• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제27권2호
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• pp.103-110
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• 2001

This study was performed to observe the effect of ultrasound(1.0MHz, $0.75W/cm^2\;and\;1.0W/cm^2$) irradiation on cultured MC3T3-E1 cell, osteoblastic like cell with respect to the proliferation, protein synthesis, and alkaline phosphatase activity of the cells. The results were as follows: 1. The proliferation of MC3T3-E1 cells was increased on ultrasound irradiated group compared with control group. 2. The protein synthesis was not apparently increased on ultrasound irradiated group compared with control group. 3. The alkaline phosphatase activity level was not apparently increased on ultrasound irradiated group compared with control group. From the above results and other literatures, we could suggest that the ultrasound with the appropriate intensity and frequency may have important roles in stimulation of cell proliferation. Therefore the ultrasound may be used in the acceleration of the bone regeneration and bone fracture healing.

• 치위생과학회지
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• 제24권3호
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• pp.181-189
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• 2024

Background: Focal adhesions (FAs) is the most important process in the first step of osseointegration between preosteoblasts and titanium (Ti). FAs improvement and pre-osteoblasts cell proliferation leads to successful Ti-based dental implants. This study aimed to confirm the applicability of rosmarinic acid (RA) as a functional substance for improving FAs and cell proliferation of MC3T3-E1 preosteoblasts on Ti surfaces during the first stage of osseointegration for successful Ti-based dental implants. Methods: We used MC3T3-E1 preosteoblasts on Ti discs incubated in a medium supplemented with or without 14 ㎍/ml to decipher the effects of RA on FAs and cell proliferation. FAs and proliferation of MC3T3-E1 cells on Ti discs were assessed via MTT assay. Actin-labeled cells and paxillin contacts were observed and imaged by fluorescent microscopy, and the associated signaling pathways were revealed through western blot analysis. Results: In RA-treated MC3T3-E1 cells on Ti discs, FAs between MC3T3-E1 preosteoblasts and Ti surfaces and the expression of focal adhesion kinase (FAK), phosphorylated FAK and paxillin proteins and filamentous-actin formation increased. RA increased the proliferation of MC3T3-E1 preosteoblasts on the Ti surface as well as the expression of Grab2, Ras, pERK1/2, and ERK1/2. In addition, the expression of secretory leukocyte protease inhibitor and thymosin b4, known as nanomolecules that enhance the interaction between implanted Ti materials and preosteoblasts in the RA-treated MC3T3-E1 preosteoblasts, increased. RA not only increased the FAs of MC3T3-E1 preosteoblasts on the Ti surface through the FAK/Paxillin signaling pathway, but also increased cell proliferation and mitosis through the FAK/Grab2/Ras/ERK1/2 signaling pathway. Conclusion: RA can be applied as an effective functional substrate to improve the FAs and proliferation of MC3T3-E1 preosteoblats on Ti surfaces, which are essential in the first step of osseointegration between implanted Ti and bone tissue for the clinical success of Ti based dental implants.

• 대한기계학회:학술대회논문집
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• 대한기계학회 2008년도 추계학술대회A
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• pp.1700-1703
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• 2008

The purpose of this study was to examine the effects of various mechanical stimuli for MC3T3-E1 cells. Among the several mechanical stimulations, we focused on compressive stain and ultrasound. In this study, we developed a bioreactor capable of applying controlled stimuli to scaffolds. PLLA/PCL scaffold was fabricated by using salt-leaching method. We performed dynamic cell culture using preosteoblasts MC3T3-E1 cells with 1MHz, 30mW/cm2 ultrasound and 10% of compressive strain. Result of CCK-8 analysis at 1, 4, 7, 10 days showed that mechanical stimuli had no significant effect for cell proliferation. However, those stimuli influenced ALP(Alkaline phopatase) activity, which is one of differentiation marker.

• 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.

• 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.

• 대한기계학회:학술대회논문집
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• 대한기계학회 2008년도 추계학술대회A
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• pp.1411-1414
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• 2008

It is reported that mechanical stimulation takes a role in improving cell growth in skeletal system. And various research groups have showed that developed bioreactor to stimulate cell-seeded and threedimensional scaffold. In this study, we designed a custom-made bioreactor capable of applying controlled compression to cell-seeded agarose gel. This device consisted of a circulation system and compression system. In circular system, culture chamber was sealed for prohibiting contamination and media solution was circulated by pump. In compression system, mechanical stimuli were controlled by LabVIEW software and mechanical transfer system. Cell-encapsulated agarose gels were cultured for up to 7 days. There were significant differences between the number of cells grown in dynamic cell culture and in static cell culture from 3 days to 7 days.

• 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.

• 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.

• 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.

• Natural Product Sciences
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• 제11권3호
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• pp.139-144
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• 2005

Bioflavone quercetin is believed to play an important role preventing bone loss by affecting osteoclastogenesis and regulating many systemic and local factors including hormones and cytokines. This study examined how quercetin acts on tumor necrosis factor-alpha ($TNF-{\alpha}$)-mediated apoptosis in MC3T3-E1 osteoblastic cells. Apoptosis assays revealed the dose-dependent acceleration of quercetin on $TNF-{\alpha}-induced$ apoptosis in MC3T3-E1 cells, which was demonstrated by the increased number of positively stained cells in the trypan blue staining and TUNEL assay, and the migration of many cells to the $sub-G_0/G_1$ phase in flow cytometric analysis. In particular, quercetin treatment alone increased the expression of p53 and p21 proteins in the cells. Consequently, this study showed that quercetin accelerates the $TNF-{\alpha}-induced$ apoptosis in MC3T3-E1 osteoblastic cells.