Sang Hyuk Park

Polymerization Shrinkage and Depth of Cure of Bulk-Fill Resin Composites and Highly Filled Flowable Resin

The aim of this study was to evaluate the polymerization behavior and depth of cure (DOC) of recently introduced resin composites for posterior use: highly filled flowable composite and composites for bulk fill. A highly filled flowable (G-aenial Universal Flo [GUF]), two bulk-fill flowables (Surefil SDR Flow [SDR] and Venus Bulk fill [VBF]), and a bulk-fill nonflowable composite (Tetric N-Ceram Bulk fill [TBF]) were compared with two conventional composites (Tetric Flow [TF], Filtek Supreme Ultra [FS]). Linear polymerization shrinkage and polymerization shrinkage stress were each measured with custom-made devices. To evaluate DOC, the composite specimen was prepared using a mold with a hole of 4 mm depth and 4 mm internal diameter. The hole was bulk filled with each of the six composites and light cured for 20 seconds, followed by 24 hours of water storage. The surface hardness was measured on the top and the bottom using a Vickers microhardness (HV) indenter. The linear polymerization shrinkage of the composite specimens after photo-initiation decreased in the following order: TF and GUF > VBF > SDR > FS and TBF (p<0.05). The polymerization shrinkage stress of the six composite groups decreased in the following order: GUF > TF and VBF > SDR > FS and TBF (p<0.05). The mean bottom surface HV of SDR and VBF exceeded 80% of the top surface HV (HV-80%). However, the bottom of GUF and TBF failed to reach HV-80%. A highly filled flowable (GUF) revealed limitations in polymerization shrinkage and DOC. Bulk-fill flowables (SDR and VBF) were properly cured in 4-mm bulk, but they shrank more than the conventional nonflowable composite. A bulk-fill nonflowable (TBF) showed comparable shrinkage to the conventional nonflowable composite, but it was not sufficiently cured in the 4-mm bulk.

Effects of moderate intensity static magnetic fields on human bone marrow-derived mesenchymal stem cells.

This study aimed to explore effects of static magnetic fields (SMFs) of moderate intensity (3-50 mT) as biophysical stimulators of proliferation and osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells (MSCs). MSCs were exposed to SMFs of three intensities: 3, 15, and 50 mT. Proliferation was assessed by cell counting and bromodeoxyuridine incorporation, and differentiation by measuring alkaline phosphatase (ALP) activity, calcium content, mineralized nodule formation, and transcripts of osteogenic markers. Moderate intensity SMFs increased cell proliferation, ALP activity, calcium release, and mineralized nodule formation in a dose- and time-dependent manner, which peaked at 15 mT. In the same manner, they upregulated expression of osteogenic marker genes such as ALP, bone sialoprotein 2 (BSP2), collagen1a1 (COL1a1), osteocalcin (OCN), osteonectin (ON), osteopontin (OPN), osterix (OSX), and runt-related transcription factor 2 (RUNX2) with peak at 15 mT after 14 or 21 days of exposure. Results demonstrate that moderate intensity SMFs promote proliferation and osteoblastic differentiation of MSCs. This effect could help to improve MSC responses during osseointegration between a dental implant and surrounding bone.

Prognostic Factors for Clinical Outcomes in Autotransplantation of Teeth with Complete Root Formation: Survival Analysis for up to 12 Years.

INTRODUCTION Tooth autotransplantation is a treatment option that has the potential to restore masticatory function and esthetics to edentulous spaces resulting from extracted teeth. The purpose of this study was to investigate the prognostic factors and clinical outcomes for autotransplanted teeth with complete root formation. METHODS Patients who had received tooth autotransplantation in the Department of Conservative Dentistry, Yonsei University Dental Hospital, Seoul, Korea, from July 2001 to August 2010 were electronically searched, and a total of 105 cases of autotransplanted teeth met the inclusion criteria. Tooth survival, inflammatory root resorption (IRR), ankylosis, and related prognostic factors were assessed by using the survival analysis that was based on clinical and radiographic examination. RESULTS The cumulative tooth survival rate was 68.2% at 12 years after the tooth autotransplantation. According to the Cox proportional hazard regression analysis, patient age, donor position, and extraoral time were significantly associated with tooth survival (P < .05). Donor extraction type was significantly associated with IRR (P < .05), and transplantation timing and initial stability were significantly associated with ankylosis (P < .05) CONCLUSIONS: Patients less than 45 years of age, maxillary donor teeth, and an extraoral time of less than 15 minutes were associated with significantly higher tooth survival. Surgical extraction of the donor tooth was associated with a significantly higher incidence of IRR. Immediate transplantation after the extraction of the recipient sites tooth and low initial stability were associated with a significantly lower incidence of ankylosis.

An Overview of Pathogen Recognition Receptors for Innate Immunity in Dental Pulp

Pathogen recognition receptors (PRRs) are a class of germ line-encoded receptors that recognize pathogen-associated molecular patterns (PAMPs). The activation of PRRs is crucial for the initiation of innate immunity, which plays a key role in first-line defense until more specific adaptive immunity is developed. PRRs differ in the signaling cascades and host responses activated by their engagement and in their tissue distribution. Currently identified PRR families are the Toll-like receptors (TLRs), the C-type lectin receptors (CLRs), the nucleotide-binding oligomerization domain-like receptors (NLRs), the retinoic acid-inducible gene-I-like receptors (RLRs), and the AIM2-like receptor (ALR). The environment of the dental pulp is substantially different from that of other tissues of the body. Dental pulp resides in a low compliance root canal system that limits the expansion of pulpal tissues during inflammatory processes. An understanding of the PRRs in dental pulp is important for immunomodulation and hence for developing therapeutic targets in the field of endodontics. Here we comprehensively review recent finding on the PRRs and the mechanisms by which innate immunity is activated. We focus on the PRRs expressed on dental pulp and periapical tissues and their role in dental pulp inflammation.

Effect of Mineral Trioxide Aggregate Surface Treatments on Morphology and Bond Strength to Composite Resin

INTRODUCTION The aim of this study was to evaluate the micromorphologic changes that accompany different surface treatments on mineral trioxide aggregate (MTA) and their effect on the bond strength to the composite resin with 4 adhesive systems. METHODS Three types of MTA cement, ProRoot MTA (WMTA) (Dentsply, Tulsa, OK), MTA Angelus (AMTA) (Angelus, Londrina, PR, Brazil), and Endocem MTA (EMTA) (Maruchi, Wonju, Korea), were prepared and stored for a week to encourage setting. Surface treatment was performed using phosphoric acid or self-etch primer, and an untreated MTA surface was prepared as a control. The surface changes were observed using scanning electron microscopy. MTA surfaces were bonded with 4 adhesive systems, including Scotchbond Multipurpose (3M ESPE, St Paul, MN), Single Bond 2 (3M ESPE), Clearfil SE BOND (Kuraray, Osaka, Japan), and AdheSE One F (Ivoclar Vivadent, Schaan, Liechtenstein), to evaluate the adhesive effectiveness of MTA followed by composite resin restoration. The shear bond strength of the polymerized specimens was tested. RESULTS For WMTA and AMTA, untreated surfaces showed an irregular crystalline plate with clusters of globular aggregate particles. For EMTA, the untreated surface presented a reticular matrix with acicular crystals. After surface treatment, superficial crystalline structures were eroded regardless of the MTA cement and adhesive system used. WMTA bonded significantly more strongly than AMTA and EMTA, regardless of the adhesive system used. In the WMTA and AMTA groups, AdheSE One F showed the highest bond strength to the composite. For EMTA, no significant differences were found across adhesive systems. CONCLUSIONS Acidic treatment of the MTA surface affected the micromorphology and the bond strength to the composite. Within the limitations of this study, using a 1-step self-etch adhesive system might result in a strong bond to WMTA when the composite resin restoration is required over MTA cement.

Tumor Necrosis Factor-α Induces Matrix Metalloproteinases-3, -10, and -13 in Human Periodontal Ligament Cells

BACKGROUND Various biologic mediators, including matrix metalloproteinases (MMPs), that are implicated in periodontal tissue breakdown can be induced by cytokines. MMPs are known to degrade periodontal ligament attachment, and bone matrix proteins and tissue inhibitors of metalloproteinase (TIMPs) inhibit the activity of MMPs. The aim of this study is to investigate the effect of tumor necrosis factor (TNF)-α on the expression of MMPs in human periodontal ligament (PDL) cells in vitro and establish which MMPs are expressed specifically in response to that stimulus. METHODS Cultured PDL cells were stimulated with TNF-α and analyzed with an MMP antibody array. Real-time polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), and western blot with cell lysate and zymography were used to measure messenger RNA (mRNA) and protein levels of MMP-3, -10, and -13. To examine TNF receptor (TNFR) expression, PDL cells were examined by flow cytometry, and expression of MMP-3, -10, and -13 was observed after blocking the TNFR with an antagonist. Results from real-time PCR, ELISA, and western blot were analyzed by paired t test. RESULTS The antibody array showed that the protein most strongly upregulated by TNF-α stimulation was MMP-3, followed by MMP-13 and MMP-10. The TNF-α receptor blocker specifically inhibited expression of MMP-3 and -13. In addition, TNF-α increased levels of MMP mRNAs in MMP-3, -13, and -10 (in decreasing order). However, ELISAs showed that MMP-13 was the most upregulated protein, followed by MMP-10 and MMP-3. Western blotting indicated that TNF-α increased MMP-3 and -13 levels but had no significant effect on the level of MMP-10, and zymography showed that TNF-α increased the activities of all forms of MMP-3 and -13, but MMP-10 was not detected. Flow cytometry demonstrated that the majority of PDL cells expressed TNFR1. CONCLUSIONS TNF-α (10 ng/mL) upregulates levels of MMP-3, -10, and -13 in human PDL cells. These results suggest that these proteins play an important role in the inflammation of PDLs.

Comparison of gene expression in human periodontal ligament cells cultured from teeth immediately after extraction and from teeth cryopreserved for 1 week.

Gene expression was compared by cDNA microarray analysis in human periodontal ligament (PDL) cells cultured from teeth immediately after extraction and from teeth cryopreserved for 1 week. Twenty healthy collateral premolar teeth without caries and restorations were obtained from 10 young patients, one maxillary and one mandibular premolar from each subject. The teeth from five patients, from two patients, and from three patients out of total 10 patients were used for cDNA microarray assay, for RT-PCR, and for real-time PCR, respectively. One premolar was used immediately after extraction (control), and another premolar was stored in liquid nitrogen at -196 degrees C for 1 week (cryopreserved) from each patient. PDL cells from these teeth were cultured separately through three passages. Total RNA was isolated and gene expression was compared between the cells from control and cryopreserved group out of each subject. The microarray data were validated using the reverse transcription-polymerase chain reaction (RT-PCR) and confirmed by quantitative real-time PCR. The cultured PDL cells from the control and cryopreserved teeth were of similar appearance under an optical microscope. In all subjects the fibroblast growth factor receptor 2 (FGFR2) gene was downregulated in the cells from the cryopreserved tooth. This study shows that cryopreservation of teeth affects the expression of the FGFR2 gene in cultured PDL cells, which is related to cell growth, cell development, and cell-cell signaling.

Human Dental Pulp Stem Cells Suppress Alloantigen-induced Immunity by Stimulating T Cells to Release Transforming Growth Factor Beta

Introduction: Human dental pulp stem cells (hDPSCs) are ideal candidates for regenerating damaged dental tissue. To examine the possibility that hDPSCs may be used to regenerate pulp, we tested their in vitro effects on acute allogeneic immune responses. Methods: A peripheral blood mononuclear cell (PBMC) proliferation assay and immunoglobulin (Ig) production assay were performed to evaluate the immunosuppressive properties of hDPSCs. Results: The mixed lymphocyte reaction was suppressed by incubation with hDPSCs. Transforming growth factor beta (TGF‐&bgr;) was the major soluble factor responsible for inhibiting the allogeneic proliferation of PBMCs. The production of IgM and IgG by allogeneic activation of responder B lymphocytes was also completely abrogated by TGF‐&bgr; released from hDPSCs via interferon gamma in response to activation of the responder T lymphocytes. Conclusions: hDPSCs inhibit acute allogeneic immune responses by their release of TGF‐&bgr; as a result of allogeneic stimulation of T lymphocytes. This study provides an insight into the potential clinical use of hDPSCs for allogeneic transplantation. Highlights:Total cell preparations enzymatically purified from dental pulp possess the generic mesenchymal stem cell phenotype.Human dental pulp stem cells have an inhibitory effect on alloantigen‐evoked peripheral blood mononuclear cell proliferation and immunoglobulin (Ig) M and IgG production.hDPSCs inhibit allogeneic adaptive hyperimmune responses by their release of transforming growth factor beta as a result of allogeneic stimulation of T lymphocytes and the production of interferon gamma.

Influence of 2 Cryopreservation Methods to Induce CCL-13 from Dental Pulp Cells

INTRODUCTION Cryopreservation preserves periodontal ligament cells but has a lower success rate with dental pulp cells (DPCs) because it causes inflammation. There are 2 well-known cryopreservation methods that reduce inflammation, slow freezing and rapid freezing, but the effects of the 2 methods on inflammation are not well-established. The purpose of this study was to compare the effects of the 2 different cryopreservation methods on CCL-13 induction from DPCs by using microarrays, real-time polymerase chain reaction (PCR), Western blotting, enzyme-linked immunosorbent assay, and confocal laser scanning microscopy (CLSM). METHODS In this study, the concentration of cryoprotectant was fixed, and the methods compared differed with respect to freezing speed. Initially we screened the DPCs of cryopreserved teeth with expression microarrays, and CCL-13 was identified as a differentially expressed gene involved in generalized inflammation. We then compared the expression of CCL-13 after exposing teeth to the 2 cryopreservation methods by using real-time PCR, Western blot, enzyme-linked immunosorbent assay, and CLSM. RESULTS Expression of CCL-13 was up-regulated significantly only in the rapid freezing group, except in measurements made by real-time PCR. CLSM analysis also confirmed this up-regulation visually. CONCLUSIONS Rapid freezing increased the expression of CCL-13 in DPCs compared with slow freezing. Understanding the inflammatory effect of cryopreservation should help to establish an optimal cryoprofile to minimize inflammation of DPCs and reduce the need for endodontic treatment.

In vitro characterization of human dental pulp stem cells isolated by three different methods

Objectives In this study, we characterized human dental pulp cells (HDPCs) obtained by different culture methods to establish the most suitable methodology for dental tissue engineering and regenerative endodontic applications. Materials and Methods HDPCs were isolated by the outgrowth method (HDPCs-OG), the enzymatic digestion method (collagenase/dispase/trypsin, HDPCs-ED), or the combination of both methods (HDPCs-Combined). The expression of mesenchymal stem cell markers (CD105, CD90, and CD73) was investigated. In vitro differentiation capacities of HDPCs into adipogenic, osteogenic, and chondrogenic lineages were compared. Differentiation markers were analyzed by quantitative reverse-transcription polymerase chain reaction (RT-PCR) and western blotting. Results Our data indicated that whole HDPCs-ED, HPDCs-OG, and HDPCs-Combined could be differentiated into adipogenic, chrondrogenic, and osteogenic cell types. However, we found that the methods for isolating and culturing HDPCs influence the differentiation capacities of cells. HDPCs-OG and HDPCs-ED were preferably differentiated into adipogenic and osteogenic cells, respectively. Differentiation markers shown by RT-PCR and western blotting analysis were mostly upregulated in the treated groups compared with the control groups. Conclusions Our findings confirmed that cell populations formed by two different culture methods and the combined culture method exhibited different properties. The results of this study could provide an insight into regenerative endodontic treatment using HDPCs.