• Journal of Periodontal and Implant Science
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• v.37 no.1
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• pp.23-33
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• 2007

파골세포에 의한 골흡수는 1) 혈관을 통한 파골세포 전구세포의 골표면 이동 및 2) 골표면에서 파골세포 전구세포로부터 파골세포 분화 두 단계를 거쳐 일어난다. Stromal cell derived factor $(SDF)-1{\alpha}$ 는 파골세포 전구세포의 화학주성인자이며 matrix metalloproteinase (MMP)-9는 파골세포 전구세포의 이동에 관여하는 단백 분해효소이다. 파골세포 전구세포의 골표면 이동에 있어서 LPS의 역할을 규명하기 위하여 E. coli 및 Actinobacillus actinomycetecomitans LPS의 1) 파골세포 전구세포 유도능, 2) LPS에 의한 파골세포 전구세포의 이동에 있어서 MMP 및 $SDF-1{\alpha}$ 의 관련성을 평가하였다. LPS에 의한 차골세포 전구세포의 RAW 세포의 이동은 matrigel 또는 type I collagen을 도포한 transwell을 이용하여 평가하였으며 MMP-9 및 $SDF-1{\alpha}$ 의 발현은 RT=PCR 또는 ELISA로 평가하였다. 각 세균의 LPS는 matrigel 또는 type I collagen을 통한 파골세포 전구세포의 이동을 증가시켰다. MMP 억제제는 각 세균의 LPS에 의한 파골세포 전구세포의 이동을 억제하였다. LPS는 파골세포 전구세포의 MMP-9의 발현을 증가시켰다. 각 세균의 LPS는 마우스 두개골에서 분리한 조골세포의 $SDF-1{\alpha}$ 의 발현을 증가시켰다. $SDF-1{\alpha}$ 을 함유한 LPS 처리 조골세포 배양상층액은 파골세포 전구세포의 이동을 증가시켰으며 anti $SDF-1{\alpha}$ Ab는 LPS처리 세포 배양상층액에 의한 파골세포 전구세포의 이동을 억제하였다. 이들 결과는 LPS가 파골세포 전구세포에서는 MMP-9을 조골세포에서는 $SDF-1{\alpha}$ 의 발현을 증가시켜 파골세포 전구세포의 이동을 촉진 시킬 수 있음을 시사한다.

• Journal of Periodontal and Implant Science
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• v.37 no.sup2
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• pp.297-310
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• 2007

치조골 흡수는 파골세포와 matrix metalloproteinases (MMPs)에 의한 골의 무기질과 유기질의 파괴로 일어나는 과정이다. 세균성 산물, 주로 내독소는 치은조직 내에서 염증세포의 유주, 사이토카인 생산, 조직파괴 효소 분비 및 파골세포 활성 등의 국소 면역반응을 유도한다. A. actinomycetemcomitans는 급진성 치주염의 원인 균주중 하나로 그 내독소는 치조골의 흡수와 관련된다. MMP-13은 세균성 산물이나 염증성 사이토카인의 자극에 의해 분비되며, 최근의 연구 결과들은 MMP-13이 치주질환의 진행과 골 흡수 과정에서 일정한 역할을 담당하는 것으로 보고하고 있으나, A. actinomycetemcomitans 내독소와 MMP-13과의 관련성에 대한 연구는 미미하다. 이에 이번 연구에서는 A. actinomycetemcomitans 내독소에 의한 MMP-13의 발현과 파골세포 형성을 세포배양을 통하여 관찰하였고, 백서 구개부 치은에 A. actinomycetemcomitans 내독소를 주입하여 흡수가 진행되고 있는 치조골에서 파골세포의 분화와 MMP-13의 발현을 TRAP 염색, 면역조직화학적 방법 등을 통해 관찰하여 다음과 같은 결과를 얻었다. MMP-13 mPNA의 발현은 A. actinomycetemcomitans 내독소 (1ug/ml)로 24시간 자극한 마우스 치주인대 섬유모세포에서 생리식염수로 자극한 세포에 비하여 약 2.6배 증가하였으며 마우스 대식세포에서는 TRAP 양성 세포가 대조군보다 더 많이 나타났다. A. actinomycetemcomitans 내독소를 주입한 백서 치주조직에서는 대조군보다 더 심한 골소실을 보였다. TRAP-양성 다핵 파골세포 유사세포는 치주염군과 대조군 모두 치조골에서 관찰되었다. TRAP-양성 다핵 파골세포 유사세포는 치주염군에서 대조군보다 유의하게 많은 숫자가 관찰되었으며, 치주염군에서 대조군보다 유의하게 많은 숫자가 관찰되었다. MMP-13 면역양성 반응은 치주염군에서 거친 골연을 갖는 치조골상에 배열된 조골세포와 그 인접한 치주인대에서 관찰되었으며 대조군에서는 MMP-13 면역 양성 반응이 치조골 표면에서만 일부 관찰되었다. 이상의 결과는 A. actinomycetemcomitans 내독속가 MMP-13의 발현을 증가시키며 파골세포의 활성을 통하여 치조골의 흡수를 유도함을 시사한다. 또한 A. actinomycetemcomitans 내독소 투여에 의한 실험적 모델은 백서에서 중등도의 골 소실을 동반한 만성 치주염 모델로 향후 치주질환 치료제의 효과를 평가하는데 유용하게 사용될 수 있으리라 기대된다.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.317-324
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• 1996

Osteoclast has been known as a primary responsible cell for the bone resorption. The activation of osteoclast, therefore, may be the key event in the regulation of bone growth and remodeling. Various factors were reported to have influence on the resorbing activity of osteoclast in organ culture. Among those factors, transforming growth factor-$\beta$ (TGF-$\beta$) has been known to have a profound effect on bone metabolism. Since a large amount of TGF-$\beta$ presents in bone tissue, it may be important for the understanding the regulatory mechanism of bone resorption to elucidate the effect of TGF-$\beta$ on the osteoclast. We have reported the dlsaggregated chick embryonic osteoclast culture as an useful assay method for determining the resorption activity of osteoclast. In this culture, we found that TGF-$\beta$ significantly enhaced the osteoclastic bone resorption activity. We also found that the timulatory effect seemed to be an indirect one that is mediated by other cells. As nordihydroguaiaretic acid significantly inhibited the TGF-$\beta$1-induced osteoclastic bone resorption, we suggest that the lipoxygenase derivative of arachidonic acid may participate in the action of TGF-$\beta$ as a paracrine or an autocrine mediator.

• Journal of Life Science
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• v.25 no.2
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• pp.223-230
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• 2015

Regulator of calcineurin 1 (RCAN1) is an endogenous calcineurin inhibitor that plays an important role in the pathogenesis of diseases related to the calcineurin-NFATc1 signaling pathway. The RCAN1-4 isoform is subject to NFATc1-dependent regulation. During receptor activator of nuclear factor kappa-B ligand (RANKL)-stimulated osteoclastogenesis, the calcineurin-NFATc1 pathway is critical. Because there is little information available on the role of RCAN1 in osteoclast differentiation, this study investigated whether changes in RCAN1 expression are related to the calcineurin-NFATc1 pathway and osteoclast differentiation. Mouse bone marrow monocytes (BMMs) were treated with 50 ng/ml of RANKL and M-CSF. Expression levels of NFATc1, calcineurin, and RCAN1 isoforms were determined using RT-PCR and Western blotting. Osteoclast differentiation was examined using tartrate-resistent acid phosphatase (TRAP) staining. To evaluate the effect of RCAN1 overexpression on osteoclastogenesis, cells were transfected with a mouse RCAN1-4 cDNA plasmid. After RANKL stimulation of BMMs, expression of NFATc1 and RCAN1 was increased at the mRNA and protein level, while calcineurin expression was unchanged. When the RCAN1-4 gene construct was transfected, the expression of RCAN1 protein was not increased despite several-fold increases in RCAN1-4 mRNA expression. Regardless of RANKL stimulation, over-expression of RCAN1-4 tended to reduce NFATc1 expression and knock-down of RCAN1 increase it. While BMMs transfected with the RCAN1-4 vector were differentiated into distinct osteoclasts, their phenotypes did not vary from those of mock controls. These results suggest that RCAN1 has a limited effect on the calcineurin-NFATc1 pathway during RANKL-stimulated osteoclast differentiation.

• Journal of Oral Medicine and Pain
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• v.30 no.3
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• pp.329-336
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• 2005

We examined the effect of PQQ, as a scavenger of superoxide, on osteoclast-like cell formation and on mature osteoclast function. To determine whether PQQ scavenges the superoxide, nitroblue tetrazolium (NBT) staining, which is a method to detect superoxide, was performed on HD-11 cells which are a chick myelomonocytic cell line having tartrate-resistant acid phosphatase (TRAP) activity in response to 1,25-dihydroxyvitamin $D_3\;[1,25(OH)_2D_3]$. Histochemical study of TRAP was also performed on HD-11 cells. PQQ inhibited the TRAP-positive multinucleated cell formation of chicken bone marrow cells was also examined. The addition of 20 ${\mu}M$ PQQ inhibited the formation of TRAP-positive multinucleated cell. When chicken osteoclasts were cultured on dentin slices, treatment of 20 ${\mu}M$ PQQ resulted in a significant decrease in dentin resorption by osteoclasts in terms of total resorption area and number of resorption pits. The present data suggest that PQQ, possibly as a scavenger of superoxide ion, inhibits the osteoclastic differentiation and bone resorption.

• Journal of Korean society of Dental Hygiene
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• v.23 no.4
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• pp.219-226
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• 2023

Objectives: This study aimed to investigate the effects of rhubarb extract on osteoclast differentiation in bone marrow-derived macrophages (BMMs). Osteoclasts are vital for bone resorption and remodeling. Osteoclast dysregulation can contribute to various bone-related disorders that directly affect oral health. Rhubarb, a medicinal plant with anti-inflammatory properties, has been shown to modulate bone metabolism. Methods: BMMs were isolated from the femurs and tibias of 5-week-old C57BL/6 mice and cultured in the presence of mouse macrophage colony-stimulating factor (M-CSF) for 3 days. Subsequently, BMMs were treated with M-CSF and receptor activator of nuclear factor-κB ligand (RANKL) to induce osteoclast differentiation. Results: Rhubarb extract effectively suppressed osteoclast differentiation in BMMs. Furthermore, rhubarb extract inhibited the mRNA expression of tartrate-resistant acid phosphatase (TRAP) and cathepsin K (CTSK), which are essential for osteoclastogenesis. Moreover, it inhibited the RANKL-induced expression of nuclear factor of activated T cell c1 (NFATc1), a crucial transcription factor in osteoclast differentiation. Conclusions: These results suggest that rhubarb extract promotes oral health by inhibiting osteoclastogenesis in BMMs. Thus, rhubarb extract shows promise as a therapeutic agent for bone-related disorders that directly affect oral health, particularly those associated with abnormal osteoclast activity. Further research and exploration of the underlying mechanisms are warranted to fully understand their potential clinical applications.

• The korean journal of orthodontics
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• v.34 no.6 s.107
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• pp.506-513
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• 2004

Osteoclast action is necessary for alveolar bone remodeling in orthodontic tooth movement. The nervous system has also been reported to be associated with bone remodeling. This study was aimed to investigate the changes of osteoclasts in the periodontal ligament (PDL) space after surgical resection of the inferior alveolar nerve (IAN). Experimental rats were divided into young and adult groups. A surgical resection procedure of the IAN was carried out in the left side of the mandible and a sham operation in the right side of the mandible. The number of osteoclasts on the bundle bone surface and the resorption activity of the osteoclasts were histomorphometrically measured. The changes in distribution of substance P (SP) immunoreactive (IR) nerve fiber were evaluated in the PDL and pulp. SP-IR nerve fiber was depleted in both the PDL and pulp of the IAN resection side in both groups, which confirmed the resection of IAN to be successfully conducted. The number of osteoclasts in the IAN resection side was significantly reduced in both the young and adult groups (p<0.01 and p<0.05), whereas the resorption activity of osteoclasts did not show any significant difference between the IAN resection side and the sham operation side in both groups (p>0.05 and p<0.05). The adult group showed that the number of osteoclasts reduced significantly (p<0.01) and the resorption activity didn't change in comparison with the young group (p>0.05). These results suggest that surgical resection of the IAN and aging reduce the population of the recruited osteoclasts within the PDL, but don't affect on the osteoclastic resorption activity.

• Journal of Periodontal and Implant Science
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• v.25 no.3
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• pp.659-667
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• 1995

파골세포는 조혈기관 단핵의 세포로부터 생성되어 골 홉수에 중요한 역할올 담당하며, 지질다당류는 그람음성균의 세포벽을 이루는 성분으로서 치주질환시 치조골 홉수에 관여한다고 알려져 왔다. 활성화된 림프구, 대식세포와 단핵세포로부터 생성되는 당단백질인 인터페론 감마는 파골세포에 의한 골홉수를 억제한다고 밝혀졌다. 이 연구 논문의 목적은 지질다당류와 인터페론 감마가 닭 골수의 미분화세포가 파골양세포로 전환되는데 어떠한 영향올 주는지를 알아보기 위함이다. 16${\sim}$18 일째의 닭의 배 (chick embryo) 에서 경골을 분리하고 횡절개하여 혈청없는 M-199 배양액에 보관했다. 이것을 9${\mu}m$ filter로 여과시켜서 이미 분화된 파골세포와 기타 다른 분화 세포를 분리했다. 여기에서 파골세포의 전구세포를 얻어 LPS와 IFN-${\gamma}$를 단독 또는 복합처리 하고나서 4일 후에 tartrate resistant acid phosphatase (TRAP) Stain을 시행하고 TRAP 양성이며 핵이 세개 이상인 다핵의 세포형성을 관찰하여 세포를 계수하여 다음과 같은 결과를 얻었다. 1. 닭에서 분리해낸 미분화세포에 0.1. 0.5. 1.0 ${\mu}/ml$ 의 LPS 농도를 처리하고 1 주일간 배양한 결과. 0.1 ${\mu}/ml$ 의 농도에서는 대조군에 비해 TRAP 양성인 파골양세포가 증가하는 경향을 보였으며, 반면에 LPS는 0.5 와 1.0 ${\mu}/ml$ 의 농도에서 세포독성을 보였다.(P<0.05) 2. IFN-${\gamma}$는 50. 500U/ml 의 농도에서 대조군에 비해 TRAP 양성인 파골양세포의 수가 감소하는 경향올 보였다 .3. INF-${\gamma}$는 LPS 에의해 유도된 TRAP 양성인 파골양세포의 형성을 감소시켰고 특히 . 250.500U/ml 의 농도에서 유의 성 있는 감소를 보였다. 위의 결과로부터 LPS는 닭의 골수세포로부터 파골양세포의 형성을 증가시키며 IFN-${\gamma}$는 LPS에의해 유도된 파골양세포수를 감소시킨다는 결론을 얻었다.

• Journal of Oral Medicine and Pain
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• v.32 no.2
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• pp.129-135
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• 2007

Ecdysteroids are known as insect molting hormone. At the same time, ecdysteroids and plant ecdysteroids (phytoecdysteorids) reveal beneficial effects on mammal. The present study was undertaken to determine the possible cellular mechanism of action of phytoecdysteroids in bone metabolism. The effects on the osteoblasts were determined by measuring cell proliferation, alkaline phosphatase (ALP) activity, and gelatinase activity. The effects on the osteoclasts were investigated by measuring tartrate-resistant acid phosphatase (TRAP)(+) multinucleated cells (MNCs) formation after culturing osteoclast precursors. Phytoecdysteroid treatment showed a increase in ALP activity of osteoblasts. Phytoecdysteroid increased the activity of gelatinase. In addition, phytoecdysteroid decreased the osteoclast generation induced by macrophage-colony stimulating factor (M-CSF) and receptor activator of NF-kB ligand (RANKL) in (M-CSF)-dependent bone marrow macrophage (MDBM) cell cultures. Taken these results, phytoecdysteroid may be a regulatory protein within the bone marrow microenvironment.

• Korean Journal of Food Science and Technology
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• v.42 no.5
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• pp.637-642
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• 2010

Bone is continuously remodeled by osteoblasts and osteoclasts. We investigated the effects of medicinal herbs, which act on bone metabolism. Fifteen kinds of medicinal herb extracts were screened for bone formation activity with osteoblastic cells, and MC3T3-E1 and bone resorption were screened with osteoclasts derived from mouse bone marrow macrophages. Among these samples, Actinidia polygama, Eucommia ulmoides Oliv., Schizonepeta tenuifolia, Sorbus commixta, and Zingiber officinale Rosc. extracts showed strong bone-forming activity accompanied with osteoblast proliferation and alkaline phosphatase activity. In addition, these extracts decreased tartrate-resistant acid phosphatase activity against osteoclast differentiation. The results indicate that these medicinal herb extracts can potentially prevent bone-related diseases such as osteoporosis by increasing osteoblast differentiation and reducing osteoclast activity.