Byung-Ock Kim

Application of rpoB and Zinc Protease Gene for Use in Molecular Discrimination of Fusobacterium nucleatum Subspecies

ABSTRACT Fusobacterium nucleatum is classified into five subspecies that inhabit the human oral cavity (F. nucleatum subsp. nucleatum, F. nucleatum subsp. polymorphum, F. nucleatum subsp. fusiforme, F. nucleatum subsp. vincentii, and F. nucleatum subsp. animalis) based on several phenotypic characteristics and DNA-DNA hybridization patterns. However, the methods for detecting or discriminating the clinical isolates of F. nucleatum at the subspecies levels are laborious, expensive, and time-consuming. Therefore, in this study, the nucleotide sequences of the RNA polymerase β-subunit gene (rpoB) and zinc protease gene were analyzed to discriminate the subspecies of F. nucleatum. The partial sequences of rpoB (approximately 2,419 bp), the zinc protease gene (878 bp), and 16S rRNA genes (approximately 1,500 bp) of the type strains of five subspecies, 28 clinical isolates of F. nucleatum, and 10 strains of F. periodonticum (as a control group) were determined and analyzed. The phylogenetic data showed that the rpoB and zinc protease gene sequences clearly delineated the subspecies of F. nucleatum and provided higher resolution than the 16S rRNA gene sequences in this respect. According to the phylogenetic analysis of rpoB and the zinc protease gene, F. nucleatum subsp. vincentii and F. nucleatum subsp. fusiforme might be classified into a single subspecies. Five clinical isolates could be delineated as a new subspecies of F. nucleatum. The results suggest that rpoB and the zinc protease gene are efficient targets for the discrimination and taxonomic analysis of the subspecies of F. nucleatum.

Clinical evaluation of a collagen matrix to enhance the width of keratinized gingiva around dental implants

Purpose The purpose of this study was to evaluate the effect of collagen matrix with apically positioned flap (APF) on the width of keratinized gingiva, comparing to the results of APF only and APF combined with free gingival graft (FGG) at the second implant surgery. Methods Nine patients were selected from those who had received treatments at the Department of Periodontics, Chosun University Dental Hospital, Gwangju, Korea. We performed APF, APF combined with FGG, and APF combined with collagen matrix coverage respectively. Clinical evaluation of keratinized gingival was performed by measuring the distance from the gingival crest to the mucogingival junction at the mid-buccal point, using a periodontal probe before and after the surgery. Results The ratio of an increase was 0.3, 0.6, and 0.6 for the three subjects in the APF cases, 3, 5, and 7 for the three in the APF combined with FGG case, and 1.5, 0.5, and 3 for the three in the APF combined with collagen matrix coverage case. Conclusions This study suggests that the collagen matrix when used as a soft tissue substitute with the aim of increasing the width of keratinized tissue or mucosa, was as effective and predictable as the FGG.

Temporal Induction of Secretory Leukocyte Protease Inhibitor (SLPI) in Odontoblasts by Lipopolysaccharide and Wound Infection

INTRODUCTION The secretory leukocyte protease inhibitor (SLPI) is a bacterial lipopolysaccharide (LPS)-induced product of macrophages that antagonizes the LPS-induced activation of a number of proinflammatory signaling factors. From our previous experiments, it was found that SLPI was expressed slightly in odontoblast-like cells (MDPC-23). Therefore, these experiments were designed to determine the function of SLPI in MDPC-23 and odontoblasts during the inflammatory response caused by infections and wounds. METHODS MDPC-23 cells were exposed to 100 ng/mL Escherichia coli LPS, and artificial wounds were induced in the right first molar of the maxillary of rats. In addition, a morphological change in the MDPC-23 cells was observed after LPS treatment. MDPC-23 cells were transfected transiently with the nuclear factor kappa-B (NF-kappaB) promoter binding vector. RESULTS The level of SLPI expression increased strongly 30 minutes after the LPS treatment. Scanning electron microscopy revealed many extensions of the cytoplasmic processes after LPS stimulation. SLPI was expressed along the dentinal tubules and odontoblasts layer in rat teeth after an artificial wound. SLPI also inhibited the LPS-induced activation of NF-kappaB in MDPC-23. CONCLUSIONS We report for the first time that SLPI is expressed temporally in infected odontoblasts and may participate in the anti-inflammatory response through NF-kappaB signaling in odontoblast-like cells.

Correlation of immunohistochemical characteristics of the craniomandibular joint with the degree of mandibular lengthening in rabbits

PURPOSE This study examined immunohistochemical changes in the craniomandibular joints of rabbits after distraction osteogenesis following mandibular corticotomy. MATERIALS AND METHODS The experimental animals (n = 8) were divided into 3 groups that underwent 2, 3.5, and 5 mm of unilateral distraction osteogenesis (groups 1, 2, and 3, respectively). After corticotomy of the left mandibular body and a 7-day healing period, a second operation was performed to expose the device. Distraction was then performed at the rate of 0.5 mm/d. A 14-day consolidation period was allowed after the distraction was complete. Changes in cartilage, osteoblast activity, and osteoclast activity were then examined. RESULTS The differentiation and proliferation of cartilage increased in groups 1 and 2, were highest in group 2, and decreased in group 3. Group 2 also showed the greatest increase in the width of the hypertrophic chondrocyte layer. Relative to the control group, osteoclast activity was only somewhat higher in groups 1 and 2 but was significantly higher in group 3. Osteoblast activity was significantly higher in groups 1 and 2 than in the control group. However, the osteoblast activity in group 3 was slightly lower than that in group 2. At the time of unilateral mandibular distraction, no degenerative changes of the temporomandibular joint were observed in groups 1 or 2, but bone resorption was observed in group 3. CONCLUSIONS The unilateral mandibular distraction of 2 or 3.5 mm was acceptable in that no degenerative changes of the temporomandibular joint were observed on either the distraction or the nondistraction sides. Five millimeters of distraction might be beyond physiologic limits.

Expression of UNCL during development of periodontal tissue and response of periodontal ligament fibroblasts to mechanical stress in vivo and in vitro

Mutations in two genes, uncoordinated (unc) and uncoordinated-like (uncl), lead to a failure of mechanotransduction in Drosophila. UNCL, the human homolog of unc and uncl, is preferentially expressed in periodontal ligament (PDL) fibroblasts compared with gingival fibroblasts. However, the precise role of UNCL in the PDL remains unclear. The aim of the present study has been to examine whether mechanical stimuli modulate the expression of UNCL in the human PDL in vivo and in vitro and to examine the roles of UNCL in the development, regeneration, and repair of the PDL. We have investigated the expression pattern of UNCL during the development of periodontal tissue and the response of PDL fibroblasts to mechanical stress in vivo and in vitro. The expression of UNCL mRNA and protein increases with PDL fibroblast differentiation from the confluent to multilayer stage but slightly decreases on mineralized nodule formation. UNCL has also been localized in ameloblasts and adjacent cells, differentiating cementoblasts, and osteoblasts of the developing tooth. Strong distinct UNCL expression has further been observed in the differentiating cementoblasts of the tooth periodontium at the site of tension after orthodontic tooth movement. Application of cyclic mechanical stress on PDL fibroblasts increases the expression of UNCL mRNA. These results indicate that UNCL plays important roles in the development, differentiation, and maintenance of periodontal tissues and also suggest a potential role of UNCL in the mechanotransduction of PDL fibroblasts.

A new method for rapid screening of bacterial species- or subspecies-specific DNA probes

A simple assay for the rapid screening of bacterial species- or subspecies-specific DNA probes for the random cloning method is presented, involving the use of genomic DNAs as probes and recombinant plasmid DNAs containing genomic DNA digested with HindIII as targets. The optimal amount of target DNAs and the concentration of digoxigenin-labeled genomic DNA probes were 20 ng and 100 ng ml(-1) (or 10 ng and 200 ng ml(-1)), respectively. The method was applied to the development of Fusobacterium nucleatum subspecies-specific probes. Our results showed that four out of 96 probes were F. nucleatum subspecies-specific, which was confirmed by Southern blot analysis. Our results indicate that the new method can be used for the rapid screening of species- or subspecies-specific probes.

Effects of decortication in the treatment of bone defect around particulate dentin-coated implants: an experimental pilot study.

OBJECTIVES The purpose of this study was to evaluate the effects of decortication during bone grafting in defect areas surrounding particulate dentin-coated implants. STUDY DESIGN Six dogs were randomly assigned to 3 groups, and each group was further divided into 2 subgroups. The subgroups consisted of dogs that received particulate dentin-coated implants after 4 or 8 weeks. The defects were treated as follows: control group, unfilled defect; experimental group 1, defect filled with Tutoplast without decortication; experimental group 2, defect filled with Tutoplast with decortication. Histologic sections and histomorphometric analysis were obtained 4 and 8 weeks after surgery. RESULTS Compared to the 4-week subgroup, statistically significant new bone formation was observed in the 8-week subgroup. In the 4-week subgroup, the area of new bone formation was larger in the group that underwent cortical bone perforation with bone grafting; however, no significant difference was detected within the 8-week subgroup. CONCLUSION According to these results, decortication increases early bone formation after implant placement.

Clinical evaluation of a biphasic calcium phosphate grafting material in the treatment of human periodontal intrabony defects

Purpose The aim of this study was to compare the clinical outcome of open flap debridement (OFD) with a biphasic calcium phosphate (BCP) graft to that of OFD without BCP graft for the treatment of intrabony periodontal defects (IBDs). Methods The study included 25 subjects that had at least one intrabony defect of 2- or 3-wall morphology and an intrabony component≥4 mm as detected radiographically. Subjects were randomly assigned to treatment with (BCP group, n=14) or without BCP (OFD group, n=11). Clinical parameters were recorded at baseline and 6 months after surgery and included the plaque index, gingival index, probing depth (PD), clinical attachment level (CAL), and gingival recession (REC). A stringent plaque control regimen was enforced for all of the patients during the 6-month observation period. Results In all of the treatment groups, significant PD reductions and CAL gains occurred during the study period (P<0.01). At 6 months, patients in the BCP group exhibited a mean PD reduction of 3.7±1.2 mm and a mean CAL gain of 3.0±1.1 mm compared to the baseline. Corresponding values for the patients treated with OFD were 2.5±0.8 mm and 1.4±1.0 mm, respectively. Compared to OFD group, the additional CAL gain was significantly greater in the patients in BCP group (P=0.028). The additional PD reduction was significant for the BCP group (P=0.048). The REC showed a significant increase in both groups, and the amount of recession was significantly smaller in the BCP group than OFD group (P=0.023). In radiographic evaluation, the height of the bone fill in the BCP group was significantly greater than OFD group. Conclusions The clinical benefits of BCP found in this study indicate that BCP may be an appropriate alternative to conventional graft materials.

Guided Tissue Regeneration of the Mixture of Human Tooth-Ash and Plaster of Paris in Dogs

This study evaluated the efficacy of periodontal tissue regeneration using guided tissue regeneration (GTR) with a mixture of tooth ash and plaster as a bone substitute material in the treatment of class II furcation defects in dogs. GTR was performed at the site of a surgically formed mandibular premolar bifurcation area in dogs. Four adult dogs were used in the animal study. The animals were in a good periodontal state without any systemic disease. A resorbable membrane (Bio-Gide®, Swiss) and bone graft material using a mixture of tooth ash and plaster were used to examine the biological activity. The defect in the control group was filled a Bio-Gide® only. For the experimental group, the defects were filled with a mixture of tooth ash and plaster and covered by a Bio-Gide®. Oral rinsing with 0.12% chlorhexidine was performed twice a day for 2 weeks. In control group, new bone formation was observed adjacent to the preexisting bone. Soft tissue ingrowth was also noted. In experimental group, new lamellar type trabecular bone formation as well as centum and PDL regeneration was observed in the base of the infrabony pocket. More bone regeneration would be expected when GTR is performed using a mixture of human tooth-ash and plaster of Paris as a bone graft material.

Orthodontic treatment for maxillary anterior pathologic tooth migration by periodontitis using clear aligner

Purpose Pathologic tooth migration (PTM) is a tooth displacement which is derived from imbalance of tooth retention force and is dominantly found out in maxillary anterior area. PTM in maxillary anterior area was tried to corrected with periodontal treatment and a clear aligner in this study and the result was evaluated clinically and radiographically. Methods For the treatment of a patient with chronic periodontal disease accompanied by maxillary anterior pathologic tooth migration, clear aligner was applied to move teeth after a series of case-related periodontal therapy. Clinically, probing depth, gingival recession, clinical attachment level and mobility were measured pre- and post-treatment, and radiographic examination was performed as well. Results Clinically, we found the decrease of the probing depth, gingival recession, clinical attachment level and mobility. And we could also acknowledge the reduction of vertical and horizontal dimension on infrabony defect radiographically. However, it is still controversial if there was an actual bone filling. Conclusions Clear aligner is an effective appliance to move teeth since it costs little in terms of expense and time. In addition, it wraps whole crowns, providing advantages to deal with crowding, spacing, and size of arch. In short, clear aligner could be a useful treatment option for PTM patient, since it provides decreased probing depth, gingival recession, clinical attachment level, mobility and esthetical restoration.