Purification and refolding of the recombinant subunit B protein of the Aggregatibacter actinomycetemcomitans cytolethal distending toxin / 대한치주과학회지

PURPOSE: Aggregatibacter actinomycetemcomitans is associated with localized aggressive periodontitis. It produces cytolethal distending toxin (CDT), which induces cell cycle arrest in the G2/M phase. The CDT holotoxin is composed of CdtA, CdtB, and CdtC. CdtB has structural homology to human DNase I and is an active component of the CDT complex acting as a DNase. In particular, the pattern homology seen in the CdtB subunit has been associated with specific DNase I residues involved in enzyme catalysis, DNA binding, and metal ion binding. So, to study the functions and regulation of recombinant CdtB, we made up a quantity of functional recombinant CdtB and tested it in relation to the metal ion effect. MATERIALS AND METHODS: We constructed the pET28a-cdtB plasmid from A. actinomycetemcomitans Y4 by genomic DNA PCR and expressed it in the BL21 (DE3) Escherichia coli system. We obtained the functional recombinant CdtB by the refolding system using the dialysis method and then analyzed the DNase activity and investigated the metal ion effect from plasmid digestion. RESULTS: The recombinant CdtB subunit was expressed as the inclusion bodies. We were able to obtain functional recombinant CdtB subunit using refolding system. We confirmed that our refolded recombinant CdtB had DNase activity and was influenced by the metal ions Mg2+ and Ca2+. CONCLUSION: We suggest that the factors influencing recombinant CdtB may contribute to CDT associated diseases, such as periodontitis, endocarditic, meningitis, and osteomyelitis.

Human Periodontal Ligament Fibroblasts Support the Osteoclastogenesis of RAW264.7 Cells / 대한치주과학회지

The fibroblasts are the principal cells in the periodontal ligament of periodontium. As the periodontal ligament fibroblasts (PDLF) show similar phenotype with osteoblasts, the PDLF are thought to play an important role in alveolar bone remodeling. Cell-to-cell contacted signaling is crucial for osteoclast formation. Recently it has been reported that PDLF enhance the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The aims of this study were to clarify the mechanism of PDLF-induced osteoclastogenesis and 2) whether we can use preosteoclast cell line instead of primary hematopoietic preosteoclast cells for studying the mechanism of PDLF-induced osteoclastogenesis. Osteoclastic differentiation of mouse macrophage cell line RAW264.7 was compared with that of mouse bone marrow-derived M-CSF dependent cell (MDBM), a well-known hematopoietic preosteoclast model, by examining, 1) osteoclast-specific gene expression such as calcitonin receptor, M-CSF receptor (c-fms), cathepsin K, receptor activator nuclear factor kappa B (RANK), 2) generation of TRAP(+) multinucleated cells (MNCs), and 3) generation of resorption pit on the OAASTM plate. RAW264.7 cultured in the medium containing of soluble osteoclast differentiation factor (sODF) showed similar phenotype with MDBM-derived osteoclasts, those are mRNA expression pattern of osteoclast-specific genes, TRAP(+) MNCs generation, and bone resorbing activity. Formation of resorption pits by osteoclastic MNCs differentiated from sODF-treated RAW264.7, was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for ODF, to the sODF-containing culture medium. The effects of PDLF on differentiation of RAW264.7 into the TRAP(+) multinucleated osteoclast-like cells were examined using coculture system. PDLF were fixed with paraformaldehyde, followed by coculture with RAW264.7, which induced formation of TRAP(+) MNCs in the absence of additional treatment of sODF. When compared with untreated and fixed PDLF (fPDLF), IL-1beta-treated, or lipopolysaccharide- treated and then fixed PDLF showed two-fold increase in the supporting activity of osteoclastogenesis from RAW264.7 coculture system. There were no TRAP(+) MNCs formation in coculture system of RAW264.7 with PDLF of no fixation. These findings suggested that we can replace the primary hematopoietic preosteoclasts for RAW264.7 cell line for studying the mechanism of PDLF-induced osteoclastogenesis, and we hypothesize that PDLF control osteoclastogenesis through ODF expression which might be enhanced by inflammatory signals.

A comparative study of periodontal conditions around mesially tipped molars by a tipping degree / 대한치주과학회지

The aim of the present study was to investigate the distribution of periodontally hopeless teeth in patients who had not been treated. Five hundred nineteen teeth of 163 patients who had visited at the Department of Periodontology, Chonbuk National University Hospital during the period from January 1999 to July 2001 were selected as a periodontally hopeless tooth. Selection criteria were as follows: 1) probing pocket depth more than 8mm, 2) tooth mobility of degree III, 3) radiographic bone loss more than 75%. The contralateral tooth to a hopeless tooth (experimental tooth) was designated as a control tooth. Between the experimental and control tooth, differences in probing pocket depth, tooth mobility and amount of radiographic bone loss were examined as well as correlation. The results showed that hopeless teeth were most frequently found in the maxillary first molar position and least frequently in the maxillary canine position. Differences between experimental and control teeth were 2mm in probing depth, 1 degree in tooth mobility, and 20% in amount of bone loss (p<0.01). The periodontal conditions between the experimental and control teeth showed correlation in general, however, statistical significances were found in posterior teeth positions. Within limitations of this study, it can be concluded that local factors may play a role in deterioration of periodontal disease along with symmetrical character of periodontal disease.

Relationship between Interdental Papilla Existence & Distance from Interdental Alveolar Crest to Contact Point in the Posterior Dentition of Korean adults / 대한치주과학회지

The anatomic structure around interproximal area plays an important role not only in the natural teeth, but also in the implant. The loss of papilla can lead to cosmetic deformity, phonetic problem, food impaction on the anterior dentition, and masticatory problem, food impaction and proximal caries on the posterior dentition. The purpose of this study was to evaluate the relationship between interdental papilla existence and distance from contact point to alveolar crest in Korean posteior dentition. 45 Korean adult patients(31males, 14 females) participated in this study. Measurements were carreid out total 126 interproximal areas, 18 first premolar, 31 second premolar, 40 first molar, and 37 second molar areas. Papilla index was recorded as suggested by Jemt. Distance between contact point and alveolar crest measrued by Florida probeR, after flap elevation. Each distance was measured 10 times by every 0.1mm unit. The results showed that the mean Papilla index 1.37 and mean distance between contact point and alveolar crest was 7.44mm. The correlation between the Papilla index and distance was high negative correlation(Pearson correlation=-0.47), and it was statistically significant(P=0.000) When the distance between contact point and alveolar crest was 5mm, the loss of papilla was appeared almost in half cases. When the distance was 6mm, the papilla loss was present 95%, when 7mm, the papilla loss was 100%.

The Scanning Electron Microscopic study on the effect during subgingival calculus removal using Nd:YAG laser / 대한치주과학회지

The purpose of this study was to evaluate in vitro the effects during subgingival calculus removal using Nd:YAG laser. The study group was consisted of 30 teeth with advanced periodontal disease extracted before the start of periodontal therapy. The specimens were divided into 8 different groups : 1) untreated control 2) scaling and root planing only 3) laser treated using 150mJ/pulse, 1sec, 5sec, contact mode 4) laser treated using 200mJ/pulse, 5sec, contact mode 5) laser treated using 150mJ/pulse, 1sec, non-contact mode 6) laser treated using 200mJ/pulse, 5sec, non-contact mode 7) laser treated using 150mJ/pulse, 1sec, contact mode with water irrigation 8) laser treated using 200mJ/pulse, 5sec, contact mode with water irrigation. All specimens were prepared for evaluation by scanning electron microscopy(SEM). Specimens from Group 2 exhibited a smear layer of scale like texture with parallel instrument tracks resulting from curet use. Specimens treated by contact mode, Group 3 and 4 featured surface changes not observed in controls such as charring, randomly distributed pitting and crater formation, and melting down of the tooth material and calculus. Specimens treated by noncontact mode, Group 5 and 6 featured similar surface changes observed in contact mode. However, the differences between contact and non-contact groups not significant. Specimens treated by contact mode with water irrigation, Group 7 and 8 featured slight surface change compared to other groups. The results suggested that Nd: YAG laser did not completely remove the subgingival calculus but was possible the application as adjunctive method.