• Journal of Acupuncture Research
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• 제21권2호
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• pp.73-87
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• 2004

전통적인 면역억제제와 면역활동제인 녹용수용추출액(DAA)은 뼈의 재생에 있어서 중요한 역할을 한다고 여겨진다. 녹용약침이 비활성 연골세포의 분화를 야기시킬수 있는 지를 결정하기 위해, 융합된 세포 배양균을 녹용약침과 함께 24, 36, 48, 72, 120 시간동안 먼저 처리하였다. 이러한 처리후에 배양액을 10-10 ~ 10-8M 1,25-(OH)2D3를 포함하는 새로운 배양액으로 바꾸어 세포들을 추가로 24시간동안 배양했다. 앞선 연구에서 더 성숙된 활성연골세포가 이러한 Vit D3 대사산물에 반응한다는 것을 보여주었기 때문에 이러한 두 번째 처리를 선택하였다. 세포성숙에 있어서 녹용약침의 전처리 효과는 ALPase의 특이활동을 측정하는 것으로 확인하였다. 기질 단백질 합성의 변화는 35SO4 결합이 proteoglycan으로의 변화되는것과 collagen 합성을 측정함으로써 증명되었다. 비활성 연골세포가 녹용약침과 함께 120시간동안 전처리되고 다시 1,25-(OH)2D3를 추가한 처치에서 ALPase 특이활동과 collagen 합성이 농도 의존적으로 증가를 일으켰다. 그러나 proteoglycan의 생성에는 영향을 미치지 않았다. 1,25-(OH)2D3와 함께 전처리 된 비활성 연골 세포는 어떠한 전처리도 받지 않았던 비활성 연골세포처럼 반응했다. 이러한 결과는 녹용약침이 직접적으로 비활성 연골 세포를 활성 연골세포로 성숙시키는데 관여한다는 것을 알려준다. 그러므로 녹용약침은 연골내 골화과정에서 연골세포의 성숙을 조절하는데 중요한 역할을 하는 것으로 나타났다.

• Journal of Acupuncture Research
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• 제23권2호
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• pp.103-111
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• 2006

Objectives : Deer antler Water Extract(DAE), prepared from the pilose antler of Cervus korean TEMMINCK var. mantchuricus Swinhoe (Nokyong), a traditional immuno-suppressive and immuno-activating Korean herbal-acupuncture, is thought to play an important role in human bone remodeling. Methods : To determine whether DAE can induce the differentiation of resting zone chondrocytes(RC) or not, confluent cell cultures were pretreated for 24, 36, 48, 72, and 120hrs with DAE. At the end of pretreatment, the media were replaced with new media containing $10^{-10}{\sim}10^{-8}M\;1,25-(OH)_2D_3$ and the cells incubated for an additional 24hrs. Results : This second treatment was chosen because prior studies had shown that only the more mature growth zone chondrocytes(GC) respond to this vitamin $D_3$ metabolite. The effect of DAE pretreatment on cell maturation was confirmed by measuring alkaline phosphatase (ALPase)-specific activity. Changes in matrix protein synthesis were examined by measuring collagen synthesis, as well as $^{35}SO_4$ incorporation into proteoglycans. When RC cells were pretreated for 120h with DAE, treatment with $1,25-(OH)_2D_3$ caused a dose-dependent increase in ALPase-specific activity and collagen synthesis, however, the proteoglycan production was not affected. RC cells pretreated with $1,25-(OH)_2D_3$ responded like RC cells that had not received any pretreatment. Conclusion : These results indicate that DAE directly regulates the maturation of RC chondrocytes into GC chondrocytes. Therefore it was indicated that DAE may play a significant role in regulating chondrocyte maturation during endochondral ossification.

• 대한치과교정학회지
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• 제25권6호
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• pp.705-714
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• 1995

Due to the great deal of effort that has gone into the study of osteoclastic differentiation and activation over the last few decades, the mechanisms of these two events have been discovered gradually. Nitric oxide($NO^-$), which is produced from arginine by a nitric oxide synthase, opened up a new area of biological research. Recently, it has been reported that $NO^-$ is produced by osteoblasts stimulated by lipopolysaccharide and several other cytokines. In this study, the effect of sodium nitroprusside(SNP), a donor of nitric oxide($NO^-$), on osteoclast-like cell formation and on mature osteoclast function was examined. To determine the mechanism of the inhibitory effects of SNP decreased not only the basal $^{45}Ca$ release but also thee bone resorption induced by PTH and 1,25-dihydroxyvitamin $D_3\;(1,25[OH]_{2}D_3)$. The inhibitory effect of SNP on bone resorption induced by PTH appeared 2 dyas after treatment, whereas SNP effect on inhibiting bone resorption induced by $1,25[OH]_{2}D_3$ appeared at the thhird days. When chicken and rat osteeoclasts were cultured on dentin slices, treatment of $300{\mu}M$ SNP resulted in a significant decrease in dentin resorption by osteoclasts in terms of total resolution area and average individual area. We also examind the effect of SNP on formation of osteoclast-like cells that is TRAP-positive multinucleated cells from chicken and rat bone marrow cells in the presence or absence of $10^{-8}\;M\;1,25[OH]_{2}D_3$. The addition of $300{\mu}M$ SNP inhibiteed the formation of TRAP-positive multinucleated cells. The present data suggest that SNP, possibly as a $NO^-$ donor, inhibits the osteoclastic differentiation and osteoclastic activity.

• The Journal of Advanced Prosthodontics
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• 제6권5호
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• pp.379-386
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• 2014

PURPOSE. These days, mesenchymal stem cells (MSCs) have received worldwide attention because of their potentiality in tissue engineering for implant dentistry. The purpose of this study was to evaluate various growth inducing factors in media for improvement of acquisition of bone marrow mesenchymal stem cells (BMMSCs) and colony forming unit-fibroblast (CFU-F). MATERIALS AND METHODS. The mouse BMMSCs were freshly obtained from female C3H mouse femur and tibia. The cells seeded at the density of $10^6$/dish in media supplemented with different density of fetal bovine serum (FBS), $1{\alpha}$, 25-dihydroxyvitamin (VD3) and recombinant human epidermal growth factor (rhEGF). After 14 days, CFU-F assay was conducted to analyze the cell attachment and proliferation, and moreover for VD3, the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was additionally conducted. RESULTS. The cell proliferation was increased with the increase of FBS concentration (P<.05). The cell proliferation was highest at the density of 20 ng/mL rhEGF compared with 0 ng/mL and 200 ng/mL rhEGF (P<.05). For VD3, although the colony number was increased with the increase of its concentration, the difference was not statistically significant (P>.05). CONCLUTION. FBS played the main role in cell attachment and growth, and the growth factor like rhEGF played the additional effect. However, VD3 did not have much efficacy compare with the other two factors. Improvement of the conditions could be adopted to acquire more functional MSCs to apply into bony defect around implants easily.

• Annals of dermatology
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• 제30권6호
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• pp.653-661
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• 2018

Background: Citron is well known for an abundance of antioxidative and anti-inflammatory ingredients such as vitamin C, polyphenol compounds, flavonoids, and limonoids. Objective: In this study, we aimed to evaluate the effects of citron essential oils on rosacea mediators in activated keratinocytes in vitro. Methods: Normal human epidermal keratinocytes (NHEKs) were stimulated with $1{\alpha}$, 25-dihydroxyvitamin $D_3$ ($VD_3$) and interleukin 33 (IL-33) with LL-37 to induce rosacea mediators such as kallikrein 5 (KLK5), cathelicidin, vascular endothelial growth factor (VEGF), and transient receptor potential vanilloid 1 (TRPV1). These mediators were analyzed by performing reverse-transcription polymerase chain reaction (PCR), quantitative real-time PCR, immunocytofluorescence and enzyme-linked immunosorbent assay after NHEKs were treated with citron seed and unripe citron essential oils. Results: The messenger RNA (mRNA) and protein levels of KLK5 and LL-37 induced by $VD_3$ were suppressed by citron seed and unripe citron essential oils. Furthermore, the mRNA and protein levels of VEGF and TRPV1 induced by IL-33 with LL-37 were also suppressed by citron essential oils. Conclusion: These results show that citron essential oils have suppressive effects on rosacea mediators in activated epidermal keratinocytes, which indicates that the citron essential oils may be valuable adjuvant therapeutic agents for rosacea.

• Tuberculosis and Respiratory Diseases
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• 제77권2호
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• pp.73-80
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• 2014

Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.

• Journal of Periodontal and Implant Science
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• 제38권sup2호
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• pp.395-404
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• 2008

Purpose: Osteoprotegerin (OPG) is a secreted glycoprotein and a member of the tumor necrosis factor (TNF) receptor family that inhibits bone resorption by suppressing osteoclastogenesis. Gingival fibroblasts (GF) play a role in periodontal disease progression, and the purpose of this experiment was to evaluate influence of osteotropic factors on the expression of osteoprotegerin mRNA in these cells. Materials and Methods: In this experiment, the influence of osteoclastogenic factors, interleukin-1 beta (IL-$1{\beta}$), TNF-$\alpha$, prostanglandin E2 ($PEG_2$). parathyroid hormone (PTH) and 1$\alpha$, 25-dihydroxyvitamin $D_3$ on the expression of osteoprotegerin mRNA in GF was studied by Northern blot hybridization. Results: As expected, $PEG_2$ tended to inhibit OPG levels and this was most prominent at 24 hours of culture with $10^{-7}M$ of $PEG_2$. TNF-$\alpha$ at 10ng/ml and also at 25ng/ml decreased OPG levels to almost 30% of the control at 24 hours. This contrasts with reports of increased OPG levels from osteoblast/stromal cells and gingival fibroblasts stimulated by TNF-$\alpha$. Decrease of OPG levels with $PEG_2$ and TNF-$\alpha$ suggests a pathway whereby these mediators exert their resorptive effects. However, OPG levels were increased almost 3-fold at 24 hours with IL-1$\beta$(1 to 15ng/ml) and increased 1.4 fold with 24-hour treatment of $10^{-7}M$ PTH. Conclusion: Increase of OPG levels suggests that these 'osteoclastogenic' factors act in more complex ways and may act to inhibit bone resorption in inflammatory periodontitis. This result supports the role of OPG as a negative feedback mechanism in osteoclastic activity.

• Biomolecules & Therapeutics
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• 제17권2호
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• pp.125-132
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• 2009

Calbindin-$D_{9k}$ (CaBP-9k), a cytosolic calcium-binding protein, is expressed in a variety of tissues, i.e., the duodenum, uterus, placenta, kidney and pituitary gland. Duodenal CaBP-9k is involved in intestinal calcium absorption, and is regulated at transcriptional and post-transcriptional levels by 1,25-dihydroxyvitamin D3, the hormonal form of vitamin D, and glucocorticoids (GCs). Uterine CaBP-9k has been implicated in the regulation of myometrial action(s) through modulation of intracellular calcium, and steroid hormones appear to be the main regulators in its uterine and placental regulation. Because phenotypes of CaBP-9k-null mice appear to be normal, other calcium-transporter genes may compensate for its gene deletion and physiological function in knockout mice. Previous studies indicate that CaBP-9k may be controlled in a tissue-specific fashion. In this review, we summarize the current information on calcium homeostasis related to CaBP-9k gene regulation by GCs, vitamin D and its receptors, and its molecular regulatory mechanism. In addition, we present related data from our current research.

• Nutrition Research and Practice
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• 제18권1호
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• pp.19-32
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• 2024

BACKGROUND/OBJECTIVES: Atherosclerosis is associated with increased inflammation in the visceral adipose tissue (VAT). Vitamin D has been reported to modulate the inflammatory responses of stromal vascular cells (SVCs) and adipocytes in adipose tissue, but the role of vitamin D in atherosclerosis biology is unclear. This study examined the effects of in vitro 1,25-dihydroxyvitamin D₃ (1,25[OH]₂D₃) treatment on the inflammatory responses of SVCs and adipocytes from atherosclerotic mice. MATERIALS/METHODS: C57BL/6J (B6) mice were divided randomly into 2 groups and fed a 10% kcal fat control diet (control group, CON) or 41% kcal fat, 0.21% cholesterol (high fat + cholesterol, HFC) diet (obese group, OB), and B6.129S7-Ldlr^tm1Her/J (Ldlr^-/-) mice were fed a HFC diet (obese with atherosclerosis group, OBA) for 16 weeks. SVCs and adipocytes isolated from VAT were pre-incubated with 1,25(OH)₂D₃ for 24 h and stimulated with lipopolysaccarides for the next 24 h. Proinflammatory cytokine production by adipocytes and SVCs, the immune cell population in SVCs, and the expression of the genes involved in the inflammatory signaling pathway in SVCs were determined. RESULTS: The numbers of total macrophages and SVCs per mouse were higher in OB and OBA groups than the CON group. The in vitro 1,25(OH)₂D₃ treatment significantly reduced macrophages/SVCs (%) in the OBA group. Consistent with this change, the production of interleukin-6 and monocyte chemoattractant protein 1 (MCP-1) by SVCs from the OBA group was decreased by 1,25(OH)₂D₃ treatment. The 1,25(OH)₂D₃ treatment significantly reduced the toll-like receptor 4 and dual-specificity protein phosphatase 1 (also known as mitogen-activated protein kinase phosphatase 1) mRNA levels in SVCs and MCP-1 production by adipocytes from all 3 groups. CONCLUSIONS: These findings suggest that vitamin D can attribute to the inhibition of the inflammatory response in VAT from atherosclerotic mice by reducing proinflammatory cytokine production.

• Childhood Kidney Diseases
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• 제8권2호
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• pp.195-204
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• 2004

목적: 저인산혈성 구루병 환아의 임상상, 각종 혈액화학 검사 수치 및 방사선 검사 소견, 치료에 대한 반응 등을 살피고 효과적인 치료에 도움이 되는 정보를 얻고자 본 연구를 시행하였다. 방법: 1983년 7월부터 2004년 2월까지 서울대학교병원 소아과에서 저인산혈성 구루병으로 진단받고 내과적인 치료를 받은 환아들의 의무기록지를 후향적으로 분석하였다. 결과: 대상환자는 56명이었고, 남녀의 비는 1:3 이었다 주소는 대부분 하지만곡과 저신장이었고, 혼자 걷기 시작하면서 발견된 경우가 가장 많았다. 진단시 평균연령은 5년 2개월 이었으며 12개월 이전에 진단 받은 경우는 없었다. 14명(25%)에서 가족력이 있었다. 진단받은 연령과 신장의 표준편차 점수는 통계적으로 의미있는 음의 상관관계가 있었다(r=-0.47, p=0.005). 생화학적 검사에서 정상 혈청 칼슘, 혈청 인의 저하, alkaline phosphatase의 상승, 세뇨관에서 인산 재흡수의 감소 및 정상 범위의 $1,25(OH)_2D_3$ 치를 보였다. 방사선 검사는 구루병에 합당하였다. 치료제로 Joulie 용액과 비타민 D 대사체를 복용하였다. 치료를 시작한 후 혈청 인과 alkaline phosphatase는 통계적으로 의미있는(P<0.05) 호전을 보였고, 방사선 검사는 구루병의 치유를 보였으나, 신장 표준편차 점수는 의미있는 변화를 보이지 않았다(.p=0.224). 치료의 부작용으로 신석회화(34%), 부갑상선호르몬의 상승(77%), 고칼슘뇨증(45%) 등이 발생하였다. 16명이 절골술을 통한 변형의 교정을 시행하였고, 그 중 4명은 하지연장을 함께 시행하였다. 결론: 증상의 시작과 진단 사이에는 수 년간의 지연이 있었다 조기치료가 환자의 예후에 가장 중요하므로, 가족력이 있는 경우 증상 발현 이전에 생화학적 검사로 조기 진단 및 치료를 하도록 해야 한다. 치료 중에는 정기적인 모니터링과 용량의 조절을 통해 합병증에 대처해야 한다.