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1.
Nanomaterials (Basel) ; 11(7)2021 Jul 14.
Article in English | MEDLINE | ID: mdl-34361212

ABSTRACT

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.

2.
Materials (Basel) ; 14(4)2021 Feb 19.
Article in English | MEDLINE | ID: mdl-33669868

ABSTRACT

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.

3.
J Telemed Telecare ; 27(8): 509-517, 2021 Sep.
Article in English | MEDLINE | ID: mdl-31903839

ABSTRACT

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.


Subject(s)
Dental Caries , Remote Consultation , Telemedicine , Dental Caries/diagnosis , France , Humans , Reproducibility of Results
4.
Article in English | MEDLINE | ID: mdl-33260512

ABSTRACT

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.


Subject(s)
Dental Pulp Cavity , Pain, Postoperative , Root Canal Filling Materials , Female , Humans , Incidence , Male , Middle Aged , Pain, Postoperative/epidemiology , Root Canal Obturation
5.
Materials (Basel) ; 12(23)2019 Nov 29.
Article in English | MEDLINE | ID: mdl-31795433

ABSTRACT

Endodontic treatment for a tooth with damaged dental pulp aims to both prevent and cure apical periodontitis. If the tooth is re-infected as a result of a poorly obturated root canal, periapical periodontitis may set-in due to invading bacteria. To both avoid any re-infection and improve the success rate of endodontic retreatment, a treated root canal should be three-dimensionally obturated with a biocompatible filling material. Recently, bioactive glass, one of the bioceramics, is focused on the research area of biocompatible biomaterials for endodontics. Root canal sealers derived from bioactive glass-based have been developed and applied in clinical endodontic treatments. However, at present, there is little evidence about the patient outcomes, sealing mechanism, sealing ability, and removability of the sealers. Herein, we have developed a bioactive glass-based root canal sealer and provided evidence concerning its physicochemical properties, biocompatibility, sealing ability, and removability. We also review the classification of bioceramics and characteristics of bioactive glass. Additionally, we describe the application of bioactive glass to facilitate the development of a new root canal sealer. Furthermore, this review shows the potential application of bioactive glass-based cement as a root canal filling material in the absence of semisolid core material.

6.
Jpn Dent Sci Rev ; 55(1): 5-11, 2019 Nov.
Article in English | MEDLINE | ID: mdl-30733839

ABSTRACT

Dental pulp is a connective tissue and has functions that include initiative, formative, protective, nutritive, and reparative activities. However, it has relatively low compliance, because it is enclosed in hard tissue. Its low compliance against damage, such as dental caries, results in the frequent removal of dental pulp during endodontic therapy. Loss of dental pulp frequently leads to fragility of the tooth, and eventually, a deterioration in the patient's quality of life. With the development of biomaterials such as bioceramics and advances in pulp biology such as the identification of dental pulp stem cells, novel ideas for the preservation of dental pulp, the regenerative therapy of dental pulp, and new biomaterials for direct pulp capping have now been proposed. Therapies for dental pulp are classified into three categories; direct pulp capping, vital pulp amputation, and treatment for non-vital teeth. In this review, we discuss current and future treatment options in these therapies.

7.
J Biomed Mater Res B Appl Biomater ; 107(1): 161-168, 2019 01.
Article in English | MEDLINE | ID: mdl-29575555

ABSTRACT

Direct pulp capping is an important procedure for preserving pulp viability. The pulp capping agent must possess several properties, including usability, biocompatibility, and the ability to induce reparative dentin formation. In this study, a novel bioactive glass-based cement was examined to determine whether the cement has the necessary properties to act as a direct pulp capping agent. Physicochemical properties of the bioactive glass-based cement and in vitro effects of the cement on odontoblast-like cells, as well as in vivo effects on the exposed dental pulp, were analyzed. The cement immersed in water stabilized at pH10, and hydroxyapatite-like precipitation was induced on the surface of the cement in simulate body fluid. There were no cytotoxic effects on the viability, alkaline phosphatase activity, or calcium deposition ability of odontoblast-like cells. In the in vivo rat study of an exposed dental pulp model, the cement induced a sufficient level of reparative dentin formation by odontoblast-like cells expressing odontoblastic markers at the exposed area of the dental pulp. These results suggest that the newly developed bioactive glass-based cement provides favorable biocompatibility with the dental pulp and may be useful as a direct pulp capping agent. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 107B: 161-168, 2019.


Subject(s)
Dental Cements , Glass/chemistry , Materials Testing , Odontoblasts/metabolism , Pulp Capping and Pulpectomy Agents , Animals , Cell Line , Dental Cements/chemistry , Dental Cements/pharmacology , Dentin/metabolism , Odontoblasts/cytology , Pulp Capping and Pulpectomy Agents/chemistry , Pulp Capping and Pulpectomy Agents/pharmacology , Rats
8.
Clin Exp Pharmacol Physiol ; 45(1): 75-83, 2018 Jan.
Article in English | MEDLINE | ID: mdl-28815657

ABSTRACT

Non-steroidal anti-inflammatory drugs (NSAIDs) exert their effects primarily by inhibiting the activity of cyclooxygenase (COX), thus suppressing prostaglandin synthesis. Some NSAIDs are known to perform functions other than pain control, such as suppressing tumour cell growth, independent of their COX-inhibiting activity. To identify NSAIDs with COX-independent activity, we examined various NSAIDs for their ability to inhibit osteoblastic differentiation using the mouse pre-osteoblast cell line MC3T3-E1. Only celecoxib and valdecoxib strongly inhibited osteoblastic differentiation, and this effect was not correlated with COX-inhibiting activity. Moreover, 2,5-dimethyl (DM)-celecoxib, a celecoxib analogue that does not inhibit COX activity, also inhibited osteoblastic differentiation. Celecoxib and DM-celecoxib inhibited osteoblastic differentiation induced by bone morphogenetic protein (BMP)-2 in C2C12 mouse myoblast cell line. Although celecoxib suppresses the growth of some tumour cells, the viability and proliferation of MC3T3-E1 cells were not affected by celecoxib or DM-celecoxib. Instead, celecoxib and DM-celecoxib suppressed BMP-2-induced phosphorylation of Smad1/5, a major downstream target of BMP receptor. Although it is well known that COX plays important roles in osteoblastic differentiation, these results suggest that some NSAIDs, such as celecoxib, have targets other than COX and regulate phospho-dependent intracellular signalling, thereby modifying bone remodelling.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Celecoxib/pharmacology , Cell Differentiation/drug effects , Osteoblasts/cytology , Osteoblasts/drug effects , Animals , Bone Morphogenetic Protein 2/metabolism , Cell Proliferation/drug effects , Cell Survival/drug effects , Cyclooxygenase 2/metabolism , Mice , Osteoblasts/metabolism , Signal Transduction/drug effects
9.
J Investig Clin Dent ; 8(3)2017 Aug.
Article in English | MEDLINE | ID: mdl-27378241

ABSTRACT

AIM: The purpose of the present study was to compare the clinical efficacy between a flowable-type nano-hybrid composite and a paste-type composite for posterior restoration. METHODS: Of 62 posterior teeth in 33 patients (mean age: 34.1 years), 31 were filled with a paste-type composite (Heliomolar [HM] group), and another 31 with a flowable nano-hybrid composite (MI FIL [MI] group). Clinical efficacy was evaluated at 2 years after the restoration. RESULTS: There were no differences for retention, surface texture deterioration, anatomical form change, deterioration of marginal adaptation, and secondary caries, while a statistical difference was found for marginal discoloration, which was significantly greater in the HM group (P < 0.05). Furthermore, color matching in the MI group was superior to that in the HM group immediately after the restoration throughout the study period. CONCLUSIONS: The present 2-year clinical evaluation of different composites showed that the flowable nano-hybrid composite could be an effective esthetic material for posterior restoration.


Subject(s)
Acrylic Resins , Composite Resins , Dental Restoration, Permanent , Polyurethanes , Adult , Aged , Female , Humans , Male , Middle Aged , Nanotechnology , Time Factors , Young Adult
10.
J Adhes Dent ; 18(2): 111-8, 2016.
Article in English | MEDLINE | ID: mdl-27022642

ABSTRACT

PURPOSE: To assess the effects of different curing stages of 4-META/MMA-TBB resin on osteoblasts and gingival keratinocytes. MATERIALS AND METHODS: The MC3T3-E1 murine pre-osteoblastic cell line and GE-1 murine gingival epithelial cell line were cultured with mixtures of Super-Bond C&B at different curing stages, and the cell viability was assessed. The alkaline phosphatase (ALP) activity of the MC3T3-E1 cells was also assessed. RESULTS: The majority of the MC3T3-E1 cells died and showed no ALP activity when cultured with 4-META/MMA-TBB resin during the initial curing phase (1 min of curing). A later curing phase of the 4-META/MMA-TBB resin (7 min of curing) showed cytotoxicity at day 1, but the toxic effect was temporary and the proliferative capacity and ALP activity in the cells were similar to control cells at day 7. Completely cured 4-META/MMA-TBB resin (after 1 or 12 h of curing) did not affect the cell viability or ALP activity of the MC3T3-E1 cells. In contrast, 4-META/MMA-TBB resin showed no effect on the GE-1 cells at any stage of curing. CONCLUSION: Although 4-META/MMA-TBB resin during the initial curing phase shows toxic effects on MC3T3-E1 cells, that cytotoxicity is minimal at later curing phases. In contrast, neither the uncured nor cured resins affected the GE-1 cells.


Subject(s)
Boron Compounds/chemistry , Gingiva/drug effects , Keratinocytes/drug effects , Methacrylates/chemistry , Methylmethacrylates/chemistry , Osteoblasts/drug effects , Resin Cements/chemistry , 3T3 Cells , Alkaline Phosphatase/analysis , Animals , Boron Compounds/pharmacology , Cell Culture Techniques , Cell Death , Cell Line , Cell Proliferation/drug effects , Cell Survival/drug effects , Gingiva/cytology , Materials Testing , Methacrylates/pharmacology , Methylmethacrylates/pharmacology , Mice , Resin Cements/pharmacology , Time Factors
11.
J Cell Biochem ; 117(11): 2552-8, 2016 11.
Article in English | MEDLINE | ID: mdl-27012464

ABSTRACT

Regulation of inflammation is important for pulp wound healing, including protective responses by odontoblast-like cells. However, methods for directly regulating pulp inflammation have not yet been described. The inflammatory response is mediated by a transcription factor, nuclear factor-κB (NF-κB), which activates inflammatory cytokines including tumor necrosis factor (TNF)-α. Macromolecular translocation inhibitor II (MTI-II) was previously demonstrated as an enhancer of the transcriptional activity of glucocorticoid-bound glucocorticoid receptor. Recently, a MTI-II peptide anti-inflammatory drug (MPAID) was bioengineered from the structure of MTI-II as an inhibitor of NF-κB transactivation. Here, we examined the effects of MTI-II and MPAID on the inflammatory responses of odontoblast-like cells. TNF-α inhibited alkaline phosphatase (ALP) activity, a marker of odontoblast/osteogenic differentiation, and induced NF-κB transcriptional activity in KN-3 cells, which are odontoblast-like cells derived from dental papilla cells of rat incisors, without affecting their viability. Exogenous expression of MTI-II suppressed the NF-κB transcriptional activity induced by TNF-α or overexpression of p65 (a main subunit of NF-κB) in the cells, whereas it failed to inhibit degradation of IκBα and nuclear translocation of p65 after TNF-α treatment, suggesting that MTI-II inhibits NF-κB transcriptional activity by modulating the duration of DNA binding by p65. MPAID also inhibited TNF-α-induced NF-κB transcriptional activity, the mRNA expression of IL-6 and IL-8, and IL-6 production. Furthermore, pretreatment of the cells with MPAID restored the inhibitory effect of TNF-α on ALP activity. These results suggest that MPAID may be able to regulate the inflammatory response and maintain a protective response of dental pulp. J. Cell. Biochem. 117: 2552-2558, 2016. © 2016 Wiley Periodicals, Inc.


Subject(s)
Anti-Inflammatory Agents/pharmacology , Inflammation/immunology , NF-kappa B/antagonists & inhibitors , Odontoblasts/immunology , Thymosin/analogs & derivatives , Animals , Apoptosis , Blotting, Western , Cell Proliferation , Cells, Cultured , Cytokines/genetics , Cytokines/metabolism , Fluorescent Antibody Technique , Inflammation/drug therapy , Inflammation/pathology , NF-kappa B/genetics , NF-kappa B/metabolism , Odontoblasts/cytology , Odontoblasts/drug effects , RNA, Messenger/genetics , Rats , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Thymosin/pharmacology
12.
J Endod ; 40(12): 1989-94, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25305237

ABSTRACT

INTRODUCTION: This study investigated the effects of Emdogain gel (EMD) on the injured open apex within periapical lesions. METHODS: Periapical lesions were induced in rats by opening the pulp chambers of the mandibular first molars and filing the apical foramen through the distal root canal with #25 K-files to make an open apex. The teeth were exposed to the oral environment for 7 days. Then we irrigated the distal root canals and divided them into EMD-treated and propylene glycol alginate-treated groups. The rats were killed 7, 14, and 28 days after treatment and examined histochemically. RESULTS: In the EMD-treated rats, more cells expressed transforming growth factor-ß1 or bone morphogenetic protein-2 at 7 days after treatment, and the regeneration of cementum and bone was observed around the root apex at 14 days after treatment. Conversely, in the propylene glycol alginate-treated group, few cells expressed transforming growth factor-ß1 or bone morphogenetic protein-2, and apical periodontal tissue recovery was rarely seen within the periapical lesions throughout the experiment. CONCLUSIONS: These results suggest that EMD does not irritate injured periapical tissue and may create a favorable environment that promotes the healing of destroyed periapical tissues.


Subject(s)
Dental Enamel Proteins/therapeutic use , Periapical Periodontitis/drug therapy , Tooth Apex/injuries , Alginates/therapeutic use , Alkaline Phosphatase/drug effects , Animals , Bone Morphogenetic Protein 2/analysis , Cell Count , Dental Cementum/drug effects , Dental Pulp Cavity/drug effects , Dental Pulp Cavity/injuries , Ectodysplasins/analysis , Fibroblasts/drug effects , Macrophages/drug effects , Male , Mandible/drug effects , Molar/drug effects , Molar/injuries , Neutrophil Infiltration/drug effects , Osteoblasts/drug effects , Osteogenesis/drug effects , Random Allocation , Rats , Regeneration/drug effects , Time Factors , Tooth Apex/drug effects , Transforming Growth Factor beta1/analysis
13.
Mol Endocrinol ; 28(9): 1460-70, 2014 Sep.
Article in English | MEDLINE | ID: mdl-25029242

ABSTRACT

Bone morphogenic proteins (BMPs) stimulate bone formation in vivo and osteoblast differentiation in vitro via a Smad signaling pathway. Recent findings revealed that the activation of nuclear factor-κB (NF-κB) inhibits BMP-induced osteoblast differentiation. Here, we show that NF-κB inhibits BMP signaling by directly targeting the Smad pathway. A selective inhibitor of the classic NF-κB pathway, BAY11-770682, enhanced BMP2-induced ectopic bone formation in vivo. In mouse embryonic fibroblasts (MEFs) prepared from mice deficient in p65, the main subunit of NF-κB, BMP2, induced osteoblastic differentiation via the Smad complex to a greater extent than that in wild-type MEFs. In p65(-/-) MEFs, the BMP2-activated Smad complex bound much more stably to the target element than that in wild-type MEFs without affecting the phosphorylation levels of Smad1/5/8. Overexpression of p65 inhibited BMP2 activity by decreasing the DNA binding of the Smad complex. The C-terminal region, including the TA2 domain, of p65 was essential for inhibiting the BMP-Smad pathway. The C-terminal TA2 domain of p65 associated with the MH1 domain of Smad4 but not Smad1. Taken together, our results suggest that p65 inhibits BMP signaling by blocking the DNA binding of the Smad complex via an interaction with Smad4. Our study also suggests that targeting the association between p65 and Smad4 may help to promote bone regeneration in the treatment of bone diseases.


Subject(s)
Bone Morphogenetic Protein 2/metabolism , Gene Expression Regulation , Smad4 Protein/metabolism , Transcription Factor RelA/metabolism , Animals , Bone Development , Bone Diseases/metabolism , Cell Differentiation/genetics , Fibroblasts/metabolism , Humans , Mice , Osteoblasts/metabolism , Osteogenesis , Protein Binding , Protein Structure, Tertiary , Recombinant Proteins/metabolism , Signal Transduction/genetics
14.
Arch Oral Biol ; 59(7): 741-8, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24814171

ABSTRACT

OBJECTIVE: Heat shock during restorative procedures can trigger damage to the pulpodentin complex. While severe heat shock has toxic effects, fever-range heat stress exerts beneficial effects on several cells and tissues. In this study, we examined whether continuous fever-range heat stress (CFHS) has beneficial effects on thermotolerance in the rat clonal dental pulp cell line with odontoblastic properties, KN-3. METHODS: KN-3 cells were cultured at 41°C for various periods, and the expression level of several proteins was assessed by Western blot analysis. After pre-heat-treatment at 41°C for various periods, KN-3 cells were exposed to lethal severe heat shock (LSHS) at 49°C for 10min, and cell viability was examined using the MTS assay. Additionally, the expression level of odontoblast differentiation makers in surviving cells was examined by Western blot analysis. RESULTS: CFHS increased the expression levels of several heat shock proteins (HSPs) in KN-3 cells, and induced transient cell cycle arrest. KN-3 cells, not pre-heated or exposed to CFHS for 1 or 3h, died after exposure to LSHS. In contrast, KN-3 cells exposed to CFHS for 12h were transiently lower on day 1, but increased on day 3 after LSHS. The surviving cells expressed odontoblast differentiation markers, dentine sialoprotein and dentine matrix protein-1. These results suggest that CFHS for 12h improves tolerance to LSHS by inducing HSPs expression and cell cycle arrest in KN-3 cells. CONCLUSIONS: The appropriate pretreatment with continuous fever-range heat stress can provide protection against lethal heat shock in KN-3 cells.


Subject(s)
Heat-Shock Proteins/metabolism , Heat-Shock Response/physiology , Odontoblasts/physiology , Adaptation, Physiological , Animals , Apoptosis/physiology , Blotting, Western , Cell Cycle/physiology , Cell Survival , Cells, Cultured , Clone Cells , Hot Temperature , In Situ Nick-End Labeling , Rats , Stress, Physiological/physiology
15.
Biol Pharm Bull ; 35(8): 1371-3, 2012.
Article in English | MEDLINE | ID: mdl-22863940

ABSTRACT

Elevation in the temperature induces heat stress to both host cells and the invading pathogen. This study aimed to determine whether continuous mild heat stress (increased temperature without causing significant damage to host cells) can increase susceptibility of biofilm formation of the opportunistic fungal pathogen Candida albicans to low concentrations of three typical antifungal agents. In this way the side effects associated with higher concentrations of the antifungal agents on host cells would be reduced. Fluconazole and micafungin at concentrations ranging from 0.0625 to 2 µg/mL and amphotericin B at concentrations ranging from 0.0625 to 1 µg/mL inhibited less than 20% of cells in biofilm formation. Biofilm formation at 39 or 41°C compared to 37°C resulted in increased susceptibility to the three agents, but especially micafungin. These data suggest that mild heat stress (39°C) would be valuable for increasing the effectiveness of low concentrations of antifungal agents against C. albicans biofilm formation. Thus, the concept of continuous mild heat stress at the site of insertion of medical devices or catheters combined with antifungal agents could be beneficial.


Subject(s)
Amphotericin B/pharmacology , Antifungal Agents/pharmacology , Biofilms/drug effects , Candida albicans/drug effects , Echinocandins/pharmacology , Fluconazole/pharmacology , Hot Temperature , Lipopeptides/pharmacology , Stress, Physiological , Micafungin
16.
Int J Dent ; 2012: 258469, 2012.
Article in English | MEDLINE | ID: mdl-22505896

ABSTRACT

We examined the effects of bone morphogenetic protein-2 (BMP-2) on growth, differentiation, and intracellular signaling pathways of odontoblast-like cells, KN-3 cells, to clarify molecular mechanisms of odontoblast differentiation during pulp regeneration process. After treatment with BMP-2, the cell morphology, growth, alkaline phosphatase (ALP) activity, and the activation and expression of BMP-induced intracellular signaling molecules, such as Smad1/5/8 and Smad6/7, as well as activities of dentin sialoprotein (DSP) and dentin matrix protein 1 (DMP1), were examined. BMP-2 had no effects on the morphology, growth, or ALP activity of KN-3 cells, whereas it induced the phosphorylation of Smad1/5/8 and expression of Smad6/7. BMP-2 also induced the expressions of DSP and DMP-1. Our results suggest that KN-3 cells may express an odontoblastic phenotype with the addition of BMP-2 through the activation of Smad signaling pathways.

17.
J Endod ; 37(7): 955-61, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21689551

ABSTRACT

INTRODUCTION: Heat stress during restorative procedures, particularly under severe starvation conditions, can trigger damage to dental pulp. In the present study, we examined effects of heat stress on odontoblastic activity and inflammatory responses in an odontoblast-like cell line (KN-3) under serum-starved conditions. METHODS: Viability, nuclear structures, and inflammatory responses of KN-3 cells were examined in culture medium containing 10% or 1% serum after exposure to heat stress at 43°C for 45 minutes. Gene expression of extracellular matrices, alkaline phosphatase activity, and detection of extracellular calcium deposition in cells exposed to heat stress were also examined. RESULTS: Reduced viability and apoptosis were transiently induced in KN-3 cells during the initial phases after heat stress; thereafter, cells recovered their viability. The cytotoxic effects of heat stress were enhanced under serum-starved conditions. Heat stress also strongly up-regulated expression of heat shock protein 25 as well as transient expression of tumor necrosis factor-alpha, interleukin-6, and cyclooxygenase-2 in KN-3 cells. In contrast, expression of type-1 collagen, runt-related transcription factor 2, and dentin sialophosphoprotein were not inhibited by heat stress although starvation suppressed ALP activity and delayed progression of calcification. CONCLUSIONS: Odontoblast-like cells showed thermoresistance with transient inflammatory responses and without loss of calcification activity, and their thermoresistance and calcification activity were influenced by nutritional status.


Subject(s)
Heat-Shock Response/physiology , Odontoblasts/physiology , Stress, Physiological/physiology , Adaptation, Physiological , Animals , Apoptosis/physiology , Calcification, Physiologic/physiology , Cell Survival , Cells, Cultured , Clone Cells , Culture Media, Serum-Free , Hot Temperature , Inflammation Mediators/metabolism , Odontoblasts/cytology , Rats
18.
J Biomed Mater Res B Appl Biomater ; 92(1): 120-8, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19802830

ABSTRACT

It is important to develop a suitable three-dimensional scaffold for the regeneration therapy of dental pulp. In the present study, the effects of hyaluronic acid (HA) sponge on responses of the odontoblastic cell line (KN-3 cells) in vitro, as well as responses of amputated dental pulp of rat molar in vivo, were examined. In vitro, KN-3 cells adhered to the stable structure of HA sponge and that of collagen sponge. In vivo, dental pulp proliferation and vessel invasion were observed in both sponges implanted at dentin defect area above amputated dental pulp, and the cell-rich reorganizing tissue was observed in the dentin defect when HA sponge was implanted as compared with collagen sponge. Expression levels of IL-6 and TNF-alpha in KN-3 cells seeded in HA sponge were nearly the same with those in the cells seeded in collagen sponge, while the numbers (0.67 x 10(3) at 1 week and 0.7 x 10(3) at 3 weeks) of granulated leukocytes that invaded into HA sponge from amputated dental pulp was significantly lower than those (1.22 x 10(3) at 1 week and 1.1 x 10(3) at 3 weeks) of collagen sponge (p < 0.01 at 1 week and p < 0.05 at 3 weeks). These results suggest that HA sponge has an appropriate structure, biocompatibility, and biodegradation for use as a scaffold for dental pulp regeneration.


Subject(s)
Dental Pulp/cytology , Hyaluronic Acid/chemistry , Odontoblasts/cytology , Animals , Base Sequence , Cell Line , DNA Primers , Microscopy, Electron, Scanning , Rats , Reverse Transcriptase Polymerase Chain Reaction
19.
J Endod ; 35(6): 858-65, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19482186

ABSTRACT

INTRODUCTION: Pulp regeneration therapy is important to overcome the limitations of conventional therapy to induce reparative dentinogenesis. In the present study, we examined the effects of controlled release of different dosages of fibroblast growth factor-2 (FGF-2) from gelatin hydrogels to regenerate the dentin-pulp complex. METHODS: After the amputation of dental pulp of rat molars, gelatin hydrogels incorporating various dosages of FGF-2 were individually implanted into dentin defects above the sites of the amputated pulps. Histologic changes as well as the expression of dentin matrix protein-1 (DMP-1) and nestin in the dentin defect area above the amputated pulp were analyzed. RESULTS: We found that controlled release of high doses of FGF-2 from gelatin hydrogels induced DMP-1-positive calcified particles in the proliferating pulp, whereas a moderate dose of FGF-2 induced DMP-1-positive dentinal bridge on the surface of the proliferating pulp. These findings indicate that the dosage of released FGF-2 has an influence on the structure of calcified tissue regenerated in dentin defects. In addition, pulp cells near calcified tissues regenerated in dentin defects were nestin-negative, suggesting that the calcified tissues might be osteodentin. CONCLUSIONS: Our results showed that the dentin regeneration on amputated pulp, not reparative dentin formation toward amputated pulp, can be regulated by adjusting the dosage of FGF-2 incorporated in biodegradable gelatin hydrogels.


Subject(s)
Dental Pulp/drug effects , Dentin, Secondary/metabolism , Dentin/drug effects , Fibroblast Growth Factor 2/pharmacology , Regeneration/drug effects , Animals , Calcification, Physiologic/drug effects , Dental Pulp/metabolism , Dental Pulp/physiology , Dental Pulp Exposure , Dentin/metabolism , Dentin/physiology , Dentin, Secondary/growth & development , Dose-Response Relationship, Drug , Drug Carriers , Extracellular Matrix Proteins/biosynthesis , Fibroblast Growth Factor 2/administration & dosage , Hydrogels , Intermediate Filament Proteins/biosynthesis , Microspheres , Nerve Tissue Proteins/biosynthesis , Nestin , Phosphoproteins/biosynthesis , Rats , Rats, Wistar , Regeneration/physiology , Specific Pathogen-Free Organisms
20.
J Endod ; 33(10): 1187-91, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17889687

ABSTRACT

To clarify mechanisms of pulp wound healing and regeneration, it is important to establish continuous odontoblast-lineage cell lines. In this study, we established the proliferating pulp progenitor cell lines from dental papilla cells of rat incisor. These cell lines showed high levels of alkaline phosphatase (ALP) activity, expression of Runx2 and dentin sialophosphoprotein (DSPP), and extracellular formation of mineralized nodules. By using the cell line with high expression level of DSPP and the prominent mineral deposition, we examined whether bacterial lipopolysaccharide (LPS) had effects on its odontoblastic properties and found that ALP activity, expression of DSPP and Runx2, and the formation of mineralized nodules were suppressed in LPS dose-dependent manner. These results indicate that our established pulp progenitor cell line exhibits odontoblastic properties, which were suppressed by LPS, suggesting that gram-negative bacterial infection might downregulate the odontoblast function.


Subject(s)
Aggregatibacter actinomycetemcomitans , Dental Pulp/drug effects , Lipopolysaccharides/pharmacology , Odontoblasts/drug effects , Stem Cells/drug effects , Alkaline Phosphatase/analysis , Animals , Calcification, Physiologic/drug effects , Calcium/metabolism , Cell Line , Cell Lineage , Clone Cells/cytology , Core Binding Factor Alpha 1 Subunit , Dental Papilla/cytology , Dental Pulp/cytology , Dose-Response Relationship, Drug , Extracellular Matrix Proteins , Extracellular Space/drug effects , Extracellular Space/metabolism , Lipopolysaccharides/administration & dosage , Odontoblasts/cytology , Phosphoproteins , Protein Precursors , Rats , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Sialoglycoproteins , Specific Pathogen-Free Organisms , Stem Cells/cytology , Time Factors
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