• Title/Summary/Keyword: Cultured bone cell

검색결과 292건 처리시간 0.031초

혈소판유래성장인자와 상피성장인자가 치주인대세포와 골수세포의 성상에 미치는 영향 (Effects of platelet-derived growth factor and epidermal growth factor on the characteristics of beagle dog's periodontal ligament and bone marrow cells)

  • 조병도;허익;박준봉;권영혁;이만섭
    • Journal of Periodontal and Implant Science
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    • 제26권2호
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    • pp.491-510
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    • 1996
  • This study was performed to evaluate the effects of platelet-derived growth factor (PDGF) and epidermal growth factor (EGF) on the characteristics of beagle dog's periodontal ligament (BPD) cells and bone marrow (BBM) cells which have the important role on the early stage of periodontal tissue regeneration in vitro. In control group, the cells ($1.5{\times}10^5$cells/ml) were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum, $50{\mu]g/ml$ ascorbic acid, and 10mM/ml ${\beta}-glycerophosphate$. In experimental groups, growth factors, PDGF or EGF(10ng/ml), were added into the above culture condition. And then each group was characterized by examining the cell proliferation rate, amount of total protein synthesis, alkaline phosphatase activity at 1, 5, 9, 13, 17th day after seeding of cells into the culture wells. The results were as follows: 1. Both BPD and BBM cells in PDGF-treated group proliferated more rapidly than non-treated cells. This finding also was observed in EGF-treated group but it was not as prominent as that of PDGF-treated group. The proliferation rates of both cells showed the time-dependent pattern during experimental periods in all three groups. 2. Amount of total protein synthesis was more increased in PDGF-treated group than in control group. But no significant difference between EGF-treated group and control group was observed throughout experimental periods even though the tendency of amount of protein synthesis was time-dependent pattern. 3. Alkaline phosphatase activity also more increased in PDGF-treated group than control group. But slight decrease tendency was seen in both cells of EGF-treated group. From the above results, PDGF appeared to enhance the proliferation and cellular activities including amount of total protein synthesis and alkaline phosphatase activity of BPD and BBM cell, but EGF did not show notable effects. The optimal application of these growth factors was thought to be useful as the adjunctive means in periodontal regeneration procedures.

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Hericium erinaceum 균사체와 자실체 열수 추출물의 몇몇 In-Vitro 및 In-Vivo 생물활성 (Some In-Vitro and In-Vivo Biological Activities of Hot Water Extracts from Fruit Body and Cultured Mycelium of Hericium erinaceum)

  • 정재현;이광호;이신영
    • KSBB Journal
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    • 제22권1호
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    • pp.22-29
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    • 2007
  • H. erinaceum은 현재 산업적으로 매우 주목되고 있는 기능성 소재들 중의 하나이다. 본 연구에서는 식용은 물론 각종 약리효과를 나타내는 H. erinaceum의 기능성 식품의 제품화 연구 일환으로, 지금까지 체계적으로 연구된 바 없는 H. erinaceum의 액체배양을 시도하고, 생성된 균사체의 생리기능성의 변화를 자실체의 물 추출물과 비교, 조사함으로써 H. erinaceum 액체배양 생성물의 생리기능 효과를 평가, 규명하고자 하였다. H. erinaceum의 자실체 및 균사체 물 추출물은 HeLa (human cervix epitheloid carcinoma; 인간 자궁 상피암세포), Raw264.7 (mouse monocyte macrophage; 생쥐 단핵 대식세포), Jurkat (human acute T cell leukemia, 인간 급성 T 백혈병 세포), KATO3 (human gastric carcinoma 인간 위장 암세포), EL4 (mouse T lymphoma 생쥐 T 림프종 세포), LyD9 (mouse bone marrow plurypotent stem cell; 생쥐 공수유래 간세포) 등 각종 암세포주에 대해 농도 의존적으로 증식 억제 효과를 나타내었다. 자실체 추출물은 균사체 추출물보다 대부분의 암세포 증식의 억제 효과가 컸으며, 가장 높은 암세포 증식 억제효과를 보인 자실체 열수 추출물의 효과는 Raw 264.7 및 EL4 세포주를 사용한 Taxol와의 비교실험 결과, 자실체 농도 0.01$\sim$10 mg/ml에서 자실체 사용량의 1/1000에 상당하는 양의 Taxol 효과와 비슷하였다. 자실체 및 균사체 열수 추출물들은 1 mg/ml 농도에서 복강세포의 nitric oxide 생산 유도능을 나타내었으며, lipopolysaccharides 자극으로 더욱 증가하였다. 반면, 비장세포의 증식유도능 및 interleukin-2 유도능에서는 각 시료 단독으로는 증식 유도 및 interleukin-2 유도능을 나타내지 않았으나 각각 lipopolysaccharide와 concanavalin A의 첨가 및 concanavalin A 자극으로 강한 증식유도능 및 interleukin-2 생산능을 나타내었고, 그 효과는 균사체 추출물에서 자실체 보다 더욱 높았다. 또 각 시료는 골수세포에 대해서도 약간의 증식효과를 보였으며, 복강세포의 탐식능을 나타내었다.

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

  • 이관훈;김정근;정진형
    • Journal of Periodontal and Implant Science
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    • 제26권1호
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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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백서 태자 두 개관에서 유래된 조골세포의 증식 및 골결절 형성양상 (THE ASPECT OF PROLIFERATION AND BONE NODULE FORMATION IN OSTEOBLAST-LIKE CELLS DERIVED FROM FETAL RAT CALVARIA IN VITRO)

  • 김시형;남순현;신홍인
    • 대한소아치과학회지
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    • 제24권1호
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    • pp.1-17
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    • 1997
  • 백서 태자 두개관을 효소처리하여 얻은 조골세포 유사세포군의 증식능 및 골결절의 형성에 미치는 dexamethasone의 영향과 형성된 골결절의 조직학적 형태 관찰을 위하여, 12 well 배양접시의 각 well 당 $4{\times}10^4cell$ 을 접종하였으며, 30일간 표준배양액으로만 배양한 군, 표준배양액에 ascorbic acid와 Na-${\beta}$-glycerophosphate를 첨가한 군, 그리고 각각에 dexa methasone을 첨가한 군등 4군으로 분리 배양하여 다음과 같은 결과를 얻었다. 1. 골세포의 증식율은 표준용액으로 배양한 군에 비해 dexamethasone 을 단독으로 투여한 군에서 약간 증가하는 경향을 보였으나 통계적인 유의성은 없었다 (p<0.05). 2. 골세포의 증식율은 ascorbic acid 에 의해 영향을 많이 받았으며, 특히 ascorbic acid와 dexamethasone을 동시에 투여했을때 더욱 현저히 세포증식율이 감소됨을 볼 수 있었다(p<0.05). 3. 배양 초기에는 세포들이 주로 섬유아세포 양으로 나타났으나, 밀생상태에 이르면서 점차 다각형 형태로 바뀌었다. 4. 배양 9일후 지방세포와 연골세포들이 관찰되었으며, 이들은 모두 dexamethasone이 첨가된 군에서 발견되었다. 5. 골결절은 ascorbic acid와 Na-${\beta}$-glycerophosphate가 첨가된 군에서만 형성되었으며, dexamethasone 을 병용첨가한 군에서 광화된 골결절이 현저히 증가됨을 관찰할 수 있었다. 6. 골결절 형성이 되지 않은 부위의 세포들은 평면형상을 이루고 있었으나, 골결절 부위를 덮고있는 세포들은 많은 세포돌기를 가진 조골세포양 세포들로 나타났으며, ascorbic acid, Na-${\beta}$-glycerophosphate 그리고 dexamethasone 을 첨가한 군에서는 침상의 돌기를 가진 광화물질들이 다발의 교원섬유들 사이에 현저히 많이 존재하였으며, 골결절 내부에는 매몰된 골세포(osteocyte)양 세포들이 관찰되었다.

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임플랜트에 연결한 영구자석이 임플랜트 주위 뼈모세포의 분포와 부착에 미치는 영향에 관한 연구 (THE EFFECT OF PERMANENT MAGNET CONNECTING WITH DENTAL IMPLANT ON DISTRIBUTION AND ATTACHMENT OF OSTEOBLAST-LIKE CELL AROUND THE DENTAL IMPLANT)

  • 오나희;최부병;권긍록;백진;이성복
    • 대한치과보철학회지
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    • 제43권4호
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    • pp.511-518
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    • 2005
  • Purpose: The purpose of this study is to find the effect of rare earth magnet's magnetic field of to the osteoblast around the implant by the means of observation number, and distribution around the implant which is connected to the permanent magnet but not, counted and compared by the number of cells attached to the surface of the implant. Material and method: The permanent magnets, made in the healing cap form, were connected to the implant future, and placed on the culture plate, The osteoblast-like cell: MC3T3-E1 were used for cell culture. As the control group, the implant were connected to normal healing cap, and cultured in the same conditions. 48 hours later, using inverted microscope, the number and distribution of osteoblast around the implant were observed, and 72 hours later, the number of the cells attached to the implant were counted. Results: As a result, the implant connected to the permanent magnet had proved to have a more concentrated cell distribution rate than the control group. The implant connected to the permanent magnet, neck area : which has about 10 gauss magnetic force, had more cells than apex area. The implant connected to the permanent magnet had proven to attach to the osteoblast more productively than control group's implant. Conclusions: This research showed that the magnetic field of the permanent magnet affected the distribution and growth rate of the osteoblast around the implant. In order to support this study, it also had need to monitor the progress of the permanent magnet specifically shown on the neck area, which has10 gauss magnetic force. So after additional research on the distribution and attachment of the cells, and further more, on bone formation, it will be concluded that the clinical applications ,such as immediate loading of implant treatment are possible.

HA-coated Zirconia의 생물학적 활성도에 관한 연구 (Biological Activities of HA-coated Zirconia)

  • 남석우;김해원;김현이;양승민;신승윤;이용무;정종평;한수부;최상묵;류인철
    • Journal of Periodontal and Implant Science
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    • 제33권1호
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    • pp.1-11
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    • 2003
  • Hydroxyapatite(HA) has been extensively used as bone graft materials and tooth implant surface coating materials because of its biocompatibility and osteoconductive properties. However, as HA is intrinsically poor in mechanical properties, zirconia($ZrO_2$) was incorporated with HA as reinforcing phases for improvement of mechanical properties. The purpose of this study was to investigate the biological activities of HA-coated zirconia through the cell proliferation test, measurements of alkaline phosphatase activity, and histologic examination. Four kinds of tested blocks were prepared according to the pore size (300-500${\mu}m$/500-700${\mu}m$) and the porosity (70%/90%). Cell proliferation and alkaline phosphatase activity was measured at 1, 7, 14 days. The number of cells proliferate after 7, 14 days were significantly increased in all groups when compared with that of the first day, but there was no significant difference between the 4 groups at each time period. At the 7 day, alkaline phosphatase activities of cells cultured in 4 groups were higher than that of the first day, but there was no significant difference between the 4 groups at each time period. The human gingival fibroblast and MG 63 cell was used to evaluate the cell cytotoxicity using MTT test. The materials tested in the current study turned out to be non-cytotoxic. In histologic examination(SEM), at 1 day there were many cells attached on the surfaces of all kinds of tested blocks. The number of cells were increased over time. At the 14 day, there were more cells proliferated than 1 day and some of the pores of blocks were partially filled with the proliferated cells. The in vitro response of osteoblast-like cells to the HA-coated zirconia showed comparable effect on transformation comparable to hydroxyapatite.

방사선조사가 MC3T3-E1 골모세포주의 TGF-${\beta}_1$ mRNA 발현과 석회화결절 형성에 미치는 영향 (Effects of irradiation on TGF-${\beta}_1$ mRNA expression and calcific nodule formation in MC3T3-E1 osteoblastic cell line)

  • 송주섭;김경아;고광준
    • Imaging Science in Dentistry
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    • 제38권3호
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    • pp.125-132
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    • 2008
  • Purpose : To investigate the effects of irradiation on transforming growth factor ${\beta}_1$ (TGF-${\beta}_1$) mRNA expression and calcific nodule formation in MC3T3-E1 osteoblastic cell line. Materials and Methods : Cells were cultured in alpha-minimum essential medium ($\alpha$-MEM) supplemented with 10% fetal bovine serum and antibiotics. When the cells reached the level of 70-80% confluence, culture media were changed with $\alpha$-MEM supplemented with 10% FBS, 5 mM $\beta$-glycerol phosphate, and $50\;{\mu}g/mL$ ascorbic acid. Thereafter the cells were irradiated with a single dose of 2, 4, 6, 8 Gy at a dose rate of 1.5 Gy/min. The expression pattern of TGF-${\beta}_1$ mRNA, calcium content and calcific nodule formation were examined on day 3, 7, 14, 21, 28, respectively, after the irradiation. Results : The amount of TGF-${\beta}_1$ mRNA expression decreased significantly on day 7 after irradiation of 4, 6, 8 Gy. It also decreased on day 14 after irradiation of 6, 8 Gy. and decreased on day 21 after irradiation of 8 Gy. The amount of calcium deposition decreased significantly on day 7 after irradiation of 4, 8 Gy (P < 0.01) and showed a decreased tendency on day 14, 21 after irradiation of 4, 6, 8 Gy. The number of calcific nodules was decreased on day 7 after irradiation of 4, 8 Gy. Conclusion: Irradiation with a single dose of 4, 6, 8 Gy influences negatively the bone formation at the molecular level by affecting the TGF-${\beta}_1$ mRNA expression that was associated with proliferation and the production of extracellular matrix in MC3T3-E1 osteoblastic cell line.

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RESPONSE OF OSTEOBLASI-LIKE CELLS ON TITANIUM SURFACE TREATMENT

  • Roh Hyun-Ki;Heo Seong-Joo;Chang Ik-Tae;Koak Jai-Young;Han Jong-Hyun;Kim Yong-Sik;Yim Soon-Ho
    • 대한치과보철학회지
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    • 제41권6호
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    • pp.699-713
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    • 2003
  • Statement of problem. Titanium is the most important material for biomedical and dental implants because of their high corrosion resistance and good biocompatibility. These beneficial properties are due to a protective passive oxide film that spontaneously forms on the surface. Purpose. The purpose of this study was to evaluate the responses of osteoblast-like cells on different surface treatments on Ti discs. Material and Methods. Group 1 represented the machined surface with no treatment. Group 2 surfaces were sandblasted with $50{\mu}m\;Al_2O_3$ under $5kgf/cm^2$ of pressure. Groups 3 and 4 were sandblasted under the same conditions. The samples were treated on a titanium oxide surface with reactive sputter depositioning and thermal oxidation at $600^{\circ}C$ (Group 3) and $800^{\circ}C$ (Group 4) for one hour in an oxygen environment. The chemical composition and microtopography were analyzed by XRD, XPS, SEM and optical interferometer. The stability of $TiO_2$ layer was studied by petentiodynamic curve. To evaluate cell response, osteoblast extracted from femoral bone marrow of young adult rat were cultured for cell attachment, proliferation and morphology on each titanium discs. Results and Conclusion. The results were as follows : 1. Surface roughness values were, from the lowest to the highest, machined group, $800^{\circ}C$ thermal oxidation group, $600^{\circ}C$ thermal oxidation group and blasted group. The Ra value of blasted group was significantly higher than that of $800^{\circ}C$ thermal oxidation group (P=0.003), which was not different from that of $600^{\circ}C$ thermal oxidation group (P<0.05). 2. The degree of cell attachment was highest in the $600^{\circ}C$ thermal oxidation group after four and eight hours (P<0.05), but after 24 hours, there was no difference among the groups (P>0.05). 3. The level of cell proliferation showed no difference among the groups after one day, three days, and seven days (P>0.05). 4. The morphology and arrangement of the cells varied with surface roughness of the discs.

Stem Cell Biology, 최근의 진보 (Recent Advancement in the Stem Cell Biology)

  • 한창열
    • Journal of Plant Biotechnology
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    • 제33권3호
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    • pp.195-207
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    • 2006
  • Stem cells are the primordial, initial cells which usually divide asymmetrically giving rise to on the one hand self-renewals and on the other hand progenitor cells with potential for differentiation. Zygote (fertilized egg), with totipotency, deserves the top-ranking stem cell - he totipotent stem cell (TSC). Both the ICM (inner cell mass) taken from the 6 days-old human blastocyst and ESC (embryonic stem cell) derived from the in vitro cultured ICM have slightly less potency for differentiation than the zygote, and are termed pluripotent stem cells. Stem cells in the tissues and organs of fetus, infant, and adult have highly reduced potency and committed to produce only progenitor cells for particular tissues. These tissue-specific stem cells are called multipotent stem cells. These tissue-specific/committed multipotent stem cells, when placed in altered environment other than their original niche, can yield cells characteristic of the altered environment. These findings are certainly of potential interest from the clinical, therapeutic perspective. The controversial terminology 'somatic stem cell plasticity' coined by the stem cell community seems to have been proved true. Followings are some of the recent knowledges related to the stem cell. Just as the tissues of our body have their own multipotent stem cells, cancerous tumor has undifferentiated cells known as cancer stem cell (CSC). Each time CSC cleaves, it makes two daughter cells with different fate. One is endowed with immortality, the remarkable ability to divide indefinitely, while the other progeny cell divides occasionally but lives forever. In the cancer tumor, CSC is minority being as few as 3-5% of the tumor mass but it is the culprit behind the tumor-malignancy, metastasis, and recurrence of cancer. CSC is like a master print. As long as the original exists, copies can be made and the disease can persist. If the CSC is destroyed, cancer tumor can't grow. In the decades-long cancer therapy, efforts were focused on the reducing of the bulk of cancerous growth. How cancer therapy is changing to destroy the origin of tumor, the CSC. The next generation of treatments should be to recognize and target the root cause of cancerous growth, the CSC, rather than the reducing of the bulk of tumor, Now the strategy is to find a way to identify and isolate the stem cells. The surfaces of normal as well as the cancer stem cells are studded with proteins. In leukaemia stem cell, for example, protein CD 34 is identified. In the new treatment of cancer disease it is needed to look for protein unique to the CSC. Blocking the stem cell's source of nutrients might be another effective strategy. The mystery of sternness of stem cells has begun to be deciphered. ESC can replicate indefinitely and yet retains the potential to turn into any kind of differentiated cells. Polycomb group protein such as Suz 12 repress most of the regulatory genes which, activated, are turned to be developmental genes. These protein molecules keep the ESC in an undifferentiated state. Many of the regulator genes silenced by polycomb proteins are also occupied by such ESC transcription factors as Oct 4, Sox 2, and Nanog. Both polycomb and transcription factor proteins seem to cooperate to keep the ESC in an undifferentiated state, pluripotent, and self-renewable. A normal prion protein (PrP) is found throughout the body from blood to the brain. Prion diseases such as mad cow disease (bovine spongiform encephalopathy) are caused when a normal prion protein misfolds to give rise to PrP$^{SC}$ and assault brain tissue. Why has human body kept such a deadly and enigmatic protein? Although our body has preserved the prion protein, prion diseases are of rare occurrence. Deadly prion diseases have been intensively studied, but normal prion problems are not. Very few facts on the benefit of prion proteins have been known so far. It was found that PrP was hugely expressed on the stem cell surface of bone marrow and on the cells of neural progenitor, PrP seems to have some function in cell maturation and facilitate the division of stem cells and their self-renewal. PrP also might help guide the decision of neural progenitor cell to become a neuron.

수종의 생약제제가 hFOB1의 염기성 인산분해 효소 활성에 미치는 영향 (Effects of Several Natural Medicines on Alkaline Phosphatase Activity in hFOB1)

  • 장길용;현아나;김윤상;유형근;신형식
    • 동의생리병리학회지
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    • 제16권5호
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    • pp.1042-1047
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    • 2002
  • Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential for periodontal tissues. Cortex Eucommiae, Eupoly phaga, Semen Cuscutae, Halloysitum Rubrum have been traditionally used as medicines for treatment of bone disease in Korea. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity in human fetal osteoblast cell line (hFOB1) with several natural medicines. hFOB1 added DMEM/F-12 were cultured with dexamethasone as a positive control, and with each natural medicine. ALP activity was measured by spectrophotometer for enzyme activity and naphthol AS-Bl staining was performed for morphometry. All of the natural medicines induced a higher ALP activity compared to negative control, especially, Cortex Eucommiae increased an ALP activity in all experimental groups (p<0.05). In naphthol AS-Bl staining, all of the natural medicines of this study increased the stained area compared to negative control. Especially, Cortex Eucommiae and Eupoly phaga showed statistical significance compared to negative control (p<0.05). These results indicate that Cortex Eucommiae, Eupoly phaga, Semen Cuscutae, Halloysitum Rubrum have an inducing ability of ALP synthesis on osteoblasts.