• International Journal of Oral Biology
• /
• v.46 no.3
• /
• pp.127-133
• /
• 2021

We previously showed that γ-glutamyltranspeptidase (GGT), an enzyme involved in glutathione metabolism, in Bacillus subtilis acts as a virulence factor for osteoclastogenesis via the RANKL-dependent pathway. Hence, it can be hypothesized that GGT of periodontopathic bacteria acts as a virulence factor in bone destruction. Because Fusobacterium nucleatum, which is a periodontopathic pathogen, has GGT with a primary structure similar to that of B. subtilis GGT (37.7% identify), the bone-resorbing activity of F. nucleatum GGT was examined here. Recombinant GGT (rGGT) of F. nucleatum was expressed in Escherichia coli and purified using the His tag of rGGT. F. nucleatum rGGT (Fn rGGT) was expressed as a precursor of GGT, and then processed to a heavy subunit and a light subunit, which is characteristic of general GGTs, including the human and B. subtilis enzymes. Osteoclastogenesis was achieved in a co-culture system of mouse calvaria-derived osteoblasts and bone marrow cells. Fn rGGT induced osteoclastogenesis to a level similar to that of B. subtilis rGGT; furthermore, osteoclastogenesis was induced in a dose-dependent manner. These results suggest that F. nucleatum GGT possesses a virulent bone-resorbing activity, which could play an important role in the pathogenesis of periodontitis.

• Archives of Craniofacial Surgery
• /
• v.24 no.6
• /
• pp.251-259
• /
• 2023

Background: Periosteum-mediated bone regeneration (PMBR) is a recognized method for mandibular reconstruction. Despite its unpredictable nature and the limited degree to which it is understood, it does not share the concerns of developmental changes to donor and recipient tissues that other treatment options do. The definitive role of the periosteum in bone regeneration in any mammal remains largely unexplored. The purpose of this study was to identify the genetic determinants of PMBR in mammals through a systematic review. Methods: Our search methodology was designed in accordance with the PRISMA (Preferred Reporting Items for Systematic Review and Meta-Analysis) guidelines. We conducted a quality assessment of each publication, and evaluated the differences in gene expression between days 7 and 15. Results: A total of four studies satisfied the inclusion criteria. The subjects and tissues examined in these studies were Wistar rat calvaria in two studies, mini-pigs in one study, and calves and mice in one study. Three out of the four studies achieved the necessary quality score of ≥ 3. Gene expression analysis showed increased activity of genes responsible for angiogenesis, cytokine activities, and immune-inflammatory responses on day 7. Additionally, genes related to skeletal development and signaling pathways were upregulated on day 15. Conclusions: The results suggest that skeletal morphogenesis is regulated by genes associated with skeletal development, and the gene expression patterns of PMBR may be characterized by specific pathways.

• Journal of Microbiology
• /
• v.45 no.6
• /
• pp.566-571
• /
• 2007

Bacterial lipopolysaccharide (LPS) is a potent stimulator of bone resorption in periodontitis. Co-culture systems of mouse calvaria-derived osteoblasts and bone marrow-derived preosteoclasts were used as an in vitro osteoclast differentiation. This study revealed that co-cultures using ddY or ICR mouse strain responded differently to LPS while responded equally to $1{\alpha},25(OH)_2D_3$. Thus, the different response to LPS indicates dissimilarity of two mouse stains in their capacity for generating osteoclasts while the two mouse strains share the similarity in response to $1{\alpha},25(OH)_2D_3$. To identify which cells between osteoblasts and preosteoclasts in the co-culture are responsible for the dissimilarity, the reciprocal co-cultures were performed between ddY and ICR mouse strains. The treatment of $1,25(OH)_2D_3$ to ddY/ICR (osteoblasts from ddY/preosteoclasts from ICR) and ICR/ddY reciprocal co-cultures also showed the similarity. In case of LPS treatment, the results of ddY/ICR were similar to ddY/ddY and the results of the other reciprocal co-culture, ICR/ddY combination, were consistent with those of ICR/ICR. It suggests that the dissimilarity between the two mouse strains may resident in osteoblasts but not in preosteoclasts. Therefore, the osteoblast is responsible for mouse strain-dependent osteoclastogenesis in response to LPS. Although mouse models will continue to provide insights into molecular mechanisms of osteoclastogenesis, caution should be exercised when using different mouse strains, especially ddY and ICR strains as models for osteoclast differentiation.

• Molecules and Cells
• /
• v.37 no.10
• /
• pp.747-752
• /
• 2014

Protein kinase C (PKC) family members phosphorylate a wide variety of protein targets and are known to be involved in diverse cellular signaling pathways. However, the role of PKC in receptor activator of NF-${\kappa}B$ ligand (RANKL) signaling has remained elusive. We now demonstrate that $PKC{\beta}$ acts as a positive regulator which inactivates glycogen synthase kinase-$3{\beta}$ (GSK-$3{\beta}$) and promotes NFATc1 induction during RANKL-induced osteoclastogenesis. Among PKCs, $PKC{\beta}$ expression is increased by RANKL. Pharmacological inhibition of $PKC{\beta}$ decreased the formation of osteoclasts which was caused by the inhibition of NFATc1 induction. Importantly, the phosphorylation of GSK-$3{\beta}$ was decreased by $PKC{\beta}$ inhibition. Likewise, down-regulation of $PKC{\beta}$ by RNA interference suppressed osteoclast differentiation, NFATc1 induction, and GSK-$3{\beta}$ phosphorylation. The administration of PKC inhibitor to the RANKL-injected mouse calvaria efficiently protected RANKL-induced bone destruction. Thus, the $PKC{\beta}$ pathway, leading to GSK-$3{\beta}$ inactivation and NFATc1 induction, has a key role in the differentiation of osteoclasts. Our results also provide a further rationale for $PKC{\beta}$'s therapeutic targeting to treat inflammation-related bone diseases.

• Journal of Periodontal and Implant Science
• /
• v.37 no.3
• /
• pp.553-562
• /
• 2007

Osteoblasts regulate osteoclastogenesis by production of various cytokines. Aggregatibacter(A) ac-tinomycetemcomitans is one of periodontopathogens which invades gingival tissue. Therefore, clarifying the effect of alive A. actinomycetemcomitans on osteoblasts is important to understand the mechanism of alveolar bone resorption in periodontitis. We investigated induction of osteoclastogenesis-inducing cytokines, adherence, and invasion by A. actinomycetemcomitans in osteoblasts. Osteoblasts were isolated from mouse calvaria and expression of cytokines was determined by RT-PCR. When the ratio of the number of A. actinomycetemcomtians to the number of osteoblasts was 10:1, 50:1 and 100:1, RANKL mRNA expression was increased. A. actinomycetemcomitans also increased expression of macrophage inflammatory protein (MIP) -1${\alpha}$, interleukin (IL)-1${\beta}$, and tumor necrosis factor (TNF)-${\alpha}$. A. actinomycetemcomitans attached to and invaded osteoblasts at ratio of 1000:1. These results suggest that A. actinomycetemcomitans increases osteoclastogenesis-inducing ability of osteoblasts by stimulating the expression of RANKL, MIP-1${\alpha}$,IL-1${\beta}$, and TNF-${\alpha}$ and that invasion of A. actinomycetemcomitans provides a means by which the bacteria escape from immune system and antibiotic therapy.

• Journal of Periodontal and Implant Science
• /
• v.37 no.3
• /
• pp.543-551
• /
• 2007

When clinicians faced with an insufficient volume of supporting bone on ideally esthetic and bio-mechanical position for dental implantation, guided bone regeneration(GBR) was indicated. Although GBR has wide application at clinic, proper time of membrane removal remains qustionable in using non-resorbable membrane, such as non-expanded polytetrafluoroethylene(PTFE), The aim of this study was to compare the effect of maintenance period of PTFE membrane on bone regeneration in rabbit calvarial defects. Eight adult New Zealand white female rabbits were used in this study. Four defects were surgically made in their calvaria. Using a trephine bur, 4 'through and through' defects were created and classified into 3 groups, which were consisted of control group(no graft), experimental group 1(autogenous bone)and experimental group 2(deproteinized bovine bone; $OCS-B^{(R)}$). The defects were covered with PTFE membrane($Cytoplast^{(R)}$). Membranes were removed after 1, 2, 4 and 8 weeks post-GBR procedure in 2 rabbits repectively, All rabbits were sacrificed after 8 week post-GBR procedure. Specimens were harvested and observed histologically. The results were as follow; 1) The use of graft material and membrane was necessary in GBR procedure. 2) When PTFE membranes were removed early, the most favorable bone regeneration was revealed in experimental group T, followed by experimental group II and control group. 3) On GBR, it is recommended that membrane should maintain for 4 weeks with autogenous graft. As well, the use of xenograft need longer maintenance period than autogenous bone. Further evaluations will be needed, such as histomorphologic research, more species and different kinds of graft materials. And on the basis of these studies, clinical researches would be required.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.38
• /
• pp.14.1-14.6
• /
• 2016

Background: In guided bone regeneration (GBR) technique, many materials have been used for improving biological effectiveness by adding on membranes. The new membrane which was constructed with chitin-fibroin-hydroxyapatite (CNF/HAP) was compared with a collagen membrane (Bio-$Gide^{(R)}$) by means of micro-computed tomography. Methods: Fifty-four rats were used in this study. A critical-sized (8 mm) bony defect was created in the calvaria with a trephine bur. The CNF/HAP membrane was prepared by thermally induced phase separation. In the experimental group (n = 18), the CNF/HAP membrane was used to cover the bony defect, and in the control group (n = 18), a resorbable collagen membrane (Bio-$Gide^{(R)}$) was used. In the negative control group (n = 18), no membrane was used. In each group, six animals were euthanized at 2, 4, and 8 weeks after surgery. The specimens were analyzed using micro-CT. Results: Bone volume (BV) and bone mineral density (BMD) of the new bone showed significant difference between the negative control group and membrane groups (P < 0.05). However, between two membranes, the difference was not significant. Conclusions: The CNF/HAP membrane has significant effect on the new bone formation and has the potential to be applied for guided bone regeneration.

• Journal of Korean Neurosurgical Society
• /
• v.47 no.3
• /
• pp.169-173
• /
• 2010

Objective : The normal anatomic relationships characteristic of the pituitary stalk area were previously thought to involve only one location. The purpose of this study was to re-evaluate the anatomic location of the pituitary stalk and possible varying locations in relation to the tuberculum sellae and dorsum sellae using morphometric evaluation and anatomic dissection of human cadaveric specimens. The surgical implications of the variations are discussed. Methods : The calvaria were removed via routine autopsy dissections, and the brains were removed from the skull while preserving the pituitary stalk. The diaphragma sellae, tuberculum sellae, and the location of the pituitary stalk were examined in 60 human cadaveric heads obtained from fresh adult cadavers. Empty sellae were excluded. Results : The openings of the diaphragma sellae averaged $6.62{\pm}1.606mm$ (range, 3-9 mm). The distance between the tuberculum sellae and the posterior part of the pituitary stalk was 1 to 8 mm. The upper face of the diaphragma sellae appeared flat in 26 (43%), concave in 24 (40%), and convex in 6 cases (10%), with a prominent tuberculum sellae in 4 cases (7%). The location of the chiasm was normal in 47 cases (78%), with a prefixed chiasm in 3 cases (5%) and a postfixed chiasm (17%) in the 10 cases. Four cadaver specimens had prominent tuberculum sellae and other parameters were not evaluated. Conclusion : When opening the chiasmatic cistern, neurosurgeons should be aware about the relationship between the pituitary stalk and the surrounding structures to prevent inadvertent injury to the pituitary stalk.

• Journal of Periodontal and Implant Science
• /
• v.34 no.4
• /
• pp.711-722
• /
• 2004

The purpose of this study is to evaluate the regenerated bone histollogically using titanium reinforced ePTFE(TR-ePTFE) membrane and to investigate cell occlusiveness, wound stabilization and tissue integration of TR-ePTFE membrane. Adult male rabbits (mean BW 2kg) and TR9W (W.L.Gore&Associate.INC,USA) were used in this study. Intramarrow penetration defects were surgically created with round carbide bur(HP long #6) on calvaria of rabbits. TR-ePTFE membrane was applied to defect. Then guided bone regeneration was carried out using TR-ePTFE membrane and resorbable suture. At 2,4,8,12 weeks after the surgery, animals were sacrificed. Nondecalcified specimens were processed for histologic analysis. The result and conclusion of this study were as follows: 1. TR-ePTFE membrane had good ability of biocompatibility and cell occlusiveness. 2. space making for guided bone regenerayion was good at TR-ePTFE membrane. 3. Tissue integration was not good at TR-ePTFE membrane. So, wound stabilization was not good. 4. At 8 weeks, 12 weeks after GBR procedure, bone formation was seen. From the above results, TR-ePTFE membrane fixed tightiy on alveolar bone might be recommended for the early bone formation.

• Journal of Periodontal and Implant Science
• /
• v.34 no.2
• /
• pp.393-410
• /
• 2004

The purpose of this study was to investigate effect of enamel matrix derivative on guided bone regeneration with intramarrow penetration in rabbits. Eight adult male rabbits (mean BW 2Kg) were used in this study. Intramarrow penetration defects were surgically created with round carbide bur(HP long #6) on calvaria of rabbits. Defects were assigned to the control group grafted with mixture of the same quantity of demineralized freeze-dried bone allograft and deproteinized bovine bone mineral. Then, guided bone regeneration was carried out using resorbable membrane and suture. Enamel matrix derivative applied to defects was assigned to the test group. And treated as same manners as the control group. At 1, 2, 3 and 8 weeks after the surgery, animals were sacrificed, specimens were obtained and stained with Hematoxylin-Eosin for light microscopic evaluation. The results of this study were as follows : 1. At 1, 2 and 3 weeks, no differences were observed between the control group and the test group in the aspect of bone formation around bone graft. 2. Proliferation of blood capillary was faster in the test group than in the control group. 3. Bone regeneration in intramarrow penetration was faster in the test group than in the control group. 4. At 8 weeks, new osteoid tissue formation around bone graft was more prominent in the test group than in the control group. From the above results, enamel matrix derivative might be considered as the osteopromotion material and effective in the guided bone regeneration with intramarrow penetration.