Sang-Min Lim

Microbial Analysis in Primary and Persistent Endodontic Infections by Using Pyrosequencing

INTRODUCTION The aim of this study was to investigate the bacterial community profile of intracanal microbiota in primary and persistent endodontic infections associated with asymptomatic chronic apical periodontitis by using GS-FLX Titanium pyrosequencing. The null hypothesis was that there is no difference in diversity of overall bacterial community profiles between primary and persistent infections. METHODS Pyrosequencing analysis from 10 untreated and 8 root-filled samples was conducted. RESULTS Analysis from 18 samples yielded total of 124,767 16S rRNA gene sequences (with a mean of 6932 reads per sample) that were taxonomically assigned into 803 operational taxonomic units (3% distinction), 148 genera, and 10 phyla including unclassified. Bacteroidetes was the most abundant phylum in both primary and persistent infections. There were no significant differences in bacterial diversity between the 2 infection groups (P > .05). The bacterial community profile that was based on dendrogram showed that bacterial population in both infections was not significantly different in their structure and composition (P > .05). CONCLUSIONS The present pyrosequencing study demonstrates that persistent infections have as diverse bacterial community as primary infections.

A comparison of nickel-titanium rotary instruments manufactured using different methods and cross-sectional areas: ability to resist cyclic fatigue

OBJECTIVE This study examined the effect of the manufacturing methods (ground, electropolished, and twisted) and the cross-sectional area (CSA) of nickel-titanium (NiTi) rotary instruments on their cyclic fatigue resistance. STUDY DESIGN A total of 80 NiTi rotary instruments (ISO 25/.06 taper) from 4 brands (K3, ProFile, RaCe, and TF) were rotated in a simulated root canal with pecking motion until fracture. The number of cycles to failure (NCF) was calculated. The CSA at 3 mm from the tip of new instruments of each brand was calculated. The correlation between the CSA and NCF was evaluated. All fractured surfaces were analyzed using a scanning electron microscope to determine the fracture mode. RESULTS The TF instruments were the most resistant to fatigue failure. The resistance to cyclic failure increased with decreasing CSA. All fractured surfaces showed the coexistence of ductile and brittle properties. CONCLUSION The CSA had a significant effect on the fatigue resistance of NiTi rotary instruments.

Comparison of Alternative Image Reformatting Techniques in Micro–Computed Tomography and Tooth Clearing for Detailed Canal Morphology

INTRODUCTION Micro-computed tomography (MCT) shows detailed root canal morphology that is not seen with traditional tooth clearing. However, alternative image reformatting techniques in MCT involving 2-dimensional (2D) minimum intensity projection (MinIP) and 3-dimensional (3D) volume-rendering reconstruction have not been directly compared with clearing. The aim was to compare alternative image reformatting techniques in MCT with tooth clearing on the mesiobuccal (MB) root of maxillary first molars. METHODS Eighteen maxillary first molar MB roots were scanned, and 2D MinIP and 3D volume-rendered images were reconstructed. Subsequently, the same MB roots were processed by traditional tooth clearing. Images from 2D, 3D, 2D + 3D, and clearing techniques were assessed by 4 endodontists to classify canal configuration and to identify fine anatomic structures such as accessory canals, intercanal communications, and loops. RESULTS All image reformatting techniques in MCT showed detailed configurations and numerous fine structures, such that none were classified as simple type I or II canals; several were classified as types III and IV according to Weine classification or types IV, V, and VI according to Vertucci; and most were nonclassifiable because of their complexity. The clearing images showed less detail, few fine structures, and numerous type I canals. Classification of canal configuration was in 100% intraobserver agreement for all 18 roots visualized by any of the image reformatting techniques in MCT but for only 4 roots (22.2%) classified according to Weine and 6 (33.3%) classified according to Vertucci, when using the clearing technique. CONCLUSIONS The combination of 2D MinIP and 3D volume-rendered images showed the most detailed canal morphology and fine anatomic structures.

Bacterial entombment by intratubular mineralization following orthograde mineral trioxide aggregate obturation: a scanning electron microscopy study

The time domain entombment of bacteria by intratubular mineralization following orthograde canal obturation with mineral trioxide aggregate (MTA) was studied by scanning electron microscopy (SEM). Single-rooted human premolars (n=60) were instrumented to an apical size #50/0.06 using ProFile and treated as follows: Group 1 (n=10) was filled with phosphate buffered saline (PBS); Group 2 (n=10) was incubated with Enterococcus faecalis for 3 weeks, and then filled with PBS; Group 3 (n=20) was obturated orthograde with a paste of OrthoMTA (BioMTA, Seoul, Korea) and PBS; and Group 4 (n=20) was incubated with E. faecalis for 3 weeks and then obturated with OrthoMTA–PBS paste. Following their treatments, the coronal openings were sealed with PBS-soaked cotton and intermediate restorative material (IRM), and the roots were then stored in PBS for 1, 2, 4, 8 or 16 weeks. After each incubation period, the roots were split and their dentin/MTA interfaces examined in both longitudinal and horizontal directions by SEM. There appeared to be an increase in intratubular mineralization over time in the OrthoMTA-filled roots (Groups 3 and 4). Furthermore, there was a gradual entombment of bacteria within the dentinal tubules in the E. faecalis inoculated MTA-filled roots (Group 4). Therefore, the orthograde obturation of root canals with OrthoMTA mixed with PBS may create a favorable environment for bacterial entombment by intratubular mineralization.

Antifungal effects of synthetic human β-defensin 3-C15 peptide

Objectives The purpose of this ex vivo study was to compare the antifungal activity of a synthetic peptide consisting of 15 amino acids at the C-terminus of human β-defensin 3 (HBD3-C15) with calcium hydroxide (CH) and Nystatin (Nys) against Candida albicans (C. albicans) biofilm. Materials and Methods C. albicans were grown on cover glass bottom dishes or human dentin disks for 48 hr, and then treated with HBD3-C15 (0, 12.5, 25, 50, 100, 150, 200, and 300 µg/mL), CH (100 µg/mL), and Nys (20 µg/mL) for 7 days at 37℃. On cover glass, live and dead cells in the biomass were measured by the FilmTracer Biofilm viability assay, and observed by confocal laser scanning microscopy (CLSM). On dentin, normal, diminished and ruptured cells were observed by field-emission scanning electron microscopy (FE-SEM). The results were subjected to a two-tailed t-test, a one way analysis variance and a post hoc test at a significance level of p = 0.05. Results C. albicans survival on dentin was inhibited by HBD3-C15 in a dose-dependent manner. There were fewer aggregations of C. albicans in the groups of Nys and HBD3-C15 (≥ 100 µg/mL). CLSM showed C. albicans survival was reduced by HBD3-C15 in a dose dependent manner. Nys and HBD3-C15 (≥ 100 µg/mL) showed significant fungicidal activity compared to CH group (p < 0.05). Conclusions Synthetic HBD3-C15 peptide (≥ 100 µg/mL) and Nys exhibited significantly higher antifungal activity than CH against C. albicans by inhibiting cell survival and biofilm.

Microbial profile of asymptomatic and symptomatic teeth with primary endodontic infections by pyrosequencing

Objectives: The purpose of this in vivo study was to investigate the microbial diversity in symptomatic and asymptomatic canals with primary endodontic infections by using GS FLX Titanium pyrosequencing. Materials and Methods: Sequencing was performed on 6 teeth (symptomatic, n = 3; asymptomatic, n = 3) with primary endodontic infections. Amplicons from hypervariable region of the small-subunit ribosomal RNA gene were generated by polymerized chain reaction (PCR), and sequenced by means of the GS FLX Titanium pyrosequencing. Results: On average, 10,639 and 45,455 16S rRNA sequences for asymptomatic and symptomatic teeth were obtained, respectively. Based on Ribosomal Database Project Classifier analysis, pyrosequencing iden- tified the 141 bacterial genera in 13 phyla. The vast majority of sequences belonged to one of the seven phyla: Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, Proteo bacteria, Spirochetes, and Synergistetes . In genus level, Pyramidobacter, Streptococcus, and Leptotrichia constituted about 50% of microbial profile in asymptomatic teeth, whereas Neisseria, Propionibacterium, and Tessaracoccus were frequently found in symptomatic teeth (69%). Grouping the sequences in operational taxonomic units (3%) yielded 450 and 1,997 species level phylotypes in asymptomatic and symptomatic teeth, respectively. The total bacteria counts were significantly higher in symptomatic teeth than that of asymptomatic teeth ( p < 0.05). Conclusions: GS FLX Titanium pyrosequencing could reveal a previously unidentified high bacterial diver- sity in primary endodontic infections. (J Kor Acad Cons Dent 2011;36(6):498-505.) ABSTRACT

Autotransplantation of mesiodens for missing maxillary lateral incisor with cone‐beam CT‐fabricated model and orthodontics

AIM Autotransplantation is a viable treatment option for a missing tooth when there is a suitable donor, especially in adolescents with remaining facial growth. This report presents the aesthetic restoration of a missing maxillary lateral incisor through orthodontic treatment and autotransplantation of a mesiodens using a CBCT-fabricated rapid-prototyping model. SUMMARY A 14-year-old male patient with a congenitally missing maxillary lateral incisor was referred from the Department of Orthodontics. The teeth were moved orthodontically to regain space for the missing lateral incisor and to close the space of the mesiodens after transplantation. A replica of the donor tooth was fabricated from a cone-beam computed tomography scan through a rapid-prototyping machine before autotransplantation surgery. The model was used to create a socket for the graft tooth, thereby shortening the extra-oral time and minimizing the damage to the root surface. After transplantation and orthodontic tooth movement, the mesiodens was finally restored with an aesthetic laminate restoration. Over 3 years, the aesthetics remained excellent, and the transplant functioned normally without any signs or symptoms of root resorption. KEY LEARNING POINT Missing anterior teeth may be replaced through a combination of orthodontics, autotransplantation with a rapid-prototyping model and prosthodontic restoration, in growing patients.

Evaluation of three obturation techniques in the apical third of mandibular first molar mesial root canals using micro-computed tomography

Background/purpose Recent studies have demonstrated a high incidence of isthmuses in mandibular first molar mesial roots, and intratubular mineralization following mineral trioxide aggregate obturation. This study assessed the filling quality of three obturation techniques in the apical 5 mm of mandibular first molar mesial root canals. Materials and methods Sixty extracted human mandibular first molar mesial roots with two separate canals that had interconnecting isthmuses, were prepared to an apical size of 40/0.06. They were allocated to three groups of 20 roots for obturation by either cold lateral compaction (CLC) or the continuous wave of condensation (CW) that used gutta-percha and AH Plus sealer, or by an orthograde canal obturation using OrthoMTA. The obturated roots were scanned by micro-computed tomography and assessed for the volumetric ratio (%) of gutta-percha, sealer, and OrthoMTA within the main canals or isthmuses in the apical 5 mm area. Measurements were analyzed statistically for differences among three obturation techniques. Results In the main canals, filled volume ratios were not significantly different among groups. Within isthmuses, the filled volume ratio for CLC was lower than in CW (P = 0.025) or OrthoMTA (P = 0.002). In isthmuses, the gutta-percha volume ratio in CLC was lower than in CW (P = 0.005), although the sealer volume ratio was higher than in CW (P = 0.049). Conclusion CLC demonstrated lower filling densities in isthmuses in the apical region than either CW or OrthoMTA. Orthograde MTA obturation showed comparable filling quality to gutta-percha with sealer.

Clinical assessment of whitening efficacy and safety of in-office tooth whitening system containing 15% hydrogen peroxide with or without light activation

Objectives: This clinical study evaluated the effect of light activation on the whitening efficacy and safety of in-office bleaching system containing 15% hydrogen peroxide gel. Materials and Methods: Thirty-three volunteers were randomly treated with (n = 17, experimental group) or without light activation (n = 16, control group), using Zoom2 white gel (15% , Discus Dental) for a total treatment time of 45 min. Visual and instrumental color measurements were obtained using Vitapan Classical shade guide and Shadepilot (DeguDent) at screening test, after bleaching, and 1 month and 3 month after bleaching. Data were analyzed using t-test, repeated measure ANOVA, and chi-squared test. Results: Zoom2 white gel produced significant shade changes in both experimental and control group when pre-treatment shade was compared with that after bleaching. However, shade difference between two groups was not statistically significant (p > 0.05). Tooth shade relapse was not detected at 3 months after bleaching. The incidence of transient tooth sensitivity was 39.4%, with being no differences between two groups. Conclusions: The application of light activation with Zoom2 white gel system neither achieved additional whitening effects nor showed more detrimental influences.

Analysis of para-chloroaniline after chemical interaction between alexidine and sodium hypochlorite using mass spectrometry: A preliminary study

The purposes of this study were firstly to investigate the any formation of precipitate after interaction between ALX and NaOCL and secondarily to analyze the PCA formation by using time of flight secondary ion mass (TOF-SIM) spectrometry. Mass spectrometry analysis was performed for the mixture of 0.5% ALX and 5.25% NaOCl. As controls, 2.5% CHX with 5.25% NaOCl and 1% PCA solutions were used. Any for- mation of precipitates in 10 tested solutions was evaluated by naked eye. Results of mass spectrum showed that the typical peak of PCA was not detected in mixed solution of ALX and NaOCl, whereas CHX/NaOCl mixture showed the same peak that found in the PCA spectrum. Precipitate formation was only observed in CHX/NaOCL mixture. The present TOF-SIM spectrometry results indicated that ALX can be a useful root canal irrigant combined with NaOCl during canal instrumentation. Further study is necessary to confirm the antimicrobial effect of ALX against endodontic pathogen before its clinical application as an endodontic irrigant.