Phytoecdysteroid가 조골세포와 파골세포의 성장과 활성에 미치는 영향

고선일,Ko, Seon-Yle 대한안면통증구강내과학회 2007 Journal of Oral Medicine and Pain Vol.32 No.2

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.

Angiopoietin-2가 조골세포와 파골세포의 성장과 활성에 미치는 영향

고선일,Ko, Seon-Yle 대한안면통증구강내과학회 2006 Journal of Oral Medicine and Pain Vol.31 No.1

The present study was undertaken to determine the possible cellular mechanism of action of angiopoietin-2 in bone metabolism. The effects on the osteoblasts were determined by measuring 1) cell viability, 2) alkaline phosphatase (ALP) activity, 3) gelatinase activity, and 4) nitric oxide production. The effects on the osteoclasts were investigated by measuring 1) tartrate-resistant acid phosphatase (TRAP)(+) multinucleated cells (MNCs) formation, and 2) resorption areas after culturing osteoclast precursors. Angiopoietin-2 treatment showed a significant increase in both the viability and ALP activity of osteoblasts. Angiopoietin-2 increased the activity of gelatinase and nitric oxide production. In addition, angiopoietin-2 decreased the osteoclast generation induced by macrophage-colony stimulating factor (M-CSF) and receptor activator of NF-kB ligand (RANKL), and inhibited osteoclastic activity in (M-CSF)-dependent bone marrow macrophage (MDBM) cell cultures. Taken these results, angiopoietin-2 may be a regulatory protein within the bone marrow microenvironment.

Effect of Pyrroloquinoline Quinone on Osteoclast Generation and Activity

고선일,한동호,김정근,Ko, Seon-Yle,Han, Dong-Ho,Kim, Jung-Keun Korean Academy of Orofacial Pain and Oral Medicine 2005 Journal of Oral Medicine and Pain Vol.30 No.3

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.

Baicalin이 조골세포의 생성 및 활성에 미치는 영향

고선일,Ko, Seon-Yle 대한안면통증구강내과학회 2008 Journal of Oral Medicine and Pain Vol.33 No.2

Baicalin is a flavonoid compound isolated from the medicinal plant Scutellaria baicalensis. It is known to affect multiple biological functions, including of antibacterial, anti-viral, anti-inflammatory and analgesic effects. Baicalin can inhibit nuclear factor-kappaB activation. It has been reported that some flavonoids possess the effects of bone metabolism. The present study was undertaken to determine the possible cellular mechanism of action of baicalin in osteoblasts. The effects on the osteoblast were determined by measuring cell proliferation, cell viability, alkaline phosphatase activity, and osteoprotegerin secretion. Baicalin has no effect on the osteoblastic cell proliferation and cell viability. Baicalin treatment showed increase in alkaline phosphatase activity and osteoprotegerin secretion of osteoblasts. Thus, baicalin may be a regulatory protein within the bone.

골조직 세포에서 Estrogen의 효과에 대한 Nitric Oxided의 역할

고선일(Seon-Yle Ko) 대한치과의사협회 2001 대한치과의사협회지 Vol.39 No.11

Endothelin-1이 HOS 세포의 증식과 활성에 미치는 영향

배문서,고선일,김정근,김세원,Bae, Moon-Seo,Ko, Seon-Yle,Kim, Jung-Keun,Kim, Se-Won 대한안면통증구강내과학회 2001 Journal of Oral Medicine and Pain Vol.26 No.4

Endothelin-1 (ET-1) is a recently discovered potent vasoconstrictive peptide. It was first identified in vascular endothelial cells. ET-1 is a 21-amino acid peptide and elicits systemic effects such as stimulation of the production of atrial natriuretic peptide and release of aldosterone and corticosterone. In this study, to examine the role of ET-1 in the bone metabolism, effect of ET-1 on the proliferation and activity of osteoblastic cells was studied using HOS cells as osteoblast model. ET-1 dose-dependently increased the cell proliferation as determined by cell counting and MTT reduction assay after 48hr treatment. Alkaline phosphatase activity was inhibited by ET-1 and showed significant inhibition by 50 and 100 nM ET-1. ET-1 increased NBT reduction by HOS cells dose-dependently showing that ET-1 may increase the superoxide production by osteoblasts. Nitrite concentration in the media of HOS cell culture without cytokine stimulation was negligible and unaffected by ET-1 after 48hr treatment. Finally, after collection and concentration of conditioned media, gelatinase activity produced by HOS cells was determined by zymography. HOS cells can produce and secrete the gelatinase (gelatinase A type as determined by molecular weight of about 65,000) into culture media, however, ET-1 had no effect on the gelatinase activity. These findings suggest that ET-1 may have diverse effects on the proliferation and differentiation of osteoblasts, therefore, it may play an important role in bone metabolism.

마우스 골수세포 배양시 transforming growth factor-β와 epidermal growth factor가 파골세포양세포의 형성에 미치는 영향

임충남,고선일,김정근,김세원,Lim, Choong-Nam,Ko, Seon-Yle,Kim, Jung-Keun,Kim, Se-Won 대한안면통증구강내과학회 2000 Journal of Oral Medicine and Pain Vol.25 No.1

Bone marrow culture systems are widely used to differentiate osteoclast-like cells in vitro using several osteotropic hormones. In this study, we isolated and cultured the mouse bone marrow cells with or without some osteotropic hormones such as parathyroid hormone(PTH), prostaglandin $E_2(PGE_2)$ and $l,25(OH)_2-vitamin$ $D_3$(Vit. $D_3$). We confirmed the formation of osteoclast-like cells morphologically and functionally by the expression of tartrate-resistant acid phosphatase(TRAP) and by their capability to resorb dentin slices. We also studied the effects of transforming growth $factor-{\beta}(TGF-{\beta})$ and epidermal growth factor(EGF) on the Vit. $D_3-induced$ osteoclast-like cell formation. In control, a few multinucleated cells were formed whereas PTH and $PGE_2$ increased the number of multinucleated cells. PTH, $PGE_2$ and Vit. $D_3$ induced the formation of TRAP-positive multinucleated cells. After culture of mouse bone marrow cells on the dentin slices with or without osteotropic hormones, giant cells with diverse morphology were found on the dentin slices under the scanning electronmicroscopy. After removing the attached cells, resorption pits were identified on the dentin slices, and the shape of resorption pits was variable. EGF increased the osteoclast-like cell formation induced by Vit. $D_3$, however, $TGF-{\beta}$ showed biphasic effect, which at low concentration, increased and at high concentration, decreased the osteoclast-like cell formation induced by Vit. $D_3$.

법랑기질 단백질 유도체가 치주인대양세포 및 조골양세포에 미치는 영향

김동운,정진형,임성빈,고선일,Kim, Dong-Woon,Chung, Chin-Hyung,Lim, Sung-Bin,Ko, Seon-Yle 대한치주과학회 2003 Journal of Periodontal & Implant Science Vol.33 No.2

Recent study on the enamel matrix derivatives explained on the effects of new bone and new attachment formation in infrabony pocket of periodontal defects. The purpose of this study was to investigate on the biological effects of enamel matrix derivatives to attachment, proliferation and activation of periodontal ligament and osteoblast cells, After treatment of osteoblast and PDL cells with various Emdogain concentration level(0.03${\mu}g$/ml, 3${\mu}g$/ml, 300${\mu}g$/ml), activation of osteogenetic factor, calcified nodule formation and measuring alkaline phosphatase activity(ALP) were performed. 1. Both osteoblast and PDL cell showed increasing initial cell attachment with 300${\mu}g$/ml Emdogain concentration. 2. At the level of 300${\mu}g$/ml, accelerated proliferation of oseoblast and PDL cell was appeared. 3. As Emdogain's concentration increased, increased ALP activation of osteoblast was shown. In case of PDL cell, Emdogain increased ALP activation prominently at the level of 300${\mu}g$/ml. 4. No statistically significant activating change were founded at all of the concentrations of Emdogain on the activating of transcript factor Runx2 for differentiating osteoblast. 5. At the level of 300${\mu}g$/ml, calcified nodule formation was increased prominently to compare with other concentration. These results indicated that Emdogain should activate initial attachment, proliferation and activation, but not on Runx2 activation and can be used for useful tool of the treatment of periodontal tissue regeneration.

숙지황과 가시오가피 복합추출물(OPB)이 난소절제 흰쥐의 골밀도 및 골대사에 미치는 영향

김정근,김세원,이병의,황현환,권종석,고선일,Kim, Jung-Keun,Kim, Se-Won,Lee, Byung-Eui,Hwang, Hyeon-Hwan,Kwon, Jong-Seok,Ko, Seon-Yle 대한안면통증구강내과학회 2007 Journal of Oral Medicine and Pain Vol.32 No.1

본 연구는 숙지황과 가시오가피 복합추출물 (OPB)이 난소절제 흰쥐의 골밀도 및 골대사 변화에 미치는 영향을 관찰하기 위하여 수행하였다. 13주된 20마리의 Sprague-Dawley 암컷 흰쥐를 대조군과 실험군으로 나누었고, OPB의 골밀도 및 골대사 변화에 대한 효과를 관찰하기 위하여 난소절제 후 3일 후부터 OPB를 100 mg/kg씩 8주 동안 경구투여 하였다. pQCT 방식으로 골밀도, 골함유량, 골강도를 측정하였고, 혈청 중 CTx (C-telopeptide of type I collagen)의 변화를 관찰하였다. 난소절제 후 대조군의 골밀도는 $-29.8{\pm}3.0%$로 현저히 감소하였으며, 이러한 감소는 OPB의 투여에 의해 $-21.4{\pm}2.3%$로 억제되었다. 골강도의 변화에서는 anti-fracture 값과 anti-twisting 값이 대조군에 비해 OPB 처리군에서 유의성 있는 차이를 나타냈다. 또한 혈청 중 골흡수 지표인 CTx 농도는 대조군에 비해 OPB 처리군에서 낮게 나타났다. 이상의 결과로 숙지황과 가시오가피의 복합 추출물인 OPB가 난소를 절제한 흰쥐에서 골 소실을 억제하므로 임상에서 난소절제 후 또는 폐경 후 골다공증의 개선제로 이용될 수 있으리라 여겨진다.

뼈흡수유도호르몬이 ROS17/2.8세포로부터 Nitric Oxide 형성에 미치는 영향

고선일,김민성,한원정,김세원,김정근 대한구강악안면방사선학회 2005 Imaging Science in Dentistry Vol.35 No.3

Purpose : We performed the present study to investigate whether osteotropic hormomes play roles on the nitric oxide (NO) production in culture of ROS17/2.8 osteoblastic cells. Materials and Methods : The osteoblastic cell line ROS17/2.8 cells were cultured in F12 medium supplemented with 5% fetal bovine serum (FBS) at 37?C in a humidified atmosphere of 5% CO₂ in air. ROS17/2.8 cells were plated in 96-well plates at a density of 2-3×10³ cells/well and grown to confluence. Then the cells were pretreated with osteotropic hormones (parathyroid hormone (PTH) 20-500 ng/mL, 1,25 dihydroxycholecalciferol (1,25[OH]₂D₃) 1-100 nM; prostaglandin E₂ (PGE₂) 20-500 ng/mL) in the medium supplemented with 0.4% FBS for 72 hours and the cells were treated with cytokines (TNFα and IFNγ) in phenol red-free F12 medium for an additional 48 hours. NO synthesis was assessed by measuring the nitrite anion concentration, the reaction product of NO, in the cell culture medium using Griess reagent. Results : PTH and 1, 25[OH]₂D₃ pretreatment induced a significant increase in NO production in the presence of TNFα and IFNγ. PGE₂ slightly induced NO production compared to the control group. But, PGE2 pretreatment did not affect in NO production in the presence of TNFα and IFNγ. Conclusions : These results suggest that the actions of osteotropic hormones in bone metabolism may be partiallymediated by NO in the presence of cytokines.