• Title/Summary/Keyword: MC3T3-E1 Cell

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Effect of Glucose at High Concentrations on the Apoptosis of the Cultured Periodontal Ligament Cells and Osteoblasts (고농도 포도당이 뼈모세포와 치주인대세포의 세포자멸사에 미치는 영향에 관한 연구)

  • Park, Sung-Ho;Jue, Seong-Suk;Hong, Jung-Pyo;Shin, Je-Won
    • Journal of Oral Medicine and Pain
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    • v.32 no.4
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    • pp.357-364
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    • 2007
  • This experiment was designed to clarify the effect of extracellular glucose on the osteoblasts and periodontal ligament cells. The cells were incubated for 24 and 48 hours with ${\alpha}$-MEM including 1,000 mg/L (control group) and 4,500 mg/L (experimental group) of glucose. Then, the expressions of caspase-3, p38 MAPK, JNK-1, and ERK-1 were examined using Elisa assay and Western blot. The results were as follows: 1. The expression of caspase-3 and p38 MAPK was increased by the high extracellular glucose in both cells. 2. The expression of caspase-3 and p38 MAPK was increased greatly in the periodontal ligament cells than the E1 cells by the high extracellular glucose. 3. The expression of JNK-1 was increased by the high extracellular glucose in both cells. 4. The expression of ERK-1 was not changed by the high extracellular glucose in both cells. These results suggest that extracellular glucose at high concentrations may inhibit the periodontal regeneration process increasing cellular apoptosis. And p38 MAPK and JNK-1 pathway may be the most responsible intracellular pathway rather than ERK-1.

THE EFFECT OF PERMANENT MAGNET CONNECTING WITH DENTAL IMPLANT ON BONE EXTRACELLULAR MATRIX FORMATION (임플랜트에 연결한 영구자석의 자력이 뼈의 세포외 기질 생성에 미치는 영향에 관한 실험실적 연구)

  • Won, In-Jae;Baik, Jin;Kwon, Kung-Rock;Lee, Sung-Bok
    • The Journal of Korean Academy of Prosthodontics
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    • v.44 no.5
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    • pp.574-583
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    • 2006
  • Statement of problem : The use of permanent magnetics is increasing in implant dentistry. Purpose : This study is to know the effect of permanent magnetics on bone matrix formation of osteoblasts. Materials and methods : The konus abutment-shaped permanent magnetics were connected to the implant fixture, and placed on the culture plate. The osteoblast-like cell Mc3T3E1 were used for cell culture. As the control group, the implants were connected to titanium healing caps, and cultured in the same conditions of experimental group. After 3. 7, 14 days, cells were cultured, and we measured and compared the amount of collagen type I, osteocalcin, which is bone matrix protein by Western immunoblotting analysis. Results: As a result of Western immunoblotting analysis for estimating the amount of bone extracellular matrix, there was no difference between osteoblast of the experimental group and the control group during 3 and 7day-osteoblast culturing. However when cells were cultured for 14days, the amount of bone extracellular matrix was increased, on the experimental group. Conclusion: From these results, magnetic field of permanent magnetics might have effect on bone formation of osteoblast, especially at initial stage of implant placement. Therefore, their clinical application for implant or bone graft could be possible.

Yam (Dioscorea batatas) Root and Bark Extracts Stimulate Osteoblast Mineralization by Increasing Ca and P Accumulation and Alkaline Phosphatase Activity

  • Kim, Suji;Shin, Mee-Young;Son, Kun-Ho;Sohn, Ho-Yong;Lim, Jae-Hwan;Lee, Jong-Hwa;Kwun, In-Sook
    • Preventive Nutrition and Food Science
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    • v.19 no.3
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    • pp.194-203
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    • 2014
  • Yam (Dioscorea batatas) is widely consumed as functional food for health promotion mainly in East Asia countries. We assessed whether yam root (tuber) or bark (peel) extracts stimulated the activity of osteoblasts for osteogenesis. MC3T3-E1 cells (mouse osteoblasts) were treated with yam root extracts (water or methanol) (study I) or bark extracts (water or hexane) (study II) within $0{\sim}10{\mu}g/mL$ during the periods of osteoblast proliferation (5~10 day), matrix maturation (11~15 day) and mineralization (16~20 day) as appropriate. In study I, both yam root water and methanol extracts increased cell proliferation as concentration-dependent manner. Cellular collagen synthesis and alkaline phosphatase (ALP) activity, both the indicators of bone matrix protein and inorganic phosphate production for calcification respectively, were also increased by yam root water and methanol extract. Osteoblast calcification as cell matrix Ca and P accumulation was also increased by the addition of yam root extracts. In study II, yam bark extracts (water and hexane) increased osteoblast proliferation and differentiation, as collagen synthesis and ALP activity and osteoblast matrix Ca and P deposition. The study results suggested that both yam root and bark extracts stimulate osteogenic function in osteoblasts by stimulating bone matrix maturation by increasing collagen synthesis, ALP activity, and matrix mineralization.

Changes in pre-osteoblast cells associated with non-precious metal cores with dental implants: Pilot test (치과용 임플란트 적용 비귀금속 코어와 관련된 전조골세포의 변화)

  • Park, Jung-Hyun;Kang, Seen-Young;Kim, Jong-Woo;Kim, Jang-Ju;Kim, Woong-Chul;Kim, Ji-Hwan
    • Journal of Technologic Dentistry
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    • v.40 no.2
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    • pp.63-69
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    • 2018
  • Purpose: The purpose of this study is to investigate the non-precious metal core materials used in the dental laboratory to fabricate the implant superstructure by CAD / CAM method. And to observe and compare the morphology and distribution of the osteoblasts in relation to implant osseointegration. Methods: In this study, the mandibular right first molar tooth model was selected as an international standard to produce a single core. Using this model, the impression was made with the silicone rubber, the tooth model was scanned, and a single core was designed and 5-axis milling was performed. The materials used were Cobalt-Chromium and Nickel-Chromium, and the cores for dental implant top structures were fabricated according to the procedures of the dental labs. After the fabrication, the marginal area of the core was separated and cell culture experiment was performed. The osteoblast cells used MC3T3-E1, which is currently widely used. For morphological analysis of osteoblasts, cells were posttreated and observed using CLSM (Confocal Laser Scanning Microscope) and compared. Results: The cell adhesion behavior of the specimen surface measured by CLSM was uniformly distributed in specimen A (Cobalt-Chromium) than in specimen B (Nickel-Chromium). The distribution and changes of the cells were different in the two specimens. Conclusion : It is possible to confirm that specimen A (Cobalt-Chromium) is suitable for the living body through adhesion and proliferation of osteoblasts related to implant osseointegration in the non-precious metal superstructure used after implantation. It is considered that it is preferable to use Co-Cr when fabricating the superstructure.

Effects of the Hot Water Extract Mixtures from Achyranthes bidentata Blume and Panax ginseng on Osteoclast and Osteoblast Differentiation (우슬과 인삼 열수추출 혼합물의 파골세포와 조골세포 분화 효과)

  • Kim, Jin Seong;Lee, Sang Won;Kim, Young Ock;Bang, Man Seok;Oh, Chung Hun;Kim, Chul Tae
    • Korean Journal of Medicinal Crop Science
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    • v.23 no.2
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    • pp.117-124
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    • 2015
  • Osteoporosis induces a bone mineral density loss due to imbalance of bone homeostasis that is achieved by osteoclasts (which are involved in bone resorption) and osteoblasts (which are involved in bone formation). Thus, this study was performed to evaluate the effects of hot water extract of the Achyranthes bidentata Blume (ABB) and Panax ginseng (Gin) on osteoclast and osteoblast differentiation. In this study, there was no cytotoxicity by ABB, 50 and $100{\mu}g/ml$ of Gin significantly decreased cell viability of RANKL-induced osteoclast in RAW264.7 cell (p < 0.01). But, it was $50{\mu}g/ml$ of ABB and Gin mixtures increased due to protective action of ABB. Furthermore, Gin contained groups (Gin, ABB and Gin mixtures) were inhibitory effects on osteoclast differentiation and bone resorption, and increased in osteoblast differentiation activity. Gin clearly inhibited RANKL-induced osteoclast differentiation by decreased calcitonin and TRAP (p < 0.01). Also, these extracts significantly increased calcium accumulation formation of osteoblastic differentiation reagents-induced osteoblast in MC3T3-E1 cell (p < 0.05). These results suggest that ABB and Gin mixtures may be a potential as drug for the treatment of osteoporosis.

Sensitivity of Escherichia coli to Seaweed (Ascophyllum nodosum) Phlorotannins and Terrestrial Tannins

  • Wang, Yuxi;Xu, Z.;Bach, S.J.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.2
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    • pp.238-245
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    • 2009
  • Pure culture experiments were conducted to assess the bacteriostatic and bactericidal effects of phlorotannins (PT) isolated from Ascophyllum nodosum (brown seaweed) on Escherichia coli O157:H7. In Exp. 1, one non-O157:H7 strain (25922) and three strains of E. coli O157:H7 (3081, EDL933 and E318N) were cultured in M9 medium with PT included at 0 (control), 25, 50 or $100{\mu}g/ml$ (n = 3). Bacterial growth was monitored by $OD_{600}$ at 0, 4, 6, 12 and 24 h, and by dilution plating at 0, 4, 6 and 24 h. All strains were inhibited (p<0.001) by PT to varying degrees. At 50 or $100{\mu}g/ml$, PT prevented growth of all four strains. At $25{\mu}g\;PT/ml$, growth of 25922, 3081, E318N and EDL933 was inhibited for 6, 12 and 24 h, respectively, but 25922 and 3081 resumed growth by 12 and 24 h. Direct plating confirmed bactericidal effects of PT on all four strains at $100{\mu}g/ml$, and on EDL933 and E318N at $50{\mu}g/ml$. In Exp. 2, strains 25922 and 3081 were incubated with no tannins or with $50{\mu}g/ml$ of PT, purified condensed tannins (CT) from Quebracho (Schinopsis balansaei), or purified tannic acid from Rhus semialata (Anacardiaceae) as hydrolysable tannins (HT). Strain 3081 was unaffected by HT or CT, but was completely inhibited (p<0.001) by PT at 4, 6 and 24 h. Strain 25922 was unaffected by HT, slightly inhibited by CT, and almost eradicated by PT at 4 and 6 h. Transmission electron microscopy revealed tannin-mediated alterations to bacterial cell walls. Phlorotannins from A. nodosum exhibit growth-inhibiting and bactericidal effects in vitro against the strains of E. coli O157:H7 investigated. Anti-E. coli efficacy of A. nodosum PT is superior to that of terrestrial tannins purified from Quebracho and from Rhus semialata.

Effects of Eisenia bicyclis Extracts on the Proliferation and Activity of Osteoblasts and Osteoclasts (대황 추출물이 조골세포와 파골세포의 성장과 활성에 미치는 영향)

  • Kim, Seoyeon;Jeon, Myeong-Jeong;Cheon, Jihyeon;Lee, Sang-Hyeon;Kong, Changsuk;Kim, Yuck Yong;Yu, Ki Hwan;Kim, Mihyang
    • Journal of Life Science
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    • v.24 no.3
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    • pp.297-303
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    • 2014
  • The effects of Eisenia bicyclis extracts on osteoblast differentiation and osteoclast formation were investigated. The proliferation of MC3T3-E1 osteoblastic cells was tested in an MTT assay. Treatment with E. bicyclis ethanol extract increased cell proliferation by approximately 128% at a concentration of 10 ${\mu}g/ml$. The ALP activities in the MC3T3-E1 cells was 179% higher when the E. bicyclis ethanol extract was processed at a concentration of 50 ${\mu}g/ml$. The proliferation of RAW 264.7 osteoclastic cells decreased significantly in response to treatment with the E. bicyclis extracts. Moreover, the proliferation of the RAW 264.7 osteoclastic cells treated with E. bicyclis hot water extract decreased by nearly 80%. In addition, the E. bicyclis extract reduced the number of tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells from osteoclastic RAW 264.7 cells. These results indicate that E. bicyclis extracts have an anabolic effect on bone through the promotion of osteoclast differentiation and suggest that the extracts could be used in the treatment of common metabolic bone diseases.

Fabrication of Calcium Phosphate Scaffolds Using Projection-based Microstereolithography and Their Effects on Osteogenesis (투영기반 마이크로 광조형 기술을 이용한 3 차원 인산칼슘 인공지지체 제작 및 골 분화 영향)

  • Seol, Young-Joon;Park, Ju-Young;Cho, Dong-Woo
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.35 no.11
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    • pp.1237-1242
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    • 2011
  • Calcium phosphates are very interesting materials for use as scaffolds for bone tissue engineering. These materials include hydroxyapatite (HA) and tricalcium phosphate (TCP), which are inorganic components of human bone tissue and are both biocompatible and osteoconductive. Although these materials have excellent properties for use as bone scaffolds, many researchers have used these materials as additives to synthetic polymer scaffolds for bone tissue regeneration, because they are difficult to manufacture three-dimensional (3D) scaffolds. In this study, we fabricated 3D calcium phosphate scaffolds with the desired inner and outer architectures using solid freeform fabrication technology. To fabricate the scaffold, the sintering behavior was evaluated for various sintering temperatures and slurry concentrations. After the fabrication of the calcium phosphate scaffolds, in-vitro cell proliferation and osteogenic differentiation tests were carried out.

A Biocompatibility Evaluation of Hydroxyapaite·Titania Surface for Dental Implant (임플란트 적용을 위한 하이드록시아파타이트·이산화티탄 표면의 생체적합성 평가)

  • Kang, Min-Kyung;Bae, Sung-Suk
    • Journal of dental hygiene science
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    • v.16 no.1
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    • pp.70-76
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    • 2016
  • The objective of this study was to fabricate hydroxyapatite (HA) containing titania layer by HA blasting and anodization method to obtain advantages of both methods and evaluated biocompatibility. To fabricate the HA containing titania layer on titanium, HA blasting treatment was performed followed by microarc oxidation (MAO) using the electrolyte solution of 0.04 M ${\beta}$-glycerol phosphate disodium salt n-hydrate and 0.4 M calcium acetate n-hydrate on the condition of various applied voltages (100, 150, 200, 250 V) for 3 minutes. The experimental group was divided according to the surface treatment procedure: SM (simple machined polishing treatment), HA, MAO, HA+MAO 100, HA+MAO 150, HA+MAO 200, HA+MAO 250. The wettability of surface was observed by contact angle measurement. Biocompatibility was evaluated by cell adhesion, and cell differentiation including alkaline phosphatase activity and calcium concentration with MC3T3-E1 cells. The porous titanium oxide containing HA was formed at 150 and 200 V. These surfaces had a more hydrophilic characteristic. Biocompatibility was demonstrated that HA titania composite layer on titanium showed enhanced cell adhesion, and cell differentiation. Therefore, these results suggested that HA containing titania layer on titanium was improved biological properties that could be applied as material for dental implant system.

Effects Of $Interferon-{\gamma}$ On The Biological Activity Of Mouse Osteoblast MC3T3/E1 Cells In Culture (($Interferon-{\gamma}$)가 마우스 조골세포의 생물학적 활성에 미치는 영향에 관한 연구)

  • Lee, Kwan-Hoon;Kim, Jung-Geun;Chung, Chin-Hyung
    • Journal of Periodontal and Implant Science
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    • v.26 no.1
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    • pp.216-229
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    • 1996
  • Interferon(IFN) is a sort of glycoproteins that are produced by activated lymphocyte, monocyte and fibroblast. IFN has anti-viral effects, immuno-defensive mechanism and regulating properties to the several kinds of cells that includes affect on the bone formation and resorption. The effect of IFN on the osteoclast & other tissue cells has been studied in a number of researchers with the limited reports on the osteoblast. The purpose of this study was to evaluate the effects of IFN on the osteoblastic function. The MC3T3/El cell(Mouse osteoblast) was incubated in ${\alpha}-minimum$ essential medium containing 10% FBS. To detect the cytotoxic effect of $IFN-{\gamma}$ on osteoblast, the cells were cultured in 96-well plate to which $IFN-{\gamma}$ of various concentrations were added for 2 days. After staining with trypan blue, total cells and living cells were counted under microscope. To determine the activity of alkaline phosphataset(ALP), various concentrations of $IFN-{\gamma}$ were treated to culture medium, and biochemical assay was performed. $IFN-{\gamma}$ and $IFN-{\gamma}$ plus cycloheximide were added to culture medium separately and then ALP activity were determined. To detect the effect of the $IFN-{\gamma}$ on the bone formation of osteoblast, long-term culture was performed, and calcified nodule formation were observed using von Kossa's staining. After the addition of $IFN-{\gamma}$ with various concentrations to the medium, no cytotoxic effect of $IFN-{\gamma}$ was detected at any concentration. The significant increase in ALP activity of osteoblast were found the concentration of $IFN-{\gamma}$ 500-2500U/ml and the culture time of 24-48 hours respectively. The enhancement of ALP activity by $IFN-{\gamma}$ of osteoblast was decreased significantly by the treatment of cycloheximide. After long-term culture of osteoblast, the nodule formation was found to be increased in number and density by the addition of 500 U/ml $IFN-{\gamma}$. These results suggest that $IFN-{\gamma}$ was affected on the bone formation of osteoblast. Forthemore this kind of study or $IFN-{\gamma}$ to osteoblast will be held continuously.

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