP control (p<0.05). The resin with 7.5% MPC+5% DMAHDM had much stronger antibacterial effects than using MPC or DMAHDM alone (p<0.05). Biofilm CFU counts on the resin with 7.5% MPC+5% DMAHDM were reduced by more than 4 orders of magnitude, compared to SBMP control. SIGNIFICANCE The use of double agents (protein-repellent MPC+antibacterial DMAHDM) in dental adhesive achieved much stronger inhibition of biofilms than using each agent alone. The novel protein-repellent and antibacterial bonding agent is promising to reduce biofilm/plaque buildup and reduce recurrent caries at the tooth-restoration margins.

Protein-repellent and antibacterial dental composite to inhibit biofilms and caries

OBJECTIVES Biofilm acids contribute to secondary caries, which is a main reason for dental restoration failures. The objectives of this study were to: (1) develop a protein-repellent and antibacterial composite, and (2) investigate the effects of combining 2-methacryloyloxyethyl phosphorylcholine (MPC) with quaternary ammonium dimethylaminohexadecyl methacrylate (DMAHDM) on composite mechanical properties and biofilm response for the first time. METHODS MPC, DMAHDM and glass particles were mixed into a dental resin composite. Mechanical properties were measured in three-point flexure. Protein adsorption onto the composites was measured by a micro bicinchoninic acid method. A human saliva microcosm model was used to grow biofilms on composites. Colony-forming unit (CFU) counts, live/dead assay, metabolic activity, and lactic acid production of biofilms were determined. RESULTS Incorporation of 3% MPC and 1.5% DMAHDM into composite achieved protein-repellent and antibacterial capabilities without compromising the mechanical properties. Composite with 3% MPC+1.5% DMAHDM had protein adsorption that was 1/10 that of a commercial composite (p<0.05). The composite with 3% MPC+1.5% DMAHDM had much greater reduction in biofilm growth than using MPC or DMAHDM alone (p<0.05). Biofilm CFU counts on composite with 3% MPC+1.5% DMAHDM were more than three orders of magnitude lower than that of commercial control. CONCLUSIONS Dental composite with a combination of strong protein-repellent and antibacterial capabilities was developed for the first time. Composite containing MPC and DMAHDM greatly reduced biofilm growth and lactic acid production, without compromising mechanical properties of the composite. CLINICAL SIGNIFICANCE Novel composite with MPC and DMAHDM greatly reduced biofilm activity and is promising to inhibit secondary caries. The dual agents of MPC plus DMAHDM may have wide applicability to other dental materials.

Development of a multifunctional adhesive system for prevention of root caries and secondary caries

OBJECTIVES The objectives of this study were to: (1) develop a novel adhesive for prevention of tooth root caries and secondary caries by possessing a combination of protein-repellent, antibacterial, and remineralization capabilities for the first time; and (2) investigate the effects of 2-methacryloyloxyethyl phosphorylcholine (MPC), dimethylaminohexadecyl methacrylate (DMAHDM), and nanoparticles of amorphous calcium phosphate (NACP) on dentin bond strength, protein-repellent properties, and dental plaque microcosm biofilm response. METHODS MPC, DMAHDM and NACP were added into Scotchbond Multi-Purpose primer and adhesive. Dentin shear bond strengths were measured. Adhesive coating thickness, surface texture and dentin-adhesive interfacial structure were examined. Protein adsorption onto adhesive resin surface was determined by the micro bicinchoninic acid method. A human saliva microcosm biofilm model was used to investigate biofilm metabolic activity, colony-forming unit (CFU) counts, and lactic acid production. RESULTS The resin with 7.5% MPC+5% DMAHDM+30% NACP did not adversely affect dentin shear bond strength (p>0.1). The resin with 7.5% MPC+5% DMAHDM+30% NACP produced a coating on root dentin with a thickness of approximately 70μm and completely sealed all the dentinal tubules. The resin with 7.5% MPC+5% DMAHDM+30% NACP had 95% reduction in protein adsorption, compared to SBMP control (p<0.05). The resin with 7.5% MPC+5% DMAHDM+30% NACP was strongly antibacterial, with biofilm CFU being four orders of magnitude lower than that of SBMP control. Significance The novel multifunctional adhesive with strong protein-repellent, antibacterial and remineralization properties is promising to coat tooth roots to prevent root caries and secondary caries. The combined use of MPC, DMAHDM and NACP may have wide applicability to bonding agents, cements, sealants and composites to inhibit caries.

Antibacterial and protein-repellent orthodontic cement to combat biofilms and white spot lesions

OBJECTIVES White spot lesions are the most undesired side-effect of fixed orthodontic treatments. The objectives of this study were to combine nanoparticles of silver (NAg) with 2-methacryloyloxyethyl phosphorylcholine (MPC) to develop a modified resin-modified glass ionomer cement (RMGI) as orthodontic cement with double benefits of antibacterial and protein-repellent capabilities for the first time. METHODS NAg and MPC were incorporated into a commercial RMGI. Another commercial orthodontic adhesive also served as control. Enamel shear bond strengths (SBS) were determined. Protein adsorption was measured via a micro bicinchoninic acid method. A dental plaque microcosm biofilm model with human saliva as inoculum was tested. Biofilms adherent on the cement samples and planktonic bacteria in the culture medium away from the cement surfaces were both evaluated for bacterial metabolic activity, colony-forming units (CFU), and lactic acid production. RESULTS Adding 0.1% NAg and 3% MPC to RMGI, and water-aging for 30 days, did not adversely affect the SBS, compared to the unmodified RMGI control (p>0.1). The modified RMGI containing 0.1% NAg and 3% MPC achieved the greatest reduction in protein adsorption, bacterial adhesion, CFU, metabolic activity and lactic acid production. The RMGI containing 0.1% NAg and 3% MPC inhibited not only the bacteria on its surface, but also the bacteria away from the surface in the culture medium. CONCLUSIONS The incorporation of double agents (antibacterial NAg+protein-repellent MPC) into RMGI achieved much stronger inhibition of biofilms than using each agent alone. The novel antibacterial and protein-repellent RMGI with substantially-reduced biofilm acids is promising as an orthodontic cement to combat white spot lesions in enamel.

Novel protein-repellent dental adhesive containing 2-methacryloyloxyethyl phosphorylcholine

OBJECTIVES Biofilms at tooth-restoration margins can produce acids and cause secondary caries. A protein-repellent adhesive resin can potentially inhibit bacteria attachment and biofilm growth. However, there has been no report on protein-repellent dental resins. The objectives of this study were to develop a protein-repellent bonding agent incorporating 2-methacryloyloxyethyl phosphorylcholine (MPC), and to investigate its resistance to protein adsorption and biofilm growth for the first time. METHODS MPC was incorporated into Scotchbond Multi-Purpose (SBMP) at 0%, 3.75%, 7.5%, 11.25%, and 15% by mass. Extracted human teeth were used to measure dentine shear bond strengths. Protein adsorption onto resins was determined by a micro bicinchoninic acid (BCA) method. A dental plaque microcosm biofilm model with human saliva as inoculum was used to measure biofilm metabolic activity and colony-forming unit (CFU) counts. RESULTS Adding 7.5% MPC into primer and adhesive did not decrease the dentine bond strength, compared to control (p>0.1). Incorporation of 7.5% of MPC achieved the lowest protein adsorption, which was 20-fold less than that of control. Incorporation of 7.5% of MPC greatly reduced bacterial adhesion, yielding biofilm total microorganism, total streptococci, and mutans streptococci CFU that were an order of magnitude less than control. CONCLUSIONS A protein-repellent dental adhesive resin was developed for the first time. Incorporation of MPC into primer and adhesive at 7.5% by mass greatly reduced the protein adsorption and bacterial adhesion, without compromising the dentine bond strength. CLINICAL SIGNIFICANCE The novel protein-repellent primer and adhesive are promising to inhibit biofilm formation and acid production, to protect the tooth-restoration margins and prevent secondary caries.

Effects of quaternary ammonium chain length on the antibacterial and remineralizing effects of a calcium phosphate nanocomposite

Composites containing nanoparticles of amorphous calcium phosphate (NACP) remineralize tooth lesions and inhibit caries. A recent study synthesized quaternary ammonium methacrylates (QAMs) with chain lengths (CLs) of 3–18 and determined their effects on a bonding agent. This study aimed to incorporate these QAMs into NACP nanocomposites for the first time to simultaneously endow the material with antibacterial and remineralizing capabilities and to investigate the effects of the CL on the mechanical and biofilm properties. Five QAMs were synthesized: DMAPM (CL3), DMAHM (CL6), DMADDM (CL12), DMAHDM (CL16), and DMAODM (CL18). Each QAM was incorporated into a composite containing 20% NACP and 50% glass fillers. A dental plaque microcosm biofilm model was used to evaluate the antibacterial activity. The flexural strength and elastic modulus of nanocomposites with QAMs matched those of a commercial control composite (n = 6; P > 0.1). Increasing the CL from 3 to 16 greatly enhanced the antibacterial activity of the NACP nanocomposite (P < 0.05); further increasing the CL to 18 decreased the antibacterial potency. The NACP nanocomposite with a CL of 16 exhibited biofilm metabolic activity and acid production that were 10-fold lesser than those of the control composite. The NACP nanocomposite with a CL of 16 produced 2-log decreases in the colony-forming units (CFU) of total microorganisms, total streptococci, and mutans streptococci. In conclusion, QAMs with CLs of 3–18 were synthesized and incorporated into an NACP nanocomposite for the first time to simultaneously endow the material with antibacterial and remineralization capabilities. Increasing the CL reduced the metabolic activity and acid production of biofilms and caused a 2-log decrease in CFU without compromising the mechanical properties. Nanocomposites exhibiting strong anti-biofilm activity, remineralization effects, and mechanical properties are promising materials for tooth restorations that inhibit caries.

A novel protein-repellent dental composite containing 2-methacryloyloxyethyl phosphorylcholine

Secondary caries due to biofilm acids is a primary cause of dental composite restoration failure. To date, there have been no reports of dental composites that can repel protein adsorption and inhibit bacteria attachment. The objectives of this study were to develop a protein-repellent dental composite by incorporating 2-methacryloyloxyethyl phosphorylcholine (MPC) and to investigate for the first time the effects of MPC mass fraction on protein adsorption, bacteria attachment, biofilm growth, and mechanical properties. Composites were synthesized with 0 (control), 0.75%, 1.5%, 2.25%, 3%, 4.5% and 6% of MPC by mass. A commercial composite was also tested as a control. Mechanical properties were measured in three-point flexure. Protein adsorption onto the composite was determined by the microbicinchoninic acid method. A human saliva microcosm biofilm model was used. Early attachment at 4 h, biofilm at 2 days, live/dead staining and colony-forming units (CFUs) of biofilms grown on the composites were investigated. Composites with MPC of up to 3% had mechanical properties similar to those without MPC and those of the commercial control, whereas 4.5% and 6% MPC decreased the mechanical properties (P<0.05). Increasing MPC from 0 to 3% reduced the protein adsorption on composites (P<0.05). The composite with 3% MPC had protein adsorption that was 1/12 that of the control (P<0.05). Oral bacteria early attachment and biofilm growth were also greatly reduced on the composite with 3% MPC, compared to the control (P<0.05). In conclusion, incorporation of MPC into composites at 3% greatly reduced protein adsorption, bacteria attachment and biofilm CFUs, without compromising mechanical properties. Protein-repellent composites could help to repel bacteria attachment and plaque build-up to reduce secondary caries. The protein-repellent method might be applicable to other dental materials.