• 제목/요약/키워드: MC3T3-E1 cells

검색결과 202건 처리시간 0.022초

유향 추출물이 MC3T3-E1 세포 활성 및 분화에 미치는 영향 (Effects of Olibanum Extracts on the Activity and Differentiation of MC3T3-E1 Cells)

  • 한상헌;김명동;유승한;유용욱;유형근;신형식
    • Journal of Periodontal and Implant Science
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    • 제31권2호
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    • pp.287-298
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    • 2001
  • Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use have been studied for their capacity of anti-bacterial, anti-inflammatory and regenerative potential of periodontal tissues. Olibanum has the effects to hemostasis, analgesic and anti-inflammatory, and it also has been traditionally used as a drug for the treatment of bone disease in oriental medicine. The purpose of the present study was to investigate the effects of Olibanum extracts on the activity and differentiation of MC3T3-E1 cells, alkaline phosphatase(ALP) synthesis, formation of bone nodules and expression of type I collagen of MC3T3-E1 cells. To examine the cellular activity, MC3T3-E1 cells were cultured with ${\alpha}-MEM(control)$ and each concentration of Olibanum for 2 days and 4 days. To compare the ALP synthesis, MC3T3-E1 cells were cultured with ${\alpha}-MEM(negative\; control)$, dexamethasone(positive control), and each concentration of Olibanum for 2 days and 4 days. To compare the bone nodule formation, MC3T3-E1 ells were cultured for 21 days, and to compare the type I collagen expression, MC3T3-E1 cells were cultured for 4 days. The cellular activity of MC3T3-E1 cells treated with $1{\mu}g/ml$ of Olibanum extracts was significantly increased at 4-day(p<0.05) to control. The activity of ALP in MC3T3-E1 cells treated with $1{\mu}g/ml$ Olibanum extracts was significantly increased at 4-day(p<0.05). All the experimental groups showed much more bone nodule formation than control groups. The group treated with $1{\mu}g/ml$ of Olibanum extracts was the highest bone nodule formation, and showed much more type I collagen expression than negative control. These results indicate that Olibanum extracts may be considered effective in the activity and differentiation of MC3T3-E1 cells.

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산양삼의 조골세포 활성화에 미치는 영향 (Effects of Wild Simulated Ginseng on the Proliferation, Differentiation and Mineralization of Osteoblastic MC3T3-E1 Cells)

  • 정진부
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2020년도 춘계학술대회
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    • pp.90-90
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    • 2020
  • Panax ginseng C.A. Meyer (P. ginseng) is known to exert a wide range of pharmacological effects both in vitro and in vivo. Although studies on ginsenoside, antioxidant activity, and anticancer effect of wild simulated ginseng (WSG) have been conducted, there is little research on the effect of WSG on bone metabolism. In this study, we investigated the potential anti-osteoporotic properties of WSG on the growth and differentiation of MC3T3-E1 cells. WSG significantly increased the viability and proliferation of MC3T3-E1 cells. WSG activated intracellular alkaline phosphatase (ALP) activity in MC3T3-E1 cells. In addition, WSG increased the mineralized nodules in MC3T3-E1 cells. Furthermore, WSG increased the expression of genes such as Runx2, ALP, OPN and OCN associated with osteoblast growth and differentiation in a dose-dependent manner.

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Chemical Constituents from Acer mandshuricum and Their Effects on the Function of Osteoblastic MC3T3-E1 Cells

  • Ding, Yan;Liang, Chun;Nguyen, Huu Tung;Choi, Eun-Mi;Kim, Jeong-Ah;Kim, Young-Ho
    • Bulletin of the Korean Chemical Society
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    • 제31권4호
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    • pp.929-933
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    • 2010
  • A new compound, 4-methoxyl 5-hydroxymethyl benzoic 3-O-$\beta$-D-glucopyranoside (1), has been isolated from the leaves and stems of Acer mandshuricum, along with nine known compounds (2-10). Their structures were determined by a variety of spectroscopic analyses. The effect of compounds 1-10 on the function of osteoblastic MC3T3-E1 cells was examined by determining alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization. Compound 1 significantly increased the function of osteoblastic MC3T3-E1 cells; $5.0\;{\mu}M$ of 1 increased ALP activity, collagen synthesis, and mineralization of MC3T3-E1 cells to 114.7, 119.5, and 108.2% (P < 0.05) of the basal value, respectively. In addition, compounds 2-10 also potently increased the function of osteoblastic MC3T3-E1 cells.

Cottonseed Extract Improves the Function of Osteoblastic MC3T3-E1 Cells

  • Choi, Eun-Mi;Lee, Kyung-Hee
    • Food Quality and Culture
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    • 제2권2호
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    • pp.85-88
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    • 2008
  • We have investigated the effects of cottonseed extract on the proliferation, differentiation and lipopolysaccharide (LPS)-induced production of local factors in murine clonal osteoblastic MC3T3-E1 cells. Ethanol extract of cotton seed ($4{\sim}63{\mu}g/mL$) significantly increased the proliferatin of MC3T3-E1 cells (p<0.05). Moreover, cottonseed extract ($10{\sim}50{\mu}g/mL$) caused a significant elevation of alkaline phosphatase (ALP) activity and collagen content in the cells. Lipopolysaccharide (LPS) is a potent stimulator of bone resorption in inflammatory diseases. We examined the effect of cottonseed extract on the LPS-induced production of tumor necrosis factor a (TNF-$\alpha$) and nitric oxide (NO) in MC3T3-E1 cells. Treatment with cottonseed extract ($10{\sim}50{\mu}g/mL$) decreased the $5{\mu}g/mL$ LPS-induced production of TNF-$\alpha$ and NO in osteoblasts, suggesting that the antiresorptive action of cottonseed extract may be mediated by decrease in these local factors. This study suggests that cottenseed may contribute to antiresorptive action against osteoblastic cells, resulting in a beneficial effect in promoting the function of osteoblastic cells.

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Nitric Oxide-Induced Autophagy in MC3T3-E1 Cells is Associated with Cytoprotection via AMPK Activation

  • Yang, Jung Yoon;Park, Min Young;Park, Sam Young;Yoo, Hong Il;Kim, Min Seok;Kim, Jae Hyung;Kim, Won Jae;Jung, Ji Yeon
    • The Korean Journal of Physiology and Pharmacology
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    • 제19권6호
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    • pp.507-514
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    • 2015
  • Nitric oxide (NO) is important in the regulation of bone remodeling, whereas high concentration of NO promotes cell death of osteoblast. However, it is not clear yet whether NO-induced autophagy is implicated in cell death or survival of osteoblast. The present study is aimed to examine the role of NO-induced autophagy in the MC3T3-E1 cells and their underlying molecular mechanism. The effect of sodium nitroprusside (SNP), an NO donor, on the cytotoxicity of the MC3T3-E1 cells was determined by MTT assay and expression of apoptosis or autophagy associated molecules was evaluated by western blot analysis. The morphological observation of autophagy and apoptosis by acridine orange stain and TUNEL assay were performed, respectively. Treatment of SNP decreased the cell viability of the MC3T3-E1 cells in dose- and time-dependent manner. SNP increased expression levels of p62, ATG7, Beclin-1 and LC3-II, as typical autophagic markers and augmented acidic autophagolysosomal vacuoles, detected by acridine orange staining. However, pretreatment with 3-methyladenine (3MA), the specific inhibitor for autophagy, decreased cell viability, whereas increased the cleavage of PARP and caspase-3 in the SNP-treated MC3T3-E1 cells. AMP-activated protein kinase (AMPK), a major autophagy regulatory kinase, was activated in SNP-treated MC3T3-E1 cells. In addition, pretreatment with compound C, an inhibitor of AMPK, decreased cell viability, whereas increased the number of apoptotic cells, cleaved PARP and caspase-3 levels compared to those of SNP-treated MC3T3-E1 cells. Taken together, it is speculated that NO-induced autophagy functions as a survival mechanism via AMPK activation against apoptosis in the MC3T3-E1 cells.

Icaritin, a Flavonoid Derived from the Herb Epimedium, Promotes Osteogenic Differentiation of MC3T3-E1 Cells

  • Park, Dan-Bi;Lee, Hee Su;Ko, Seong-Hee
    • International Journal of Oral Biology
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    • 제42권4호
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    • pp.163-168
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    • 2017
  • Osteoporosis is a metabolic bone disease that is characterized by low bone mass resulting from an increase in bone resorption relative to bone formation. The most current therapies for osteoporosis have focused on inhibiting bone resorption by osteoclasts. The purpose of this study is to develop new anabolic agents for treatment of osteoporosis that have fewer risks compared to conventional therapies. We searched the natural products that were derived from the traditional Asian medicines which have been used for treatment of bone related diseases. Icaritin is a flavonoid glycoside derived from the herb Epimedium which has beneficial effects on bone formation. To determine the effect of icaritin on bone formation, we examined the effect of icaritin on MC3T3-E1 cell proliferation and differentiation. For determining the effects of icaritin on proliferation, we performed the MTT assay using MC3T3-E1 cells. To evaluate whether icaritin could promote the osteogenic differentiation of MC3T3-E1 cells, alkaline phosphatase (ALP) activity and mRNA expressions of Runx2, osteocalcin (OCN), RANKL, and osteoprotegerin (OPG) were determined. Icaritin increased MC3T3-E1 cell proliferation. Icaritin increased the ALP activity of MC3T3-E1 cells on 72 hour culture in osteogenic media. mRNA expression of Runx2 was increased after 24 hour culture with icaritin. mRNA expression of osteocalcin was increased after 72 hour culture with icaritin. In addition, icaritin increased the mRNA expressions of OPG and RANKL. However, icaritin increased the mRNA expression of OPG much more than that of RANKL, and then, it increased the OPG/RANKL ratio. These results suggest that icaritin promotes osteogenic differentiation of osteoblasts and decreases osteoclast formation regulated by osteoblasts.

Sodium Fluoride가 조골세포주 MC3T3-E1의 제 1 형 ${\alpha}2$ 교원질 mRNA에 미치는 영향에 관한 연구 (THE EFFECTS OF SODIUM FLUORIDE ON TYPE I $\alpha$ 2 COLLAGEN RIBONUCLEIC ACID (mRNA) LEVEL IN MURIN OSTEOBLAST LIKE (MC3T3-E1) CELLS)

  • 황정빈;정규림;박영국
    • 대한치과교정학회지
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    • 제23권3호
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    • pp.415-425
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    • 1993
  • Fluoride is one of the most potent stimulators of bone formation in vivo. But its direct effects on osteoblast is not yet clear This study was to investigate the effects of Sodium fluoride on alkaline phosphatase(ALP) activity, cAMP formation responsive to parathormone(PTH) and type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level in Murin osteoblast-like (MC3T3-E1) cells. The cells were cultured in $\alpha-Minimal$ essential medium $(\alpha-MEM)$ supplemente with $10\%$ fetal bovine serum (FBS) and then changed to $0.1\%$ FBS with various concentration of Sodium fluoride. The ALP activity was assayed by the method of Lowry with disodium phenyl phosphated as substrate. cAMP formation was measured by Radioimmuno Assay(RIA). Type I $\alpha$ 2 collagen ribonucleic acid(mRNA) expression was studied by Nothern blot analysis. The results were as follows: 1. cAMP level was increased by PTH in MC3T3-E1 cells. 2. Sodium fluoride showed the tendency of inhibitory effects on cAMP responsiveness to PTH in MC3T3-E1 cells. 3. Sodium fluoride increased ALP activity at cocentration of $2{\mu}M,\;4{\mu}M,\;and\;10{\mu}M$ significantly different from control at the 0.001 level. ALP activity revealed maximum value at $10{\mu}M$ in this study. 4. Nothern blot analysis of Sodium fluoride treated cells, using Type I $\alpha$ 2 collagen prove, revealed significant increase at $10{\mu}M$ in MC3T3-E1 cells.

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MC3T3-E1세포의 ALP activity에 대한 IGF-I의 영향 (The Effect of IGF-1 on ALP Activity of MC3T3-E1 Cell)

  • 이후정;이재목;최병주;유현모;서조영
    • Journal of Periodontal and Implant Science
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    • 제27권4호
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    • pp.669-684
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    • 1997
  • Polypeptide growth factors belong to a class of potent biologic mediators which regulate cell differentiation, proliferation, migration and metabolism. IGF-I is polypeptides secreted by skeletal cells and is considered as regulators of bone formation. The purpose of this study is to evaluate the effects of IGF-I on bone nodule formation and alkaline phosphatase activity of MC3T3-E1 cells. MC3T3-E1 cells were seeded at $1{\times}10^4$ cells/well, $1{\times}10^5$ cells/well in alpha-modified Eagle medium containing 10% fetal bovine serum, 10 mM ${\beta}-glycerophosphate$ and $5O{\mu}g/ml$ of ascorbic acid. Before 48 hours of indicated time, medium were changed with serum free medium. After 24 hours, 0.1, 1, 10 ng/ml IGF-I were added to the cells and cultured for 3, 7, 14, 21, 28 days. And histochemical analysis was done and ALP activity was measured and was expressed as nmol/min/mg of protein. The bone nodule formation in MC3T3-E1 cells of IGF-I was seen at 21, 28 days, but there were no difference between control group and experimental groups. The ALP activity decreased when it is compare to control 2 group except for 1 ng/ml, 10 ng/ml IGF-I of 21-day-groups and 1 ng/ml IGF-I of 28-day-groups. Dose response effects of IGF-I of ALP activity in MC3T3-E1 cells were seen the highest ALP activity at 1ng/ml until 21days and the highest ALP activity at 10 ng/ml of 28 daygroups. The peak times were seen at 7-day group, 14-day group on control group and experimental group respectively, and 1 ng/ml group was the highest ALP activity, From the above results, IGF-I was not seen notable effect on bone nodule formation and decreased ALP activity of MC3T3-E1 cells but the use of IGF-I to mediate biological stimulation of MC3T3-E1 cells shows promise for future therapeutic application.

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Glycyrrhiza uralensis (licorice) extracts increase cell proliferation and bone marker enzyme alkaline phosphatase activity in osteoblastic MC3T3-E1 cells

  • Cho, Young-Eun;Kwun, In-Sook
    • Journal of Nutrition and Health
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    • 제51권4호
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    • pp.316-322
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    • 2018
  • Purpose: The Glycyrrhiza uralensis species (Leguminosae) as a medicinal biocompound, and one of its root components, isoliquritigenin (ISL), which is a flavonoid, has been reported to have anti-tumor activity in vitro and in vivo. However, its function in bone formation has not been studied yet. In this study, we tested the effect of Glycyrrhiza uralensis (ErLR) and baked Glycyrrhiza uralensis (EdLR) extracts on osteoblast proliferation, alkaline phosphatase (ALP) activity, and bone-related gene expression in osteoblastic MC3T3-E1 cells. Methods: MC3T3-E1 cells were cultured in various levels of ErLR (0, 5, 10, 15, $20{\mu}g/mL$), EdLR (0, 5, 10, 15, $20{\mu}g/mL$), or ISL (0, 5, 10, 15, $20{\mu}M$) in time sequences (1, 5, and 20 days). Also, isoliquritigenin (ISL) was tested for comparison to those two biocompound extracts. Results: MTT assay results showed that all three compounds (ErLR, EdLR, and ISL) increased osteoblastic-cell proliferation in a concentration-dependent manner for one day. In addition, both ErLR and EdLR compounds elevated the osteoblast proliferation for 5 or 20 days. Extracellular ALP activity was also increased as ErLR, EdLR, and ISL concentration increased at 20 days, which implies the positive effect of Glycyrrhiza species on osteoblast mineralization. The bone-related marker mRNAs were upregulated in the ErLR-treated osteoblastic MC3T3-E1 cells for 20 days. Bone-specific transcription factor Runx2 gene expression was also elevated in the ErLR- and EdLR-treated osteoblastic MC3T3-E1 cells for 20 days. Conclusion: These results demonstrated that Glycyrrhiza uralensis extracts may be useful for preventing osteoporosis by increasing cell proliferation, ALP activity, and bone-marker gene expression in osteoblastic cells.

Unique cartilage matrix-associated proteins에 의한 MC3T3-E1 조골세포에서의 고혈당 스트레스 완화 효과 (Unique Cartilage Matrix-Associated Protein Alleviates Hyperglycemic Stress in MC3T3-E1 Osteoblasts)

  • 주현영;박나래;김정은
    • 생명과학회지
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    • 제33권11호
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    • pp.851-858
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    • 2023
  • Unique cartilage matrix-associated protein (UCMA)은 γ-카르복실화(Gla) 잔기가 풍부한 간외 비타민 K 의존 단백질이다. UCMA는 조골세포 분화를 촉진하고 뼈 형성을 강화한다고 보고되고 있지만 고혈당 스트레스 하에서 조골세포에 미치는 영향에 대해서는 아직 알려진 바가 없다. 본 연구에서는 고혈당 조건하에서의 MC3T3-E1 조골세포에서 UCMA 효과를 조사하기 위해 MC3T3-E1 조골세포를 높은 포도당에 노출한 후 재조합 UCMA 단백질을 처리하였다. MC3T3-E1 세포에서 활성 산소종(ROS)의 생성은 고혈당 조건하에서 증가했으나 UCMA 단백질 처리 후 감소했음을 CellROX 및 MitoSOX 염색으로 확인하였다. 또한 고혈당 조건에서 UCMA 단백질을 함께 처리한 MC3T3-E1 세포에서 정량적 중합효소 연쇄반응 결과, 항산화 유전자인 nuclear factor erythroid 2-related factor 2 와 superoxide dismutase 1 발현이 증가하였다. 동일 조건하에서 UCMA 단백질 처리에 의해 heme oxygenase-1 발현 감소와 함께 세포질에서 핵으로의 전위가 감소되었고, 미토콘드리아 분열에 관여하는 dynamin-related protein 1 발현이 증가하였으며, AKT 신호 활성은 억제되었다. 종합적으로 UCMA는 고혈당에 노출된 조골세포에서 ROS 생성을 완화하고, 항산화 유전자 발현을 증가시키고, 미토콘드리아 역학에 영향을 미치며, AKT 신호를 조절하는 것으로 보인다. 본 연구는 UCMA의 세포 메커니즘에 대한 이해를 돕고, 대사 장애 관련한 골 합병증에 대한 새로운 치료제로서의 잠재적 사용 가능성을 제시하고 있다.