Yun Chan Hwang

Effect of Biodentine and Bioaggregate on odontoblastic differentiation via mitogen-activated protein kinase pathway in human dental pulp cells.

AIMnTo compare the mineralization inductive capacity of Biodentine and Bioaggregate with Mineral trioxide aggregate (MTA) and to investigate possible signaling pathways of mineralization in human dental pulp cells (HDPCs).nnnMETHODOLOGYnViability of HDPCs in response to Biodentine, Bioaggregate, and MTA was measured using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide. To investigate their potential to induce odontoblast differentiation, expression of dentine sialophosphoprotein (DSPP) and dentine matrix protein1 (DMP1) mRNA level was evaluated by RT-PCR. For the mineralized nodule assay, Alizarin red staining was performed. To determine the role of MAPK signaling in the odontoblastic differentiation of HDPCs, activated MAPKs were investigated by Western blot and the effect of MAPK inhibitor was examined by Alizarin red S staining. The results were statistically analysed using one-way anova and the Bonferroni test.nnnRESULTSnThe effects of MTA, Biodentine, and Bioaggregate on cell viability were similar. Biodentine and Bioaggregate enhanced DSPP and DMP1 mRNA expression compared to the control group, but to the same extent as MTA (P < 0.05). MTA, Biodentine, and Bioaggregate increased the area of calcified nodules compared to the control (P < 0.01). MTA, Biodentine, and Bioaggregate increased phosphorylation of extracellular signal-regulated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK). MAPK inhibitors attenuated mineralized nodule formation, which was increased by MTA, Biodentine, and Bioaggregate, respectively (P < 0.01).nnnCONCLUSIONnBiodentine and Bioaggregate stimulated odontoblastic differentiation and mineralization nodule formation by activating the MAPK pathway as did MTA. This suggests that the new materials could be useful for regenerative endodontic procedures.

Transcriptional Factor ATF6 is Involved in Odontoblastic Differentiation

ATF6 is an endoplasmic reticulum (ER) membrane-bound transcription factor that regulates various cellular functions. The purpose of this study was to investigate the role of ATF6 in odontoblast differentiation. Rat tooth germs were isolated, changes in gene expression were evaluated over time, and localization of ATF6 was determined by immunohistochemistry. Human dental pulp cells (HDPCs) were cultured with 50 µg/mL ascorbic acid and 5 mmol/L β-glycerophosphate or 100 ng/mL bone morphogenetic protein 2 to induce differentiation. Translocation of ATF6 was observed by immunofluorescence and confocal microscopy. Overexpression of ATF6 was performed with an adenoviral vector. Matrix mineralization was evaluated by alizarin red staining. Immunoreactivity to anti-ATF6 was observed in the odontoblastic layer of the molar tooth germ, and expressions of ATF6, dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP1) increased gradually during tooth germ development. When HDPCs were cultured in differentiation media, ATF6, DSPP, and DMP1 expression increased with the expression of unfolded protein response (UPR) markers, BiP and CHOP. Immunofluorescence results showed that ATF6 protein moved from cytoplasm to nucleus when cells were exposed to differentiation media. Notably, overexpression of ATF6 increased DSPP and DMP1 expression, alkaline phosphatase (ALP) activity, and matrix mineralization in HDPC cultures. Inhibition of ATF6 decreased ALP activity and mineralization. These results suggest that ER membrane-bound transcriptional factor ATF6 may be involved in odontoblastic differentiation.

Physical and chemical properties of experimental mixture of mineral trioxide aggregate and glass ionomer cement

5 DSRI, 6 2nd stage of BK21, Chonnam National University School of Dentistry, Gwangju, Korea Objectives: The purpose of this study was to determine the setting time, compressive strength, solubility, and pH of mineral trioxide aggregate (MTA) mixed with glass ionomer cement (GIC) and to compare these properties with those of MTA, GIC, IRM, and SuperEBA. Materials and Methods: Setting time, compressive strength, and solubility were determined according to the ISO 9917 or 6876 method. The pH of the test materials was determined using a pH meter with specified electrode for solid specimen. Results: The setting time of MTA mixed with GIC was significantly shorter than that of MTA. Compressive strength of MTA mixed with GIC was significantly lower than that of other materials at all time points for 7 days. Solubility of 1 : 1 and 2 : 1 specimen from MTA mixed with GIC was significantly higher than that of other materials. Solubility of 1 : 2 specimen was similar to that of MTA. The pH of MTA mixed with GIC was 2-4 immediately after mixing and increased to 5-7 after 1 day. Conclusions: The setting time of MTA mixed with GIC was improved compared with MTA. However, other properties such as compressive strength and pH proved to be inferior to those of MTA. To be clinically fea- sible, further investigation is necessary to find the proper mixing ratio in order to improve the drawbacks of MTA without impairing the pre-existing advantages and to assess the biocompatibility. ABSTRACT

Simvastatin inhibits the expression of inflammatory cytokines and cell adhesion molecules induced by LPS in human dental pulp cells.

AIMnTo investigate the effect of simvastatin on lipopolysaccharide (LPS)-stimulated inflammatory cytokines, cell adhesion molecules and nuclear factor-κB (NF-κB) transcription factors in human dental pulp cells (HDPCs).nnnMETHODOLOGYnThe effect of LPS and simvastatin on human dental pulp cell (HDPCs) viability was measured using a 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium bromide (MTT) assay. The expression of inflammatory cytokines and cell adhesion molecules was evaluated by reverse-transcription polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. NF-κB transcription factors were evaluated by Western blot analysis. Statistical analysis was performed with analysis of variance (anova).nnnRESULTSnThe viability of cells exposed to different concentrations of E.xa0coli LPS, P.xa0gingivalis LPS and simvastatin was not significantly different compared with that of control cells (Pxa0>xa00.05). LPS significantly increased interleukin (IL)-1β (Pxa0<xa00.05) and IL-6 mRNA expression (Pxa0<xa00.05) and vascular cell adhesion molecule-1 (VCAM-1) (Pxa0<xa00.05) and intercellular adhesion molecule-1 (ICAM-1) protein expression (Pxa0<xa00.05) in HDPCs. Treatment with simvastatin significantly attenuated LPS-stimulated production of IL-1β, IL-6, VCAM-1 and ICAM-1 (Pxa0<xa00.05). Treatment with simvastatin decreased LPS-induced expression of p65 and phosphorylation of IκB and also significantly decreased the phosphorylation of p65 and IκB in the cytoplasm and the level of p65 in the nucleus (Pxa0<xa00.05).nnnCONCLUSIONSnSimvastatin has a suppressing effect on LPS-induced inflammatory cytokine, cell adhesion molecules and NF-κB transcription factors in HDPCs. Therefore, simvastatin might be a useful candidate as a pulp-capping agent in vital pulp therapy.

SHP is Involved in BMP2-induced Odontoblast Differentiation:

Small Heterodimer Partner (SHP) interacts with diverse transcription factors such as Runx2 and regulates many cellular events including differentiation, proliferation, and energy metabolism. SHP is reported to be a positive regulator of BMP2-induced bone formation. This study aimed to clarify the role of SHP in odontoblast differentiation and matrix mineralization. Rat tooth germs were isolated, and gene expression was determined by RT-PCR and real-time PCR. Localization of SHP protein expression was identified by immunofluorescent analysis. Primary human dental pulp cells (HDPCs) were cultured with BMP2 and/or Ad-siSHP. Matrix mineralization was evaluated by Alizarin red staining. Transient transfection experiment was performed with the SHP or Dlx5 expressional plasmids and the DSPP gene. In tooth germs from post-natal days 3 to 9, BMP-2 and SHP expression increased with DSPP and DMP1 mRNA expression. In an immunostaining study, SHP was expressed in odontoblasts and surrounding osteoblasts. When HDPCs were cultured with BMP2 in mineralization-inducing medium, SHP expression also increased with an increase in DSPP expression. Down-regulation of SHP by Ad-siSHP inhibited matrix mineralization. In transient transfection experiments, overexpression of SHP was shown to enhance DSPP promoter activity through interactions between SHP and Dlx5. These results suggest that SHP may mediate BMP2 signaling to promote mineralization of the dentin matrix.

Ketoprofen inhibits expression of inflammatory mediators in human dental pulp cells

INTRODUCTIONnConventional root canal treatment is the treatment of choice for the irreversible pulpitis caused by bacterial infection. More recently, vital pulp therapy has been proposed as an alternative for management of inflamed dental pulp. Ketoprofen is an anti-inflammatory agent commonly used as a component of mouth rinse for oral lesions. Here, we examined the effect and mechanisms of action of ketoprofen on the expression of inflammatory mediators induced by the lipopolysaccharide (LPS) in dental pulp cells.nnnMETHODSnHuman dental pulp cells were exposed to LPS or LPS + ketoprofen, and reverse-transcription polymerase chain reaction was used to detect interleukin-1β and tumor necrosis factor α. The effect of these treatments on mitogen-activated protein kinase pathways was assessed by Western blots for extracellular signal-regulated kinase and c-Jun N-terminal kinase.nnnRESULTSnLPS induced interleukin-1β and tumor necrosis factor α in dental pulp cells. Ketoprofen effectively inhibited interleukin-1β and tumor necrosis factor α production in LPS-stimulated dental pulp cells. Notably, ketoprofen inhibited phosphorylation of extracellular signal-regulated kinase and c-Jun N-terminal kinase.nnnCONCLUSIONSnKetoprofen inhibited expression inflammatory mediators in dental pulp cells stimulated with LPS. The inhibitory effect of ketoprofen on inflammatory cytokines is associated with inhibition of the mitogen-activated protein kinase pathway.

Treatment of non-vital immature teeth with amoxicillin-containing triple antibiotic paste resulting in apexification

A recent treatment option for non-vital immature teeth in young patients is revascularization with triple antibiotic paste (TAP). However, tooth discoloration was reported with the use of conventional minocycline-containing TAP. In this case report, amoxicillin-containing TAP was used for revascularization of non-vital immature teeth to prevent tooth discoloration. At the 1 yr follow up, the teeth were asymptomatic on clinical examination and showed slight discoloration of the crown due to mineral trioxide aggregate (MTA) filling rather than amoxicillin-containing TAP. Radiographic examination revealed complete resolution of the periapical radiolucency, and closed apex with obvious periodontal ligament space. However, the root growth was limited, and the treatment outcome was more like apexification rather than revascularization. These results may be due to unstable blood clot formation which could not resist the condensation force of MTA filling, whether or not a collagen matrix was in place. These cases showed that although revascularization was not successful, apexification could be expected, resulting in the resolution of the periapical radiolucency and the closure of the apex. Therefore, it is worthwhile attempting revascularization of non-vital immature teeth in young patients.

Autogenous tooth transplantation for replacing a lost tooth: case reports

The autogenous tooth transplantation is an alternative treatment replacing a missing tooth when a suitable donor tooth is available. It is also a successful treatment option to save significant amount of time and cost comparing implants or conventional prosthetics. These cases, which required single tooth extraction due to deep caries and severe periodontal disease, could have good results by transplanting non-functional but sound donor tooth to the extraction site.

Behaviour of human dental pulp cells cultured in a collagen hydrogel scaffold cross-linked with cinnamaldehyde

AIMnTo investigate the effects of the cross-linking agent cinnamaldehyde (CA) on differentiation of human dental pulp cells (hDPCs) cultured in a collagen hydrogel, which may be useful as a scaffold for regenerative endodontic therapy.nnnMETHODOLOGYnThe odontogenic potential of hDPCs exposed to CA was examined using alkaline phosphatase (ALP) activity, Alizarin red S staining and real-time polymerase chain reaction for odontogenic gene expression. The morphological features of hDPCs cultured in CA-treated collagen were evaluated by scanning electron microscopy. Determination of cell numbers for evaluating proliferation was assessed by optical and fluorescence microscopy. To assess the mechanical properties of collagen treated with CA, setting time, compressive strength and surface roughness were measured. Statistical analysis was performed using Students t-test compared with control (Pxa0=xa00.05).nnnRESULTSnCA per se did not increase ALP activity, calcium nodule formation and expression of odontogenic-related markers (Pxa0>xa00.05). On the contrary, the proliferation and odontogenic differentiation of hDPCs cultured in a collagen scaffold was promoted in the presence of CA (Pxa0<xa00.05). The setting time was significantly shortened, and the compressive strength and surface roughness were increased by treatment with CA (Pxa0<xa00.05).nnnCONCLUSIONSnCross-linking of collagen scaffolds by CA had beneficial effects with respect to attachment, proliferation and differentiation of hDPCs. Consequently, the application of cross-linking agents such as CA may represent a new strategy for dentine-pulp complex regeneration.

Application of orthodontic forces prior to autotransplantation – case reports

AIMnThis case report describes the successful autotransplantation of mandibular molars after application of orthodontic forces and discusses the advantages of this technique, that is, pre-application of an orthodontic force for autotransplantation.nnnSUMMARYnAfter clinical and radiographic examination, autotransplantation was planned with the patients written informed consent. An orthodontic force was applied, and the surgical procedure was performed after tooth mobility had increased. Root canal treatment was performed within 2 weeks of autotransplantation. At the 1-year follow-up, the transplanted teeth revealed asymptomatic and healthy periodontal conditions.nnnKEY LEARNING POINTSnAutotransplantation is the surgical movement of a tooth from its original location to another site. The pre-application of orthodontic force technique was recently introduced for autogenous tooth transplantation. Pre-application of an orthodontic force may be a useful treatment option for autotransplantation.