• Title/Summary/Keyword: Alkaline phosphatase and DNA synthetic activity

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The Effects of Static Magnetic Field and Pulsed Electromagnetic Field on Alkaline Phosphatase and DNA synthetic Activity of ME3T3-E1 Cells (정자기장과 맥동전자기장이 MC3T3-E1 세포의 ALP 및 DNA 활성도에 미치는 영향)

  • Son, Jeong-Hee;Bae, Seong-Min;Sung, Jae-Hyun
    • The korean journal of orthodontics
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    • v.27 no.4 s.63
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    • pp.623-632
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    • 1997
  • The purpose of this study was to evaluate the effects of magnetic field on cellular activity of MC3T3-E1 cells. The cellular activity was monitored by alkaline phosphatase and DNA synthetic activity in control, static magnetic field and pulsed electromagnetic field groups. A static magnetic field was applied to the cell by placing one, two, three, foue, and five samarium-cobalt magnets above and below each cell plate for 24hours per day. A pulsed electromagnetic field with a frequency of 100 herz was applied for 10 hours per day. After 10 days of magnetic field exposure, there were increase of alkaline phosphatase activity in static magnetic field groups consisted of one, two and three magnetic groups. Alkaline phosphatase activities were not significantly increased in four and five magnetic groups. Application of pulsed electromagnetic field did not result in significant increase in alkaline phosphatase activity compared to control. DNA synthetic activity in both static and pulsed electromagnetic field group were not significantly different from that in control group. The result of this study suggest that magnetic field could have effect on the metabolism of bone cells related to the cellular metabolic process.

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THE STUDY ON THE EFFECTS OF THE INSULIN-LIKE GROWTH FACTOR-I ON THE BIOLOGICAL ACTIVITY OF THE HUMAN PERIODONTAL LIGAMENT CELLS (Insulin-like growth factor-I 이 치주인대세포의 생물학적 활성도에 미치는 영향에 대한 연구)

  • Kim, Seong-Jin;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.24 no.2
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    • pp.219-237
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    • 1994
  • The ultimate goal of clinical periodontal therapy is to achieve regeneration of a healthy connective tissue reattachment. Conventional therapy including scaling, root planing, gingival curettage, gingivectomy and flap procedures of various types results primarily in repair rather than regeneration of the periodontium. In order for periodontal regeneration to occur, progenitor periodontal ligament cells must migrate to the denuded root surface, attach to it, proliferate and mature into an organized and functional fibrous attachment apparatus. Polypeptide growth factors belong to a class of potent biologic mediators which regulate cell differentiation, proliferation, migration and metabolism. Insulin-like growth factor-I (IGF- I ) of these factors appear to have an important role in periodontal wound healing and bone formation. The purpose of this study is to evaluate the effects of IGF- I on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were obtained from periodontal tissue explants culture of the first premolar tooth extracted for the orthodontic treatment. Cells were cultured in Dulbecco's modified Eagle medium(DMEM) with 10% fetal bovine serum. Fourth to seventh passage cells were plated in 24 well tissue culture plates and medium changed to serum-free medium prior to addition of growth factors. Cell proliferation was measured by the incorporation of $[^3H]-thymidine$ into DNA, Protein synthesis was determined by measurement of $[^3H]-proline$ incorporation into collagenase-digestible protein(CDP) and noncollagenous protein(NCP) according to the method of Peterkofsky and Diegelmann (1971), And alkaline phosphatase activity was measured as one parameter of osteoblastic differentiation. The results were as follows : The DNA synthetic activity was increased in a dose-dependent manner with IGF- I except for 0.1ng/ml concentration of IGF- I At the concentration of 10, 100ng/ml, IGF- I significantly increased the DNA synthetic activity(P<0.05) The total protein, collagen and noncollagen synthesis was increased in a dose-dependent manner with IGF- I except for 0.1ng/ml concentration of IGF- I. At the concentration of 1, 10, 100ng/ml, IGF- I significantly increased the total protein, collagen and noncollagen synthesis activity(P<0.95, P<0.001). The % of collagen was not effected according to the concentration of IGF- I. The alkaline phosphatase activity was increased in a dose-, time-dependent manner with IGF- I (10, 100ng/ml). In conclusions, the present study shows that IGF- I has a potentiality to enhance the DNA synthesis of periodontal ligament cells with including the increase of the total protein and collagen synthetic activity. The use of IGF- I to mediate biological stimulation of periodontal ligament cells shows promise for future therapeutic applications.

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The Effects of 1,25- Dihydroxyvitamin $D_3$ on Expression of IGF-I Gene and Cellular Proliferation in MC3T3-E1 Cells (골아세포의 IGF-I 유전자 발현 및 세포증식에 대한 1,25-dihydroxyvitamin $D_3$의 영향)

  • Choi, Hee-Dong;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.30 no.1
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    • pp.39-52
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    • 2000
  • Polypeptide growth factor belong to a class of potent biologic mediator which regulate cell differentiation, proliferation, migration and metabolism. 1,25-dihydroxyvitamin $D_3$ decrease cell proliferation, and stimulate alkaline phosphatase activity which express in osteoblast during cell differentiation period. IGF-I is known to stimulate cell proliferation and differentiation too. 1,25-dihydroxyvitamin $D_3$ is known to increase IGF-I binding sites and IGF binding protein which inhibite the effect of IGF. The purpose of this study is to evaluate potential role of IGF-I as mediator that control the action of 1,25-dihydroxyvitamin $D_3$. MC3T3-E1 cell were seeded $5{\times}10^5/ml$ at 100mm culture plate in ${\alpha}-MEM$ containing 10% fetal bovine serum. After 48 hour incubation period, medium were changed ${\alpha}-MEM$ containing 5% fetal bovine serum. After 24 hours, $10^{-9}M$ 1,25-dihydroxyvitamin $D_3$ added. Total mRNA was extracted at 0, 6, 24, 48, 72 hour. PRPCR method was programed for the detection of IGF-I mRNA. In the both groups of 1,25-dihydroxy vitamin $D_3$ treated and control, alternative splicing form of IGF-I, IGF-IA and IGF-IB were expressed. In the 1,25-dihydroxyvitamin $D_3$ treated group, IGF-I mRNA expression was matained until 24 hour, there after expression was decresed. MC3T3-E1 cell were seeded $2.5{\times}10^4/ml$ at 24well plate in ${\alpha}-MEM$ containing 10% fetal bovine serum. After 48 hour incubation period, medium were changed ${\alpha}-MEM$ containing 3% fetal bovine serum. After 24 hours, $10^{-9}M$ 1,25-dihydroxyvitamin $D_3$ and 10 ng/ml IGF-I were added separately or together. Cell were cultured for 1 and 3 days, $2{\mu}Ci/ml\;[^3H]$ -thymidine was added for the last 24h of culture of each days. ${[^3H]}$-thymidine incorporation in to DNA was measured and expressed counter per minute(CPM). DNA synthetic activity was significantly decreased by 1,25-dihydroxyvitamin $D_3$ both at 1 day and 3 day, and in the combination group of 1,25-dihydroxyvitamin $D_3$ and IGF-I, DNA synthetic activity was also decreased both at 1 day and 3 days. IGF-I did not affect the DNA synthetic activity compared to control group both at 1 day and 3 day. From the above results, 1,25-dihydroxyvitamin $D_3$ was potent inhibitor of cell proliferaton in MC3T3-E1 cells. It assumed that the effect of 1,25-dihydroxyvitamin $D_3$ on osteoblast proliferation may be mediated in part by decreased level of IGF-I.

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THE STUDY ON EFFECTS OF THE PLATELET-DERIVED GROWTH FACTOR-AA, BB ON THE CELLULAR ACTIVITY OF THE HUMAN PERIODONTAL LIGAMENT CELLS (Platelet-derived growth factor-AA, BB가 치주인대세포의 세포활성에 미치는 영향에 대한 연구)

  • Oh, Sang-Deok;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.24 no.2
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    • pp.303-320
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    • 1994
  • Current acceptable methods For promotin gperiodontal regeneration are base on removal of diseased soft tissue, root treatment, guided tissue regeneration, inteoduction of new graft materials and biological mediators. Platelet-derived growth factor(PDGF) is one of polypeptide growth factor. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of PDGF-AA, BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/10% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Author measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis and alkaline phosphatase activity according to the concentration of PDGF-AA and BB(0, 0.1, 1, 10, 100ng/ml). The results were as follows : The DNA synthetic activity was increased dose dependently by PDGF-AA and BB. The maximum mitogenic effect was at the 100ng/ml of PDGF-AA and 10ng/ml of PDGF-BB. The total protein, collagen and noncollagen systhesis was increased dose dependently by PDGF-AA and BB. The % of collagen was slightly decresed according to the concentration of PDGF-AA and BB. The effect of PDGF-AA and BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. The effect of PDGF-AA and BB on alkaline phosphatase activity did not show any significant, meanwhile the alkaline phosphatase activity of 14 days group showed significnat increase. In conclusion, PDGF-AA and BB may have important roles in stimulation of DNA synthesis in human periodontal ligament cells, which means an increase in collagen-synthesizing cells, and may be useful for clinical application in periodontal regenerative procedures.

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