Deog-Gyu Seo

Heavy Metal Analysis of Ortho MTA and ProRoot MTA

INTRODUCTION Recently, several kinds of mineral trioxide aggregate (MTA)-based products have been introduced in endodontics. Ortho MTA (BioMTA, Seoul, Republic of Korea) is one of those products, which was developed for retrograde filling, perforation repair, orthograde root canal obturation, and direct pulp capping. The inclusion of heavy metals in MTA-based materials is of concern because they come into direct contact with hard and soft tissues. The aim of this study was to investigate and compare the levels of arsenic (As), chromium (Cr), hexavalent chromium (Cr(6+)), and lead (Pb) in Ortho MTA and ProRoot MTA. METHODS One gram of each MTA was digested using a mixture of hydrochloric and nitric acids and filtered. The As, Cr, and Pb in the resulting filtrates were analyzed by inductively coupled plasma-optical emission spectrometry. The level of Cr(6+) was measured by the methods suggested in the Korean Standard L 5221. The results were statistically analyzed using the Mann-Whitney U test. RESULTS The concentration of As in ProRoot MTA was 1.16 ppm, but As was not detected in Ortho MTA. Cr(6+) and Pb were not detected in either MTA. Ortho MTA contained significantly less Cr than ProRoot MTA (P < .05). CONCLUSIONS Ortho MTA and ProRoot MTA meet the ISO specification 9917-1 regarding the safety limits of As and Pb and are safe biomaterials when the purity of As, Cr(6+), and Pb is considered.

Prognostic Factors for Clinical Outcomes According to Time after Direct Pulp Capping

INTRODUCTION Direct pulp capping is a treatment option for teeth with carious-exposed pulp. Because pulp capping studies have exhibited fluctuations in success rates according to different follow-up times, investigating the clinical pulpal survival rate and the potential factors contributing to the survival with respect to time is necessary. METHODS A total of 175 patients treated between November 2007 and August 2010 met the inclusion criteria. During the follow-up, we investigated 7 clinical variables with respect to the survival of the pulp capping treatment: sex, age, maxilla versus mandible, tooth position, capping materials, temporary filling materials, and exposure site. Survival analysis was performed using the Kaplan-Meier method and the Cox proportional hazard regression model. RESULTS The Kaplan-Meier survival curves and log-rank tests revealed that only age, exposure site, and capping material had significant effects on the pulpal survival rate (P< .05). A Cox regression model showed that mineral trioxide aggregate was the sole factor affecting the survival of the treated pulps (P< .05). In the analyses performed separately according to time, there was no conspicuous factor that affected the survival rate before 100 days. However, after 100 days, the type of pulp capping material was the single most important factor influencing the survival rate (P< .05). CONCLUSIONS The results of this study indicated that careful patient selection and the type of pulp capping material should be taken into consideration when performing a pulp capping treatment.

Evaluation of the shear bond strength of resin cement to Y-TZP ceramic after different surface treatments.

The purpose of this study was to evaluate the effect of various surface treatments on the shear bond strength of Y-TZP (Yttria-Tetragonal Zirconia Polycrystal) ceramics with zirconia primer and two different resin cements both containing 10-methacryloyloxydecyl dihydrogen phosphate (MDP). Zirconia blocks (LAVA, 3M ESPE, St. Paul, MN) were polished and assigned to five groups according to the surface treatment: (1) no further treatment (control); (2) airborne abrasion with Al2 O3 particles; (3) Z-PRIME Plus (Bisco, Schaumburg, IL) applied on polished zirconia; (4) Z-PRIME Plus applied on zirconia after airborne abrasion; and (5) tribochemical silica-coating performed with the CoJet system (3M ESPE) followed by application of ESPE®-Sil (3M ESPE). Each group was further divided into one of two resin cements: Panavia F2.0 (Kuraray, Kurashiki, Okayama, Japan) and Clearfil SA Luting (Kuraray). Resin cement placed inside a gel-cap was polymerized on the zirconia surface. Shear bond strength was tested with a universal testing machine at 0.5 mm/min. One-way analysis of variance and paired t-test were done. (p < 0.05), and scanning electron microscope (SEM) images were taken. Zirconia primer applied after airborne abrasion significantly increased the shear bond strength resulting in the highest value for both resin cements. Control groups for both cements showed the weakest value for shear bond strength. No significant differences were found between the shear bond strengths of the individual resin cements applied to zirconia surfaces treated the same way. In conclusion, the combined surface treatment of airborne abrasion followed by a zirconia primer is recommended for zirconia bonding with Panavia F2.0 and Clearfil SA Luting cements.

Identification of Porphyromonas gingivalis lipopolysaccharide-binding proteins in human saliva

Porphyromonas gingivalis causes periodontal diseases and its lipopolysaccharide (LPS) is considered as a major virulence factor responsible for pathogenesis. Since initial recognition of P. gingivalis LPS (Pg.LPS) in the oral cavity might be crucial for the host response, we identified Pg.LPS-binding proteins (Pg.LPS-BPs) using Pg.LPS-immobilized beads and a high-resolution mass spectrometry. LPS purified from P. gingivalis was conjugated onto N-hydroxysuccinimidyl-Sepharose(®) 4 Fast Flow beads. Notably, Pg.LPS-conjugated beads could stimulate Toll-like receptor 2 (TLR2) as determined by a TLR2-depdendent reporter expression system using CHO/CD14/TLR2. In addition, the Pg.LPS-conjugated beads induced the production of inflammatory mediators such as nitric oxide and interferon-gamma-inducible protein-10 in the macrophage cell-line, RAW 264.7. These results imply that Pg.LPS retained its immunological properties during the conjugation process. Then, the Pg.LPS-conjugated beads were mixed with a pool of saliva obtained from nine human subjects to capture Pg.LPS-BPs and molecular identities were determined by LTQ-Orbitrap hybrid fourier transform mass spectrometry. Pg.LPS-BPs captured at high frequencies included alpha-amylase, cystatin, prolactin-inducible protein, lysozyme C, immunoglobulin components, serum albumin, lipocalin-1, and submaxillary gland androgen-regulated protein 3B. These proteins are known to be involved in bacterial adhesion and colonization, anti-microbial functions or modulation of immune responses.

Lipoteichoic acid of Streptococcus mutans interacts with Toll-like receptor 2 through the lipid moiety for induction of inflammatory mediators in murine macrophages

Streptococcus mutans is a pathogenic Gram-positive bacterium that is closely associated with dental caries and subsequent pulpal inflammation. Although lipoteichoic acid (LTA) is considered a major virulence factor of Gram-positive bacteria, little is known about the innate immunity to S. mutans LTA. In this study, we purified LTA from S. mutans (Sm.LTA) through n-butanol extraction, hydrophobic interaction column chromatography, and ion-exchange column chromatography to investigate its immunological properties using murine macrophages. The Sm.LTA preparation had no detectable contamination with endotoxins, proteins, or nucleic acids. Upon exposure to Sm.LTA, the murine macrophage cell-line RAW 264.7 cells produced TNF-α and nitric oxide (NO) in a dose-dependent manner. Sm.LTA preferentially bound to and activated CHO/CD14/TLR2 cells rather than CHO/CD14/TLR4 cells, which are stable transfectants expressing CD14 and TLR2 or CD14 and TLR4, respectively. Sm.LTA could not induce TNF-α or NO production in macrophages derived from TLR2-deficient mice whereas it dose-dependently induced those inflammatory mediators in wild-type macrophages. TLR2-dependent induction of NO by Sm.LTA was also confirmed in RAW 264.7 cells using specific antibodies blocking TLR2. Furthermore, Sm.LTA deacylated by alkaline hydrolysis neither stimulated TLR2 nor induced TNF-α or NO production, suggesting that Sm.LTA lipid moieties are crucial for the immuno-stimulatory activity of Sm.LTA. Unlike Staphylococcus aureus LTA, which has potent immuno-stimulating activity, Sm.LTA showed a modest induction of NO production comparable to LTAs of other oral bacteria Enterococcus faecalis and Lactobacillus plantarum. In conclusion, our results suggest that the Sm.LTA interacts with TLR2 through the lipid moiety for the induction of inflammatory mediators in macrophages.

Differential profiles of salivary proteins with affinity to Streptococcus mutans lipoteichoic acid in caries-free and caries-positive human subjects

Streptococcus mutans is a representative oral pathogen that causes dental caries and pulpal inflammation. Its lipoteichoic acid (Sm.LTA) is known to be an important cell-wall virulence factor involved in bacterial adhesion and induction of inflammation. Since Sm.LTA-binding proteins (Sm.LTA-BPs) might play an important role in pathogenesis and host immunity, we identified the Sm.LTA-BPs in the saliva of caries-free and caries-positive human subjects using Sm.LTA-conjugated beads and LTQ-Orbitrap hybrid Fourier transform mass spectrometry. Sm.LTA was conjugated to N-hydroxysuccinimidyl-Sepharose(®) 4 Fast Flow beads (Sm.LTA-beads). Sm.LTA retained its biological properties during conjugation, as determined by the expression of nitric oxide and interferon-γ-inducible protein 10 in a murine macrophage cell line and activation of Toll-like receptor 2 (TLR2) in CHO/CD14/TLR2 cells. Sm.LTA-BPs were isolated from pooled saliva prepared from 10 caries-free or caries-positive human subjects each, electrophoresed to see their differential expression in each group, and further identified by high-resolution mass spectrometry. A total of 8 and 12 Sm.LTA-BPs were identified with statistical significance in the pooled saliva from the caries-free and caries-positive human subjects, respectively. Unique Sm.LTA-BPs found in caries-free saliva included histone H4, profilin-1 and neutrophil defensin-1, and those in caries-positive saliva included cystatin-C, cystatin-SN, cystatin-S, cystatin-D, lysozyme C, calmodulin-like protein 3 and β-actin. The Sm.LTA-BPs found in both groups were hemoglobin subunits α and β, prolactin-inducible protein, protein S100-A9, and SPLUNC2. Collectively, we identified Sm.LTA-BPs in the saliva of caries-free and caries-positive subjects, which exhibit differential protein profiles.

Alpha-amylase is a human salivary protein with affinity to lipopolysaccharide of Aggregatibacter actinomycetemcomitans

Aggregatibacter actinomycetemcomitans lipopolysaccharide (Aa.LPS) is a major virulence factor associated with aggressive periodontitis. Although the recognition of Aa.LPS is potentially initiated by salivary proteins in the oral cavity, Aa.LPS-binding proteins (Aa.LPS-BPs) in saliva are poorly characterized. The purpose of this study was to capture and identify Aa.LPS-BPs in human saliva using a LTQ-Orbitrap hybrid Fourier transform mass spectrometry. Aa.LPS conjugated onto N-hydroxysuccinimidyl-Sepharose(®) 4 Fast Flow beads (Aa.LPS-beads) activated Toll-like receptor 4 and produced nitric oxide and Interferon gamma-inducible protein-10, implying that the conjugation process did not alter the biological properties of Aa.LPS. Aa.LPS-BPs were subsequently isolated from the nine human saliva samples from healthy individuals with the Aa.LPS-beads followed by identification with the mass spectrometry. Aa.LPS-BPs include α-amylase, serum albumin, cystatin, lysozyme C, submaxillary gland androgen-regulated protein 3B, immunoglobulin subunits, polymeric immunoglobulin receptor, deleted in malignant brain tumors 1, prolactin-inducible protein, lipocalin-1, and basic salivary proline-rich protein 2. Specific binding was validated using a pull-down assay with α-amylase which was captured at the highest frequency. Alpha-amylase demonstrated to interfere with the adherence and biofilm formation of A. actinomycetemcomitans. Even heat-inactivated α-amylase showed the interference to the same extent. Conclusively, we identified unique Aa.LPS-BPs that provide useful information to understand bacterial pathogenesis and host innate immunity in the oral cavity.

A biometric study of C‐shaped root canal systems in mandibular second molars using cone‐beam computed tomography

AIM To investigate the configuration of C-shaped canals in mandibular second molars, canal wall thickness and the orientation of the thinnest area at 1-mm intervals from the canal orifice to the apex by using cone-beam computed tomographic (CBCT) images. METHODOLOGY Three-dimensional CBCT images of 92 Korean mandibular second molars having C-shaped root canals were analysed to determine their configuration using a modification of Meltons classification, as well as the thinnest walls and their location. Associations between configuration type and distance from the canal orifice to the apex, as well as associations between the directional orientation of the thinnest root wall and distance from the canal orifice to the apex, were assessed by Fishers exact test. Because serial measurements of minimum wall thicknesses were correlated with individual teeth, a mixed-effects analysis was applied. RESULTS The most common configuration types were Meltons type I in the coronal region and Meltons type III in the apical region. Mean thicknesses of the thinnest root canal walls were 1.39 ± 0.38, 0.85 ± 0.25 and 0.77 ± 0.20 mm in the coronal, middle and apical regions, respectively. The thicker the root canal walls at the orifice region, the greater the decrease in thickness towards the apical region (P < 0.05), with the linguo-central root area being the thinnest. The pattern of decreasing thickness from the orifice to the apex formed a nonlinear cubic curve. CONCLUSIONS The most prevalent configuration types were Meltons type I (coronal region) and type III (apical region). The linguo-central root area was the thinnest in C-shaped root canals of Korean mandibular second molars. These anatomical variations should be considered during surgical or nonsurgical endodontic procedures.

Minimum-intensity projection for in-depth morphology study of mesiobuccal root

OBJECTIVE This micro-computed tomography (MCT) study investigated the utility of thin-slab minimum-intensity projection (TS-MinIP) technique as an adjunct to 3-dimensional (3D) modeling for in-depth morphology study. STUDY DESIGN One hundred one extracted maxillary first molars were scanned for microtomographic analysis (SkyScan). Two-dimensional TS-MinIP and 3D images of mesiobuccal (MB) roots were produced and analyzed to record the number and configurations of the canals, the incidence and location of accessory canals, loop, and intercanal connections, and number of foramina. RESULTS Multiple-canal MB roots were present in 76.2%, and all of the roots had intercanal communications. Weine type III configuration was the most common in the multiple-canal roots. Accessory canals were found in 78.2% of the roots. Configurations that were nonclassifiable were found in 10.9% of the MB roots. CONCLUSIONS MB root canal anatomy was complex, and MinIP may serve as an adjunct to 3D modeling for in-depth morphology study.

Performance of universal adhesives on bonding to leucite-reinforced ceramic.

BackgroundThis study aimed to investigate the microshear bond strength of universal bonding adhesives to leucite-reinforced glass-ceramic.MethodsLeucite-reinforced glass-ceramic blocks were polished and etched with 9.5% hydrofluoric acid for 1 min. The specimens were assigned to one of four groups based on their surface conditioning (n = 16): 1) NC: negative control with no further treatment; 2) SBU: Single Bond Universal (3M ESPE); 3) ABU: ALL-BOND Universal (Bisco); and 4) PC: RelyX Ceramic Primer and Adper Scotchbond Multi-Purpose Adhesive (3M ESPE) as a positive control. RelyX Ultimate resin cement (3M ESPE) was placed on the pretreated ceramic and was light cured. Eight specimens from each group were stored in water for 24 h, and the remaining eight specimens were thermocycled 10,000 times prior to microshear bond strength evaluation. The fractured surfaces were examined by stereomicroscopy and scanning electron microscopy (SEM).ResultsAfter water storage and thermocycling, the microshear bond strength values decreased in the order of PC > SBU and ABU > NC (P < 0.05). Thermocycling significantly reduced the microshear bond strength, regardless of the surface conditioning used (P < 0.05). Cohesive failure in the ceramic and mixed failure in the ceramic and resin cement were observed in the fractured specimens. The percentage of specimens with cohesive failure after 24 h of water storage was: NC (50%), SBU (75%), ABU (75%), and PC (87%). After thermocycling, the percentage of cohesive failure in NC decreased to 25%; however, yet the percentages of the other groups remained the same.ConclusionsAlthough the bond strength between resin and hydrofluoric acid-etched glass ceramic was improved when universal adhesives were used, conventional surface conditioning using a separate silane and adhesive is preferable to a simplified procedure that uses only a universal adhesive for cementation of leucite-reinforced glass-ceramic.