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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.

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.

(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.

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.

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.

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.

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.

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 infection of Enterococcus faecalis and its interacting microorganisms in the root canal could cause persistent apical periodontitis (AP). Antibacterial root canal sealer has favorable prospects to inhibit biofilms. The purpose of this study was to investigated the antibacterial effect of root canal sealer containing dimethylaminododecyl methacrylate (DMADDM) on persistent AP in beagle dogs for the first time. Persistent AP was established by a two-step infection with Enterococcus faecalis and multi-bacteria (Enterococcus faecalis, Lactobacillus acidophilus, Actinomycesnaeslundii, Streptococcus gordonii). Root canal sealer containing DMADDM (0%, 1.25%, 2.5%) was used to complete root canal filling. The volume of lesions and inflammatory grade in the apical area were evaluated by cone beam computer tomography (CBCT) and hematoxylin-eosin staining. Both Enterococcus-faecalis- and multi-bacteria-induced persistent AP caused severe apical destruction, and there were no significant differences in pathogenicity between them. DMADDM-modified sealer significantly reduced the volume of periapical lesion and inflammatory grade compared with the control group, among them, the therapeutic effect of the 2.5% group was better than the 1.25% group. In addition, E.faecalis-induced reinfection was more sensitive to the 2.5% group than multi-bacteria reinfection. This study shows that root canal sealer containing DMADDM had a remarkable therapeutic effect on persistent AP, especially on E. faecalis-induced reinfection.

Candida albicans (C. albicans) biofilm is a common etiological factor in denture stomatitis. The purpose of this study was to investigate the effects of incorporating 2-methacryloyloxyethyl phosphorylcholine (MPC) as a protein repellent into a new high-impact denture acrylic (HIPA) resin on the surface roughness, solution pH, and C. albicans biofilm adhesion to the denture base. The new acrylic denture resin base was formulated by mixing MPC into HIPA resin at mass fractions of 1.5%, 3%, and 4.5%. Surface roughness was measured using a Mitutoyo surface roughness tester. C. albicans biofilm growth and viability were assessed via colony forming unit counts. The pH of the biofilm growth medium was measured using a digital pH meter. Adding MPC to the HIPA resin at percentages of 1.5% and 3% increased the roughness values significantly (P &lt, 0.05), while adding 4.5% MPC resulted in no difference in roughness values to that of the control group (P &gt, 0.05). All experimental groups demonstrated neutral pH values (pH @ 7) and were not significantly different from each other (P &gt, 0.05). Incorporating 2-methacryloyloxyethyl phosphorylcholine at 4.5% resulted in a significant (@1 log) colony-forming unit reduction compared with the control group with 0% MPC (P &lt, 0.05). A fungal-retarding denture acrylic resin was developed through the incorporation of MPC for its protein-repelling properties. This newly developed denture acrylic material has the potential to prevent oral microbial infections, such as denture stomatitis.

The enamel demineralization is common in fixed orthodontics. Plaque accumulation around the bracket plays a critical role and could cause various degrees of white spot lesions (WSLs) on the surface of teeth. The 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer is a biological polymer with protein repellent and an anti-bacterial adhesion effects. In this study, the enamel shear bond strength (SBS) and protein repellent property in vitro of self-etching orthodontic adhesive with MPC were evaluated. It was found that the self-etching adhesive with 0-7.5%MPC met the orthodontic clinical requirement on the SBS values at three different points of time, The incorporation of 7.5%MPC significantly reduced the bacterial adhesion and total microorganism of the yield biofilm. Moreover, the MTT assay showed that the amount of plaque metabolism in 7.5%MPC was the lowest among the groups. To conclude, the novel protein repellent self-etching adhesive was able to inhibit biofilm formation efficiently and minimize enamel demineralization.

Polyetheretherketone (PEEK) is an important high-performance thermoplastic. Its excellent strength, stiffness, toughness, fatigue resistance, biocompatibility, chemical stability and radiolucency have made PEEK attractive in dental and orthopedic applications. However, PEEK has an inherently hydrophobic and chemically inert surface, which has restricted its widespread use in clinical applications, especially in bonding with dental resin composites. Cutting edge research on novel methods to improve PEEK applications in dentistry, including oral implant, prosthodontics and orthodontics, is reviewed in this article. In addition, this article also discusses innovative surface modifications of PEEK, which are a focus area of active investigations. Furthermore, this article also discusses the necessary future studies and clinical trials for the use of PEEK in the human oral environment to investigate its feasibility and long-term performance.

Decays in the roots of teeth is prevalent in seniors as people live longer and retain more of their teeth to an old age, especially in patients with periodontal disease and gingival recession. The objectives of this study were to develop a biocompatible nanocomposite with nano-sized calcium fluoride particles (Nano-CaF2), and to investigate for the first time the effects on osteogenic and cementogenic induction of periodontal ligament stem cells (hPDLSCs) from human donors.Nano-CaF2 particles with a mean particle size of 53 nm were produced via a spray-drying machine.Nano-CaF2 was mingled into the composite at 0%, 10%, 15% and 20% by mass. Flexural strength (160 &plusmn, 10) MPa, elastic modulus (11.0 &plusmn, 0.5) GPa, and hardness (0.58 &plusmn, 0.03) GPa for Nano-CaF2 composite exceeded those of a commercial dental composite (p &lt, 0.05). Calcium (Ca) and fluoride (F) ions were released steadily from the composite. Osteogenic genes were elevated for hPDLSCs growing on 20% Nano-CaF2. Alkaline phosphatase (ALP) peaked at 14 days. Collagen type 1 (COL1), runt-related transcription factor 2 (RUNX2) and osteopontin (OPN) peaked at 21 days. Cementogenic genes were also enhanced on 20% Nano-CaF2 composite, promoting cementum adherence protein (CAP), cementum protein 1 (CEMP1) and bone sialoprotein (BSP) expressions (p &lt, 0.05). At 7, 14 and 21 days, the ALP activity of hPDLSCs on 20% Nano-CaF2 composite was 57-fold, 78-fold, and 55-fold greater than those of control, respectively (p &lt, 0.05). Bone mineral secretion by hPDLSCs on 20% Nano-CaF2 composite was 2-fold that of control (p &lt, 0.05). In conclusion, the novel Nano-CaF2 composite was biocompatible and supported hPDLSCs. Nano-CaF2 composite is promising to fill tooth root cavities and release Ca and F ions to enhance osteogenic and cementogenic induction of hPDLSCs and promote periodontium regeneration.

Caries-related biofilms and associated complications are significant threats in dentistry, especially when biofilms grow over dental restorations. The inhibition of cariogenic biofilm associated with the onset of carious lesions is crucial for preventing disease recurrence after treatment. This in vitro study defined optimized parameters for using a photosensitizer, toluidine blue O (TBO), activated via a red light-emitting diode (LED)-based wireless device to control the growth of cariogenic biofilms. The effect of TBO concentrations (50, 100, 150, and 200 &mu, g/mL) exposed to light or incubated in the dark was investigated in successive cytotoxicity assays. Then, a mature Streptococcus mutans biofilm model under sucrose challenge was treated with different TBO concentrations (50, 100, and 150 &mu, g/mL), different light energy doses (36, 108, and 180 J/cm2), and different incubation times before irradiation (1, 3, and 5 min). The untreated biofilm, irradiation with no TBO, and TBO incubation with no activation represented the controls. After treatments, biofilms were analyzed via S. mutans colony-forming units (CFUs) and live/dead assay. The percentage of cell viability was within the normal range compared to the control when 50 and 100 &mu, g/mL of TBO were used. Increasing the TBO concentration and energy dose was associated with biofilm inhibition (p &lt, 0.001), while increasing incubation time did not contribute to bacterial elimination (p &gt, 0.05). Irradiating the S. mutans biofilm via 100 &mu, g/mL of TBO and &asymp, 180 J/cm2 energy dose resulted in &asymp, 3-log reduction and a higher amount of dead/compromised S. mutans colonies in live/dead assay compared to the control (p &lt, 0.001). The light energy dose and TBO concentration optimized the bacterial elimination of S. mutans biofilms. These results provide a perspective on the determining parameters for highly effective photo-killing of caries-related biofilms and display the limitations imposed by the toxicity of the antibacterial photodynamic therapy&rsquo, s chemical components. Future studies should support investigations on new approaches to improve or overcome the constraints of opportunities offered by photodynamic inactivation of caries-related biofilms.

Oral biofilm accumulation at the tooth&ndash, restoration interface often leads to recurrent dental caries and restoration failure. The objectives of this study were to: (1) develop a novel bioactive crown cement containing dimethylaminohexadecyl methacrylate (DMAHDM) and nano-sized amorphous calcium phosphate (NACP), and (2) investigate the mechanical properties, anti-biofilm activity, and calcium (Ca2+) and phosphate (PO43&minus, ) ion release of the crown cement for the first time. The cement matrix consisted of pyromellitic glycerol dimethacrylate and ethoxylated bisphenol-A dimethacrylate monomers and was denoted PEHB resin matrix. The following cements were tested: (1) RelyX luting cement (commercial control), (2) 55% PEHB + 45% glass fillers (experimental control), (3) 55% PEHB + 20% glass + 25% NACP + 0% DMAHDM, (4) 52% PEHB + 20% glass + 25% NACP + 3% DMAHDM, (5) 51% PEHB + 20% glass + 25% NACP + 4% DMAHDM, (6) 50% PEHB + 20% glass + 25% NACP + 5% DMAHDM. Mechanical properties and ion release were measured. Streptococcusmutans (S. mutans) biofilms were grown on cements, and colony-forming units (CFUs) and other biofilm properties were measured. The novel bioactive cement demonstrated strong antibacterial properties and high levels of Ca2+ and PO43&minus, ion release to remineralize tooth lesions. Adding NACP and DMAHDM into the cement did not adversely affect the mechanical properties and dentin bonding strength. In conclusion, the novel NACP + DMAHDM crown cement has excellent potential for restoration cementation to inhibit caries by suppressing oral biofilm growth and increasing remineralization via Ca2+ and PO43&minus, ions. The NACP + DMAHDM composition may have wide applicability to other biomaterials to promote hard-tissue formation and combat bacterial infection.

The objectives were to investigate a novel combination of gene-knockout with antimicrobial dimethylaminohexadecyl methacrylate (DMAHDM) composite in regulating oral biofilm from a cariogenic state toward a non-cariogenic state. A tri-species biofilm model included cariogenic Streptococcus mutans (S. mutans), and non-cariogenic Streptococcus sanguinis (S. sanguinis) and Streptococcus gordonii (S. gordonii). Biofilm colony-forming-units (CFUs), lactic acid and polysaccharide production were measured. TaqMan real-time-polymerase-chain reaction was used to determine the percentage of each species in biofilm. The rnc gene-knockout for S. mutans with DMAHDM composite reduced biofilm CFU by five logs, compared to control (p &lt, 0.05). Using parent S. mutans, an overwhelming S. mutans percentage of 68.99% and 69.00% existed in biofilms on commercial composite and 0% DMAHDM composite, respectively. In sharp contrast, with a combination of S. mutans rnc knockout and DMAHDM composite, the cariogenic S. mutans percentage in biofilm was reduced to only 6.33%. Meanwhile, the non-cariogenic S. sanguinis + S. gordonii percentage was increased to 93.67%. Therefore, combining rnc-knockout with bioactive and therapeutic dental composite achieved the greatest reduction in S. mutans, and the greatest increase in non-cariogenic species, thereby yielding the least lactic acid-production. This novel method is promising to obtain wide applications to regulate biofilms and inhibit dental caries.

Osteomyelitis and post-operative infections are major problems in orthopedic, dental and craniofacial surgeries. It is highly desirable for a tissue engineering construct to kill bacteria, while simultaneously delivering stem cells and enhancing cell function and tissue regeneration. The objectives of this study were to: (1) develop a novel injectable calcium phosphate cement (CPC) scaffold containing antibiotic ornidazole (ORZ) while encapsulating human periodontal ligament stem cells (hPDLSCs), and (2) investigate the inhibition efficacy against Staphylococcus aureus (S. aureus) and the promotion of hPDLSC function for osteogenesis for the first time. ORZ was incorporated into a CPC-chitosan scaffold. hPDLSCs were encapsulated in alginate microbeads (denoted hPDLSCbeads). The ORZ-loaded CPCC+hPDLSCbeads scaffold was fully injectable, and had a flexural strength of 3.50 ± 0.92 MPa and an elastic modulus of 1.30 ± 0.45 GPa, matching those of natural cancellous bone. With 6 days of sustained ORZ release, the CPCC+10ORZ (10% ORZ) scaffold had strong antibacterial effects on S. aureus, with an inhibition zone of 12.47 ± 1.01 mm. No colonies were observed in the CPCC+10ORZ group from 3 to 7 days. ORZ-containing scaffolds were biocompatible with hPDLSCs. CPCC+10ORZ+hPDLSCbeads scaffold with osteogenic medium had 2.4-fold increase in alkaline phosphatase (ALP) activity and bone mineral synthesis by hPDLSCs, as compared to the control group (p < 0.05). In conclusion, the novel antibacterial construct with stem cell delivery had injectability, good strength, strong antibacterial effects and biocompatibility, supporting osteogenic differentiation and bone mineral synthesis of hPDLSCs. The injectable and mechanically-strong CPCC+10ORZ+hPDLSCbeads construct has great potential for treating bone infections and promoting bone regeneration.

Secondary caries often occurs at the tooth-composite margins. This study developed a novel bioactive composite containing DMAHDM (dimethylaminohexadecyl methacrylate) and NACP (nanoparticles of amorphous calcium phosphate), inhibiting caries at the enamel restoration margins in an in vitro saliva-derived biofilm secondary caries model for the first time. Four composites were tested: (1) Heliomolar nanocomposite, (2) 0% DMAHDM + 0% NACP, (3) 3% DMAHDM + 0% NACP, (D) 3% DMAHDM + 30% NACP. Saliva-derived biofilms were tested for antibacterial effects of the composites. Bovine enamel restorations were cultured with biofilms, Ca and P ion release of nanocomposite and enamel hardness at the enamel restoration margins was measured. Incorporation of DMAHDM and NACP into composite did not affect the mechanical properties (p &gt, 0.05). The biofilms&rsquo, CFU (colony-forming units) were reduced by 2 logs via DMAHDM (p &lt, 0.05). Ca and P ion release of the nanocomposite was increased at cariogenic low pH. Enamel hardness at the margins for DMAHDM group was 25% higher than control (p &lt, 0.05). With DMAHDM + NACP, the enamel hardness was the greatest and about 50% higher than control (p &lt, 0.05). Therefore, the novel composite containing DMAHDM and NACP was strongly antibacterial and inhibited enamel demineralization, resulting in enamel hardness at the margins under biofilms that approached the hardness of healthy enamel.

(1) Background: The objective of this study was to develop a novel dental nanocomposite containing dimethylaminohexadecyl methacrylate (DMAHDM), 2-methacryloyloxyethyl phosphorylcholine (MPC), and nanoparticles of calcium fluoride (nCaF2) for preventing recurrent caries via antibacterial, protein repellent and fluoride releasing capabilities. (2) Methods: Composites were made by adding 3% MPC, 3% DMAHDM and 15% nCaF2 into bisphenol A glycidyl dimethacrylate (Bis-GMA) and triethylene glycol dimethacrylate (TEGDMA) (denoted BT). Calcium and fluoride ion releases were evaluated. Biofilms of human saliva were assessed. (3) Results: nCaF2+DMAHDM+MPC composite had the lowest biofilm colony forming units (CFU) and the greatest ion release, however, its mechanical properties were lower than commercial control composite (p &lt, 0.05). nCaF2+DMAHDM composite had similarly potent biofilm reduction, with mechanical properties matching commercial control composite (p &gt, 0.05). Fluoride and calcium ion releases from nCaF2+DMAHDM were much more than commercial composite. Biofilm CFU on composite was reduced by 4 logs (n = 9, p &lt, 0.05). Biofilm metabolic activity and lactic acid were also substantially reduced by nCaF2+DMAHDM, compared to commercial control composite (p &lt, 0.05). (4) Conclusions: The novel nanocomposite nCaF2+DMAHDM achieved strong antibacterial and ion release capabilities, without compromising the mechanical properties. This bioactive nanocomposite is promising to reduce biofilm acid production, inhibit recurrent caries, and increase restoration longevity.

Quaternary ammonium methacrylates (QAMs) are useful antimicrobial compounds against oral bacteria. Here, we investigated the effects of two QAMs, dimethylaminododecyl methacrylate (DMADDM) and dimethylaminohexadecyl methacrylate (DMAHDM), on biofilm formation, survival and development of tolerance by biofilm, and survival and development of tolerance against QAMs after prolonged starvation. Enterococcus faecalis (E. faecalis), Streptococcus gordonii (S. gordonii), Lactobacillus acidophilus (L. acidophilus), and Actinomyces naeslundii (A. naeslundii) were used. Minimum inhibitory concentration (MIC) of QAMs against multispecies biofilm was determined. Biofilm formed under sub-MIC was observed by crystal violet staining and confocal laser scanning microscopy (CLSM). Metabolic activity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactic acid production measurement. Development of tolerance was determined by MIC values before and after exposure to QAMs or after prolonged starvation. It was found that E. faecalis and S. gordonii could survive and form biofilm under sub-MIC of QAMs. Lactic acid production from biofilms formed under sub-MIC was significantly higher than control specimens ( p < 0.05 ). The exposure to sub-MIC of QAMs promoted biofilm formation, and prolonged starvation or prolonged contact with sub-MIC helped bacteria develop tolerance against killing by QAMs.

Polymeric materials are the first choice for restoring tooth cavities, bonding tooth-colored fillings, sealing root canal systems, and many other dental restorative applications. However, polymeric materials are highly susceptible to bacterial attachment and colonization, leading to dental diseases. Many approaches have been investigated to minimize the formation of biofilms over polymeric restorative materials and at the tooth/material interfaces. Among them, contact-killing compounds have shown promising results to inhibit dental biofilms. Contact-killing compounds can be immobilized within the polymer structure, delivering a long-lasting effect with no leaching or release, thus providing advantages compared to release-based materials. This review discusses cutting-edge research on the development of contact-killing compounds in dental restorative materials to target oral pathogens. Contact-killing compounds in resin composite restorations, dental adhesives, root canal sealers, denture-based materials, and crown cements have all demonstrated promising antibacterial properties. Contact-killing restorative materials have been found to effectively inhibit the growth and activities of several oral pathogens related to dental caries, periodontal diseases, endodontic, and fungal infections. Further laboratory optimization and clinical trials using translational models are needed to confirm the clinical applicability of this new generation of contact-killing dental restorative materials.

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.

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.

According to the American Dental Association Survey of Dental Services Rendered (published in 2007), 15 million root canal treatment procedures are performed annually. Endodontic therapy relies mainly on biomechanical preparation, chemical irrigation and intracanal medicaments which play an important role in eliminating bacteria in the root canal. Furthermore, adequate obturation is essential to confine any residual bacteria within the root canal and deprive them of nutrients. However, numerous studies have shown that complete elimination of bacteria is not achieved due to the complex anatomy of the root canal system. There are several conventional antibiotic materials available in the market for endodontic use. However, the majority of these antibiotics and antiseptics provide short-term antibacterial effects, and they impose a risk of developing antibacterial resistance. The root canal is a dynamic environment, and antibacterial and antibiofilm materials with long-term effects and nonspecific mechanisms of action are highly desirable in such environments. In addition, the application of acidic solutions to the root canal wall can alter the dentin structure, resulting in a weaker and more brittle dentin. Root canal sealers with bioactive properties come in direct contact with the dentin wall and can play a positive role in bacterial elimination and strengthening of the root structure. The new generation of nanostructured, bioactive, antibacterial and remineralizing additives into polymeric resin-based root canal sealers are discussed in this review. The effects of these novel bioactive additives on the physical and sealing properties, as well as their biocompatibility, are all important factors that are presented in this article.

OBJECTIVES. This represents the first report on the development of metformin-containing dental resins. The objectives were to use the resin as a carrier to deliver metformin locally to stimulate dental cells for dental tissue regeneration, and to investigate the effects on odontogenic differentiation of dental pulp stem cells (DPSCs) and mineral synthesis. METHODS. Metformin was incorporated into a resin at 20% by mass as a model system. DPSC proliferation attaching on resins was evaluated. Dentin sialophosphoprotein (DSPP), dentin matrix phosphoprotein 1 (DMP-1), alkaline phosphatase (ALP) and runt-related transcription factor 2 (Runx2) genes expressions were measured. ALP activity and alizarin red staining (ARS) of mineral synthesis by the DPSCs on resins were determined. RESULTS. DPSCs on metformin-containing resin proliferated well (mean ± SD; n = 6), and the number of cells increased by 4 folds from 1 to 14 days (p > 0.1). DSPP, ALP and DMP-1 gene expressions of DPSCs on metformin resin were much higher than DPSCs on control resin without metformin (p < 0.05). ALP activity of metformin group was 70% higher than that without metformin at 14 days (p < 0.05). Mineral synthesis by DPSCs on metformin-containing resin at 21 days was 9 folds that without metformin (p < 0.05). SIGNIFICANCE. A novel metformin-containing resin was developed, achieving substantial enhancement of odontoblastic differentiation of DPSCs and greater mineral synthesis. The metformin resin is promising for deep cavities and perforated cavities to stimulate DPSCs for tertiary dentin formation, for tooth root coatings with metformin release for periodontal regeneration, and for root canal fillings with apical lesions to stimulate bone regeneration.

Novel strategies utilizing magnetic nanoparticles (MNPs) and magnetic fields are being developed to enhance bone tissue engineering efficacy. This article first reviewed cutting-edge research on the osteogenic enhancements via magnetic fields and MNPs. Then the current developments in magnetic strategies to improve the cells, scaffolds and growth factor deliveries were described. The magnetic-cell strategies included cell labeling, targeting, patterning, and gene modifications. MNPs were incorporated to fabricate magnetic composite scaffolds, as well as to construct delivery systems for growth factors, drugs and gene transfections. The novel methods using magnetic nanoparticles and scaffolds with magnetic fields and stem cells increased the osteogenic differentiation, angiogenesis and bone regeneration by 2-3 folds over those of the controls. The mechanisms of magnetic nanoparticles and scaffolds with magnetic fields and stem cells to enhance bone regeneration were identified as involving the activation of signaling pathways including MAPK, integrin, BMP and NF-κB. Potential clinical applications of magnetic nanoparticles and scaffolds with magnetic fields and stem cells include dental, craniofacial and orthopedic treatments with substantially increased bone repair and regeneration efficacy.

Hard tissue repair and regeneration cost hundreds of billions of dollars annually worldwide, and the need has substantially increased as the population has aged. Hard tissues include bone and tooth structures that contain calcium phosphate minerals. Smart biomaterial-based tissue engineering and regenerative medicine methods have the exciting potential to meet this urgent need. Smart biomaterials and constructs refer to biomaterials and constructs that possess instructive/inductive or triggering/stimulating effects on cells and tissues by engineering the material’s responsiveness to internal or external stimuli or have intelligently tailored properties and functions that can promote tissue repair and regeneration. The smart material-based approaches include smart scaffolds and stem cell constructs for bone tissue engineering; smart drug delivery systems to enhance bone regeneration; smart dental resins that respond to pH to protect tooth structures; smart pH-sensitive dental materials to selectively inhibit acid-producing bacteria; smart polymers to modulate biofilm species away from a pathogenic composition and shift towards a healthy composition; and smart materials to suppress biofilms and avoid drug resistance. These smart biomaterials can not only deliver and guide stem cells to improve tissue regeneration and deliver drugs and bioactive agents with spatially and temporarily controlled releases but can also modulate/suppress biofilms and combat infections in wound sites. The new generation of smart biomaterials provides exciting potential and is a promising opportunity to substantially enhance hard tissue engineering and regenerative medicine efficacy., Bone repair: a new generation of smart biomaterials Smart biomaterials that are able to instruct bone repair can overcome some of the shortcomings of bone grafting and meet the growing need for hard tissue regeneration in ageing populations. Hockin Xu at the University of Maryland in Baltimore, United States, and colleagues review recent advances in the development of smart biomaterials for repairing and regenerating damaged bones and teeth. They highlight scaffolds that closely mimic natural bone tissue and immunomodulatory biomaterials that can prevent infection and promote cell survival for guiding and enhancing bone regeneration. Furthermore, scaffolds with shape-memory capability and materials that provide tailored spatio-temporal delivery of drugs or bioactive agents in response to internal or external stimuli, hold great promise not only for bone tissue engineering but also for dentistry.

OBJECTIVES: A nanoparticle-doped adhesive that can be controlled with magnetic forces was recently developed to deliver drugs to the pulp and improve adhesive penetration into dentin. However, it did not have bactericidal and remineralization abilities. The objectives of this study were to: (1) develop a magnetic nanoparticle-containing adhesive with dimethylaminohexadecyl methacrylate (DMAHDM), amorphous calcium phosphate nanoparticles (NACP) and magnetic nanoparticles (MNP); and (2) investigate the effects on dentin bond strength, calcium (Ca) and phosphate (P) ion release and anti-biofilm properties. METHODS: MNP, DMAHDM and NACP were mixed into Scotchbond SBMP at 2%, 5% and 20% by mass, respectively. Two types of magnetic nanoparticles were used: acrylate-functionalized iron nanoparticles (AINPs); and iron oxide nanoparticles (IONPs). Each type was added into the resin at 1% by mass. Dentin bonding was performed with a magnetic force application for 3 minutes, provided by a commercial cube-shaped magnet. Dentin shear bond strengths were measured. Streptococcus mutans biofilms were grown on resins, and metabolic activity, lactic acid and colony-forming units (CFU) were determined. Ca and P ion concentrations in, and pH of biofilm culture medium were measured. RESULTS: Magnetic nanoparticle-containing adhesive using magnetic force increased the dentin shear bond strength by 59% over SBMP Control (p < 0.05). Adding DMAHDM and NACP did not adversely affect the dentin bond strength (p > 0.05). The adhesive with MNP+DMAHDM+NACP reduced the Streptococcus mutans biofilm CFU by 4 logs. For the adhesive with NACP, the biofilm medium became a Ca and P ion reservoir. The biofilm culture medium of the magnetic nanoparticle-containing adhesive with NACP had a safe pH of 6.9, while the biofilm medium of commercial adhesive had a cariogenic pH of 4.5. CONCLUSIONS: Magnetic nanoparticle-containing adhesive with DMAHDM and NACP under a magnetic force yielded much greater dentin bond strength than commercial control. The novel adhesive reduced biofilm CFU by 4 logs and increased the biofilm pH from a cariogenic pH 4.5 to 6.9, and therefore is promising to enhance the resin-tooth bond, strengthen tooth structures, and suppress secondary caries at the restoration margins.

Enamel demineralization is one of the most undesired side effects of fixed orthodontic treatment, which will lead to white spot lesions (WSLs) on tooth surfaces. The development of WSLs is due to prolonged accumulation of bacterial plaque and associated acid production. Self-etching adhesives have been used in orthodontic treatments with several advantages over the more traditional acid-etch method. However, current self-etching adhesives in orthodontic treatments have no antibacterial activity. The objectives of this study were to develop a self-etching and antibacterial orthodontic adhesive, and to investigate its enamel bond strength and antibacterial properties. A novel quaternary ammonium monomer dimethylaminohexadecyl methacrylate (DMAHDM) was incorporated into a commercial self-etching adhesive (Adper Easy One, 3M). It showed that the 5% DMAHDM appeared to be optimal in obtaining the strongest antibacterial function without compromising the enamel bond strength both at 15 min and after 30 days of immersion plus thermal cycling.

Biofilms at the tooth-restoration bonded interface can produce acids and cause recurrent caries. Recurrent caries is a primary reason for restoration failures. The objectives of this study were to synthesize a novel bioactive dental bonding agent containing dimethylaminohexadecyl methacrylate (DMAHDM) and 2-methacryloyloxyethyl phosphorylcholine (MPC) to inhibit biofilm formation at the tooth-restoration margin and to investigate the effects of water-aging for 6 months on the dentin bond strength and protein-repellent and antibacterial durability. A protein-repellent agent (MPC) and antibacterial agent (DMAHDM) were added to a Scotchbond multi-purpose (SBMP) primer and adhesive. Specimens were stored in water at 37 °C for 1, 30, 90, or 180 days (d). At the end of each time period, the dentin bond strength and protein-repellent and antibacterial properties were evaluated. Protein attachment onto resin specimens was measured by the micro-bicinchoninic acid approach. A dental plaque microcosm biofilm model was used to test the biofilm response. The SBMP + MPC + DMAHDM group showed no decline in dentin bond strength after water-aging for 6 months, which was significantly higher than that of the control (P 0.1). In conclusion, a bonding agent with MPC and DMAHDM achieved a durable dentin bond strength and long-term resistance to proteins and oral bacteria. The novel dental bonding agent is promising for applications in preventive and restorative dentistry to reduce biofilm formation at the tooth-restoration margin. A strong, long-lasting bonding agent developed by researchers in the US and China can protect tooth restorations from bacteria and proteins. Tooth restorations often fail due to recurrent decay which starts when bacteria invade the interface between the tooth and the restoration material, producing acids. To combat this problem, Ning Zhang at the University of Maryland, Baltimore, and Capital Medical University, Beijing, and co-workers infused the commercial Scotchbond Multi-Purpose primer and adhesive with two chemicals known to inhibit the formation of bacterial biofilms. The team stored bonded teeth in warm water for six months, after which they found no decline in the bond strength, or its resistance to oral proteins and bacteria. They hope that by incorporating the same protective chemicals into cements, composites and sealants, failure rates in tooth restorations could be reduced.

Background Compelling evidence indicates that metformin, a low-cost and safe orally administered biguanide prescribed to millions of type 2 diabetics worldwide, induces the osteoblastic differentiation of mesenchymal stromal cells (MSCs) through the 5' adenosine monophosphate (AMP)-activated protein kinase (AMPK) pathway. As a highly hydrophilic cationic compound, metformin uptake is facilitated by cell membrane organic cation transporters (OCTs) of the solute carrier 22A gene family. We hypothesized that to effectively enhance osteogenic differentiation, and ultimately bone regeneration, metformin must gain access into functional OCT-expressing MSCs. Methods Data was obtained through immunoblotting, cellular uptake, mineralization and gene expression assays. Results We demonstrate for the first time that functional OCTs are expressed in human-derived MSCs from umbilical cord Wharton's jelly, an inexhaustible source of nonembryonic MSCs with proven osteogenic potential. A clinically relevant concentration of metformin led to AMPK activation, enhanced mineralized nodule formation and increased expression of the osteogenic transcription factor Runt-related transcription factor 2 (RUNX2). Indeed, targeting OCT function through pharmacological and genetic approaches markedly blunted these responses. Conclusions Our findings indicate that functional OCT expression in UC-MSCs is a biological prerequisite that facilitates the intracellular uptake of metformin to induce an osteogenic effect. Future pre-clinical studies are warranted to investigate whether the expression of functional OCTs may serve as a potential biomarker to predict osteogenic responses to metformin.

Introduction Metformin is a first-line drug for treating type 2 diabetes that regulates the differentiation of mesenchymal stem cells. Its effects on human dental pulp cells (DPCs) remain unknown. This study aimed to investigate the effects of metformin on the proliferation and differentiation of DPCs. Methods A live/dead viability assay kit was used to examine the effects of metformin on the cell viability of DPCs. Cell proliferation was analyzed using a cell counting kit (CCK-8; Dojindo, Tokyo, Japan). Levels of phosphorylated and unphosphorylated adenosine 5′-monophosphate-activated protein kinase (AMPK) were quantified by Western blot analysis in response to metformin and the AMPK signaling inhibitor Compound C (EMD Chemicals, San Diego, CA). The effects of Compound C on the metformin-induced odontoblast differentiation of DPCs were determined by alkaline phosphatase activity assay and von Kossa staining, and the expression of odontoblastic markers was evaluated by reverse-transcription polymerase chain reaction analysis. Results DPCs exhibited mesenchymal stem cell characteristics using flow cytometry. Different doses of metformin were shown to be cytocompatible with DPCs, yielding >90% cell viability. None of the concentrations of metformin up to 50 μmol/L affected cell proliferation. The Western blot assay showed that DPCs express functional organic cation transporter 1, a transmembrane protein that mediates the intracellular uptake of metformin. Metformin significantly activated the AMPK pathway in a dose-dependent manner. In addition, it stimulated alkaline phosphatase activity; enhanced mineralized nodule formation; and increased the expression of odontoblastic markers including dentin sialophosphoprotein, dentin matrix protein 1, runt-related transcription factor 2, and osteocalcin. Moreover, pretreatment with Compound C, a specific AMPK inhibitor, markedly reversed metformin-induced odontoblastic differentiation and cell mineralization. Conclusions This study shows that metformin can induce DPC differentiation and mineralization in an AMPK-dependent manner and that this well-tolerated antidiabetic drug has potential in regenerative endodontics as well as in other regenerative applications.

Osteomyelitis is caused by Staphylococcus aureus (S. aureus), with associated progressive bone loss. This study developed for the first time a calcium phosphate cement (CPC) for delivery of doxycycline (DOX) and human platelet lysate (hPL) to fight against S. aureus infection and enhance the osteogenesis of human periodontal ligament stem cells (hPDLSCs). Chitosan-containing CPC scaffolds were fabricated in the absence (CPCC) or presence of DOX (CPCC+DOX). In addition, hPL was encapsulated in alginate microbeads and incorporated into CPCC+DOX (CPCC+DOX+ hPL). Flexural strength of CPCC+DOX + hPL was (5.56 ± 0.55) MPa, lower than (8.26 ± 1.6) MPa of CPCC+DOX (p 0.05), but exceeding the reported strength of cancellous bone. CPCC+DOX and CPCC+DOX + hPL exhibited strong antibacterial activity against S. aureus, reducing biofilm CFU by 4 orders of magnitude. The hPDLSCs encapsulated in microbeads were co-cultured with the CPCs. The hPDLSCs were able to be released from the microbeads and showed a high proliferation rate, increasing by about 8 folds at 14 days for all groups. The hPL was released from the scaffold and promoted the osteogenic differentiation of hPDLSCs. ALP activity was 28.07 ± 5.15 mU/mg for CPCC+DOX + hPL, higher than 17.36 ± 2.37 mU/mg and 1.34 ± 0.37 mU/mg of CPCC+DOX and CPCC, respectively (p 0.05). At 7 days, osteogenic genes (ALP, RUNX2, COL-1, and OPN) in CPCC+DOX + hPL were 3-10 folds those of control. The amount of hPDLSC-synthesized bone mineral with CPCC+DOX + hPL was 3.8 folds that of CPCC (p 0.05). In summary, the novel CPC + DOX + hPL-hPDLSCs scaffold exhibited strong antibacterial activity, excellent cytocompatibility and hPDLSC osteogenic differentiation, showing a promising approach for treatment and prevention of bone infection and enhancement of bone regeneration.

Patients with dry mouth often have an acidic oral environment lacking saliva to provide calcium (Ca) and phosphate (P) ions. There has been no report on tooth remineralization in acidic pH4 and CaP ion-lacking solutions. The objective of this study was to develop a novel method of combining poly(amido amine) (PAMAM) with adhesive containing nanoparticles of amorphous calcium phosphate (NACP) for dentin remineralization in pH4 and CaP-lacking solution for the first time. Demineralized dentin was tested in four groups: (1) dentin control, (2) dentin with PAMAM, (3) dentin with NACP adhesive, (4) dentin with PAMAM + NACP adhesive. Dentin samples were examined by scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDS) and hardness testing. Increasing the NACP filler level in adhesive from 0 to 40 wt% did not negatively affect the dentin bond strength (p > 0.1). NACP adhesive released CaP ions and neutralized the acid. PAMAM alone failed to achieve dentin remineralization in lactic acid. NACP alone induced slight dentin remineralization in lactic acid (p > 0.1). In contrast, the novel PAMAM + NACP group in the pH4 and CaP-lacking solution completely remineralized the predemineralized dentin, increasing its hardness which approached that of healthy dentin (p > 0.1). In conclusion, dentin remineralization via PAMAM + NACP adhesive in pH4 and CaP-lacking acid was achieved for the first time, when conventional remineralization methods such as PAMAM or NACP did not work. The novel PAMAM + NACP method is promising to increase the longevity of the composite-tooth bond, inhibit caries, remineralize lesions and protect tooth structures, even for patients with dry mouth and an acidic oral environment. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2414-2424, 2018.

Metformin, a first-line antidiabetic drug used by millions of patients, has been shown to have potential osteogenic properties. The present study was performed to test the hypothesis that clinically relevant doses of metformin promote the osteogenic differentiation and mineralization of induced pluripotent stem cell-derived mesenchymal stem cells (iPSC-MSCs). iPSC-MSCs were treated with metformin (10 μm) to assess cell viability, osteogenic differentiation, mineralization and activation of the LKB1/AMP-activated protein kinase (AMPK) signalling pathway, a surrogate marker of metformin action. To determine its potential application in MSC-based bone and periodontal tissue engineering, iPSC-MSCs were also treated with metformin when seeded on to calcium phosphate cement (CPC) scaffolds. Immunoblotting and cellular uptake assays showed that iPSC-MSCs express functional organic cation transporter-1 (OCT-1), a transmembrane protein that mediates the intracellular uptake of metformin. Although metformin treatment did not impair iPSC-MSC viability, it significantly stimulated alkaline phosphatase activity, enhanced mineralized nodule formation and increased expression of osteogenic markers, including Runt-related transcription factor 2 (RUNX2) and osterix. Inhibition of LKB1 activity, a common upstream AMPK kinase, markedly reversed metformin-induced AMPK activation, RUNX2 expression and nuclear localization. Moreover, metformin substantially increased mineralized nodule formation of iPSC-MSC seeded on CPC scaffolds. Collectively, functional OCT-expressing iPSC-MSCs responded to metformin by inducing an osteogenic effect in part mediated by the LKB1/AMPK pathway. Considering the widespread use of metformin in diabetics, this work may lead to novel tissue-engineering platforms where autogenous OCT-expressing iPSC-MSCs might be used to enhance bone and periodontal regeneration in diabetic patients prescribed with daily doses of metformin.

Angiogenesis is a limiting factor in regenerating large bone defects. The objective of this study was to investigate angiogenic and osteogenic effects of co-culture on calcium phosphate cement (CPC) scaffold using human umbilical vein endothelial cells (hUVECs) and mesenchymal stem cells (MSCs) from different origins for the first time. hUVECs were co-cultured with four types of cell: human umbilical cord MSCs (hUCMSCs), human bone marrow MSCs (hBMSCs) and MSCs from induced pluripotent stem cells (hiPSC-MSCs) and embryonic stem cells (hESC-MSCs). Constructs were implanted in 8 mm cranial defects of rats for 12 weeks. CPC without cells served as control 1. CPC with hBMSCs served as control 2. Microcapillary-like structures were successfully formed on CPC in vitro in all four co-cultured groups. Microcapillary lengths increased with time (p 0.05). Osteogenic and angiogenic gene expressions were highly elevated and mineralization by co-cultured cells increased with time (p 0.05). New bone amount and blood vessel density of co-cultured groups were much greater than controls (p 0.05) in an animal study. hUVECs co-cultured with hUCMSCs, hiPSC-MSCs and hESC-MSCs achieved new bone and vessel density similar to hUVECs co-cultured with hBMSCs (p 0.1). Therefore, hUCMSCs, hiPSC-MSCs and hESC-MSCs could serve as alternative cell sources to hBMSCs, which require an invasive procedure to harvest. In conclusion, this study showed for the first time that co-cultures of hUVECs with hUCMSCs, hiPSC-MSCs, hESC-MSCs and hBMSCs delivered via CPC scaffold achieved excellent osteogenic and angiogenic capabilities in vivo. The novel co-culture constructs are promising for bone reconstruction with improved angiogenesis for craniofacial/orthopaedic applications. Copyright © 2017 John WileySons, Ltd.

Enamel demineralization is destructive, esthetically compromised, and costly complications for orthodontic patients. Nano-sized amorphous calcium phosphate (NACP) has been explored to address this challenge. The 20% NACP-loaded ortho-cement notably exhibited favorable behavior on reducing demineralization of enamel around brackets in a caries model designed to simulate the carious attack. The 20% NACP-loaded ortho-cement markedly promotes higher calcium and phosphate release at a low pH, and the mineral loss was almost two fold lower and carious lesion depth decreased the by 1/3. This novel approach is promising co-adjuvant route for prevention of dental caries dissemination in millions of patients under orthodontic treatment.