Nanoparticles Of Amorphous Calcium Phosphate Research Articles

Novel Remineralizing and Antibiofilm Low-Shrinkage-Stress Nanocomposites to Inhibit Salivary Biofilms and Protect Tooth Structures.

Recurrent caries remain a persistent concern, often linked to microleakage and a lack of bioactivity in contemporary dental composites. Our study aims to address this issue by developing a low-shrinkage-stress nanocomposite with antibiofilm and remineralization capabilities, thus countering the progression of recurrent caries. In the present study, we formulated low-shrinkage-stress nanocomposites by combining triethylene glycol divinylbenzyl ether and urethane dimethacrylate, incorporating dimethylaminododecyl methacrylate (DMADDM), along with nanoparticles of calcium fluoride (nCaF2) and nanoparticles of amorphous calcium phosphate (NACP). The biofilm viability, biofilm metabolic activity, lactic acid production, and ion release were evaluated. The novel formulations containing 3% DMADDM exhibited a potent antibiofilm activity, exhibiting a 4-log reduction in the human salivary biofilm CFUs compared to controls (p < 0.001). Additionally, significant reductions were observed in biofilm biomass and lactic acid (p < 0.05). By integrating both 10% NACP and 10% nCaF2 into one formulation, efficient ion release was achieved, yielding concentrations of 3.02 ± 0.21 mmol/L for Ca, 0.5 ± 0.05 mmol/L for P, and 0.37 ± 0.01 mmol/L for F ions. The innovative mixture of DMADDM, NACP, and nCaF2 displayed strong antibiofilm effects on salivary biofilm while concomitantly releasing a significant amount of remineralizing ions. This nanocomposite is a promising dental material with antibiofilm and remineralization capacities, with the potential to reduce polymerization-related microleakage and recurrent caries.

Novel Bioactive Nanocomposites Containing Calcium Fluoride and Calcium Phosphate with Antibacterial and Low-Shrinkage-Stress Capabilities to Inhibit Dental Caries.

Composites are commonly used for tooth restorations, but recurrent caries often lead to restoration failures due to polymerization shrinkage-stress-induced marginal leakage. The aims of this research were to: (1) develop novel low-shrinkage-stress (L.S.S.) nanocomposites containing dimethylaminododecyl methacrylate (DMADDM) with nanoparticles of calcium fluoride (nCaF2) or amorphous calcium phosphate (NACP) for remineralization; (2) investigate antibacterial and cytocompatibility properties. Nanocomposites were made by mixing triethylene glycol divinylbenzyl ether with urethane dimethacrylate containing 3% DMADDM, 20% nCaF2, and 20% NACP. Flexural strength, elastic modulus, antibacterial properties against Streptococcus mutans biofilms, and cytotoxicity against human gingival fibroblasts and dental pulp stem cells were tested. Nanocomposites with DMADDM and nCaF2 or NACP had flexural strengths matching commercial composite control without bioactivity. The new nanocomposite provided potent antibacterial properties, reducing biofilm CFU by 6 logs, and reducing lactic acid synthesis and metabolic function of biofilms by 90%, compared to controls (p < 0.05). The new nanocomposites produced excellent cell viability matching commercial control (p > 0.05). Bioactive L.S.S. antibacterial nanocomposites with nCaF2 and NACP had excellent bioactivity without compromising mechanical and cytocompatible properties. The new nanocomposites are promising for a wide range of dental restorations by improving marginal integrity by reducing shrinkage stress, defending tooth structures, and minimizing cariogenic biofilms.

Resin Loaded with Nanoparticles and Remineralizing Agents on the Bond Integrity and Microleakage of Tooth Color Restoration Bonded to the Bleached Enamel Surface

Aims: Assessment of remineralizing agent-modified resin adhesives i.e., Nanoparticles of amorphous calcium phosphate (NACP), Casein phosphopeptide-amorphous calcium phosphate (CPP- ACP), and Sodium Fluoride (NaF) on the shear bond strength (SBS) and microleakage of tooth color restoration bonded to the bleached enamel. Materials and Methods: Eighty premolar buccal enamel surfaces were bleached using a 10% carbamide peroxide gel. After 2 weeks samples were etched and then randomly allocated into four groups (n = 20) Group 1: No remineralizating agent + ER, Group 2: NaF + ER, Group 3: CPP-ACP + ER, and Group 4: NACP + ER. Composite restoration was performed followed by immersion in distilled water. Microleakage, SBS, and failure mode assessment were achieved. The mean and standard deviation (SD) of SBS and marginal sealing were compared using the Tukey post hoc test and One-way ANOVA. Results: Group 1 samples displayed the maximum value of marginal leakage. However, Group 4 samples exhibited the minimum scores of microleakage. Regarding SBS, Group 4 unveiled the highest outcomes of bond integrity. Whereas, Group 1 presented the lowest value of SBS. Conclusion: NACP-modified resin adhesives proved to be a better and safer alternative in improving the bond strength and microleakage scores of tooth color restorative material to bleached enamel.

Dual-functional adhesive containing amorphous calcium phosphate nanoparticles and dimethylaminohexadecyl methacrylate promoted enamel remineralization in a biofilm-challenged environment

ObjectiveThe 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. MethodsArtificial 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. ResultsEnamel of NACP+DMAHDM group demonstrated excellent remineralization effectiveness. And the NACP+DMAHDM adhesive released a great number of Ca2+ and PO43- ions, increased pH to 5.81 via acid neutralization, decreased production of lactic acid, and reduced CFU count of S. mutans (P < 0.05). SignificanceThe NACP+DMAHDM adhesive would be applicable to preventing secondary caries, strengthening enamel-adhesive interface, and extending the lifespan of composite restoration.

Effects of thermal cycling on mechanical and antibacterial durability of bioactive low-shrinkage-stress nanocomposite

ObjectivesRecent 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. MethodsThe 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. ResultsAfter 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. ConclusionsBioactive 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. Clinical significanceThe 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.

Novel bioactive adhesive containing dimethylaminohexadecyl methacrylate and calcium phosphate nanoparticles to inhibit metalloproteinases and nanoleakage with three months of aging in artificial saliva

ObjectivesThe 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. MethodsDMAHDM 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. ResultsAdding 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). SignificanceDMAHDM+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.

Evaluation of the ability of adhesives with antibacterial and remineralization functions to prevent secondary caries in vivo.

The bonding interface of dental filling therapy is the weak point in resisting secondary caries. Adhesives containing nanoparticles of amorphous calcium phosphate (NACP) and dimethylaminohexadecyl methacrylate (DMAHDM) have been demonstrated in vitro to prevent bacteria from producing acid and to promote tooth remineralization. The present study aimed to evaluate the efficacy of adhesive with NACP and DMAHDM to prevent secondary caries in vivo. Artificial cavities were created on the first molar on both sides of the maxillary in a rat model. One side was treated with adhesive containing NACP + DMAHDM, while on the other side, a commercial adhesive served as control. After 24days of cariogenic feeding, the degree of secondary caries was evaluated by micro-CT and a modified Keyes scoring method. Quantitative real-time PCR (qPCR) and colony-forming unit (CFU) counts were used to evaluate the antibacterial efficacy of the materials. Biocompatibility was also investigated. In the rat model, the adhesive with NACP + DMAHDM showed excellent biocompatibility and effectively decreased the amount of bacteria. The experimental group demonstrated excellent remineralization effectiveness, with a lower modified Keyes score and mineral loss of 34.16 ± 2.13 vol% μm, compared with 77.44 ± 7.22 vol% μm in the control group, according to micro-CT (P < 0.05), showing excellent capacity to inhibit secondary caries. The NACP-DMAHDM-containing adhesive exhibited good performance in preventing secondary caries in vivo. Adhesives containing NACP and DMAHDM have great potential for use in clinical dentistry to prevent secondary caries by inhibiting bacterial growth and promoting remineralization.

Minimally-invasive dentistry via dual-function novel bioactive low-shrinkage-stress flowable nanocomposites

The objectives of this in vitro study were to develop a novel low-shrinkage-stress flowable nanocomposite with antibacterial properties through the incorporation of dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP), and investigate the mechanical and oral biofilm properties, to be used in minimally-invasive techniques. MethodsThe light-cured low-shrinkage-stress flowable resin was formulated by mixing urethane dimethacrylate (UDMA) and triethylene glycol divinylbenzyl ether (TEG-DVBE) at a 1:1 mass ratio. Different mass fractions of glass, and either 5% DMAHDM or 20%NACP or both were incorporated. Paste flowability, ultimate micro tensile strength and surface roughness were evaluated. The antibacterial response of DMAHDM resin was assessed by using biofilms of human saliva-derived microcosm model. Virtuoso flowable composite was used as a control. Results(45% resin+5% DMAHDM+20% NACP+30% glass) formula yielded the needed outcomes. It had flow rate within the range of ISO requirement. The micro tensile strength was (39.1 ± 4.3) MPa, similar to (40.1 ± 4.0) MPa for commercial control (p > 0.05). The surface roughness values of the novel composite (0.079 ± 0.01) µm similar to commercial composite (0.09 ± 0.02) µm (p > 0.05). Salivary microcosm biofilm colony forming unit values were reduced by 5–6 logs (p < 0.05). Biofilm metabolic activity was also substantially reduced, compared to control composite (p < 0.05). SignificanceThe novel bioactive flowable nanocomposite achieved strong antibacterial activities without compromising the mechanical properties. It is promising to be used as pit and fissure sealants, and as fillings in conservative cavities to inhibit recurrent caries and increase restoration longevity.

Novel nanostructured resin infiltrant containing calcium phosphate nanoparticles to prevent enamel white spot lesions

ObjectivesThe 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. MethodsNACP 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. ResultsIncorporating 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. SignificanceThe novel ICON+30% NACP infiltrant is promising to inhibit enamel WSLs, protect the enamel and increase its hardness.

Novel low-shrinkage-stress bioactive nanocomposite with anti-biofilm and remineralization capabilities to inhibit caries

Background/purposeA 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 methodsThe 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.ResultsAdding 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 < 0.05). Adding 5% DMAHDM did not compromise the Ca and P ion release. The low-shrinkage-stress composite maintained a high degree of conversion of approximately 70%, while reducing the shrinkage stress by 37%, compared to a commercial control (p < 0.05).ConclusionThe 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.

Dentin remineralization in acidic solution without initial calcium phosphate ions via poly(amido amine) and calcium phosphate nanocomposites after fluid challenges.

A previous study showed that the combination of poly(amido amine) (PAMAM) and rechargeable composites with nanoparticles of amorphous calcium phosphate (NACP) induced dentin remineralization in an acidic solution with no initial calcium (Ca) and phosphate (P) ions, mimicking the oral condition of individuals with dry mouths. However, the frequent fluid challenge in the oral cavity may decrease the remineralization capacity. Therefore, the objective of the present study was to investigate the remineralization efficacy on dentin in an acid solution via PAMAM + NACP after fluid challenges for the first time. The NACP nanocomposite was stored in a pH 4 solution for 77days to exhaust its Ca and P ions and then recharged. Demineralized dentin samples were divided into four groups: (1) control dentin, (2) dentin coated with PAMAM, (3) dentin with recharged NACP composite, and (4) dentin with PAMAM + recharged NACP. PAMAM-coated dentin was shaken in phosphate-buffered saline for 77days to desorb PAMAM from dentin. Samples were treated in pH 4 lactic acid with no initial Ca and P ions for 42days. After 77days of fluid challenge, PAMAM failed to prevent dentin demineralization in lactic acid. The recharged NACP nanocomposite raised the pH to above 6.5 and re-released more than 6.0 and 4.0mmol/L Ca and P ions daily, respectively, which inhibited further demineralization. In contrast, the PAMAM + NACP combined method induced great dentin remineralization and restored the dentin microhardness to 0.54 ± 0.04 GPa, which approached that of sound dentin (P = 0.426, P > 0.05). The PAMAM + NACP combination achieved dentin remineralization in an acid solution with no initial Ca and P ions, even after severe fluid challenges. The novel PAMAM + NACP has a strong and sustained remineralization capability to inhibit secondary caries, even for individuals with dry mouths.

Novel calcium phosphate ion-rechargeable and antibacterial adhesive to inhibit dental caries.

This study aimed to develop an antibacterial and calcium (Ca) and phosphate (P) rechargeable adhesive and investigate the effects of dimethylaminododecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP) on dentin bonding, biofilm response, and repeated Ca and P ion recharge and re-release capability for the first time. Pyromellitic glycerol dimethacrylate (PMGDM), ethoxylated bisphenol A dimethacrylate (EBPADMA), 2-hydroxyethyl methacrylate (HEMA), and bisphenol A glycidyl dimethacrylate (BisGMA) formed the adhesive (PEHB). Three groups were tested: (1) Scotchbond (SBMP, 3M) control, (2) PEHB + 30% NACP, and (3) PEHB + 30% NACP + 5% DMAHDM. Specimens were tested for dentin shear bond strength, and Ca and P ion release, recharge, and re-release. Biofilm lactic acid production and colony-forming units (CFU) on resins were analyzed. The four groups had similar dentin shear bond strengths (p > 0.1). Adhesive with DMAHDM showed significant decrease in metabolic activity, lactic acid production, and biofilm CFU (p < 0.05). The adhesives containing NACP released high levels of Ca and P ions initially and after being recharged. This study developed the first Ca and P ion-rechargeable and antibacterial adhesive, achieving strong antibacterial activity and Ca and P ion recharge and re-release for long-term remineralization. Considering the restoration-tooth bonded interface being the weak link and recurrent caries at the margins being the primary reason for restoration failures, this novel calcium phosphate-rechargeable and antibacterial adhesive is promising for a wide range of tooth-restoration applications to inhibit caries.

A comprehensive review of the antibacterial activity of dimethylaminohexadecyl methacrylate (DMAHDM) and its influence on mechanical properties of resin-based dental materials.

The repetitive restorative cycle should be avoided, aiming at the smallest number of restorations’ replacements to ensure greater tooth longevity. Antibacterial materials associated with the control of caries etiological factors can help improve restoration's durability. This review aimed to analyze the results of in vitro studies that added Dimethylaminohexadecyl methacrylate (DMAHDM), an antibacterial monomer, to restorative materials. The PubMed, SCOPUS, Web of Science, and Biblioteca Virtual em Saúde databases were screened for studies published between 2015 and 2020. After full-text reading, 24 articles were included in the final sample. DMAHDM has demonstrated antibacterial efficacy against several bacteria related to dental caries and periodontal diseases, causing a transition in the biofilm balance without inducing resistance. When DMAHDM was included in acrylic resin, the material cytotoxicity increased, and changes in mechanical properties were observed. In contrast, resin composites had their mechanical properties maintained in most studies; however, toxicity was not examined. The association between DMAHDM and 2-methacryloyloxyethyl phosphorylcholine or silver nanoparticles improved the antibacterial effect. Besides, the association with nanoparticles of amorphous calcium phosphate or nanoparticles of calcium fluoride can provide remineralization capacity. There is a lack of information on the cytotoxicity and bacteria resistance induction, and further studies are needed to address this.

Remineralization effectiveness of adhesive containing amorphous calcium phosphate nanoparticles on artificial initial enamel caries in a biofilm-challenged environment.

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 Antibacterial and Remineralizing Effects of Biomaterials Combined with DMAHDM Nanocomposite: A Systematic Review.

Researchers have developed novel nanocomposites that incorporate additional biomaterials with dimethylaminohexadecyl methacrylate (DMAHDM) in order to reduce secondary caries. The aim of this review was to summarize the current literature and assess the synergistic antibacterial and remineralizing effects that may contribute to the prevention of secondary caries. An electronic search was undertaken in MEDLINE using PubMed, Embase, Scopus, Web of Science and Cochrane databases. The initial search identified 954 papers. After the removal of duplicates and screening the titles and abstracts, 15 articles were eligible for this review. The amalgamation of 2-methacryloyloxyethyl phosphorylcholine (MPC) and silver nanoparticles (AgNPs) with DMAHDM resulted in increased antibacterial potency. The addition of nanoparticles of amorphous calcium phosphate (NACP) and polyamidoamine dendrimers (PAMAM) resulted in improved remineralization potential. Further clinical studies need to be planned to explore the antibacterial and remineralizing properties of these novel composites for clinical success.

Long-term antibacterial activity and cytocompatibility of novel low-shrinkage-stress, remineralizing composites

A low-shrinkage-stress (LSS), antibacterial and remineralizing nanocomposite was recently developed; however, validation of its long-term antibacterial potency in modulating human salivary-derived biofilm is an unmet need. This study aimed to evaluate the antibacterial effect of the bioactive LSS composite before and after aging in acidic solution for 90 days using a multi-species biofilm model, and to evaluate its cytotoxicity. The LSS composite consisted of urethane dimethacrylate (UDMA) and triethylene glycol divinylbenzyl ether (TEG-DVBE), 3% dimethylaminohexadecyl methacrylate (DMAHDM) and 20% nanoparticles of amorphous calcium phosphate (NACP). Biofilm colony-forming units (CFU), lactic acid production, and confocal laser scanning microscopy (3D biofilm) were evaluated before and after three months of aging. Cytotoxicity was assessed against human gingival fibroblasts (HGF). The new LSS composite presented the lowest biofilm CFU, lactic acid and biofilm biomass, compared to controls (n = 6, p < 0.05). Importantly, the new composite exhibited no significant difference in antibacterial performance before and after 90-day-aging, demonstrating long-term antibacterial activity (p > 0.1). The LSS antibacterial and remineralizing composite presented a low cell viability at original extract that has increased with further dilutions. In conclusion, this study spotlighted that the new bioactive composite not only had a low shrinkage stress, but also down-regulated the growth of oral biofilms, reduced acid production, maintained antibacterial activity after the 90-day-aging, and did not compromise the cytocompatibility.

Low-shrinkage-stress nanocomposite: An insight into shrinkage stress, antibacterial, and ion release properties.

The aims are: (a) To develop the first low-shrinkage-stress nanocomposite with antibacterial and remineralization capabilities through the incorporation of dimethylaminododecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP); (b) to investigate the effects of the new composite on biofilm inhibition, mechanical properties, shrinkage stress, and calcium (Ca) and phosphate (P) ion releases. The low-shrinkage-stress resin consisted of urethane dimethacrylate and triethylene glycol divinylbenzyl ether. Composite was formulated with 3% DMAHDM and 20% NACP. Mechanical properties, shrinkage stress, and degree of conversion were evaluated. Streptococcus mutans biofilm growth on composites was assessed. Ca and P ion releases were measured. The shrinkage stress of the low-shrinkage-stress composite containing 3% DMAHDM and 20% NACP was 36% lower than that of traditional composite control (p < 0.05), with similar degrees of conversion of 73.9%. The new composite decreased the biofilm colony-forming unit by 4 log orders and substantially reduced biofilm lactic acid production compared to control composite (p < 0.05). Incorporating DMAHDM to the low-shrinkage-stress composite did not adversely affect the Ca and P ion release. A novel bioactive nanocomposite was developed with low shrinkage stress, strong antibiofilm activity, and high levels of ion release for remineralization, without undermining the mechanical properties and degree of conversion.

Pronounced Effect of Antibacterial Bioactive Dental Composite on Microcosm Biofilms Derived From Patients With Root Carious Lesions

Resin composites are the material of choice for dental restorative treatment in oral health care. However, the inherent composition of this class of material commonly results in microbial adherence and colonization, which carries the potential risk of recurrent carious lesions around dental restorations. The high risk of resin composites failure complicates the treatment of root caries, defined as the onset of tooth decay over the prone root surface of a tooth. The restorative treatment of root caries among high caries risk individuals, especially for senior patients, is a challenging, painful, and costly. The dysbiotic microbiota colonizes the composite’s surfaces and forms polymicrobial biofilms that are difficult to be dislodged by regular tooth brushing. This study assesses the antibiofilm performance of a surface contact killing antibacterial dental resin composites on the growth of microcosm biofilms using dental plaque sampled from patients with active root carious lesions as an inoculum. The designed formulations contain dimethylaminohexadecyl methacrylate (DMAHDM), a tailored quaternary ammonium monomer with an alkyl chain length of 16, at 3–5 wt.% in a base resin with and without 20 wt.% nanoparticles of amorphous calcium phosphate (NACP). Biofilms were grown on the tested resin composites using a 48 h plaque-derived microcosm biofilm model. Dental plaque collected from active root carious lesions was used as an inoculum to emulate the microbiota present in those lesions. The biofilm growth was assessed via the colony-forming unit (CFU) counts in four culture media, metabolic behavior, lactic acid production, and confocal microscopy. The percentage of reacted double bonds of the formulations was also investigated. The dental resin composites formulated with 3–5 wt.% DMAHDM and 20 wt.% NACP were effective at eradicating surface-attached biofilms from the total microbial load and each relevant cariogenic group: total streptococci, mutans streptococci, and lactobacilli. The metabolic activities and lactic acid production of the plaque-derived microcosm biofilms were reduced by 80–95%, respectively. Fewer viable microorganisms were observed over resin composites containing DMAHDM and NACP. Besides, all the experimental formulations demonstrated an acceptable degree of conversion values. This new strategy fits with ongoing dental caries preventive and minimally invasive approaches by preventing biofilm growth over-restored carious root lesions and improving the lifespan of dental restorations.

Rechargeable adhesive with calcium phosphate nanoparticles inhibited long-term dentin demineralization in a biofilm-challenged environment

ObjectivesThis study aims to investigate the long-term demineralization-inhibition capability of a rechargeable adhesive with nanoparticles of amorphous calcium phosphate (NACP) on dentin in a biofilm-challenged environment. MethodsThe NACP adhesive was immersed in a pH 4 solution to exhaust calcium (Ca) and phosphate (P) ions and then recharged with Ca and P ions. Dentin samples were demineralized underStreptococcus mutans biofilms for 24 h and randomly divided into two groups: (1) dentin control, (2) dentin with recharged NACP adhesives. Each day, all the samples were immersed in brain heart infusion broth with 1% sucrose (BHIS) for 4 h, and then in artificial saliva (AS) for 20 h. This cycle was repeated for 10 days. The pH of BHIS, the Ca and P ions content of the BHIS and AS were measured daily. After 10 days, the lactic acid production and colony-forming units of the biofilms were tested. The changes of remineralization/demineralization were also analyzed. ResultsDentin in the control group showed further demineralization. The recharged NACP adhesive neutralized acids, increasing the pH to above 5, and released large amounts of Ca and P ions each day. The recharged NACP adhesive decreased the production of lactic acid (P < 0.05), inhibited dentin demineralization and sustained the dentin hardness in the biofilm-challenged environment, showing an excellent long-term demineralization-inhibition capability. ConclusionsThe NACP adhesive could continuously inhibit dentin demineralization in a biofilm-challenged environment by recharging with Ca and P ions. SignificanceThe rechargeable NACP adhesive could provide long-term dentin bond protection.

Novel Nanocomposite Inhibiting Caries at the Enamel Restoration Margins in an In Vitro Saliva-Derived Biofilm Secondary Caries Model.

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 > 0.05). The biofilms’ CFU (colony-forming units) were reduced by 2 logs via DMAHDM (p < 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 < 0.05). With DMAHDM + NACP, the enamel hardness was the greatest and about 50% higher than control (p < 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.