Controlled cell proliferation and immortalization of human dental pulp stem cells with a doxycycline-inducible expression system.

Human dental pulp stem cells are a potentially useful resource for cell-based therapies and tissue repair in dental and medical applications. However, the primary culture of isolated dental pulp stem cells has notably been limited. A major requirement of an ideal human dental pulp stem cell culture system is the preservation of efficient proliferation and innate stemness over prolonged passaging, while also ensuring ease of handling through standard, user-friendly culture methods. In this study, we have engineered a novel human dental pulp stem cell line, distinguished by the constitutive expression of telomerase reverse transcriptase (TERT), and the conditional expression of the R24C mutant cyclin-dependent kinase 4 (CDK4R24C) and Cyclin D1. We have named this cell line Tet-off K4DT hDPSCs. Furthermore, we have conducted a comprehensive comparative analysis of their biological attributes in relation to a previously immortalized human dental pulp stem cells, hDPSC-K4DT, which were immortalized by the constitutive expression of CDK4R24C, Cyclin D1 and TERT. In Tet-off K4DT cells, the expression of the K4D genes can be precisely suppressed by the inclusion of doxycycline. Remarkably, Tet-off K4DT cells demonstrated an extended cellular lifespan, increased proliferative capacity, and enhanced osteogenic differentiation potential when compared to K4DT cells. Moreover, Tet-off K4DT cells had no observable genomic aberrations and also displayed a sustained expression of stem cell markers even at relatively advanced passages. Taken together, the establishment of this new cell line holds immense promise as powerful experimental tool for both fundamental and applied research involving dental pulp stem cells.

Bioactive gelatin-sheets as novel biopapers to support prevascularization organized by laser-assisted bioprinting for bone tissue engineering.

Despite significant advances in the management of patients with oral cancer, maxillofacial reconstruction after ablative surgery remains a clinical challenge. In bone tissue engineering, biofabrication strategies have been proposed as promising alternatives to solve issues associated with current therapies and to produce bone substitutes that mimic both the structure and function of native bone. Among them, laser-assisted bioprinting (LAB) has emerged as a relevant biofabrication method to print living cells and biomaterials with micrometric resolution onto a receiving substrate, also called 'biopaper'. Recent studies have demonstrated the benefits of prevascularization using LAB to promote vascularization and bone regeneration, but mechanical and biological optimization of the biopaper are needed. The aim of this study was to apply gelatin-sheet fabrication process to the development of a novel biopaper able to support prevascularization organized by LAB for bone tissue engineering applications. Gelatin-based sheets incorporating bioactive glasses (BGs) were produced using various freezing methods and crosslinking (CL) parameters. The different formulations were characterized in terms of microstructural, physical, mechanical, and biological properties in monoculture and coculture. Based on multi-criteria analysis, a rank scoring method was used to identify the most relevant formulations. The selected biopaper underwent additional characterization regarding its ability to support mineralization and vasculogenesis, its bioactivity potential andin vivodegradability. The biopaper 'Gel5wt% BG1wt%-slow freezing-CL160 °C 24 h' was selected as the best candidate, due to its suitable properties including high porosity (91.69 ± 1.55%), swelling ratio (91.61 ± 0.60%), Young modulus (3.97 × 104± 0.97 × 104Pa) but also its great cytocompatibility, osteogenesis and bioactivity properties. The preorganization of human umbilical vein endothelial cell using LAB onto this new biopaper led to the formation of microvascular networks. This biopaper was also shown to be compatible with 3D-molding and 3D-stacking strategies. This work allowed the development of a novel biopaper adapted to LAB with great potential for vascularized bone biofabrication.

New Implications of Metabolites and Free Fatty Acids in Quality Control of Crossbred Wagyu Beef during Wet Aging Cold Storage.

Efficient cold-chain delivery is essential for maintaining a sustainable global food supply. This study used metabolomic analysis to examine meat quality changes during the "wet aging" of crossbred Wagyu beef during cold storage. The longissimus thoracic (Loin) and adductor muscles (Round) of hybrid Wagyu beef, a cross between the Japanese Black and Holstein-Friesian breeds, were packaged in vacuum film and refrigerated for up to 40 days. Sensory evaluation indicated an increase in the umami and kokumi taste owing to wet aging. Comprehensive analysis using gas chromatography-mass spectrometry identified metabolite changes during wet aging. In the Loin, 94 metabolites increased, and 24 decreased; in the Round, 91 increased and 18 decreased. Metabolites contributing to the umami taste of the meat showed different profiles during wet aging. Glutamic acid increased in a cold storage-dependent manner, whereas creatinine and inosinic acid degraded rapidly even during cold storage. In terms of lipids, wet aging led to an increase in free fatty acids. In particular, linoleic acid, a polyunsaturated fatty acid, increased significantly among the free fatty acids. These results provide new insight into the effects of wet aging on Wagyu-type beef, emphasizing the role of free amino acids, organic acids, and free fatty acids generated during cold storage.

Lipopolysaccharide-mediated ATP signaling regulates interleukin-6 mRNA expression via the P2-purinoceptor in human dental pulp cells.

Dental pulp cells play a crucial role in maintaining the balance of the pulp tissue. They actively respond to bacterial inflammation by producing proinflammatory cytokines, particularly interleukin-6 (IL-6). While many cell types release adenosine triphosphate (ATP) in response to various stimuli, the mechanisms and significance of ATP release in dental pulp cells under inflammatory conditions are not well understood. This study aimed to investigate ATP release and its relationship with IL-6 during the inflammatory response in immortalized human dental pulp stem cells (hDPSC-K4DT) following lipopolysaccharide (LPS) stimulation. We found that hDPSC-K4DT cells released ATP extracellularly when exposed to LPS concentrations above 10 µg/mL. ATP release was exclusively attenuated by N-ethylmaleimide, whereas other inhibitors, including clodronic acid (a vesicular nucleotide transporter inhibitor), probenecid (a selective pannexin-1 channel inhibitor), meclofenamic acid (a selective connexin 43 inhibitor), suramin (a nonspecific P2 receptor inhibitor), and KN-62 (a specific P2X7 antagonist), did not exhibit any effect. Additionally, LPS increased IL-6 mRNA expression, which was mitigated by the ATPase apyrase enzyme, N-ethylmaleimide, and suramin, but not by KN-62. Moreover, exogenous ATP induced IL-6 mRNA expression, whereas ATPase apyrase, N-ethylmaleimide, and suramin, but not KN-62, diminished ATP-induced IL-6 mRNA expression. Overall, our findings suggest that LPS-induced ATP release stimulates the IL-6 pathway through P2-purinoceptor, indicating that ATP may function as an anti-inflammatory signal, contributing to the maintenance of dental pulp homeostasis.

Effect of Bioactive Glasses and Basic Fibroblast Growth Factor on Dental Pulp Cells.

Ideal regeneration of hard tissue and dental pulp has been reported with the use of a combination of bioactive glass and basic fibroblast growth factor (bFGF). However, no previous study has investigated the molecular mechanisms underlying the processes induced by this combination in dental pulp cells. This study aimed to examine the cellular phenotype and transcriptional changes induced by the combination of bioactive glass solution (BG) and bFGF in dental pulp cells using phase-contrast microscopy, a cell counting kit-8 assay, alkaline phosphatase staining, and RNA sequence analysis. bFGF induced elongation of the cell process and increased the number of cells. Whereas BG did not increase ALP activity, it induced extracellular matrix-related genes in the dental pulp. In addition, the combination of BG and bFGF induces gliogenesis-related genes in the nervous system. This is to say, bFGF increased the viability of dental pulp cells, bioactive glass induced odontogenesis, and a dual stimulation with bioactive glass and bFGF induced the wound healing of the nerve system in the dental pulp. Taken together, bioactive glass and bFGF may be useful for the regeneration of the dentin-pulp complex.

Influence of New Sleeve Composite on Fracture Behavior of Anterior Teeth with Flared Root Canals.

We evaluated the fracture strength and failure mode of non-ferrule teeth with flared root canals that were restored using new experimental sleeve composites. Fifty endodontically treated anterior teeth with flared root canals were restored with direct restorations utilizing different techniques. Group A had teeth (non-ferrule) restored using commercialized MI glass fiber post + Gradia Core as core build-up. Group B had teeth (non-ferrule) restored with commercialized i-TFC glass fiber post + sleeve system. In Group C, the teeth (non-ferrule) were restored with an experimental sleeve composite with commercialized MI glass fiber post and Gradia Core. Group D, teeth (non-ferrule), were restored using custom-made tapered E-glass filling post and Gradia Core. Group E, teeth (with ferrule), were restored with commercialized MI glass fiber post + Gradia Core. After core construction, all specimens underwent direct composite crown restoration and were loaded until fracture using a universal testing machine. Average fracture loads were compared, and the failure modes were observed. Group C exhibited significantly greater fracture strength than other groups (p < 0.05). Favorable fracture teeth ratio of group C was more than that of the other groups. Thus, the new experimental sleeve composite could be clinically useful for core construction of non-ferrule teeth.

In Vivo Application of Silica-Derived Inks for Bone Tissue Engineering: A 10-Year Systematic Review.

As the need for efficient, sustainable, customizable, handy and affordable substitute materials for bone repair is critical, this systematic review aimed to assess the use and outcomes of silica-derived inks to promote in vivo bone regeneration. An algorithmic selection of articles was performed following the PRISMA guidelines and PICO method. After the initial selection, 51 articles were included. Silicon in ink formulations was mostly found to be in either the native material, but associated with a secondary role, or to be a crucial additive element used to dope an existing material. The inks and materials presented here were essentially extrusion-based 3D-printed (80%), and, overall, the most investigated animal model was the rabbit (65%) with a femoral defect (51%). Quality (ARRIVE 2.0) and risk of bias (SYRCLE) assessments outlined that although a large majority of ARRIVE items were "reported", most risks of bias were left "unclear" due to a lack of precise information. Almost all studies, despite a broad range of strategies and formulations, reported their silica-derived material to improve bone regeneration. The rising number of publications over the past few years highlights Si as a leverage element for bone tissue engineering to closely consider in the future.

2-hydroxyethyl methacrylate-derived reactive oxygen species stimulate ATP release via TRPA1 in human dental pulp cells.

Extracellular ATP (adenosine triphosphate) and transient receptor potential ankyrin 1 (TRPA1) channels are involved in calcium signaling in odontoblasts and dental pain. The resin monomer 2-hydroxyethyl methacrylate (HEMA), used in dental restorative procedures, is related to apoptotic cell death via oxidative stress. Although the TRPA1 channel is highly sensitive to reactive oxygen species (ROS), the effect of HEMA-induced ROS on ATP release to the extracellular space and the TRPA1 channel has not been clarified in human dental pulp. In this study, we investigated the extracellular ATP signaling and TRPA1 activation by HEMA-derived ROS in immortalized human dental pulp cells (hDPSC-K4DT). Among the ROS-sensitive TRP channels, TRPA1 expression was highest in undifferentiated hDPSC-K4DT cells, and its expression levels were further enhanced by osteogenic differentiation. In differentiated hDPSC-K4DT cells, 30 mM HEMA increased intracellular ROS production and ATP release, although 3 mM HEMA had no effect. Pretreatment with the free radical scavenger PBN (N-tert-butyl-α-phenylnitrone) or TRPA1 antagonist HC-030031 suppressed HEMA-induced responses. These results suggest that ROS production induced by a higher dose of HEMA activates the TRPA1 channel in human dental pulp cells, leading to ATP release. These findings may contribute to the understanding of the molecular and cellular pathogenesis of tertiary dentin formation and pain in response to dental biomaterials.

Effects of rice fermented extracts, "Sake Lees", on the functional activity of odontoblast-like cells (KN-3 cells).

This study was designed to investigate the effects of Sake Lees extracts (SLE, Sake Kasu) on the functional activity of odontoblastic cells and tooth pulp of the rats. For in vitro studies, a rat clonal odontoblast-like cell line, KN-3 cells were cultured. SLE significantly decreased KN-3 cell proliferation, but showed no significant cytotoxicity. SLE effects on several protein productions of KN-3 cells were compared with PBS. SLE and PBS increased alkaline phosphatase (ALP), dentin sialoprotein (DSP), and osterix in a day-course dependent manner, while SLE increased the induction of ALP on day 9-21 and DSP on day 15-21. SLE also increased Runx2 expression on day 3 and 9 compared to PBS. Alizarin Red stainings revealed that SLE showed a subtle increase in mineralization of KN-3 cells on day 15 and 21. A histological investigation was conducted to assess if SLE induced reparative dentin formation after direct capping at the exposed tooth pulp in rats, suggesting that SLE could increase the reparative dentin formation more than PBS. These findings suggest that Sake Lees could have functional roles in the alterations of odontoblastic activity, which might influence the physiology of the tooth pulp.

Development of a Dengue Virus Serotype-Specific Non-Structural Protein 1 Capture Immunochromatography Method.

Four serotypes of dengue virus (DENV), type 1 to 4 (DENV-1 to DENV-4), exhibit approximately 25-40% of the difference in the encoded amino acid residues of viral proteins. Reverse transcription of RNA extracted from specimens followed by PCR amplification is the current standard method of DENV serotype determination. However, since this method is time-consuming, rapid detection systems are desirable. We established several mouse monoclonal antibodies directed against DENV non-structural protein 1 and integrated them into rapid DENV detection systems. We successfully developed serotype-specific immunochromatography systems for all four DENV serotypes. Each system can detect 104 copies/mL in 15 min using laboratory and clinical isolates of DENV. No cross-reaction between DENV serotypes was observed in these DENV isolates. We also confirmed that there was no cross-reaction with chikungunya, Japanese encephalitis, Sindbis, and Zika viruses. Evaluation of these systems using serum from DENV-infected individuals indicated a serotype specificity of almost 100%. These assay systems could accelerate both DENV infection diagnosis and epidemiologic studies in DENV-endemic areas.

Evaluation of New Hollow Sleeve Composites for Direct Post-Core Construction.

The preset shape and diameter of a prefabricated FRC post rarely follows the anatomy of the root canal. To solve this problem, a new hollow sleeve composite (HSC) system for post-core construction was developed and characterized. A woven fiber was impregnated with two types of resins: Bis-GMA or PMMA, and rolled into cylinders with outer diameter of 2 mm and two different inner diameters, namely 1.2 or 1.5 mm. The commercial i-TFC system was used as a control. Dual-cure resin composite was injected into these sleeves. Additionally, conventional solid fiber post was used as the inner part of the sleeve. The three-point bending test was used to measure the mechanical properties of the specimens and the fracture surface was examined using an electron microscope (SEM). The HSC (1.5 mm, Bis-GMA) revealed a statistically similar flexural modulus but higher flexural strength (437 MPa) compared to i-TFC (239 MPa; ANOVA, p < 0.05). When a fiber post was added inside, all values had a tendency to increase. After hydrothermal accelerated aging, the majority of specimens showed a significant (p < 0.05) decrease in flexural strength and modulus. SEM fracture analysis confirmed that the delamination occurred at the interface between the outer and inner materials. The HSC system provided flexibility but still high mechanical values compared to the commercial system. Thus, this system might offer an alternative practical option for direct post-core construction.

Physicochemical Properties, Cytocompatibility, and Biocompatibility of a Bioactive Glass Based Retrograde Filling Material.

The ideal retrograde filling material that is easy to handle, has good physicochemical properties, and is biocompatible has not yet been developed. The current study reports the development of a novel bioactive glass based powder for use as a retrograde filling material that is capable of altering the consistency and hardening rate of mixtures when mixed with existing bioactive glass based cement. Furthermore, its physicochemical properties, in vitro effects on human cementoblast-like cells, and in vivo effects on inflammatory responses were evaluated. The surface of the hardened cement showed the formation of hydroxyapatite-like precipitates and calcium and silicate ions were eluted from the cement when the pH level was stabilized at 10.5. Additionally, the cement was found to be insoluble and exhibited favorable handling properties. No adverse effects on viability, proliferation, and expression of differentiated markers were observed in the in vitro experiment, and the cement was capable of inducing calcium deposition in the cells. Moreover, the cement demonstrated a lower number of infiltrated inflammatory cells compared to the other materials used in the in vivo mouse subcutaneous implantation experiment. These findings suggest that the retrograde filling material composed of bioactive glass and the novel bioactive glass based powder exhibits favorable physicochemical properties, cytocompatibility, and biocompatibility.

Odontoblast differentiation is regulated by an interplay between primary cilia and the canonical Wnt pathway.

Primary cilium is a protruding cellular organelle that has various physiological functions, especially in sensory reception. While an avalanche of reports on primary cilia have been published, the function of primary cilia in dental cells remains to be investigated. In this study, we focused on the function of primary cilia in dentin-producing odontoblasts. Odontoblasts, like most other cell types, possess primary cilia, which disappear upon the knockdown of intraflagellar transport protein 88. In cilia-depleted cells, the expression of dentin sialoprotein, an odontoblastic marker, was elevated, while the deposition of minerals was slowed. This was recapitulated by the activation of canonical Wnt pathway, also decreased the ratio of ciliated cells. In dental pulp cells, as they differentiated into odontoblasts, the ratio of ciliated cells was increased, whereas the canonical Wnt signaling activity was repressed. Our results collectively underscore the roles of primary cilia in regulating odontoblastic differentiation through canonical Wnt signaling. This study implies the existence of a feedback loop between primary cilia and the canonical Wnt pathway.

Effects of Both Fiber Post/Core Resin Construction System and Root Canal Sealer on the Material Interface in Deep Areas of Root Canal.

This study aimed to examine the resin polymerization of a fiber post/core resin construction system and the interface between resin and root canal sealers, which are important for root canal sealing. We used the i-TFC Luminus fiber post and i-TFC Luminus LC flow (i-TFC-L), the GC fiber post and Unifil Core EM (GCF), and the FiberKor post and Build-It FR (FKP) as core construction systems, and Nishika Canal Sealer BG (CS-BG), Metaseal Soft (META), and Nishika Canal Sealer EN (CS-EN) as sealers. The light transmission of fiber posts (n = 5), the polymerization of core resin (n = 5), and the adhesion between the sealer and core resin (n = 10) were evaluated. The i-TFC Luminus fiber post light transmission was significantly higher than that of other posts. Without shielding, i-TFC-L showed a significantly greater amount of polymerized resin than the other systems. With shielding, although i-TFC-L showed a significantly greater amount of polymerized resin immediately after light irradiation, polymerized resin was significantly greater in GCF and FKP after 10 min. All systems adhered to CS-BG and META but not to CS-EN. These results indicate that resin polymerization in the cavity differs among fiber post/core resin construction systems and that the adhesion of the resin and sealer depends on the property of the sealer.

Concordance study between regular face-to-face dental diagnosis and dental telediagnosis using fluorescence.

BACKGROUND: Teledentistry consultations are an effective way to increase access to care. Whether it be for a screening, referral or even an adapted treatment plan for a certain number of patients whose access to care is complicated, demonstrating the reliability of remote consultations is essential in allowing the technique to become generalised. AIM: This study aimed to determine if teledentistry consultations using fluorescence are of the same quality as regular consultations in the diagnosis of caries. METHODS: Patients were seen in consultation in the dental care centre at the Montpellier University Hospital (France) and in the centre at Kyushu Dental University Hospital (Japan). The protocol was broken down into three parts: the regular consultation, the recording of videos with the Soprocare camera and the remote consultation. The regular consultation and the remote consultation were blinded and carried out by two different dentists. The recording of videos was carried out by a third dentist. The carious diagnosis was based on the International Caries Detection and Assessment System: a clinical rating system for the detection and assessment of caries. RESULTS: One hundred and ninety-five patients met the predefined inclusion criteria. Most patients had at least one surface at stage 3 or higher (73%) with a higher proportion amongst French patients (81% compared to 66%). However, they had good dental hygiene, given that dental hygiene was only deemed unsatisfactory for 10.8% (19% for French patients and 2% for Japanese patients). The odontogram (presence/absence of each tooth) seemed to be correctly identified during the remote consultation (reinterpretation). Out of the 195 patients, 168 (86.2%) were identified without error. CONCLUSIONS: Teledentistry consultations can represent acceptable diagnostic performance with regard to the detection of dental caries. The Soprocare camera enables an early diagnosis of carious lesions with optimal efficiency. Several areas still need to be improved, however, so that the use of the camera during remote consultations is as coherent and effective as possible, especially with regard to the organisational aspects of remote consultations.

Effect of Bioactive Glass-Based Root Canal Sealer on the Incidence of Postoperative Pain after Root Canal Obturation.

The purpose of this study is to evaluate the effect of a bioactive glass-based root canal sealer, Nishika Canal Sealer BG (CS-BG), on the incidence of postoperative pain (PP) after root canal obturation (RCO). Eleven dentists performed pulpectomy or infected root canal treatments for 555 teeth. During RCO, CS-BG was used. After RCO, the rate of PP and the factors affecting PP (pain during RCO and pain immediately after RCO) were analyzed. PP was observed in eight teeth (1.5%), and within 7 days after RCO, there were no teeth with pain. In these teeth with PP, there was a significant difference in the occurrence of pain during RCO, but not in the occurrence of pain immediately after RCO, when compared with pulpectomy and infected root canal treatment. These clinical results show that CS-BG has an excellent biocompatibility, and can suppress the distress of patients during RCO.

A small nuclear acidic protein (MTI-II, Zn2+-binding protein, parathymosin) attenuates TNF-α inhibition of BMP-induced osteogenesis by enhancing accessibility of the Smad4-NF-κB p65 complex to Smad binding element.

Pro-inflammatory cytokines prevent bone regeneration in vivo and activation of nuclear factor-κB (NF-κB) signaling has been proposed to lead to suppression of bone morphogenetic protein (BMP)-induced osteogenesis via direct binding of p65 to Smad4 in vitro. Application of a small nuclear acidic protein (MTI-II) and its delivered peptide, MPAID (MTI-II peptide anti-inflammatory drug) has been described to elicit therapeutic potential via strong anti-inflammatory action following the physical association of MTI-II and MPAID with p65. However, it is unclear whether MTI-II attenuates tumor necrosis factor (TNF)-α inhibition of BMP-induced osteogenesis. Herein, we found that TNF-α-mediated suppression of responses associated with BMP4-induced osteogenesis, including expression of the osteocalcin encoding gene Ocn, Smad binding element (SBE)-dependent luciferase activity, alkaline phosphatase activity, and alizarin red S staining were largely restored by MTI-II and MPAID in MC3T3-E1 cells. Mechanistically, MTI-II and MPAID did not inhibit nuclear translocation of p65 or disassociate Smad4 from p65. Further, results from chromatin immunoprecipitation (ChIP) analyses revealed that Smad4 enrichment in cells over-expressing MTI-II and treated with TNF-α was equivalent to that in cells without TNF-α treatment. Alternatively, Smad4 enrichment was considerably decreased following TNF-α treatment in control cells. Moreover, p65 enrichment in the Id-1 promoter SBE was detected only when cells over-expressing MTI-II were stimulated with TNF-α. Overall, our study concludes that MTI-II restored TNF-α-inhibited suppression of BMP-Smad-induced osteogenic differentiation by enhancing accessibility of the Smad4-p65 complex to the SBE rather than by liberating Smad4 from p65.

A novel inhibitor of NF-κB-inducing kinase prevents bone loss by inhibiting osteoclastic bone resorption in ovariectomized mice.

Musculoskeletal diseases and disorders, including osteoporosis and rheumatoid arthritis are diseases that threaten a healthy life expectancy, and in order to extend the healthy life expectancy of elderly people, it is important to prevent bone and joint diseases and disorders. We previously reported that alymphoplasia (aly/aly) mice, which have a loss-of-function mutation in the Nik gene involved in the processing of p100 to p52 in the alternative NF-κB pathway, show mild osteopetrosis with a decrease in the osteoclast number, suggesting that the alternative NF-κB pathway is a potential drug target for ameliorating bone diseases. Recently, the novel NF-κB-inducing kinase (NIK)-specific inhibitor compound 33 (Cpd33) was developed, and we examined its effect on osteoclastic bone resorption in vitro and in vivo. Cpd33 inhibited the receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis accompanied by a decrease in the expression of nfatc1, dc-stamp, and cathepsin K, markers of osteoclast differentiation, without affecting the cell viability, in a dose-dependent manner. Cdp33 specifically suppressed the RANKL-induced processing of p100 to p52 but not the phosphorylation of p65 or the degradation or resynthesis of IκBα in osteoclast precursors. Cpd33 also suppressed the bone-resorbing activity in mature osteoclasts. Furthermore, Cdp33 treatment prevented bone loss by suppressing the osteoclast formation without affecting the osteoblastic bone formation in ovariectomized mice. Taken together, NIK inhibitors may be a new option for patients with a reduced response to conventional pharmacotherapy or who have serious side effects.

Expression and function of dopamine in odontoblasts.

Dopamine (DA) is produced from tyrosine by tyrosine hydroxylase (TH). A recent study has reported that DA promotes the mineralization of murine preosteoblasts. However, the role of DA in odontoblasts has not been examined. Therefore, in this investigation, we researched the expression of TH and DA in odontoblasts and the effects of DA on the differentiation of preodontoblasts (KN-3 cells). Immunostaining showed that TH and DA were intensely expressed in odontoblasts and preodontoblasts of rat incisors and molars. KN-3 cells expressed D1-like and D2-like receptors for DA. Furthermore, DA promoted odontoblastic differentiation of KN-3 cells, whereas an antagonist of D1-like receptors and a PKA signaling blocker, inhibited such differentiation. However, antagonists of D2-like receptors promoted differentiation. These results suggested that DA in preodontoblasts and odontoblasts might promote odontoblastic differentiation through D1-like receptors, but not D2-like receptors, and PKA signaling in an autocrine or paracrine manner and plays roles in dentinogenesis.

Adhesive bonding of alumina air-abraded Ag-Pd-Cu-Au alloy with 10-methacryloyloxydecyl dihydrogen phosphate.

The aim of this paper is to study changes in the Ag-Pd-Cu-Au alloy surfaces by alumina air-abrasion process and effect of those changes on the adhesive bonding characteristic. Surface roughness, surface composition and chemical state of the alumina air-abraded alloys were analyzed by a confocal laser scanning microscope, an energy dispersive X-ray spectroscopy and an X-ray photoelectron spectroscopy. The results showed that the alumina air-abrasion changed the alloy surface by mechanical roughening, alumina remain and copper oxidation. Effect of the changes in the alloy surface on the adhesive bonding characteristic was examined by using a methyl methacrylate/tri-n-butylborane derivative (MMA/TBB) resin cement with the 10-methacryloyloxydecyl dihydrogen phosphate (MDP) contained primer. The shear bond strength test results indicated that the surface oxidation by the abrasion is the main contributor that improved the adhesive bonding rather than other effects such as mechanical roughening or alumina remain.