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Interleukin-10 이 $interleukin-1{\beta}$로 유도되는 골흡수에 미치는 효과
유윤정,강윤선,이승일,Yu, Yun-Jung,Kang, Yun-Sun,Lee, Syng-Ill 대한치주과학회 1994 Journal of Periodontal & Implant Science Vol.24 No.2
The cytokines released by osteoblasts induce bone resorption via the differentiation of osteoclast precursors. In this process, $interleukin-1{\beta}$($IL-1{\beta}$)-induced bone resorption is mediated by granulocyte macrophage-colony stimulation factor(GM-CSF), interleukin-6 (IL-6), and tumor necrosis factor ${\alpha}$($TNF-{\alpha}$) released from osteoblasts. Since these cytokines (GM-CSF, IL-6, $TNF-{\alpha}$) are produced by not only osteoblasts but also monocytes, and interleukin-10(I1-10) inhibits the secretion of these cytokines from monocytes, it may be speculated that IL 10 could modulate the production of GM-CSF, IL-6, and $TNF-{\alpha}$ by osteoblasts, then control $IL-1{\beta}-induced$ bone resorption. Therefore, the aims of the present study were to examine the effects of IL-10 on bone resorption. The sixten or seventeen-day pregnant ICR mice were injected with $^{45}Ca$ and sacrificed one day after injection. Then fetal mouse calvaria prelabeled with $^{45}Ca$ were dissected out. In order to confirm the degree of bone resorption, mouse calvaria were treated with Lipopolysaccharide(LPS), $TNF-{\alpha}$, $IL-1{\alpha}$, IL-8, $IL-1{\beta}$, and $IL-1{\alpha}$, Then, IL-10 and $interferon-{\gamma}$ ($IFN-{\gamma}$) were added to calvarial medium, in an attempt to evaluate the effect of $IL-1{\beta}-induced$ bone resorption. In addition, osteoclasts formation in bone marrow cell cultures, and the concentration of IL-6, $TNF-{\alpha}$, and GM-CSF produced from mouse calvarial cells were investigated in response to $IL-1{\beta}$ alone and simultaneously adding f $IL-1{\beta}$ and IL-10. The degree of bone resorption was expressed as the ratio of $^{45}Ca$ release(the treated/the control). The osteoclasts in bone marrow cultures were indentified by tartrate resistant acid phosphatase(TRAP) stain and the concentration of the cytokines was quantified using enzyme linked immunosorbent method. As results of these studies, bone resorption was induced by LPS(1 ng/ml ; the ratio of $^{45}Ca$ release, $1.14{\pm}0.07$). Also $IL-1{\beta}$(1 ng/ml), $IL-1{\alpha}$(1 ng/ml), and $TNF-{\alpha}$(1 ng/ml) resulted in bone resorption(the rations of $^{45}Ca$ release, $1.61{\pm}0.26$, $1.77{\pm}0.03$, $1.20{\pm}0.15$ respectively), but IL-8 did not(the ratio of $^{45}Ca$ release, $0.93{\pm}0.21$). The ratios of $^{45}Ca$ release in response to IL-10(400 ng/ml) and $IFN-{\gamma}$(100 ng/ml) were $1.24{\pm}0.12$ and $1.08{\pm}0.04$ respectively, hence these cytokines inhibited $IL-1{\beta}$(1 ng/ml)-induced bone resorption(the ratio of $^{45}Ca$ release $1.65{\pm}0.24$). While $IL-1{\beta}$(1 ng/ml) increased the number of TRAP positive multinulcleated cells in bone marrow cultures($20{\pm}11$), simultaneously adding $IL-1{\beta}$(1 ng/ml) and IL-10(400 ng/ml) decreased the number of these cells($2{\pm}2$). Nevertheless, IL-10(400 ng/ml) did not affect the IL-6, GM-CSF, and $TNF-{\alpha}$ secretion from $IL-1{\beta}$(1 ng/ml)-activated mouse calvarial cells. From the above results, it may be suggested that IL-10 inhibites $IL-1{\beta}-induced$ osteoclast differntiation and bone resorption. However, the inhibitory effect of IL-10 on the osteoclast formation seems to be mediated not by the reduction of IL-6, GM-CSF, and $TNF-{\alpha}$ production, but by other mechanisms.
Calcium sulfate와 혈소판 유래성장인자의 혼합사용이 치주인대세포에 미치는 영향
김준성,최성호,유윤정,채중규,김종관,조규성,Kim, Jun-Seong,Choi, Seong-Ho,Yu, Yun-Jung,Chai, Jung-Kiu,Kim, Chong-Kwan,Cho, Kyoo-Sung 대한치주과학회 1997 Journal of Periodontal & Implant Science Vol.27 No.4
It was well known that calcium sulfate was biocompatible, resorbed rapidly in the body, had potential as a good barrier membrane. Platelet-derived growth factor(PDGF) was one of polypeptide growth factor that had been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purpose of this study was to evaluate the effects of a combination of calcium sulfate and PDGF on periodontal ligament cells in vitro to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the premolar tooth extracted for the orthodontic treatment. Cells were cultured in ${\alpha}-MEM$ contained with 20% FBS, at the $37^{\circ}C$, 100% of humidity, 5% $Co_2$ incubator. Cells were inoculated and cultured into 96 well culture plate with $1{\times}10^4cells/well$ of ${\alpha}-MEM$ for 1 day. After discarding the medium, those cells were cultured in ${\alpha}-MEM$ contained with 10% FBS alone(control group), in calcium sulfate(calcium sulfate group), in calcium sulfate treated with 15ng/ml of PDGF-BB(calcium sulfate+PDGF group), in ${\alpha}-MEM$ contained with 10% FBS treated with 15ng/ml of PDGF-BB(PDGF group) for 1, 2, 3 day respectively. And then each group was characterized by examining of the cell counting, MTT assay, collagen synthesis. The results were as follows. 1. In the analysis of cell proliferation by cell counting, both calcium sulfate group and calcium sulfate plus PDGF group showed no stastically significant difference compared to control group, but there was stastically significant difference between PDGF group and calcium sulfate group at 1, 2 day(P<0.05). 2. In the analysis of cell proliferation by MTT assay in calcium sulfate extracts, both calcium sulfate group and calcium sulfate plus PDGF group showed no stastically significant difference compared to control group, but there was stastically significant difference between PDGF group and calcium sulfate group at 2, 3 day, and between calcium sulfate plus PDGF group and calcium sulfate group at 2 day(P<O.05). 3. In the analysis of cell proliferation by MTT assay in transwell, both control group and PDGF group showed stastically significant difference compared to both calcium sulfate group and calcium sulfate plus PDGF group at 1 day, but there was no stastically significant difference compared to both calcium sulfate group and calcium sulfate plus PDGF group at 2, 3 day(P(0.05). 4. In the analysis of collagen synthesis by immunoblotting assay in calcium sulfate extracts, high level was detected on calcium sulfate group at 3 day, on calcium sulfate plus PDGF group at 1 day, on PDGF group at 2 day. On the basis of these results, calcium sulfate was biocompatible on the periodontal ligament cells and might have potential possibility as a vehicle of PDGF in the periodontal tissue regeneration.
Fluoride, Bisphosphonate 및 Gallium이 상아질 흡수 억제에 미치는 영향
백은영,유윤정,노병덕,최윤정,이승종 大韓齒科保存學會 1997 Restorative Dentistry & Endodontics Vol.22 No.2
Replacement resorption is followed by the delayed replantation of an avulsed tooth. Currently no effective treatment is substantiated for replacement resorption. The purpose of this study was to investigate the effect of stannous fluoride, bisphosphonate(etidronate disodium) and gallium nitrate, which have been shown to reduce dentin resorption, on human dentin. Osteoclasts were collected from tibeas of chich embryo. The cells were well agitated to prevent adhesion and seeded onto the sliced human dentin wafers which had been soaked in either culture media(control), or several different concentrations of stannous fluoride, etidronate disodium(1-hydroxyethylidene-1, 1-bisphosphonate disodium), and gallium nitrate. Resorption was measured by counting the number of resorptive pit produced by the cells. Results are ad follows. Stannous fluoride and etidronate disodium showed statistically significant reduction of dentin resorption(p<0.05) but the effect of stannous fluoride seemed to be its high cytotoxicity. Etidronate disodium did not show cytotoxicities in all experimented concentrations. Gallium nitrate did not show differences in resorption either between different concentrations of from the control group.
Interleukin-1β에 의하여 치주인대세포에서 유리된 cytokine이 파골세포형성에 미치는 영향
이종갑,곽월아,유윤정,이승일,김태선 大韓小兒齒科學會 1996 大韓小兒齒科學會誌 Vol.23 No.1
Tooth movement is induced by bone remodeling during orthodontic treatment. Bone remodeling is regulated by various cytokines. Especially interleukin-1 (IL-1β), a cytokine present in periodontal ligaments of experimentally moved teeth, elicits bone resorption. In these processes, IL-1-induced bone resorption is mediated by interleukin-6 (IL-6) and granulocyte macrophage-colony stimulating fector (GM-CSF) secreted from osteoblasts. Periodontal ligament cells, which function as an anchorage for tooth, lie between alveolar bone and cementum. Therefore cytokines produced in the periodontal ligament (PDL) cells may also directly affect alveolar bone resorption in orthodontic tooth movement. Here I have examined whether PDL cells express IL-1β,interleukin-6 (IL-6) and granulocyte macrophage-colony stimulating factor (GM-CSF) mRNA and secrete those cytokines in response to IL-1β. Finally I have investigated whether IL-6 produced from PDL cells induces osteoclast formation in mouse bone marrow cell cultures. The expression of mRNA was estimated by polymerase chain reaction (PCR). The concentration of cytokines was quantified using enzyme linked immunosorbent method and the osteoclasts in bone marrow cultures were identified by tartrate resistant acid phosphatase (TRAP) stain. As results of these studies, IL-1βstimulated the expression of IL-1β, IL-6 and GM-CSF mRNA in PDL cells. 0.05 ng/ml IL-1βalso induced maximum production of Il-6 and GM-CSF in these cells. After an addition of IL-1β(0.05 ng/ml), IL-6 production increased from 2 hours to 8 hours and GM-CSF production also increased from 4 hours to 8 hours. IL-6 (100 ng/ml) increased the number of TRAP positive multinucleated cells in the presence of soluble interleukin-6 receptor (sIL-6R, 100 ng/ml). These results suggest that IL-1βmay stimulate alveolar bone resorption by inducing IL-6 and GM-CSF production in PDL cells which enhance osteoclast differentiation and IL-6 enhances osteoclast formation in the presence of sIL-6R. And this process by IL-1βmay be closely associated with alveolar bone resorption induced by orthodontic force.
Rat의 간으로부터 추출한 수용성 단백질중에 있는 High Affinity Insulin Receptor Protein
김기한,유주현,유윤정,배동훈,이기호 연세대학교 산업기술연구소 1990 논문집 Vol.22 No.1
We confirmed that there were 350 kd, 320 kd, 290 kd, 210 kd and 160 kd protein in insulin soluble receptor extracted from rat liver membrane. These insulin soluble receptors had ??-insulin specific binding activity. The ??-insulin binding activity was affected by the concentration of unlabeled insulin. In intract membrane, only 350 kd protein bound ??-insulin in contrast to the soluble receptor protein. With the GSH treatment before cross-linking, ??-insulin binding activity decreased but after cross-linking, GSH treatment dissociated the insulin receptor proteins into α-subunit(-135 kd) and β-subunit(-95 kd). These findings might be thought that the SH residues of insulin receptor were important to bind ??-insulin.