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Implant-related infections are a primary reason for implant failures that affect millions of patients. It is of paramount importance to develop novel implants that possess the dual functions of osteogenesis-promotion and antibacterial activity. The objectives of this study were to: (1) develop novel dual-functional titanium (Ti) implants by combining oxygen non-thermal plasma and covalent bonding of antibacterial organosilicon quaternary ammonium monomers; (2) investigate the physicochemical properties, bioactivity and antibacterial effects of the modified implants for the first time.Surface characteristics of the modified Ti surfaces were tested. Adherence and viability of rat bone marrow-derived stem cells (rBMSCs) on the surface were evaluated. Metabolic activity of biofilm on the surfaces were measured. The stability of the dual-function after 5000 thermal cycles was also evaluated.The presence of chemical bonding between Ti and organosilicon monomers demonstrated covalent immobilization of the antibacterial agents. The water contact angle of the treated Ti surfaces decreased from 70.98 ± 3.68° to 59.86 ± 4.91°. The adhesion and proliferation of rBMSCs on the modified Ti were increased by 40%, compared to control group (P 0.05). The metabolic level of biofilms on modified Ti were reduced by more than half, compared to control (P 0.05). The modified Ti implants exhibited cell-promotion and antibacterial stability after thermal cycles.The new dual-functional Ti implant is promising to promote osteogenesis while simultaneously preventing infections. Furthermore, the novel surface modification and processing methods have applicability to enhancing a wide range of other implants to improve bioactivity and combat infections.

Peri-implantitis are a major problem causing implant failure these days. Accordingly, anti-infection during the early stage and subsequent promotion of osseointegration are two main key factors to solve this issue. Micro-arc oxidation (MAO) treatment is a way to form an oxidation film on the surface of metallic materials. The method shows good osteogenic properties but weak antibacterial effect. Therefore, we developed combined strategies to combat severe peri-implantitis, which included the use of a novel compound, PD, comprising dendrimers poly(amidoamine) (PAMAM) loading dimethylaminododecyl methacrylate (DMADDM) as well as MAO treatment. Here, we explored the chemical properties of the novel compound PD, and proved that this compound was successfully synthesized, with the loading efficiency and encapsulation efficiency of 23.91% and 31.42%, respectively. We further report the two-stage double benefits capability of PD + MAO: (1) in the first stage, PD + MAO could decrease the adherence and development of biofilms by releasing DMADDM in the highly infected first stage after implant surgery both in vitro and in vivo; (2) in the second stage, PD + MAO indicated mighty anti-infection and osteoconductive characteristics in a rat model of peri-implantitis in vivo. This study first reports the two-staged, double benefits of PD + MAO, and demonstrates its potential in clinical applications for inhibiting peri-implantitis, especially in patients with severe infection risk., Graphical abstract Image 1, Highlights • PAMAM-DMADDM shows toxicity reduction and osteoconductive properties in vitro. • Micro-arc oxidation treatment shows osteoconductive but weak antibacterial effect. • PAMAM-DMADDM + Micro-arc oxidation treatment shows multifunctional properties.

Background Enterococcus faecalis (E. faecalis) is the most recovered species from the root canals after failed root canal treatment. Calcium phosphate bone cement (CPC) scaffold is promising for applications in endodontic treatment as a kind of root canal sealer. Graphene oxide (GO) has been extensively considered as a kind of promising nano-materials for antibacterial applications. In the present study, an injectable CPC-chitosan paste containing GO was developed for promising endodontic therapy. The antibacterial properties of this paste against E. faecalis biofilms as well as the support for human dental pulp stem cells (hDPSCs) were investigated. Methods CPC-chitosan composite with or without GO injectable scaffold was fabricated. The hDPSC growth and viability on scaffolds were investigated by live/dead assay. Antibacterial effects against E. faecalis biofilms were determined in clinical detin block samples. Results The antibacterial CPC-chitosan-GO disks had excellent hDPSC support with the percentages of live cells at around 90%. CPC-chitosan-GO also had greater antibacterial activity on E. faecalis than that of CPC-chitosan control using detin block models (p Conclusions The injectable CPC-chitosan-GO paste had strong effects on inhibition E. faecalis and hDPSC support, which could fill the void of adjusting paste to the defect and shaping in situ for promising endodontic therapy.

The limited durability of dentin bonding harshly shortens the lifespan of resin composites restorations. The controlled, dynamic movement of materials through non-contacting forces provides exciting opportunities in adhesive dentistry. We, herein, describe comprehensive investigations of a new dental adhesive with superparamagnetic iron oxide nanoparticles (SPIONs) sensitive to magnetic fields for bonding optimization. This contribution outlines a roadmap of (1) designing and tuning of an adhesive formulation containing SPIONs to enhance penetrability into etched dentin guided by magnetic-field; (2) employing a clinically relevant model of simulated hydrostatic pulpal pressure on the microtensile bond to dentin; and (3) investigating a potential antibacterial effect of the formulated adhesives, and their biocompatibility. SPION-concentration-dependency chemical and mechanical behavior was shown via the degree of conversion, ultimate tensile strength, and micro shear bond strength to dentin. The effects of SPIONs carried on a dental adhesive on the bonding strength to dentin are studied in depth by combining experiments with in vitro simulated model. The results show that under the guided magnetic field, 0.07 wt.% of SPIONs-doped adhesive increased the bond strength that surpasses the reduction caused by hydrostatic pulpal pressure. Using a magnetic guide workflow during the bonding procedures, SPIONs-doped adhesives improved dentin's adhesion without changing adhesives’ physicochemical properties. This outcome addresses the key challenge of poor resin infiltration of dentin's conventional total etching during the bonding procedure. The real-time magnetic motion of dental adhesives may open new paths to enhance resin-based restorations’ longevity. Statement of significance In this study, dental adhesives containing superparamagnetic iron oxide nanoparticles (SPIONs) were developed to enhance penetrability into dentin guided by a magnetic field. The adhesives were screened for physical, chemical, antibacterial properties, and cytotoxicity. For the first time, simulated pulpal pressure was used concurrently with the magnetic field to simulate a clinical setting. This approach showed that it is feasible to overcome pulpal pressure jeopardization on bond strength when SPIONs and a magnetic field are applied. The magnetic-responsive adhesives had great potential to improve bond strength, opening new paths to enhance resin-based restorations' longevity without affecting adhesives' biological properties. The use of magnetic-responsive particles and magnetically assisted motion is a promising strategy to improve the sealing ability of dental adhesives.

Cervical composites treating root carious and noncarious cervical lesions usually extend subgingivally. The subgingival margins of composites present poor plaque control, enhanced biofilm accumulation, and cause gingival irritation. A potential material to restore such lesions should combine agents that interfere with bacterial biofilm development and respond to acidic conditions. Here, we explore the use of new bioresponsive bifunctional dental composites against mature microcosm biofilms derived from subgingival plaque samples. The designed formulations contain 2 bioactive agents: dimethylaminohexadecyl methacrylate (DMAHDM) at 3 to 5 wt.% and 20 wt.% nanosized amorphous calcium phosphate (NACP) in a base resin. Composites with no DMAHDM and NACP were used as controls. The newly formulated 5% DMAHDM–20% NACP composite was analyzed by micro-Raman spectroscopy and transmission electron microscopy. The wettability and surface-free energy were also assessed. The inhibitory effect on the in vitro biofilm growth and the 16S rRNA gene sequencing of survival bacterial colonies derived from the composites were analyzed. Whole-biofilm metabolic activity, polysaccharide production, and live/dead images of the biofilm grown over the composites complement the microbiological assays. Overall, the designed formulations had higher contact angles with water and lower surface-free energy compared to the commercial control. The DMAHDM-NACP composites significantly inhibited the growth of total microorganisms, Porphyromonas gingivalis, Prevotella intermedia/nigrescens, Aggregatibacter actinomycetemcomitans, and Fusobacterium nucleatum by 3 to 5-log ( P < 0.001). For the colony isolates from control composites, the composition was typically dominated by the genera Veillonella, Fusobacterium, Streptococcus, Eikenella, and Leptotrichia, while Fusobacterium and Veillonella dominated the 5% DMAHDM–20% NACP composites. The DMAHDM-NACP composites contributed to over 80% of reduction in metabolic and polysaccharide activity. The suppression effect on plaque biofilms suggested that DMAHDM-NACP composites might be used as a bioactive material for cervical restorations. These results may propose an exciting path to prevent biofilm growth and improve dental composite restorations’ life span.

Objective Fluoride (F)-releasing restoratives typically are either weak mechanically or release only low levels of F ions. The objectives of this study were to: (1) develop a novel photo-cured nanocomposite with strong mechanical properties and high levels of sustained F ion release via a two-step “co-precipitation + spray-drying” technique to synthesize CaF2 nanoparticles (nCaF2); and (2) investigate the effect of spray-drying treatment after co-precipitation of nCaF2 on mechanical properties and F ion release of composite. Methods Two types of CaF2 particles were synthesized: A co-precipitation method yielded CaF2cp; “co-precipitation + spray-drying” yielded nCaF2cpsd. Composites were fabricated with fillers of: (1) 0% CaF2 + 70% glass; (2) 10% CaF2cp + 60% glass; (3) 15% CaF2cp + 55% glass; (4) 20% CaF2cp + 50% glass; (5) 10% nCaF2cpsd + 60% glass; (6) 15% nCaF2cpsd + 55% glass; and (7) 20% nCaF2cpsd + 50% glass. A commercial F-releasing nanocomposite served as control. Results The nCaF2cpsd had much smaller particle size (median = 32 nm) and narrower distribution (22–57 nm) than CaF2cp (median = 5.25 μm, 162 nm–67 μm). The composite containing nCaF2cpsd had greater flowability, flexural strength, elastic modulus and hardness than CaF2cp composite and commercial control composite. At 84-day immersion in water, the nanocomposites containing 20% nCaF2cpsd had 65 times higher cumulative F release, and 77 times greater long-term F-release rate, than commercial control. Conclusions A novel two-step “co-precipitation + spray-drying” technique of synthesizing nCaF2 was developed. The photo-cured nanocomposite containing 20% nCaF2cpsd possessed strong mechanical properties and excellent long-term F-release ability, and hence is promising for dental restoration applications to inhibit secondary caries.

Dental caries is closely associated with acid-producing bacteria, and Streptococcus mutans is one of the primary etiological agents. Bacterial accumulation and dental demineralization lead to destruction of bonding interface, thus limiting the longevity of composite. The present study investigated remineralization effectiveness of adhesive containing nanoparticles of amorphous calcium phosphate (NACP) in a stimulated oral biofilm environment. The enamel blocks were immersed in demineralization solution for 72 h to imitate artificial initial carious lesion and then subjected to a Streptococcus mutans biofilm for 24 h. All the samples then underwent 4-h demineralization in brain heart infusion broth with sucrose (BHIS) and 20-h remineralization in artificial saliva (AS) for 7 days. The daily pH of BHIS after 4-h incubation, lactic acid production, colony-forming unit (CFU) count, and content of calcium (Ca) and phosphate (P) in biofilm were evaluated. Meanwhile, the remineralization effectiveness of enamel was analyzed by X-ray diffraction (XRD), surface microhardness testing, transverse microradiography (TMR) and scanning electron microscopy (SEM). The NACP adhesive released abundant Ca and P, achieved acid neutralization, reduced lactic acid production, and lowered CFU count (P < 0.05). Enamel treated with NACP adhesive demonstrated the best remineralization effectiveness with remineralization value of 52.29 ± 4.79% according to TMR. Better microhardness recovery of cross sections and ample mineral deposits were also observed in NACP group. The NACP adhesive exhibited good performance in remineralizing initial enamel lesion with cariogenic biofilm. The NACP adhesive is promising to be applied for the protection of bonding interface, prevention of secondary caries, and longevity prolonging of the restoration.

The objectives were to: (1) develop a new nanostructured dental adhesive with antibiofilm and remineralization properties using dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of calcium fluoride (nCaF2); (2) investigate the effects on dentin bond strength and Streptococcus mutans biofilms. Bisphenol A glycidyl dimethacrylate, triethylene glycol dimethacrylate, 2-hydroxyethyl methacrylate, and methacryloyloxy ethyl phthalate formed the adhesive. Biofilm colony-forming units (CFU), metabolic activities, biomass, and acid-production were measured. Adding nCaF2 and DMAHDM into adhesive did not compromise the dentin bond strength (p > 0.1). The nCaF2 + DMAHDM-containing adhesive decreased biofilm CFU by 4 logs, with tenfold reduction in acid-production, compared to control (p

Staphylococcus aureus (S. aureus) is a major pathogen for osteomyelitis. Calcium phosphate bone cement (CPC) paste is promising for orthopedic uses. Nanostructured graphene oxide (GO) showed antibacterial effect on Gram-positive bacteria. However, there has been no report of incorporating GO into CPC. The objectives of this study were to (a) develop an injectable and mechanically strong CPC-chitosan paste containing GO and (b) investigate the inhibition of S. aureus infection and the promotion of human umbilical cord mesenchymal stem cells (hUCMSCs) for bone regeneration. Injectable CPC-chitosan-GO paste was fabricated. Flexural strength, elastic modulus, and work-of-fracture of the CPC-chitosan and CPC-chitosan-GO bars were evaluated. Antibacterial effects against S. aureus biofilms were determined. hUCMSC growth and viability on disks were investigated. CPC-chitosan-GO bars had a flexural strength of 7.2 ± 1.6 MPa, matching that of CPC-chitosan control without GO. CPC-chitosan-GO had strong antibacterial effects on S. aureus, with an inhibition zone of 55.2 ± 2.5 mm, greater than that of CPC-chitosan control (30.1 ± 2.0 mm) (p0.05). CPC-chitosan-GO had potent antibacterial activity on S. aureus biofilms in vitro (p0.05). The injectable and antibacterial CPC-GO paste had no toxic effect, yielding excellent hUCMSC growth and viability on disks. The CPC-chitosan-GO had injectability, good strength, strong antibacterial effects, and excellent stem cell attachment and growth. CPC-chitosan-GO is promising for dental, craniofacial, and orthopedic applications to control infections and good biocompatibility to support stem cell viability to enhance bone regeneration.

Recurrent caries is a primary reason for restoration failure caused by biofilm acids. The objectives of this study were to: (1) develop a novel multifunctional composite with antibacterial function and calcium (Ca) and phosphate (P) ion release, and (2) investigate the effects on enamel demineralization and hardness at the margins under biofilms.Dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP) were incorporated into composite. Four groups were tested: (1) Commercial control (Heliomolar), (2) Experimental control (0% DMAHDM + 0% NACP), (3) antibacterial group (3% DMAHDM + 0% NACP), (D) antibacterial and remineralizing group (3% DMAHDM + 30% NACP). Mechanical properties and Ca and P ion release were measured. Colony-forming units (CFU), lactic acid and polysaccharide of Streptococcus mutans (S. mutans) biofilms were evaluated. Demineralization of bovine enamel with restorations was induced via S. mutans, and enamel hardness was measured. Data were analyzed via one-way and two-way analyses of variance and Tukey's multiple comparison tests.Adding DMAHDM and NACP into composite did not compromise the mechanical properties (P0.05). Ca and P ion release of 3% DMAHDM + 30% NACP was increased at cariogenic low pH. Biofilm lactic acid and polysaccharides were greatly decreased via DMAHDM, and CFU was reduced by 4 logs (P0.05). Under biofilm acids, enamel hardness at the margins was decreased to about 0.5 GPa for control; it was about 1 GPa for antibacterial group, and 1.3 GPa for antibacterial and remineralizing group (P0.05).The novel 3% DMAHDM + 30% NACP composite had strong antibacterial effects. It substantially reduced enamel demineralization adjacent to restorations under biofilm acid attacks, yielding enamel hardness that was 2-fold greater than that of control composites. The novel multifunctional composite is promising to inhibit recurrent caries.

Objective Pit and fissure sealants with antibacterial and remineralization properties have broad application prospects in caries prevention. The objectives of this study were to: (1) develop a novel pit and fissure sealant containing CaF2 nanoparticles (nCaF2) and dimethylaminohexadecyl methacrylate (DMAHDM); and (2) investigate the effects of nCaF2 and DMAHDM on biofilm response and fluoride (F) ion release for the first time. Methods Helioseal F was used as a control. Bioactive sealants were formulated with DMAHDM and nCaF2. Flow properties, enamel shear bond strength, hardness and F ion releases were measured. Streptococcus mutans (S. mutans) biofilms were grown on sealants. Biofilm metabolic activity, lactic acid production, colony-forming units (CFU), and pH of biofilm culture medium were measured. Results Adding 5% DMAHDM and 20% nCaF2 did not reduce the paste flow and enamel bond strength, compared to control (p Significance The new bioactive pit and fissure sealant with nCaF2 and DMAHDM achieved high fluoride release and strong antibacterial performance. This novel fluoride-releasing and antibacterial sealant is promising to inhibit caries and promote the remineralizaton of enamel and dentin.

Recurrent dental caries is one of the main reasons for resin composite restoration failures. This study aimed to: (1) develop a bioactive, low-shrinkage-stress, antibacterial and remineralizing composite and evaluate the sustainability of its antibacterial effect against Streptococcus mutans (S. mutans) biofilms; and (2) evaluate the remineralization and cariostatic potential of the composite containing nanoparticles of amorphous calcium phosphate (NACP) and dimethylaminohexadecyl methacrylate (DMAHDM), using dentin hardness measurement and a biofilm-induced recurrent caries model. The antibacterial and remineralizing low-shrinkage-stress composite consisted of urethane dimethacrylate (UDMA) and triethylene glycol divinylbenzyl ether (TEG-DVBE), 3% DMAHDM and 20% NACP. S. mutans biofilm was used to evaluate antibiofilm activity, before and after 3 months of composite aging in acidic solution. Human dentin was used to develop a recurrent caries biofilm-model. Adding DMAHDM and NACP into low shrinkage-stress composite did not compromise the flexural strength. The low-shrinkage-stress composite with DMAHDM achieved substantial reductions in biofilm colony-forming units (CFU), lactic acid production, and biofilm biomass (p0.05). The low-shrinkage-stress DMAHDM+NACP composite exhibited no significant difference in antibacterial performance before and after 3 months of aging, demonstrating long-term antibacterial activity. Under S. mutans biofilm acidic attack, dentin hardness (GPa) was 0.24 ± 0.04 for commercial control, and 0.23 ± 0.03 for experimental control, but significantly higher at 0.34 ± 0.03 for DMAHDM+NACP group (p0.05). At an instrumental compliance of 0.33 μm/N, the polymerization shrinkage stress of the new composite was 36% lower than that of a traditional composite (p0.05). The triple strategy of antibacterial, remineralization and lower shrinkage-stress has great potential to inhibit recurrent caries and increase restoration longevity. Statement of Significance Polymerization shrinkage stress, masticatory load over time as well as biochemical degradation can lead to marginal failure and secondary caries. The present study developed a new low-shrinkage-stress, antibacterial and remineralizing dental nanocomposite. Polymerization shrinkage stress was greatly reduced, biofilm acid production was inhibited, and tooth dentin mineral and hardness were preserved. The antibacterial composite possessed a long-lasting antibiofilm effect against cariogenic bacteria S. mutans. The new bioactive nanocomposite has the potential to suppress recurrent caries at the restoration margins, protects tooth structures, and increases restoration longevity.

Short-term studies on calcium-phosphate (CaP) ion-rechargeable composites were reported. The long-term rechargeability is important but unknown. The objectives of this study were to investigate nanocomposite with strong antibacterial and ion-recharge capabilities containing dimethylaminododecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP), and evaluate long-term ion-recharge by testing for 12 cycles (taking 6 months to complete) for the first time. Three groups were tested: (1) Heliomolar control; (2) Resin+20%NACP+50%glass; (3) Resin+3%DMAHDM+20%NACP+50%glass. Biofilm acid and colony-forming units (CFU) were measured. Ion-recharge was tested for 12 cycles. NACP-DMAHDM composite reduced biofilm acid, and reduced CFU by 4 logs. High levels of ion releases were maintained throughout 12 cycles of recharge, maintaining steady-state releases without reduction in 6 months (p>0.1), representing long-term remineralization potential. Bioactive nanocomposite demonstrated long-term ion-rechargeability for the first time, showed remineralization and potent anti-biofilm functions, with promise for tooth restorations to combat caries.

Titanium (Ti) has achieved extensive applications due to its excellent biocompatibility and mechanical properties. Plasma can enhance surface hydrophilia of Ti with decreased carbon contamination. The traditional conditions using a single gas plasma was for longer treatment time and more prone to being contaminated. We designed and developed novel and universal apparatus and methods with a special clamping device of non-thermal atmospheric plasma (NTAP) treatment using mixed gas for Ti surface activation. We systematically and quantitatively investigated the effective effects of NTAP-Ti. The surface water contact angle decreased by 100%, the carbon content decreased by 80% and oxygen content increased by 50% in the novel NTAP-Ti surfaces. NTAP treatment accelerated the attachment, spread, proliferation, osteogenic differentiation and mineralization of MC3T3-E1 mouse preosteoblasts in vitro. The percentage of bone-to-implant contact increased by 25–40%, and the osteoclasts and bone resorption were suppressed by 50% in NTAP-Ti in vivo. In conclusion, NTAP-Ti substantially enhanced the physical and biological effects and integration with bone. The novel and universal apparatus and methods with a special clamping device using gas mixtures are promising for implant activation by swiftly and effectively changing the Ti surface to a hydrophilic one to enhance dental and orthopedic applications.

Antibacterial quaternary ammonium monomers (QAMs) are used in resins. The rnc gene in Streptococcus mutans (S. mutans) plays a key role in resisting antibiotics. The objectives of this study were to investigate for the first time: (1) the effects of rnc deletion on S. mutans biofilms and acid production; (2) the combined effects of rnc deletion with dimethylaminohexadecyl methacrylate (DMAHDM) on biofilm-inhibition efficacy.Parent S. mutans strain UA159 (ATCC 700610) and the rnc-deleted S. mutans were used. Bacterial growth, minimum inhibitory concentration (MIC), and minimal bactericidal concentration (MBC) were measured to analyze the bacterial susceptibility of the parent and rnc-deleted S. mutans against DMAHDM, with the gold-standard chlorhexidine (CHX) as control. Biofilm biomass, polysaccharide and lactic acid production were measured.The drug-susceptibility of the rnc-deleted S. mutans to DMAHDM or CHX was 2-fold higher than parent S. mutans. The drug-susceptibility did not increase after 10 passages (p0.05). Deleting the rnc gene increased the biofilm susceptibility to DMAHDM or CHX by 2-fold. The rnc-deletion in S. mutans reduced biofilm biomass, polysaccharide and lactic acid production, even at no drugs. DMAHDM was nearly 40 % more potent than the gold-standard CHX. The combination of rnc deletion+DMAHDM treatment achieved the greatest reduction in biofilm biomass, polysaccharide synthesis, and lactic acid production.Gene modification by deleting the rnc in S. mutans reduced the biofilm growth and acid production, and the rnc deletion+DMAHDM method showed the greatest biofilm-inhibition efficacy, for the first time. The dual strategy of antibacterial monomer+bacterial gene modification shows great potential to control biofilms and inhibit caries.

Infection is a main cause of implant failure. Early implant-related infections often occur in the first 4 weeks post-operation. Inhibiting bacterial adhesion and biofilm formation at the early stage and promoting subsequent implant osseointegration are important for implant success. Our previous studies demonstrated that dimethylaminododecyl methacrylate (DMADDM) provided dental materials with antibacterial effects. In the present study, DMADDM and hydroxyapatite (HA) are loaded on to the titanium (Ti) surface via poly dopamine (PDA) self-polymerization. This local DMADDM-delivery Ti is referred as Ti-PHD. Here we report the two-staged capability of Ti-PHD: (1) in the first stage, releasing DMADDM during the high-infection-risk initial period post-implantation for 4 weeks; (2) then in the second stage, enhancing osteogenesis and promoting osseointegration. Ti-PHD has a porous surface with higher average roughness and greater hydrophilicity than pure Ti. Its biocompatibility is verified in vitro and in vivo. During the first 4 weeks of release, both DMADDM remaining on Ti surface and DMADDM released into the soaking medium greatly reduced the adherence and growth of pathogens. This is further confirmed by the prevention of bone destruction in a rat osteomyelitis model. After releasing DMADDM for 4 weeks, Ti-PHD promotes osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) and new bone formation around the implants in vivo. This article represents the first report on the two-staged, time-dependent antibacterial and osteogenesis effects of Ti-PHD, demonstrating its potential for clinical applications to inhibit implant-associated infections. STATEMENT OF SIGNIFICANCE: The present study develops a two-staged time-dependent system for local dimethylaminododecyl methacrylate (DMADDM) delivery via Ti implant (referred to as Ti-PHD). DMADDM and hydroxyapatite (HA) are loaded on to the Ti surface with poly dopamine (PDA). Ti-PHD can release DMADDM during the high-risk period of infection in the first stage, and then promote osseointegration and new bone formation in the second stage. This bioactive and therapeutic Ti is promising to inhibit infections and enhance implant success.

The objectives of this study were to: (1) develop a multifunctional adhesive via dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP); and (2) investigate its ability to provide metalloproteinases (MMPs) deactivation and remineralization for long-term dentin bonding durability.DMAHDM and NACP were incorporated into Adper™ Single Bond 2 Adhesive (SB2) at mass fractions of 5% and 20%, respectively. Degree of conversion and contact angle were measured. Endogenous MMP activity of the demineralized dentin beams, Masson's trichrome staining, nano-indentation, microtensile bond strength and interfacial nanoleakage analyses were investigated after 24 h and 3 months of storage aging in artificial saliva.Adding DMAHDM and NACP did not compromise the degree of conversion and contact angle of SB2 (p 0.05). DMAHDM and NACP incorporation reduced the endogenous MMP activity by 53 %, facilitated remineralization, and increased the Young's modulus of hybrid layer by 49 % after 3 months of aging in artificial saliva, compared to control. For SB2 Control, the dentin bond strength decreased by 38 %, with greater nanoleakage expression, after 3 months of aging (p 0.05). However, DMAHDM+NACP group showed no loss in bond strength, with much less nanoleakage, after 3 months of aging (p 0.05).DMAHDM+NACP adhesive greatly reduced MMP-degradation activity in demineralized dentin, induced remineralization at adhesive-dentin interface, and maintained the dentin bond strength after aging, without adversely affecting polymerization and dentin wettability. This new adhesive has great potential to help eliminate secondary caries, prevent hybrid layer degradation, and increase the resin-dentin bond longevity.

Recent studies developed low-shrinkage-stress composite with remineralizing and antibacterial properties to combat secondary caries and increase restoration longevity. However, their long-term durability in thermal cycling is unclear. The objectives of this study were to develop an antibacterial, remineralizing and low-shrinkage-stress composite, and to investigate its durability in thermal cycling for 20,000 cycles, equivalent to two years of clinical life.The resin consisted of urethane dimethacrylate (UDMA) and triethylene glycol divinylbenzyl ether (TEG-DVBE). Composites were made with 5% dimethylaminohexadecyl methacrylate (DMAHDM) and 20% of nanoparticles of amorphous calcium phosphate (NACP). Composites were thermal cycled at 5°C and 55°C for 20,000 cycles. A human salivary biofilm model was used to evaluate antibiofilm activity before and after thermal cycling.After 20,000 cycles, the flexural strength of bioactive low-shrinkage-stress composite matched commercial control with no antibacterial activity (p 0.05). Surface roughness was clinically acceptable at less than 0.2 μm. UV+NACP+DMAHDM composite reduced the total microorganisms, total streptococci, and mutans streptococci by 2-5 logs, compared to commercial composite. Biofilm lactic acid production was reduced by 11 folds. The antibacterial performance was maintained after thermal cycling, with no decrease after 20,000 cycles.Bioactive low-shrinkage-stress composite possessed good mechanical properties that matched commercial composite both before and after thermal cycling. The new composite had potent antibacterial activity, which was maintained and did not decrease after thermal cycling.The new bioactive low-shrinkage-stress composite could reduce polymerization shrinkage stress and release calcium and phosphate ions, with good mechanical properties and strong antibacterial function that were durable after thermal cycling. These properties indicate great potential for inhibiting recurrent caries and increasing the restoration longevity.

Tissue engineering is promising for dental and craniofacial regeneration. The objectives of this study were to develop a novel xeno-free alginate-fibrin-platelet lysate hydrogel with human periodontal ligament stem cells (hPDLSCs) for dental regeneration, and to investigate the proliferation and osteogenic differentiation of hPDLSCs using hPL as a cell culture nutrient supplement.hPDLSCs were cultured with Dulbecco's modified eagle medium (DMEM), DMEM + 10% fetal bovine serum (FBS), and DMEM + hPL (1%, 2.5%, and 5%). hPDLSCs were encapsulated in alginate-fibrin microbeads (Alg+Fib), alginate-hPL microbeads (Alg+hPL), or alginate-fibrin-hPL microbeads (Alg+Fib+hPL). hPDLSCs encapsulated in alginate microbeads were induced with an osteogenic medium containing hPL or FBS. Quantitative real-time polymerase chain reaction (qRT-PCR), alkaline phosphatase (ALP) activity, ALP staining, and alizarin red (ARS) staining was investigated.hPDLSCs were released faster from Alg+Fib+hPL than from Alg+hPL. At 14 days, ALP activity was 44.1 ± 7.61 mU/mg for Alg+Fib+hPL group, higher than 28.07 ± 5.15 mU/mg of Alg+Fib (p0.05) and 0.95 ± 0.2 mU/mg of control (p0.01). At 7 days, osteogenic genes (ALP, RUNX2, COL1, and OPN) in Alg+Fib+hPL and Alg+Fib were 3-10 folds those of control. At 21 days, the hPDLSC-synthesized bone mineral amount in Alg+Fib+hPL and Alg+Fib was 7.5 folds and 4.3 folds that of control group, respectively.The 2.5% hPL was determined to be optimal for hPDLSCs. Adding hPL into alginate hydrogel improved the viability of the hPDLSCs encapsulated in the microbeads. The hPL-based medium enhanced the osteogenic differentiation of hPDLSCs in Alg+Fib+hPL construct, showing a promising xeno-free approach for delivering hPDLSCs to enhance dental, craniofacial and orthopedic regenerations.

(1) Background: Vascularization remains a critical challenge in bone tissue engineering. The objective of this study was to prevascularize calcium phosphate cement (CPC) scaffold by co-culturing human periodontal ligament stem cells (hPDLSCs) and human umbilical vein endothelial cells (hUVECs) for the first time, (2) Methods: hPDLSCs and/or hUVECs were seeded on CPC scaffolds. Three groups were tested: (i) hUVEC group (hUVECs on CPC), (ii) hPDLSC group (hPDLSCs on CPC), (iii) co-culture group (hPDLSCs + hUVECs on CPC). Osteogenic differentiation, bone mineral synthesis, and microcapillary-like structures were evaluated, (3) Results: Angiogenic gene expressions of co-culture group were 6–9 fold those of monoculture. vWF expression of co-culture group was 3 times lower than hUVEC-monoculture group. Osteogenic expressions of co-culture group were 2–3 folds those of the hPDLSC-monoculture group. ALP activity and bone mineral synthesis of co-culture were much higher than hPDLSC-monoculture group. Co-culture group formed capillary-like structures at 14–21 days. Vessel length and junction numbers increased with time, (4) Conclusions: The hUVECs + hPDLSCs co-culture on CPC scaffold achieved excellent osteogenic and angiogenic capability in vitro for the first time, generating prevascularized networks. The hPDLSCs + hUVECs co-culture had much better osteogenesis and angiogenesis than monoculture. CPC scaffolds prevacularized via hPDLSCs + hUVECs are promising for dental, craniofacial, and orthopedic applications.

The objective of this study was to develop a novel nanostructured resin infiltrant containing nanoparticles of amorphous calcium phosphate (NACP) to treat enamel white spot lesions (WSLs). Physical properties and the therapeutic effect of the new resin infiltrant were investigated for the first time.NACP was incorporated into ICON (Icon caries infiltrant, DMG, Germany) with different mass fractions. Cytotoxicity, degree of conversion, surface hardness, calcium (Ca) and phosphorus (P) ions release concentrations were tested. After application to the demineralized enamel samples, the color changes were determined. Surface and cross-sectional hardness were measured, scanning electron microscopy (SEM) images were taken on the cross-section of samples to observe microstructure changes after 14-day pH cycling.Incorporating 10%-30% of NACP did not compromise the biocompatibility and physical properties of the resin infiltrant. ICON + 30% NACP group had long-lasting and high level of Ca and P ion release. After 14-day pH cycling, enamel surface hardness of ICON + 30% NACP group was 1.83 ± 0.21 GPa, significantly higher than the control group (1.32 ± 0.18 GPa) (p 0.05). ICON + 30NACP group had the highest cross-sectional enamel hardness among all groups (p 0.05), especially at 50 μm and 100 μm depth. SEM images showed that apparent enamel prism and inter-prism gaps in negative control were masked by mineral deposition in ICON + 30% NACP group.The novel ICON+30% NACP infiltrant is promising to inhibit enamel WSLs, protect the enamel and increase its hardness.

The cariogenic biofilm on enamel, restoration, and bonding interface is closely related to dental caries and composite restoration failure. Enamel remineralization at adhesive interface is conducive to protecting bonding interface and inhibiting secondary caries. This study intended to assess the remineralization efficiency of adhesive with dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP) on initial caries lesion of biofilm-coated enamel.Artificial initial carious lesion was created via 72-hour immersion in demineralization solution and cariogenic biofilm was formed after 24-hour culture of Streptococcus mutans (S. mutans). Specimens were then divided into 4 groups: enamel control, enamel treated with NACP, DMAHDM and NACP+DMAHDM respectively. Samples next underwent 7-day cycling, 4 h in BHIS (brain heart infusion broth containing 1 % sucrose) and 20 h in AS (artificial saliva) per day. The pH of BHIS was tested daily. So did the concentration of calcium and phosphate in BHIS and AS. Live/dead staining, colony-forming unit (CFU) count, and lactic acid production of biofilms were measured 7 days later. The enamel remineralization efficiency was evaluated by microhardness testing and transverse microradiography (TMR) quantitatively.Enamel of NACP+DMAHDM group demonstrated excellent remineralization effectiveness. And the NACP+DMAHDM adhesive released a great number of CaThe NACP+DMAHDM adhesive would be applicable to preventing secondary caries, strengthening enamel-adhesive interface, and extending the lifespan of composite restoration.

Background/purpose A common reason for dental composite restoration failure is recurrent caries at the margins. Our objectives were to: (1) develop a novel low-shrinkage-stress, antibacterial and remineralizing resin composite; (2) evaluate the effects of dimethylaminohexadecyl methacrylate (DMAHDM) on mechanical properties, biofilm inhibition, calcium (Ca) and phosphate (P) ion release, degree of conversion, and shrinkage stress on the new low-shrinkage-stress resin composite for the first time. Material and methods The resin consisted of urethane dimethacrylate (UDMA) and triethylene glycol divinylbenzyl ether (TEG-DVBE) with high resistance to salivary hydrolytic degradation. Composites were made with 0%–8% of DMAHDM for antibacterial activity, and 20% of nanoparticles of amorphous calcium phosphate (NACP) for remineralization. Mechanical properties and Streptococcus mutans biofilm growth on composites were assessed. Ca and P ion releases, degree of conversion and shrinkage stress were evaluated. Results Adding 2–5% DMAHDM and 20% NACP into the low-shrinkage-stress composite did not compromise the mechanical properties (p > 0.05). The incorporation of DMAHDM greatly reduced S. mutans biofilm colony-forming units by 2–5 log and lactic acid production by 7 folds, compared to a commercial composite (p Conclusion The bioactive low-shrinkage-stress composite reduced the polymerization shrinkage stress, without compromising other properties. Increasing the DMAHDM content increased the antibacterial effect in a dose-dependent manner.

Millions of people worldwide suffer from a toothache due to tooth cavity, and often permanent tooth loss. Dental caries, also known as tooth decay, is a biofilm-dependent infectious disease that damages teeth by minerals loss and presents a high incidence of clinical restorative polymeric fillings (tooth colored fillings). Until now, restorative polymeric fillings present no bioactivity. The complexity of oral biofilms contributes to the difficulty in developing effective novel dental materials. Nanotechnology has been explored in the development of bioactive dental materials to reduce or modulate the activities of caries-related bacteria. Nano-structured platforms based on calcium phosphate and metallic particles have advanced to impart an anti-caries potential to restorative materials. The bioactivity of these platforms induces prevention of mineral loss of the hard tooth structure and antibacterial activities against carries-related pathogens. It has been suggested that this bioactivity could minimize the incidence of caries around restorations (CARS) and increase the longevity of such filling materials. The last few years witnessed growing numbers of studies on the preparation evaluations of these novel materials. Herein, the caries disease process and the role of pathogenic caries-related biofilm, the increasing incidence of CARS, and the recent efforts employed for incorporation of bioactive nanoparticles in restorative polymer materials as useful strategies for prevention and management of caries-related-bacteria are discussed. We highlight the status of the most advanced and widely explored interaction of nanoparticle-based platforms and calcium phosphate compounds with an eye toward translating the potential of these approaches to the dental clinical reality., Graphical abstract Image 1, Highlights • Current progress and future applications of functional nanoparticles and remineralizing compounds incorporated in dental direct restorative materials. • Overview of the antibacterial and remineralizing mechanisms presenting direct and indirect implications on the tooth mineral loss. • These investigations, although in the initial phase of evidence are necessary and their results are encouraging and open the doors to future clinical studies that will allow the therapeutic value of nanotechnology-based restorative materials to be established.

Objective Periodontal tissue destruction and tooth loss are increasingly a worldwide problem as the population ages. Periodontitis is caused by bacterial infection and biofilm plaque buildup. Therefore, the objectives of this study were to: (1) develop a near-infrared light (NIR)-triggered core-shell nanostructure of upconversion nanoparticles and TiO2 (UCNPs@TiO2), and (2) investigate its inhibitory effects via antibacterial photodynamic therapy (aPDT) against periodontitis-related pathogens. Methods The core β-NaYF4:Yb3+,Tm3+ were synthesized via thermal decomposition and further modified with the TiO2 shell via a hydrothermal method. The core-shell structure and the upconversion fluorescence-induced aPDT treatment via 980 nm laser were studied. Three periodontitis-related pathogens Streptococcus sanguinis (S. sanguinis), Porphyromonas gingivalis (P. gingivalis) and Fusobacterium nucleatum (F. nucleatum) were investigated. The killing activity against planktonic bacteria was detected by a time-kill assay. Single species 4-day biofilms on dentin were tested by live/dead staining, colony-forming units (CFU), and metabolic activity. Results The hexagonal shaped UCNPs@TiO2 had an average diameter of 39.7 nm. UCNPs@TiO2 nanoparticles had positively charged (+12.4 mV) surface and were biocompatible and non-cytotoxic. Under the excitation of NIR light (980 nm), the core NaYF4:Yb3+,Tm3+ UCNPs could emit intense ultraviolet (UV) light, which further triggered the aPDT function of the shell TiO2 via energy transfer, thereby realizing the remarkable antibacterial effects against planktons and biofilms of periodontitis-associated pathogens. NIR-triggered UCNPs@TiO2 achieved much greater reduction in biofilms than control (p Significance Upconversion fluorescence-based aPDT achieved strong inhibiting effects against all three species of periodontitis-related pathogens. This novel nanotechnology demonstrated a high promise to inhibit periodontitis, with exciting potential to combat other oral infectious diseases such as deep endodontic infections.

Oral diseases such as tooth caries, periodontal diseases, endodontic infections, etc., are prevalent worldwide. The heavy burden of oral infectious diseases and their consequences on the patients' quality of life indicates a strong need for developing effective therapies. Advanced understandings of such oral diseases, e.g., inflammatory periodontal lesions, have raised the demand for antibacterial therapeutic strategies, because these diseases are caused by viruses and bacteria. The application of antimicrobial photodynamic therapy (aPDT) on oral infectious diseases has attracted tremendous interest in the past decade. However, aPDT had a minimal effect on the viability of organized biofilms due to the hydrophobic nature of the majority of the photosensitizers (PSs). Therefore, novel nanotechnologies were rapidly developed to target the delivery of hydrophobic PSs into microorganisms for the antimicrobial performance improvement of aPDT. This review focuses on the state-of-the-art of nanomaterials applications in aPDT against oral infectious diseases. The first part of this article focuses on the cutting-edge research on the synthesis, toxicity, and therapeutic effects of various forms of nanomaterials serving as PS carriers for aPDT applications. The second part discusses nanomaterials applications for aPDT in treatments of oral diseases. These novel bioactive nanomaterials have demonstrated great potential to serve as carriers for PSs to substantially enhance the PDT therapeutic effects. Furthermore, the novel aPDT applications not only have exciting therapeutic potential to inhibit bacterial plaque-initiated oral diseases, but also have a wide applicability to other biomedical and tissue engineering applications.

Peri-implantitis is the most common risk factor for dental implant failure. Nanostructured ceria (nano-CeO2) has anti-inflammatory and antibacterial functions, and different shapes of ceria enclosed by specific crystal planes could be an effective approach to enhance intrinsic catalysis. In the present study, the authors developed a novel implant surface-modification strategy by coating different shapes of nano-CeO2 onto titanium (Ti) surfaces to enhance their antibacterial and anti-inflammatory properties. The objectives of the study were to: (1) develop novel Ti surfaces modified with different shapes of nano-CeO2 (nanorod, nanocube and nano-octahedron) for peri-implantitis prevention; (2) investigate and compare the inhibition efficacy of different shapes of CeO2-modified surfaces against biofilms of peri-implantitis-related pathogens; and (3) evaluate the different CeO2-modified surfaces on cell inflammatory response in vitro and in vivo. The results showed that nanorod CeO2-modified Ti had more bacteria attachment of Streptococcus sanguinis in the early stage, compared with other CeO2-modified Ti (p 0.1). Nanocube and nano-octahedron CeO2-modified Ti exerted much better anti-inflammatory effects and ROS-scavenging ability than nanorod CeO2 in vitro (p Statement of Significance Peri-implantitis is the most common risk factor for dental implant failure. Nanostructured ceria (nano-CeO2) has anti-inflammatory and antibacterial functions, and different shapes of ceria enclosed by specific crystal planes could be an effective approach to enhance intrinsic catalysis. In the present study, we developed a novel implant surface-modification strategy by coating different shapes of nano-CeO2 onto titanium surfaces to enhance their antibacterial and anti-inflammatory properties for dental implants. In addition, we found that the nano-octahedron CeO2 coating on titanium would have great therapeutic potential for alleviating and eliminating peri-implantitis.

Objective Secondary 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. Methods Microcapsules 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 K IC and self-healing efficiency. A dental plaque microcosm biofilm model was used with human saliva as inoculum. Results The 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 IC 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. Significance This 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.

Persistent apical periodontitis, mainly caused by microorganisms infections, represents a critical challenge for endodontists. Dimethylaminododecyl methacrylate (DMADDM) is a well-studied and potent antibacterial agent used in various studies described in the literature. The aim of this study is to develop a novel antibacterial root canal sealer by incorporating DMADDM into EndoREZ and investigate the properties of the resulting material. Different mass fractions (0, 1.25%, 2.5%, and 5%) of DMADDM were incorporated into EndoREZ and the cytotoxicity, apical sealing ability and solubility of the resulting material were evaluated. Furthermore, a direct contact test, determination of colony-forming units, a crystal violet assay, scanning electronic microscopy and live/dead bacteria staining were performed to evaluate the antibacterial effect of the sealer to multispecies bacteria (Enterococcus faecalis, Streptococcus gordonii, Actinomyces naeslundii, and Lactobacillus acidophilus), in planktonic cells or biofilms. Fluorescence in situ hybridization and quantitative real-time polymerase chain reaction were carried out to assess the composition of the multispecies biofilms. No difference on the cytotoxicity, apical sealing ability and solubility between sealers containing DMADDM (1.25%, 2.5%) and EndoREZ (0%) could be determined. However, when the mass fraction of DMADDM increased to 5%, significantly different properties were found compared to the 0% (p E. faecalis could be decreased in multispecies microecology in sealers containing DMADDM. Therefore, a EndoREZ sealer material containing DMADDM could be considered useful in clinical applications for preventing and treating persistent apical periodontitis.

Objective Calcium phosphate cements (CPCs) mimic nanostructured bone minerals and are promising for dental, craniofacial and orthopedic applications. Vascularization plays a critical role in bone regeneration. This article represents the first review on cutting-edge research on prevascularization of CPC scaffolds to enhance bone regeneration. Methods This article first presented the prevascularization of CPC scaffolds. Then the co-culture of two cell types in CPC scaffolds was discussed. Subsequently, to further enhance the prevascularization efficacy, tri-culture of three different cell types in CPC scaffolds was presented. Results (1) Arg–Gly–Asp (RGD) incorporation in CPC bone cement scaffold greatly enhanced cell affinity and bone prevascularization; (2) By introducing endothelial cells into the culture of osteogenic cells (co-culture of two different cell types, or bi-culture) in CPC scaffold, the bone defect area underwent much better angiogenic and osteogenic processes when compared to mono-culture; (3) Tri-culture with an additional cell type of perivascular cells (such as pericytes) resulted in a substantially enhanced prevascularization of CPC scaffolds in vitro and more new bone and blood vessels in vivo, compared to bi-culture. Furthermore, biological cell crosstalk and capillary-like structure formation made critical contributions to the bi-culture system. In addition, the pericytes in the tri-culture system substantially promoted stability and maturation of the primary vascular network. Significance The novel approach of CPC scaffolds with stem cell bi-culture and tri-culture is of great significance in the regeneration of dental, craniofacial and orthopedic defects in clinical practice.

Fixed orthodontic treatments often lead to enamel demineralization and cause white spot lesions (WSLs). The aim of this study was to evaluate the mineralization degree of 2 types of WSLs based on ICDAS index and compare the remineralizing efficacy of 3 oral hygiene practices after 1 month and 3 months. 80 mild demineralized and 80 severe demineralized enamel specimens were randomized into three treatments: fluoride toothpaste (FT), fluoride varnish plus fluoride toothpaste (FV+FT), and CPP-ACP plus fluoride toothpaste (CPP-ACP+FT). Microhardness tester, DIAGNODent Pen 2190, and scanning electron microscope were used to evaluate the changes of mineralization degree. Both qualitative and quantitative indicators suggested that the mild and severe white spot lesions were different in the degree of mineralization. Severe WSLs demineralized much more seriously than mild lesions even after 3 months of treatment. Despite the variation in severity, both lesions had the same variation trend after each measure was applied: FT had weak therapeutic effect, FV + FT and CPP-ACP + FT were effective for remineralization. Their remineralizing efficacy was similar after 1 month, and combined use of CPP-ACP plus F toothpaste was more effective after 3 months. In order to fight WSLs, early diagnosis was of great importance, and examination of the tooth surface after air-dry for 5 seconds was recommended. Also, when WSLs were found, added remineralizing treatments were required.

The objectives of this studywere to: (1) develop a novel bioactive nanocomposite for Class V restorations with subgingival margins to inhibit periodontal pathogens; and (2) investigate if the bioactive nanocomposite could inhibit multi-species periodontal biofilms with a potency as strong as that against single species biofilms.Nanocomposite was fabricated using dimethylaminohexadecyl methacrylate (DMAHDM), 2-methacryloyloxyethyl phosphorylcholine (MPC) and nanoparticles of amorphous calcium phosphate (NACP). Biofilms with 1, 3, 6 and 9 species of periodontal pathogens were grown on the composites and tested for live/dead staining, colony-forming units (CFU), metabolic activity, and biofilm matrix polysaccharide production.The bioactive composite reduced protein adsorption by an order of magnitude (p 0.05) and greatly reduced biofilm viability. It decreased the biofilm CFU by more than 3 orders of magnitude for all four types of periodontal biofilms, compared to control composite. With increasing the biofilm species from 1 to 9, the antibacterial efficacy of DMAHDM composite decreased; the CFU reduction folds decreased from 947 folds to 44 folds. In contrast, the MPC + DMAHDM composite maintained a CFU reduction folds of greater than 3000, showing a similar antibacterial potency from 1 to 9 species in the biofilms (p 0.1).Dual agents MPC + DMAHDM achieved the greatest inhibition in biofilm, without decreasing its antibacterial potency when the biofilm species was increased from 1 to 9. A single agent became less effective when the biofilm species was increased from 1 to 9.The multifunctional MPC + DMAHDM composite is promising for root caries treatment and Class V restorations with subgingival margins to effectively inhibit multispecies periodontal biofilms, combat periodontitis and protect the periodontium.

Introduction Alginate/gelatin hydrogel (Alg-Gel) scaffold has been applied in tissue engineering, but the research on its application in dental tissues regeneration is still lacking. We investigated the effect of this scaffold on human dental pulp stem cells (hDPSCs). Methods hDPSCs were cultured in both Alg-Gel and 3D-printed Alg-Gel scaffolds. Cell growth and adhesion were compared using fluorescein isothiocyanate–phalloidin staining and scanning electron microscopic micrographs. Changes in the proliferation in hDPSCs cultured in the complete culture medium containing aqueous extracts of the Alg-Gel or 3D-printed Alg-Gel scaffolds were examined using Cell Counting Kit-8 assay and flow cytometry analysis. Cells were cultured in the mineralization medium containing aqueous extracts of the Alg-Gel or 3D-printed Alg-Gel scaffolds for 7 or 14 days, and the differentiation of cells was shown by alizarin red S staining and alkaline phosphatase staining. The messenger RNA and protein expression of mineralization-related genes were detected with real-time polymerase chain reaction and Western blotting. Elemental analysis was used to test the material extract composition. Results More cells were grown and adhered to the 3D-printed Alg-Gel scaffolds than the Alg-Gel scaffolds. The aqueous extracts of 3D-printed scaffolds can promote cell proliferation, and compared with Alg-Gel scaffolds, the extracts of 3D-printed scaffolds were more effective. Compared with the negative control group, 3D-printed Alg-Gel scaffold and Alg-Gel scaffold aqueous extracts promoted osteogenic/odontoblastic differentiation of hDPSCs with the enhanced formation of bone-like nodules and the alkaline phosphatase staining. The expression of mineralization-related genes was also up-regulated. 3D-printed scaffold aqueous extract contained more calcium and phosphorus ions than the Alg-Gel scaffold. Conclusions These findings suggest that compared with the Alg-Gel scaffold, 3D-printed Alg-Gel is more suitable for the growth of hDPSCs, and the scaffold extracts can better promote cell proliferation and differentiation.

Objectives The objective of the present study was to investigate long-term dentin remineralization via the combination of poly(amido amine) (PAMAM) with a novel rechargeable adhesive containing nanoparticles of amorphous calcium phosphate (NACP). Methods The NACP adhesive was immersed in lactic acid at pH 4 to exhaust its calcium (Ca) and phosphate (P) ion release, and then recharged with Ca and P ions. Dentin samples were pre-demineralized with 37% phosphoric acid, and then divided into four groups: (1) dentin control, (2) dentin treated with PAMAM, (3) dentin with recharged NACP adhesive, (4) dentin with PAMAM + recharged NACP adhesive. In group (2) and (4), the PAMAM-coated dentin was immersed in phosphate-buffered saline with vigorous shaking for 77 days to accelerate any detachment of the PAMAM macromolecules from the demineralized dentin. Samples were treated with a cyclic remineralization/demineralization regimen for 21 days. Results After 77 days of fluid flow challenge, the immersed PAMAM still retained its nucleation template function. The recharged NACP adhesive possessed sustained ion re-release and acid-neutralization capability, both of which did not decrease with repeated recharge and re-release cycles. The immersed PAMAM with the recharged NACP adhesive achieved long-term dentin remineralization, and restored dentin hardness to that of healthy dentin. Conclusions The PAMAM + NACP adhesive completely remineralizes pre-demineralized dentin even after long-term fluid challenges and provides long-term remineralization to protect tooth structures. Clinical significance The novel PAMAM + NACP adhesive provides long-term bond protection and caries inhibition to increase the longevity of resin-based restorations.

Bulk fracture and secondary caries are the two main problems causing failures and shortening the lifetime of dental resinous restorations. This article reviews recent research on self-healing dental and biomedical materials. This includes the development of self-healing dental resin composites and adhesives, combining self-healing with calcium phosphate nanoparticles in the resins for tooth lesion remineralization, and adding antibacterial monomer into self-healing resins to suppress oral biofilm grows and acid production. Furthermore, since the oral environment experiences saliva and drinks, this paper also reviews research on the self-healing of dental resins while being submerged in an aqueous environment, and the effect of long-term water-aging time from 1 day to 6 months on the self-healing capability. The new class of materials have demonstrated excellent self-healing efficacy in various material systems including bonding agents, composites and cements. They could heal cracks, regain load-bearing ability, inhibit oral pathogens, reduce or eliminate biofilm acids, raise biofilm pH to protect the teeth, and regenerate lost tooth minerals. Furthermore, their effects were indicated to be durable and long-lasting. While most of the recent publications on self-healing dental resins are from our group, this article also reviews publications from other researchers. The novel class of dental materials with triple benefits of self-healing, antibacterial and remineralization capabilities offer the much-needed improvements to address the two main reasons for restoration failures: fracture and secondary caries. They are expected to have potential for a wide range of dental and biomedical applications to overcome the current challenges and prolong the restoration life.

Superparamagnetic iron oxide nanoparticles (IONPs) are promising bioactive additives to fabricate magnetic scaffolds for bone tissue engineering. To date, there has been no report on osteoinductivity of IONP-incorporated calcium phosphate cement (IONP-CPC) scaffold on stem cells using an exterior static magnetic field (SMF). The objectives of this study were to: (1) develop a novel magnetic IONP-CPC construct for bone tissue engineering, and (2) investigate the effects of IONP-incorporation and SMF application on the proliferation, osteogenic differentiation and bone mineral synthesis of human dental pulp stem cells (hDPSCs) seeded on IONP-CPC scaffold for the first time. The novel magnetic IONP-CPC under SMF enhanced the cellular performance of hDPSCs, yielding greater alkaline phosphatase activities (about 3-fold), increased expressions of osteogenic marker genes, and more cell-synthesized bone minerals (about 2.5-fold), compared to CPC control and nonmagnetic IONP-CPC. In addition, IONP-CPC induced more active osteogenesis than CPC control in rat mandible defects. These results were consistent with the enhanced cellular performance by magnetic IONP in media under SMF. Moreover, nano-aggregates were detected inside the cells by transmission electron microscopy (TEM). Therefore, the enhanced cell performance was attributed to the physical forces generated by the magnetic field together with cell internalization of the released magnetic nanoparticles from IONP-CPC constructs.

Objectives (1) To develop a novel bioactive root canal sealer with antibiofilm and remineralization properties using dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP); (2) investigate the effects on E. faecalis biofilm inhibition, sealer flow and sealing ability, compared with an epoxy-resin-based sealer AH Plus; and (3) investigate the calcium (Ca) and phosphate (P) ion release from the sealers. Methods A series of dual-cure endodontic sealers were formulated with DMAHDM and NACP at 5% and 20% by mass, respectively. Flow properties and sealing ability of the sealers were measured. Colony-forming units (CFU), live/dead assay, and polysaccharide production of biofilms on sealers were determined. Ca and P ion releases from the sealers were measured. Results The new sealer containing 20% NACP and 5% DMAHDM yielded a paste flow of (28.99 ± 0.69) mm, within the range of ISO recommendations. The sealing properties of the sealer with 5% DMAHDM and 20% NACP were similar to a commercial control (p > 0.05). The sealer with DMAHDM decreased E. faecalis biofilm CFU by more than 4 orders of magnitude, compared to AH plus and experimental controls. The sealer with 20% NACP and 5% DMAHDM had relatively high levels of Ca and P ion release necessary for remineralization. Conclusions A new bioactive endodontic sealer was developed with strong antibiofilm activity against E. faecalis biofilms and high levels of Ca and P ion release for remineralization, without compromising the paste flow and sealing properties. Clinical significance The bioactive antibacterial and remineralizing root canal sealer is promising to inhibit E. faecalis biofilms to prevent endodontic treatment failure and secondary endodontic infections, while releasing high levels of Ca and P ions that could remineralize and strengthen the tooth structures and potentially prevent future root fractures and teeth extractions.

The aims of this study were to investigate the effects of water aging for up to 6months on the mechanical and anti-biofilm properties of a novel antibacterial glass ionomer cement (GIC) containing dimethylaminododecyl methacrylate (DMADDM).GIC specimens (n=180) which contained DMADDM (0wt.%, 1.1wt.% or 2.2wt.%) were prepared. The mechanical properties surface roughness, microhardness and the surface charge density of ammonium groups were measured before and after water aging for 3 and 6months at 37°C. Further six months aged specimens (n=216) were worn by 6 volunteers in their oral cavities for 24h and 72h. Biofilm formation was analyzed and rated by fluorescence microscopy (FM) and by scanning electron microscopy (SEM). Biofilm viability was analyzed by FM.Water aging did not show any adverse effects on the surface roughness and hardness of the material. The surface charge density of the GIC samples containing DMADDM decreased due to the aging procedure, however, was still higher than that of the GIC without DMADDM. In situ biofilm formation was significantly reduced after 24h on DMADDM containing GIC (p0.05). FM results showed a higher ratio of red/green fluorescence on GIC-DMADDM samples.Incorporating DMADDM into GIC affected the material properties in a tolerable manner even after 6months of storage in water. The new GIC is a promising material to affect the biofilm formation on the surface of restorations.

The dentin bonding often fails over time, leading to secondary caries and restoration failure. The objectives of this study were to develop an adhesive with dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP), and investigate the effects of storage in artificial saliva for six months on the bonding durability, antibacterial activity, ion release and biofilm pH properties for the first time.DMAHDM was added at 5% (by mass) to Scotchbond Primer and Adhesive (SBMP). NACP was added at 10%, 20%, and 30% to SBMP adhesive. Dentin bonding durability, antibacterial activity against Streptococcus mutans biofilms, and calcium (Ca) and phosphate (P) ion liberation properties were investigated after 1 day and 6months of storage in artificial saliva.Dentin bond strength (n = 50) had 25% loss after 6 months of aging for SBMP control. However, SBMP + DMAHDM+10NACP and SBMP + DMAHDM+20NACP showed no loss in bond strength after storage in artificial saliva for 6 months. The DMAHDM + NACP incorporation method dramatically reduced the biofilm metabolic activity and acid production, and decreased the biofilm CFU by four orders of magnitude, compared to SBMP control, even after 6 months of aging (p0.05). DMAHDM + NACP had long-lasting Ca and P ion releases, and raised the biofilm pH to 6.8, while the control group had a cariogenic biofilm pH of 4.5.Incorporating DMAHDM + NACP in bonding agent yielded potent and long-lasting antibacterial activity and ions liberation ability, and much higher long-term dentin bond strength after 6-month of aging. The new bonding agent is promising to inhibit caries at the restoration margins and increase the resin-dentin bonding longevity.The novel bioactive adhesive is promising to protect tooth structures from biofilm acids and secondary caries. NACP and DMAHDM have great potential for applications to a wide range of dental materials to reduce plaque and achieve therapeutic effects.

Secondary caries at the restoration-tooth margins is a main reason for dental restoration failures. Gene-modification for Streptococcus mutans (S. mutans) and composites containing dimethylaminohexadecyl methacrylate (DMAHDMA) and nanoparticles of amorphous calcium phosphate (NACP) all have the potential to suppress bacterial acids and promote remineralization. However, there has been no report of their effects on marginal caries-inhibition and enamel hardness. The objective of this study was to investigate the effects of gene-modification and DMAHDM–NACP composite restoration on enamel demineralization and hardness at the margins under biofilm acids for the first time. Parent S. mutans and rnc gene-deleted S. mutans were tested side by side. The bioactive composite contained 3% DMAHDM and 30% NACP. Mechanical properties and calcium (Ca) and phosphate (P) ion releases were measured. Colony-forming units (CFU), MTT, lactic acid and polysaccharide of biofilms were evaluated. Demineralization of bovine enamel with composite restorations was induced via biofilms, then enamel hardness was measured. The dual strategy of combining rnc-deletion with DMAHDM+30NACP: (1) achieved the strongest biofilm-inhibition, with the greatest reduction in biofilm CFU by 6 logs; (2) decreased biofilm lactic acid and polysaccharide production by more than 80%; (3) achieved enamel hardness that was 140% higher than that of a commercial fluoride-releasing composite under 30 days of biofilm acids. Therefore, the novel dual approach of rnc gene-deletion and DMAHDM+NACP nanocomposite is promising to inhibit secondary caries at the margins and increase the longevity of tooth restorations.

Periodontitis is a common bacteria-borne inflammatory disease that can damage the supporting structures of teeth, eventually leading to tooth loss and systemic inflammations. The present study developed novel nanoparticles (ZIF-8:Ce) by doping cerium (Ce) into zeolitic imidazolate framework-8 (ZIF-8) to simultaneously obtain antibacterial and anti-inflammatory capabilities. The objectives of this study were to: (1) develop novel ZIF-8 nanoparticles doped with different Ce/(Ce + Zn) molar ratios of 1%, 5% and 10% to combat periodontitis; (2) investigate the inhibition efficacy of different nanoparticles against biofilms of periodontal pathogens; and (3) evaluate the effects of different ZIF-8:Ce on inflammatory secretion and macrophage polarization in vitro. The results showed that Ce doping at a ratio from 1% to 10% did not compromise the regular and uniform structure of ZIF-8. ZIF-8:Ce possessed a sustained Zn2+ and Ce3+/Ce4+ release and favorable superoxide dismutase/catalase activities without cytotoxicity at a concentration below 30 μg mL-1. ZIF-8 exhibited an excellent anti-biofilm function against periodontal pathogens. Although 10% Ce doping into ZIF-8 slightly reduced the antibacterial effects, the CFU reduction still remained at approximately 2 orders of magnitude. More importantly, ZIF-8:Ce exhibited increasing anti-inflammatory effects with increasing amounts of Ce doping. In addition, ZIF-8:Ce10% exerted much better anti-inflammatory effects by inhibiting pro-inflammatory factor expression via restraining the NF-kB/p65 subunit translocation (p < 0.05). Meanwhile, ZIF-8:Ce10% elevated the polarization of the M2 phenotype of macrophages and promoted anti-inflammatory cytokine secretion. Therefore, the novel ZIF-8:Ce nanoparticles provided a unique insight into the development of effective anti-inflammatory and antibacterial platforms for treating periodontitis.

Recurrent caries often occurs and is a primary reason for the failure of dental composite restorations. The objectives of this study were to: (1) develop a bioactive composite containing dimethylaminohexadecyl methacrylate (DMAHDM), (2) investigate its antibacterial effects and suppression on biofilm growth, and (3) investigate its ability to modulate biofilm species composition for the first time. DMAHDM was incorporated into a composite at mass% of 0%, 0.75%, 1.5%, 2.25% and 3%. A commercial composite Heliomolar served as a comparative control. A biofilm model consisting of Streptococcus mutans (S. mutans), Streptococcus sanguinis (S. sanguinis) and Streptococcus gordonii (S. gordonii) was tested by growing biofilms for 48 h and 72 h on composites. Colony-forming units (CFUs), metabolic activity and live/dead staining were evaluated. Lactic acid and polysaccharide productions were measured to assess biofilm cariogenicity. TaqMan real-time polymerase chain reaction was used to determine the proportion of each species in the biofilm. DMAHDM-containing composite had a strong anti-biofilm function, reducing biofilm CFU by 2–3 orders of magnitude, compared to control composite. Biofilm metabolic activity, lactic acid and polysaccharides were decreased substantially, compared to control (p < 0.05). At 72 h, the cariogenic S. mutans proportion in the biofilm on the composite with 3% DMAHDM was 19.9%. In contrast, an overwhelming S. mutans proportion of 92.2% and 91.2% existed in biofilms on commercial control and 0% DMAHDM, respectively. In conclusion, incorporating DMAHDM into dental composite: (1) yielded potent anti-biofilm properties; (2) modulated the biofilm species composition toward a non-cariogenic tendency. The new DMAHDM composite is promising for applications in a wide range of tooth cavity restorations to modulate oral biofilm species and combat caries.

Resin cements with remineralizing and antibacterial properties are favorable for inhibition of caries. The objectives of this study were: (1) to investigate the capability of the novel dimethylaminohexadecyl-methacrylate (DMAHDM) and nano-sized amorphous calcium phosphate (NACP) containing cement to reduce saliva microcosm biofilm, and (2) to investigate the long-term ion release, recharge, and re-release of DMAHDM-NACP cement.Pyromellitic glycerol dimethacrylate (PMGDM) and ethoxylated bisphenol-A-dimethacrylate (EBPADMA) were used to make PEHB monomer. Five cements were fabricated: (1) PEHB+0%NACP+0%DMAHDM (experimental control); (2) PEHB+25%NACP+0%DMAHDM, (3) PEHB+25%NACP+0%DMAHDM; (4) PEHB+25%NACP+3%DMAHDM; (5) PEHB+25%NACP+5%DMAHDM. RelyX luting cement was used as commercial control. Colony-forming units (CFU), lactic acid production, metabolic activities, and minimum inhibitory concentration (MIC) were performed. Long-term Calcium (Ca) and phosphate (P) ion release, recharge, and re-release were assessed.Compared to experimental and commercial controls, the NACP-DMAHDM cement significantly reduced CFU biofilm by 2-3 orders of magnitude, metabolic activities from 0.24±0.06 AIncorporating DMAHDM and NACP into resin-based crown cement provides strong antibacterial action against saliva microcosm biofilm and presents a high level of Ca and P ion recharge abilities, exhibiting long-term Ca and P ion release and remineralization potential.Resin based cement containing NACP and DMAHDM were developed with remineralizing and potent antibacterial effects. This cement formulation showed ion release and remineralization potential and are promising formulations to inhibit the incidence of recurrent caries and could promote remineralization and be sustainable for the long term.