대한치과교정학회지 (The korean journal of orthodontics)

  • 제34권6호
  • /
  • Pages.514-525
  • /
  • 2004
  • /
  • 2234-7518(pISSN)
  • /
  • 2005-372X(eISSN)
대한치과교정학회 (The Korean Association Of Orthodontists)
권호 표지

니코틴이MG63 조골세포주의 오스테오칼신과오스테오프로 테제린의 생성 및 mRNA 발현에 미치는 영향

Effects of nicotine on the formation of osteocalcin and osteoprotegerin and synthesis of its mRNA in MG63 osteoblast-like cell

  • 양준호 (부산대학교 치과대학 교정학교실) ;
  • 박수병 (부산대학교 치과대학 교정학교실)
  • Yang, Jun-Ho (Orthodontic Department of Dental College, Busan National University) ;
  • Park, Soo-Byung (Orthodontic Department of Dental College, Busan National University)
  • 발행 : 2004.12.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

흡연이 골형성 세포의 기능을 억제한다는 것은 이미 잘 알려져 있으나 대다수의 연구가 주로 역학조사에 기초하여 이루어져 왔다. 본 연구의 목적은 흠연과 골형성 세포의 상관관계를 실험적으로 평가하기 위한 것이다. 본 연구를 위해 인간 골육종 세포인 MG63 조골제포주를 이용하였으며, 니코틴이 조골세포의 증식, 염기성 인산분해효소의 활성, 오스테오칼신곽 오스테오프로테제린의 생성 그리고 mRNA발현에 미치는 영향을 관찰하였다 MG63조골세포를 니코틴 농도가 각각 $1.0{\mu}M$,\;1.0mM, 2.5mM, 5.0mM, 7.5mM, 그리고 10.0mM,이 함유된 배지에서 1일, 2일, 3일 그리고 6일 동안 배양 실험을 하여 다음과 같은 결과를 얻었다. MG63 조골세포의 증식이 저농도의 니코틴에서는 일시적으로 활성화되었지만 5.0mM 이상의 고농도에서는 억제되었다. 염기성 인산분해효소의 활성은 니코틴 농도가 증가함에 따라 감소하였다. 오스테오칼신 생성양은, 배양 1일 후, 5.0mM 이하의 농도에서 증가하였으나 7.5mM 이상의 고농도에서는 감소하였다. MG63 조골세포를 3일 배양한 경우, 오스테오칼신 생성양은 1.0mM의 저농도에서도 감소하였다 니코틴이 오스테오칼신 단백질 생성과 오스테오칼신 mRNA생성에 미치는 영향은 1일과 3일에서 다소 차이가 있었다. 오스테오프로테제린의 생성양은 모든 실험군에서 니코틴을 함유하지 않은 대조군보다 감소하였다. 하지만 mRNA 수치는 단백질 생성과 상반되게 7.5mM (3일), 5.0mM (6일) 이상의 고농도에서 증가하였다.

The purpose of this study was to evaluate the correlation between nicotine and the activity of bone forming cell. MG63 osteoblast-like cells were used for this study. Several factors were examined including the proliferation of cell, alkaline phosphatase activity, the formation of osteocalcin and osteoprotegerin. and the synthesis of its mRNA. MG63 osteoblast-like cells were incubated for 1, 2, 3 and 6 days with nicotine added to the culture medium in 1.0 ${\mu}M$, 1.0mM, 2.5mM, 5.0mM, 7.5mM, and 10.0mM concentrations. The proliferation of MG63 osteoblast-like cells was temporarily activated at the low nicotine concentrations. At high concentrations (>5.0 mM), however. it was suppressed. Alkaline phosphatase activity was suppressed in a dose-dependent manner as the concentration of nicotine increased. Osteocalcin decreased in a dose-dependent manner at high nicotine concentrations of more than 7.5mM and the same result was show when the osteoblasts were treated with low concentrations for longer than 3 days. There was a difference in the influence of nicotine on the synthesis of osteocalcin mRNA and formation of osteocalcin itself at 1 and 3 days. Generally, osteoprotegrin notably declined in all experimental groups. However, the level of its mRNA increased at high nicotine concentrations of more than 7.5mM after 3 days and more than 5.0mM after 6days.

키워드

참고문헌

  1. Bergstrom J, Eliasson S, Preber H. Cigarette smoking and periodontal boneloss. J Periodontol 1991;62:242-6 https://doi.org/10.1902/jop.1991.62.4.242
  2. Daniell HW. Postmenopaussal tooth loss. Contribution to edentulism by osteoporosis and cigarette smoking. Arch Intern Med 1983;143:1678-82 https://doi.org/10.1001/archinte.143.9.1678
  3. Cuff MJ, McQuade MJ,Scheidt MJ,Sutherland DE, Van Dyke TE. The presence of nicotine on root surfaces of penodontally diseased teeth in smokers. J PeriodontoI 1989:60: 564-9
  4. Raulln LA. McPherson JC 3rd, McQuade MJ, Hanson BS. The effect of nicotine on the attachment of human fibroblasts to glass andhuman rootsurfaces in vitro. J PenodontoI 1988;59:318-25
  5. Stevenson JC, Lees B, Devenport M, Cust MP, Ganger KF. Determinants of bone density in normal women.risk factors for future osteoporosis? BMJ 1989:298:924-8
  6. Hollinger JO, Schmitt JM, Hwang K. Soleymani P, Buck D. Impact of nicotine on bone healing. J Blomed Mater Res 1999:45:294-301
  7. Aloia JF, Cohn SH, Vaswani A, Yeh JK, Yuen K, ElliS K. Risk factors for postmenopausal osteoporosis. Am J Med 1985;78:95100.
  8. Buck DL, Griffith DA, Mills MJ. Histologic evidence for lipid during human tooth movement. AmJ Orthod 1973;64:619-24 https://doi.org/10.1016/0002-9416(73)90292-3
  9. Schwartz AM. Tissue changes incidental to orthodontic tooth movement. IntJ Orthod 1931;18:331-52
  10. Price PA, Poser JW,Raman N. Primary structure of thegamrna-carboxy glutamic acid-containing protein from bovine bone. Proc Natl Acad SciUSA 1976;73:3374-5 https://doi.org/10.1073/pnas.73.10.3374
  11. Lian JB, Gundberg CM. Osteocalcin: biochemical considerations andclinical applications. Clin Orthop Rei Res 1988;226:267-91
  12. Price PA, Urist MR, Otawara Y. Matrix glaprotein, a new -carboxyglutamic acid containing protein which is associated with the organic matrix of bone. Biochem Biophys Res Commun 1983; 117:765-71 https://doi.org/10.1016/0006-291X(83)91663-7
  13. Lacey DL, Timms E, Tan HL,Kelley MJ, Dunstan CR, Burgess T, Elliot R, Colombero A, Elliot G, Scully S, Hsu H, Sullivan J, Hawkins N, Davy E, Capparelli C, Eli A, Qian YX, Kaufman S, Sarosi I, Shalhoub V, Senaldi G, Guo J, Delaney J, Boyle WJ. Osteoprotegerin ligand is a cytokine thatregulates osteoclast differentiation andactivation. Cell 1998;93:165-76 https://doi.org/10.1016/S0092-8674(00)81569-X
  14. Simonet WS, Lacey DL, Dunstan CR, Kelleym M,Chang MS, Luthy R, Nguyer HQ, Wooden S, Bennett L, Boone T, Shimomoto G, De. Rose. M. Osteoproteqerln.a novel secreted protein involved in theregulation of bonedensity. Cell 1997;89:303-19
  15. Takahashi N, Udagawa N, Suda T. A new member of tumor necrosis factor ligand family, ODF/OPGL/TRANCE/RANKL, regulates osteoclast differentiation and function. Biochem Biophys Res Commun 1999;256:449-55 https://doi.org/10.1006/bbrc.1999.0252
  16. Ramp WK, Lenz LG, Galvin RJS. Nicotine inhibits collagen synthesis and alkaline phosphatase activity, but stimulate DNA synthesis in osteoblast-like cells. Proc Soc Exp Bial Med 1991; 19736-43
  17. Hoffmann 0, Adams JD. Carcinogenic tobacco-specific Nnitrosamines in snuff and in the saliva of snuff dippers. Cancer Res 1981:41 :4305-8
  18. Pindborg JJ. Tobacco andgingivitis. Statistical examination of the significance of tobacco in thedevelopment of ulceromembranous gingivitis and in the formation of calculus. J Dent Res 1947;26:261-4 https://doi.org/10.1177/00220345470260030901
  19. Sheiham A. Periodontal disease and oral cleanliness in tobacco smokers. J Periodontol 1971 :42:259-63
  20. Feldman RS, Bravacos JS, Rose CL. Association between smoking different tobacco products and periodontal disease indexes. J Periodontol 1983; 54:481-7 https://doi.org/10.1902/jop.1983.54.8.481
  21. Lee CK, Fulp C, Bombick BR, Doolittle DJ. Inhibition of mutagenicity of N-nitrosamines by tobacco smoke and its constituents. Mut Res 1996;367:83-92 https://doi.org/10.1016/0165-1218(95)00078-X
  22. Bridget, Turnbull. Smoking and periodontal disease. J new Zealand SocPeriodontoI 1995:79:10-5
  23. Peacock ME, Sutherland DE, Schuster GS, Brennan WA, O'Neal RB, Strong SL, Van Dyke TE. The effectof nicotine on reproduction and attachment of human gingival fibroblast in vitro. J PeriodontoI 1993;64:658-65 https://doi.org/10.1902/jop.1993.64.7.658
  24. Bordin S, Page R, Narayanan A. Heterogeneity of normal human diploid fibroblasts:isolation an characterization of one phenotype. Science 1984;223 :171-3 https://doi.org/10.1126/science.6691142
  25. Fang MA, Frost PJ, lida-Klein A, Hahn TJ. Effects of nicotine on cellular function in UMR 106-01 osteoblast-like cells. Bone 1991 ;12:283-6 https://doi.org/10.1016/8756-3282(91)90077-V
  26. Walker LM, Preston MR, Magnay JL, Thomas PB, EI Haj AJ. Nicotinic regulation of c-fos andosteopontin expression in human derived osteoblast-like cells and human trabecular bone organi culture. Bone 2001 ;28:603-8 https://doi.org/10.1016/S8756-3282(01)00427-6
  27. Henderson JS, Johnson RB. The effects of smokeless tobacco extract on bone nodule formation and mineralization by chick osteoblasts in vitro. Archs oral Bioi 1995:40:615-21
  28. Laroche M, Lasne Y, Felez A, Moulinier L, Bon e, Cantagrel A. Osteoclacin andsmoking. Rev Rhum Ed Fr 1994;61:433-6
  29. 이지영, 황성명, 시험관에서 Osteocalcin 및 Tumor necrosis factor-가 생쥐골수세포의 Tartrate-resistant acid phosphatase 양성세포 형성에 미치는 영향. 서울대학교 논문집, 1994
  30. Lian JB, Dunn K, Key LL. Invitro degradation of boneparticles by human monocytes is decreased with the depletion of thevitamin K-dependent bone protein from the matrix. Endocrinology. 1986;118:1636-42 https://doi.org/10.1210/endo-118-4-1636