• Journal of Korean Dental Science
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• v.8 no.2
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• pp.74-81
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• 2015

Purpose: In osteoclast differentiation, actin-rich membrane protrusions play a crucial role in cell adhesion. Odontogenic ameloblast-associated protein (Odam) contributes to cell adhesion by inducing actin rearrangement. Odam-mediated RhoA activity may play a significant role in multinucleation of osteoclasts. However, the precise function of Odam in osteoclast cell adhesion and differentiation remains largely unknown. Here, we identify a critical role for Odam in inducing osteoclast adhesion and differentiation. Materials and Methods: The expression of Odam in osteoclasts was evaluated by immunohistochemistry. Primary mouse bone marrow and RAW264.7 cells were used to test the cell adhesion and actin ring formation induced by Odam. Result: Odam was expressed in osteoclasts around alveolar bone. Odam transfection induced actin filament rearrangement and cell adhesion compared with the control or collagen groups. Overexpression of Odam promoted actin stress fiber remodeling and cell adhesion, resulting in increased osteoclast fusion. Conclusion: These results suggest that Odam expression in primary mouse osteoclasts and RAW264.7 cells promotes their adhesion, resulting in the induction of osteoclast differentiation.

• Toxicological Research
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• v.13 no.1_2
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• pp.55-60
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• 1997

Gamma irradiation was applied to Angelicae gigantis radix and Aloe to improve their hygienic quality. The effective dose of irradiation was 7 kGy in Angelicae gigantis radlx and 5 kGy in Aloe for the sterilization of all contaminated microorganisms tested. After 8 months of storage at room temperature, no growth of microorganisms was observed in the irradiated products. The safety of these products were evaluated by Salmonella typhimurium reversion assay and in vivo micronucleus assay using mouse bone marrow cells. They were negative in the bacterial reversion assay with S. typhimurium TA 98, TA100, TA1535 and TA1537. In the in vivo mouse micronucleus assay, they did not show any clastogenic effect at all doses tested. These results indicate that the gamma irradiation of Angelicae gigantis radix at 12 kGy and of Aloe at 10 kGy have no genotoxic effects under these experimental conditions.

• Toxicological Research
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• v.14 no.2
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• pp.211-216
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• 1998

In order to evaluate the mutagenic potential of SDK(skin decontamination kit) produced by Agency for Defense Development(ADD), were performed Salmonella typhimurium reversion assay, chromosomal aberration test on chinese hamster ovarian cells and in vivo micronucleus assay using mouse bone marrow cells according to the established regulation of Korean Food and Drug Administration. In the reverse mutation test using Salmonella typhimurium TA98, TA100, TA1535 and TA1537 did not in-crease the number of revertant at any of the concentration tested in this study. SDK did not increase the number of cells having structural or numerical chromosome aberration in cytogenetic test. In mouse micronucleus test, no significant increase in the occurrence oj micro nucleated polychromatic erythrocytes were observed in ICR male mice intraperitoneally administered with SDK. These results indicate that SDK has no mutagenic effects under these experimental conditions.

• Journal of Environmental Health Sciences
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• v.16 no.1
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• pp.55-65
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• 1990

This study was carried out to investigate the effects on sister chromatid exchanges (SCEs) and chromosome aberrations in PHA or LPS stimulated mouse spleen and bone marrow lymphocytes after an acute whole body irradiation. Frequencies of sister chromatid exchanges were significantly increased with the increased dose(from zero to 400tad) but there was no differences between B-cell and T-cell. By times, the maximum induced SCE levels was observed at 12 hours after irradiation and then returned to base level at one day in 100rad group and three day in 400rad group. There was a significant difference in chromosome aberration with increasing exposure. X-ray irradiated chromosome aberration was long lived relative to SCE. This results show that counting the incidence of SCE may not provide a sensitive system for detecting X-ray exposure.

• YAKHAK HOEJI
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• v.43 no.5
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• pp.614-622
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• 1999

The standard operation procedure of mouse lymphoma L5178Y $tk^{+/-}-3.7.2C$ gene mutation assay (MOLY) has been regarded as a sensitive in vitro mammalian cell gene mutation assay that is capable of detecting clastogens as well as mutagens. Using MOLY, one of natural sweetner, stevioside (5mg/ml) and its aglycon, steviol ($340{\;}\mu\textrm{g}/ml$) were evaluated the mutagenicity. Stevioside and steviol did not induce mutagenicity in MOLY. On the other hand, stevioside (250mg/kg, B.W.) and steviol (200mg/kg, B.W.) were also evaluated their ability to induce micronuclei in regenerating hepatocytes and bone marrow cells of ddY mice. From these results, stevioside and steviol did not induce any mutagenic effect both MOLY and in vivo micronucleus test.

• Neonatal Medicine
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• v.15 no.1
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• pp.22-31
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• 2008

Purpose : Bronchopulmonary dysplasia (BPD) is characterized by arrested vascular and alveolar growth in the premature lung. Considering the consequences of arrested lung growth, the idea of administering bone marrow cells to enhance the inborn repair mechanism is promising as this may reduce the morbidity and mortality of BPD. We followed enhanced green fluorescent protein (EGFP)-labeled bone marrow cells (BMC) injected intraperitoneally into non-EGFP mice in order to determine their fate after transplantation. Methods : An angiogenesis inhibitor, SU1498, was injected subcutaneously on day 3 in non-EGFP C57BL/6 newborn mice to create a model of arrested alveolar development. On the following day, $1{\times}10^6$ BMCs isolated from major histocompatibility complex (MHC)- matched syngenic EGFP mice were injected intraperitoneally to non-EGFP BPD mice. Morphometric analysis, immunostaining, and confocal microscopy were performed to determine the fate of EGFP-positive stem cells in the injured lung. Results : SU1498 injection reduced alveolar surface area and mean alveolar volume in newborn mice. BMC injection resulted in recovery of lung structure comparable to controls. EGFP-positive BMCs were identified in the lungs of the recipient mice after intraperitoneal injection. The injected EGFP cells were co-stained with endothelial and epithelial cells of the developing lung as determined by confocal microscopy. Conclusion : Our results illustrated that EGFP-positive BMCs engrafted and trans-differentiated into epithelial and endothelial cells after intraperitoneal injection in a mouse model of arrested alveolar development.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.5
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• pp.353-359
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• 2010

Introduction: This study examined the effect of cyclosporine A (CsA) on the allogenic cranial bone graft in the mice. Materials and Methods: Twenty eight 12-week-old male ICR mice weighing 40 g were used. The experimental group was injected subcutaneously with CsA (10 mg/kg/day) diluted in Caster oil for 7 days prior to the graft until sacrifice. The control group was injected with the same solution without CsA. Two full-thickness bone defects with a diameter of 3 mm were made with a trephine bur in the parietal bone lateral to the sagittal suture. A calvarial defect of a mouse was grafted with allogenic calvarial bone disc from another mouse. The experimental and control groups were injected with CsA and the solution without CsA in the same manner before surgery, respectively. The mice were sacrificed at 1 week, 2 weeks and 4 weeks after the bone graft, respectively. Results: In the experimental group, fibrous connective tissues and small amounts of inflammatory cells were observed. At 2 weeks after the allograft in the experimental group, new bone formation in fibrous collagenous tissue and around the allogenic bone was noted. At 4 weeks after the allograft, new bone formation was active along and at the periphery of the mature allogenic bone. The proliferation of blood vessels increased in bone marrow. In the control group, fibrous tissues and inflammatory cells were observed around the allogenic bone and existing bone at 1 week. At 2 weeks after the allograft, the proliferation of blood vessels accompanied by inflammatory cells were scattered in the fibrous connective tissues. New bone formation around the allogenic and existing bone could be observed. At 4 weeks after the allograft, inflammatory cells were severely infiltrated around the allogenic bone. Osteoclasts were scattered along the allogenic bone and induced bone resorption. Conclusion: These results suggest that the daily administration of CsA (10 mg/kg/day) induces efficient immunosuppression without serious complications, and this protocol might be useful for the experimental model of allogenic bone grafts.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.1
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• pp.23-26
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• 2010

The vascular changes in periodontal tissues cause local hypoxia which seems to affect the periodontal tissue cells. Abrupt changes in oxygen availability within the periodontium have been suggested to have a regulatory role in alveolar bone remodeling during tooth movement, bone growth or fracture healing. The purpose of this study was to study the effects of hypoxia on formation of osteoclast responsible for bone resorption, in vitro. Primary mouse bone marrow cells were cultured in normoxic (20% $O_2$) and hypoxic (1% $O_2$) conditions and assayed for cellular proliferation. The results obtained were as follows : 1. Reducing oxygen tension increased the formation of multinucleated osteoclasts. 2. Hypoxic stimulus increased the size of mature osteoclasts.

• Food Science and Biotechnology
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• v.14 no.1
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• pp.94-98
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• 2005

Bone resorption surrounding tooth root causes tooth loss in periodontitis patients. Osteoclast has bone resorption activity. Effects of Artemisia asiatica on bone resorption induced by periodontopathogens, Porphyromonas gingivalis and Treponema denticola, were examined using co-culture systems of mouse osteoblasts and bone marrow cells. Addition of A. asiatica ethanol extract to bacterial sonicate abolished bacteria-induced osteoclastogenesis. To determine inhibitory mechanism of A. asiatica against osteoclastogenesis, effects of A. asiatica on expressions of osteoclastogenesis-inducing factors such as receptor activator of NF-${\kappa}B$ ligand (RANKL), prostaglandin $E_2\;(PGE_2)$, interleukin (IL)-1, and tumor necrosis factor (TNF)-${\alpha}$, in osteoblasts were examined. A. asiatica suppressed expressions of RANKL, $PGE_2$, IL-$1{\beta}$, and TNF-${\alpha}$ increased by each bacterial sonicate. These results suggest inhibitory action of A. asiatica against osteoclastogenesis is associated with down-regulations of RANKL, $PGE_2$ IL-$1{\beta}$, and TNF-${\alpha}$ expressions.

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