Intercellular crosstalk in adult dental pulp is mediated by heparin-binding growth factors Pleiotrophin and Midkine.

BACKGROUND: In-depth knowledge of the cellular and molecular composition of dental pulp (DP) and the crosstalk between DP cells that drive tissue homeostasis are not well understood. To address these questions, we performed a comparative analysis of publicly available single-cell transcriptomes of healthy adult human DP to 5 other reference tissues: peripheral blood mononuclear cells, bone marrow, adipose tissue, lung, and skin. RESULTS: Our analysis revealed that DP resident cells have a unique gene expression profile when compared to the reference tissues, and that DP fibroblasts are the main cell type contributing to this expression profile. Genes coding for pleiotrophin (PTN) and midkine (MDK), homologous heparin-binding growth-factors, possessed the highest differential expression levels in DP fibroblasts. In addition, we identified extensive crosstalk between DP fibroblasts and several other DP resident cells, including Schwann cells, mesenchymal stem cells and odontoblasts, mediated by PTN and MDK. CONCLUSIONS: DP fibroblasts emerge as unappreciated players in DP homeostasis, mainly through their crosstalk with glial cells. These findings suggest that fibroblast-derived growth factors possess major regulatory functions and thus have a potential role as dental therapeutic targets.

Mice Lacking PLAP-1/Asporin Show Alteration of Periodontal Ligament Structures and Acceleration of Bone Loss in Periodontitis.

Periodontal ligament-associated protein 1 (PLAP-1), also known as Asporin, is an extracellular matrix protein expressed in the periodontal ligament and plays a crucial role in periodontal tissue homeostasis. Our previous research demonstrated that PLAP-1 may inhibit TLR2/4-mediated inflammatory responses, thereby exerting a protective function against periodontitis. However, the precise roles of PLAP-1 in the periodontal ligament (PDL) and its relationship to periodontitis have not been fully explored. In this study, we employed PLAP-1 knockout mice to investigate its roles and contributions to PDL tissue and function in a ligature-induced periodontitis model. Mandibular bone samples were collected from 10-week-old male C57BL/6 (WT) and PLAP-1 knockout (KO) mice. These samples were analyzed through micro-computed tomography (µCT) scanning, hematoxylin and eosin (HE) staining, picrosirius red staining, and fluorescence immunostaining using antibodies targeting extracellular matrix proteins. Additionally, the structure of the PDL collagen fibrils was examined using transmission electron microscopy (TEM). We also conducted tooth extraction and ligature-induced periodontitis models using both wild-type and PLAP-1 KO mice. PLAP-1 KO mice did not exhibit any changes in alveolar bone resorption up to the age of 10 weeks, but they did display an enlarged PDL space, as confirmed by µCT and histological analyses. Fluorescence immunostaining revealed increased expression of extracellular matrix proteins, including Col3, BGN, and DCN, in the PDL tissues of PLAP-1 KO mice. TEM analysis demonstrated an increase in collagen diameter within the PDL of PLAP-1 KO mice. In line with these findings, the maximum stress required for tooth extraction was significantly lower in PLAP-1 KO mice in the tooth extraction model compared to WT mice (13.89 N ± 1.34 and 16.51 N ± 1.31, respectively). In the ligature-induced periodontitis model, PLAP-1 knockout resulted in highly severe alveolar bone resorption, with a higher number of collagen fiber bundle tears and significantly more osteoclasts in the periodontium. Our results demonstrate that mice lacking PLAP-1/Asporin show alteration of periodontal ligament structures and acceleration of bone loss in periodontitis. This underscores the significant role of PLAP-1 in maintaining collagen fibrils in the PDL and suggests the potential of PLAP-1 as a therapeutic target for periodontal diseases.

In Silico Finite Element Analysis of Implant-Supported CAD-CAM Resin Composite Crowns.

PURPOSE: The aim of this study was to evaluate the mechanical behavior of an implant-supported crown made using computer-aided design and computer-aided manufacturing (CAD-CAM) resin composite (RC) blocks in the posterior region. MATERIAL AND METHODS: Four commercially available CAD-CAM RC blocks were used in this study: Cerasmart 300 (CS300; GC, Tokyo, Japan), Katana Avencia P Block (KAP; Kuraray Noritake Dental, Niigata, Japan); KZR HR3 Gamma Theta (HR3; Yamakin, Osaka, Japan), and Estelite P block (ESP; Tokuyama Dental, Tokyo, Japan). Katana Zirconia STML (ST; Kuraray Noritake Dental) was used as the control group. The elastic moduli of each material were determined by a three-point bending test. After the CAD models were designed, two different loading scenarios (oblique, vertical) were created. 3D finite element analysis was conducted with the prepared models. RESULTS: The elastic modulus of the material utilized for the implant restorations did not cause any change in the stresses transmitted to the implant or peripheral bone. An important difference was detected in the abutment-crown junction area. The minimum von Mises value at the abutment-crown interface was obtained in ST, which has the closest elastic modulus to the titanium abutment. CONCLUSIONS: The 3D finite element model designed in this study was used to demonstrate that implant-supported crowns fabricated with four different CAD-CAM RCs showed no critical stress concentrations in the bone or implant under all loading conditions. These results suggest that CAD-CAM RC blocks could be used as an alternative material for implant-supported restorations in the posterior region in terms of stress distribution.

Mechanical behavior of endodontically treated teeth: A three-dimensional finite element analysis using displacement vector.

PURPOSE: This study aimed to analyze the effects of core materials, remaining tooth structures, and interfacial bonding on stress distribution in endodontically treated teeth using finite element analysis (FEA). MATERIALS AND METHODS: Three-dimensional FEA was conducted using a reverse engineering technique based on maxillary premolars scanned by micro-computed tomography. Six models were generated with or without ferrules and with one of the following three abutment systems: metal core, resin core, or resin core with fiber posts. In each model, bonding and debonding were assumed in the dentin and surrounding structures: bonded and debonded models. The maximum principal stress values were recorded, and stress distribution of the entire restored teeth and dentin was generated. Furthermore, the distribution of the displacement vector of the debonded models was generated. RESULTS: In comparing the bonded and debonded models, the debonded models showed larger values for tensile stresses than those in bonded models for all abutment models. The models without ferrules rotated around the center of the abutment, whereas those with ferrules did not show remarkable displacement in the analysis. CONCLUSION: FEA assuming fracture of adhesive interface proved to be an effective method to clarify the significance of ferrules. It prevents stress concentration in dentin by reducing the rotation of the abutment, even when the adhesive fails.

Poly(lactic acid/caprolactone) bilayer membrane blocks bacterial penetration.

BACKGROUND AND OBJECTIVE: The clinical outcomes of guided tissue regeneration (GTR) or guided bone regeneration (GBR) procedures can be impaired if a bacterial infection develops at the surgical site. Membrane exposure is one of the causes of the onset of bacterial infection. Previously, we have fabricated a poly(lactic acid/caprolactone) (PLCL) bilayer membrane composed of a porous layer and a compact layer. The compact layer acts as a barrier against connective tissue and epithelial cells, and we hypothesized that it could also be an effective barrier against bacterial cells. The objective of this study was to evaluate the ability of the PLCL bilayer membrane to block bacterial cell penetration, which would be useful for preventing postoperative infections. METHODS: Porphyromonas gingivalis, Streptococcus mutans, and multispecies bacteria collected from human saliva were used in this study. Bacteria were seeded directly on the compact layer of a PLCL bilayer membrane, and bacterial adhesion to the membrane, as well as penetration into the membrane's structure, were assessed. Bacterial adhesion was evaluated by the number of colonies formed at 6, 24, and 72 h, and penetration was observed using a scanning electron microscope at 24 and 72 h. Commercially available membranes, composed of poly(lactic-co-glycolic acid) or type I collagen, were used as controls. RESULTS: P. gingivalis, S. mutans, and the multispecies bacteria obtained from human saliva adhered onto all the membranes after only 6 h of incubation. However, fewer adherent cells were observed for the PLCL bilayer membrane compared with the controls for all experimental periods. The PLCL membrane was capable of blocking bacterial penetration, and no bacterial cells were observed in the structure. In contrast, bacteria penetrated both the control membranes and were observed at depths of up to 80 µm after 72 h of incubation. CONCLUSION: Membrane characteristics may influence how bacterial colonization occurs. The PLCL membrane had reduced bacterial adhesion and blocked bacterial penetration, and these characteristics could contribute to a favorable outcome for regenerative treatments. In the event of membrane exposure at GTR/GBR surgical sites, membranes with an efficient barrier function, such as the PLCL bilayer membrane, could simplify the management of GTR/GBR complications.

Fabrication of pH-Responsive Zn2+-Releasing Glass Particles for Smart Antibacterial Restoratives.

The on-demand release of antibacterial components due to pH variations caused by acidogenic/cariogenic bacteria is a possible design for smart antibacterial restorative materials. This study aimed to fabricate pH-responsive Zn2+-releasing glass particles and evaluate their solubilities, ion-releasing characteristics, and antibacterial properties in vitro. Three kinds of silicate-based glass particles containing different molar ratios of Zn (PG-1: 25.3; PG-2: 34.6; PG-3: 42.7 mol%) were fabricated. Each particle was immersed in a pH-adjusted medium, and the solubility and concentration of the released ions were determined. To evaluate the antibacterial effect, Streptococcus mutans was cultured in the pH-adjusted medium in the presence of each particle, and the bacterial number was counted. The solubility and concentration of Zn2+ released in the medium increased with a decrease in medium pH. PG-3 with a greater content of Zn demonstrated higher concentrations of released Zn2+ compared with PG-1 and PG-2. PG-2 exhibited bactericidal effects at pH 5.1, whereas PG-3 demonstrated bactericidal effects at pH values of 5.1 and 6.1, indicating that PG-3 was effective at inhibiting S. mutans even under slightly acidic conditions. The glass particle with 42.7 mol% Zn may be useful for developing smart antibacterial restoratives that contribute to the prevention of diseases such as caries on root surfaces with lower acid resistance.

Large three-dimensional cell constructs for tissue engineering.

Much research has been conducted on fabricating biomimetic biomaterials in vitro. Tissue engineering approaches are often conducted by combining cells, scaffolds, and growth factors. However, the degradation rate of scaffolds is difficult to control and the degradation byproducts occasionally limit tissue regeneration. To overcome these issues, we have developed a novel system using a thermo-responsive hydrogel that forms scaffold-free, three-dimensional (3D) cell constructs with arbitrary size and morphology. 3D cell constructs prepared using bone marrow-derived stromal stem cells (BMSCs) exhibited self-organizing ability and formed bone-like tissue with endochondral ossification. Endothelial cells were then introduced into the BMSC construct and a vessel-like structure was formed within the constructs. Additionally, the bone formation ability was promoted by endothelial cells and cell constructs could be freeze-dried to improve their clinical application. A pre-treatment with specific protein protectant allowed for the fabrication of novel bone substitutes composed only of cells. This 3D cell construct technology using thermo-responsive hydrogels was then applied to other cell species. Cell constructs composed of dental pulp stem cells were fabricated, and the resulting construct regenerated pulp-like tissue within a human pulpless tooth. In this review, we demonstrate the approaches for the in vitro fabrication of bone and dental pulp-like tissue using thermo-responsive hydrogels and their potential applications.

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.

Application of a direct pulp capping cement containing S-PRG filler.

OBJECTIVES: To evaluate new pulp capping cements containing surface pre-reacted glass ionomer (S-PRG) filler and to investigate ion release kinetics and pH shift of eluates from the cement. MATERIALS AND METHODS: Molars of Wistar rats were directly pulp capped using three kinds of cement containing S-PRG filler and mineral tri-oxide aggregate (MTA) was used as a control. After 1, 2, or 4 weeks, histological evaluation was performed and differences of tertiary dentin formation were analyzed. Release of Sr2+, BO33-, SiO32-, Na+, and Al3+ ions was determined by inductively coupled plasma-atomic emission spectrometry, and F- ion release was measured using a fluoride ion selective electrode. The pH of the eluate from each cement after mixing was measured with a pH electrode. RESULTS: One of S-PRG cements promoted tertiary dentin formation to the same extent as the control (p > 0.05) and it showed a tendency of less inflammatory response. This cement released more BO33- and SiO32-, but less Sr2+, Na+, and F- than other S-PRG specimens. Each cement recovered nearly neutral compared with glass ionomer cement. CONCLUSIONS: S-PRG cement induced tertiary dentin formation based on multiple ion releases, suggesting that it is suitable as a pulp capping material. CLINICAL RELEVANCE: This new material can be an alternative pulp capping agent to MTA.

In Silico Analysis of the Biomechanical Stability of Commercially Pure Ti and Ti-15Mo Plates for the Treatment of Mandibular Angle Fracture.

PURPOSE: To investigate the influence of different materials and fixation methods on maximum principal stress (MPS) and displacement in reconstruction plates using in silico 3-dimensional finite element analysis (3D-FEA). MATERIALS AND METHODS: Computer-assisted designed (CAD) models of the mandible and teeth were constructed. Champy and AO/ASIF plates and fixation screws were designed with CAD software. 3D-FEA was performed by image-based CAE software. Maximum and minimum values of biomechanical stability, MPS, and displacement distribution were compared in Champy and AO/ASIF plates made from commercially pure titanium grade 2 (cp-Ti) and a titanium-and-molybdenum (14.47% wt) alloy (Ti-15Mo). RESULTS: For plates fixed on a model of a fractured left angle of the mandible, the maximum and minimum values of MPS in the cp-Ti-constructed Champy plate, upper AO/ASIF plate, and lower AO/ASIF plate were 19.5 and 20.3%, 15.2 and 25.3%, and 21.4 and 4.6% lower, respectively, than those for plates made from Ti-15Mo. In the same model, the maximum and minimum values of displacement in the cp-Ti-constructed Champy plate, upper AO/ASIF plate, and lower AO/ASIF plate were 1.6 and 3.8%, 3.1 and 2.7%, and 5.4 and 10.4% higher, respectively, than those for plates made from Ti-15Mo. CONCLUSIONS: This in silico 3D-FEA shows that Ti-15Mo plates have greater load-bearing capability.

Inhibition of matrix metalloproteinases and toxicity of gold and platinum nanoparticles in L929 fibroblast cells.

This study evaluated the inhibition of matrix metalloproteases (MMPs) and cellular responses elicited by gold (Au) and platinum (Pt) nanoparticles (NPs). The interaction of MMP-1 and NPs was evaluated using an MMP assay kit. The cultured L929 cells were exposed to various concentrations of NPs. The cellular responses to NPs were examined using a cytotoxicity assay (that evaluated cell viability and lactic dehydrogenase production), real-time polymerase chain reaction (RT-qPCR), and transmission electron microscopy. Both types of NPs, when used at concentrations above 10 µg ml(-1), inhibited MMP-1 activity. No cytotoxic effects were found when the cells were exposed to AuNPs. In contrast, PtNPs, at both 100 and 400 µg ml(-1), induced cytotoxicity. No inflammatory responses (production of interleukin-6 and tumor necrosis factor-alpha) to NPs were identified by RT-qPCR. The negative surface charge of NPs (COOH(-)) binds to the Zn(2+) of the MMP active center by chelation, leading to MMP inhibition. Gold nanoparticles are plausible candidates for MMP inhibitors in resin-bonding materials because they effectively inhibit MMP-1 activity without cytotoxic or inflammatory effects.

Dentistry in the 21st century: challenges of a globalising world.

Oral health is - literally - vital to good general health, not least because the mouth is the sentinel of the body. Dentistry, the Cinderella of health care, faces immense challenges of globalisation. Governments, having spent freely on everything from defence to social security, face mountains of debts which make budget cutbacks essential. Simultaneously, most developed countries have to pay increasing costs of caring for rapidly ageing populations. Dentistry is being pulled two ways: wealthy members of society demand high-end expensive treatment, much of it cosmetic rather than necessary to deal with disease, whereas many millions of poor people in developing countries cannot afford basic dental treatment and may never see a dentist. Too many governments and dentists persist with the expensive and destructive regime of 'drill and fill (and bill)'. International advances in care may not reach the clinician's chair because treatment guidelines and payments are set locally. An international symposium to celebrate Mikako Hayashi becoming Professor of Restorative Dentistry and Endodontology at Osaka University concluded that dentistry should move from an increasingly un-affordable curative model to a cost-effective evidence-based preventive model. The goal is to help people retain healthy natural teeth throughout their lives, as an essential part of enhancing their general health.

CAD-CAM resin composites: Effective components for further development.

OBJECTIVE: This paper summarizes the effective components of computer-aided design and computer-aided manufacturing (CAD-CAM) resin composites that contribute to achieving greater mechanical properties and further development. METHODS: In silico multi-scale analysis, in silico nonlinear dynamic finite element analysis (FEA), and artificial intelligence (AI) were used to explore the effective components of CAD-CAM resin composites. The effects of the filler diameter and silane coupling ratio on the mechanical properties of CAD-CAM resin composites have been clarified through multi-scale analysis. The effects of the filler contents, and filler and monomer compositions have been investigated by AI algorithms. The fracture behavior of CAD-CAM composite crown was analyzed using in silico non-linear dynamic FEA. The longevity of CAD-CAM composite crown was assessed through step-stress accelerating life testing (SSALT). RESULTS: As the filler diameter decreases, there is an increase in elastic moduli and compressive strengths at the macroscale. At the nanoscale, a decrease in the filler diameter results in a decrease in the maximum value of the maximum principal strain. When the silane coupling ratio decreases, there is a decrease in the elastic modulus and compressive strength. According to the exhaustive search and feature importance analysis based on the AI algorithm, the combination of certain components was narrowed down to achieve a flexural strength of 269.5 MPa. The in silico non-linear FEA successfully detected the sign of the initial crack of the CAD-CAM composite molar crown. The SSALT revealed that CAD-CAM resin composite molar crowns containing nanofillers with a high fraction of resin matrix demonstrated great longevity. SIGNIFICANCE: This paper summarized the effective components of CAD-CAM resin composites for their further development. The integration of in vitro and in silico approaches will expedite the advancement of CAD-CAM resin composites, offering benefits such as time efficiency and reduction of material waste for researchers and manufacturers.

In silico nonlinear dynamic finite-element analysis for biaxial flexural strength testing of CAD/CAM materials.

PURPOSE: The aim of this study was to establish and assess the validity of in silico models of biaxial flexural strength (BFS) tests to reflect in vitro physical properties obtained from two commercially available computer-aided design/computer-aided manufacturing (CAD/CAM) ceramic blocks and one CAD/CAM resin composite block. METHODS: In vitro three-point bending and BFS tests were conducted for three CAD/CAM materials (n = 10): Katana Zirconia ST10 (raw material: super-translucent multilayered zirconia, ST10; Kuraray Noritake Dental, Niigata, Japan), Katana Zirconia HT10 (raw material: highly translucent multilayered zirconia, HT10; Kuraray Noritake Dental), and Katana Avencia N (AN; Kuraray Noritake Dental). Densities, flexural moduli, and fracture strains were obtained from the in vitro three-point bending test and used as an input for an in silico nonlinear finite element analysis. The maximum principal stress (MPS) distribution was obtained from an in silico BFS analysis. RESULTS: The elastic moduli of AN, HT10, and ST10 were 6.513, 40.039, and 32.600 GPa, respectively. The in silico fracture pattern of ST10 observed after the in silico evaluation was similar to the fracture pattern observed after the in vitro testing. The MPS was registered in the center of the tensile surface for all three specimens. The projections of the supporting balls were in the form of a triple asymmetry. CONCLUSIONS: The in silico approach established in this study provided an acceptable reflection of in vitro physical properties, and will be useful to assess biaxial flexural properties of CAD/CAM materials without wastage of materials.

Poly(lactic acid/caprolactone) bilayer membrane achieves bone regeneration through a prolonged barrier function.

Guided bone regeneration (GBR) is a treatment strategy used to recover bone volume. Barrier membranes are a key component of GBR protocols, and their properties can impact treatment outcomes. This study investigated the efficacy of an experimental, slow-degrading, bilayer barrier membrane for application in GBR using in vivo animal models. A synthetic copolymer of poly(lactic acid/caprolactone) (PLCL) was used to prepare a slow-degrading bilayer membrane. The biodegradability of PLCL was evaluated by subcutaneous implantation in a rat model. The barrier function of the PLCL membrane was investigated in a rat calvaria defect model and compared with commercially available membranes composed of type I collagen (Col) and poly(lactic-co-glycolic acid) (PLGA). An alveolar bone defect model in beagle dogs was used to simulate GBR protocols to evaluate the bone regeneration ability of the experimental PLCL membrane. The PLCL membrane showed slow biodegradation, resulting in an efficient and prolonged barrier function compared with commercial materials. In turn, this barrier function enabled the space-making ability of PLCL membrane and facilitated bone regeneration. In the alveolar bone defect model, significantly greater regeneration was achieved by treatment with PLCL membrane compared with Col and PLGA membranes. Additionally, a continuous alveolar ridge contour was observed in PLCL-treated bone defects. In conclusion, the PLCL bilayer membrane is a promising biomaterial for use in GBR given its slow degradation and prolonged barrier function.

Effect of Heparan Sulfate on Vasculogenesis and Dentinogenesis of Dental Pulp Stem Cells.

INTRODUCTION: Heparan sulfate (HS) is a major component of dental pulp tissue. We previously reported that inhibiting HS biosynthesis impedes endothelial differentiation of dental pulp stem cells (DPSCs). However, the underlying mechanisms by which exogenous HS induces DPSC differentiation and pulp tissue regeneration remain unknown. This study explores the impact of exogenous HS on vasculogenesis and dentinogenesis of DPSCs both in vitro and in vivo. METHODS: Human-derived DPSCs were cultured in endothelial and odontogenic differentiation media and treated with HS. Endothelial differentiation of DPSCs was investigated by real-time polymerase chain reaction and capillary sprouting assay. Odontogenic differentiation was assessed through real-time polymerase chain reaction and detection of mineralized dentin-like deposition. Additionally, the influence of HS on pulp tissue was assessed with a direct pulp capping model, in which HS was delivered to exposed pulp tissue in rats. Gelatin sponges were loaded with either phosphate-buffered saline or 101-102 µg/mL HS and placed onto the pulp tissue. Following a 28-day period, tissues were investigated by histological analysis and micro-computed tomography imaging. RESULTS: HS treatment markedly increased expression levels of key endothelial and odontogenic genes, enhanced the formation of capillary-like structures, and promoted the deposition of mineralized matrices. Treatment of exposed pulp tissue with HS in the in vivo pulp capping study induced formation of capillaries and reparative dentin. CONCLUSIONS: Exogenous HS effectively promoted vasculogenesis and dentinogenesis of DPSCs in vitro and induced reparative dentin formation in vivo, highlighting its therapeutic potential for pulp capping treatment.

Development of Antibacterial Resin Composites Incorporating Poly(METAC) Clusters.

This study examined the antibacterial effects and physical properties of a novel resin composite incorporating poly[{2-(methacryloyloxy)ethyl}trimethylammonium chloride] (poly(METAC)), a methacrylate cationic polymer comprising quaternary ammonium compounds (QACs). Resin composites incorporating poly(METAC) were fabricated by adding 6 wt.% METAC aqueous solution to a commercially available resin composite. The FE-SEM/EDS and Raman spec-troscopy analyses showed that METAC was assembled and polymerized in the resin composites after curing. The antibacterial effect was evaluated by inoculating Streptococcus mutans or Strepto-coccus sobrinus suspensions on the surface of cured resin composites, and the experimental resin composites incorporating poly(METAC) clusters exhibited bactericidal effects even after 28 days of ageing. The physical properties of the experimental resin composites were within the ISO-stipulated ranges. Newly fabricated resin composites containing the QAC-based poly(METAC) cluster ex-hibited long-term bactericidal effects against oral bacteria on their surfaces and demonstrated ac-ceptable physical properties for clinical use.

Novel pit and fissure sealant with nano-CaF2 and antibacterial monomer: Fluoride recharge, microleakage, sealing ability and cytotoxicity.

Conventional resin-based sealants release minimal fluoride ions (F) and lack antibacterial activity. The objectives of this study were to: (1) develop a novel bioactive sealant containing calcium fluoride nanoparticles (nCaF2) and antibacterial dimethylaminohexadecyl methacrylate (DMAHDM), and (2) investigate mechanical performance, F recharge and re-release, microleakage, sealing ability and cytotoxicity. Helioseal F served as commercial control. The initial F release from sealant containing 20% nCaF2 was 25-fold that of Helioseal F. After ion exhaustion and recharge, the F re-release from bioactive sealant did not decrease with increasing number of recharge and re-release cycles. Elastic modulus of new bioactive sealant was 44% higher than Helioseal F. The new sealant had excellent sealing, minimal microleakage, and good cytocompatibility. Hence, the nanostructured sealant had substantial and sustained F release and antibacterial activity, good sealing ability and biocompatibility. The novel bioactive nCaF2 sealant is promising to provide long-term F ions for caries prevention.

Influence of MDPB-containing primer on Streptococcus mutans biofilm formation in simulated Class I restorations.

PURPOSE: To evaluate the activity of a methacryloyloxydodecylpyridinium bromide (MDPB)-containing self-etching primer (Clearfil Protect Bond) against Streptococcus mutans and its ability to reduce biofilm formation on standardized experimental Class I restorations in vitro. MATERIALS AND METHODS: Forty experimental Class I round restorations were prepared on enamel-dentin slabs using different adhesive strategies: group 1 = MDPB-containing adhesive system (Clearfil Protect Bond); group 2 = MDPB-free self-etching adhesive system (Clearfil SE Bond); group 3: MDPB-containing self-etching primer in combination with a fluoride-free bonding agent; group 4: MDPB-free self-etching primer in combination with a fluoride-containing bonding agent; group 5: a three-step etch-and-rinse adhesive system (Adper Scotchbond Multi Purpose). A Streptococcus mutans biofilm was grown for 48 h on the restoration surfaces and subsequently evaluated using scanning electron microscopy on three different areas: enamel, composite, and interface surfaces. Statistical analysis was performed by multiple ANOVA after data transformation. RESULTS: Specimens in groups 2, 4 and 5 showed greater biofilm formation than those in groups 1 and 3 (p < 0.001) on all investigated substrates (enamel, composite, and interface areas). CONCLUSIONS: Specimens prepared with an MDPB-containing primer exhibited significant decreases in biofilm formation on Class I restorations in vitro. Further in vitro and in vivo studies are required to clarify the role of quaternary ammonium compounds in reducing bacterial biofilm formation on restoration surfaces.

Quantitative evaluation of the degradation amount of the silane coupling layer of CAD/CAM resin composites by water absorption.

PURPOSE: Degradation of silane coupling layers by water ingress in computer-aided design/computer-aided manufacturing (CAD/CAM) of resin composites has been reported qualitatively. In this study, we quantitatively evaluated how water absorption of CAD/CAM resin composites affects the silane coupling layer by in vitro and in silico methods. METHODS: A Katana Avencia block (KAB) and an experimental matrix block composed of only a matrix resin were used to evaluate the effect of water immersion for seven days on the elastic modulus. X-ray photoelectron spectroscopy (XPS) with fluorine-labeling of the KAB was performed to evaluate the atomic percentage of F1s, which represents the hydrolysis amount by water immersion. In silico analysis of the three-dimensional model of the KAB was performed to determine the coupling ratios before and after water immersion. RESULTS: The elastic modulus of the KAB was 8.2 GPa before and 6.9 GPa after immersion in water. The atomic percentages of F1s in the after- and before-immersion groups were 14.31% and 11.52%, respectively, suggesting that hydrolysis of the silane coupling layer occurred during water immersion. From in silico analysis of the three-dimensional model of the KAB, the coupling ratio was predicted to be 78.2% before water immersion. After water immersion, the coupling ratio was predicted to be 68.4%. CONCLUSIONS: The in vitro and in silico approaches established in this study were able to predict the silane coupling ratios of CAD/CAM resin composites, and they showed that the silane coupling ratio decreased by water absorption.