Restorative Dentistry and Endodontics

  • 제27권6호
  • /
  • Pages.600-611
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  • 2002
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  • 2234-7658(pISSN)
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  • 2234-7666(eISSN)
대한치과보존학회 (The Korean Academy of Conservative Dentistry)
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클로로필린이 첨가된 탈구치아 보존액에서의 치주인대 세포 생활력에 대한 연구

In Vitro Assessment on Viability of Human Periodontal Ligament Cells after Storage in Chlorophyllin-added Medium

  • 정원균 (연세대학교 대학원 치의학과) ;
  • 김진 (연세대학교 대학원 치의학과) ;
  • 이은주 (연세대학교 대학원 치의학과) ;
  • 이승종 (연세대학교 대학원 치의학과)
  • 발행 : 2002.11.01
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초록

본 연구의 목적은 클로로필린이 치주인대 세포 생활력에 미치는 영향을 평가하는데 있다. 치주인대 세포는 건강한 사람에서 발치한 소구치의 치주인대 조직으로부터 채취하여 배양하였다. 비교 기준을 설정하기 위하여, HBSS 내에서 $37^{\ circ}C$로 보관한 치주인대 세포수의 50%가 생존하는데 소요되는 시간을 MTT 분석법에 의해 측정하였다. 그 결과는 6시간이었다. HBSS에 클로로필린 10, 100, 500 nM이 각각 첨가된 3군의 실험 보존액과 F-medium, ViaSpan, Likorol 등 모두 6군의 실험 보존액을 96-well plate에 접종한 후, 치주인대 세포를 동일한 수로 분주하였다. 이를 6시간동안 보관한 후, 각 실험군 보존액에서의 세포 생활력을 MTT 분석법으로 측정하였다 또한, HBSS와 F-medium 및 클로로필린 500 nM이 첨가된 HBSS군의 세포들에 대해 유식세포 계측을 시행하여 각각의 세포주기를 분석하였다. 실험 결과, 클로로필린 500 nM이 첨가된 HBSS에서 보관한 세포들이 가장 높은 생활력을 나타내었으며, 클로로필린이 첨가되지 않은 HBSS에 비해서 유의하게 양호한 세포 생활력 유지 능력을 보였다. 클로로필린으로 처치한 세포들은 클로로필린의 농도가 커짐에 따라 세포 생활력이 증가하는 양상을 나타내었다. 유식세포 계측 결과, HBSS와 F-medium 및 클로로필린 500 nM이 첨가된 HBSS에서 보관한 세포의 77~80%가 G0-G1 단계의 세포 주기로 측정되어, 대부분의 세포들이 안정된 세포 대사 상태를 보이는 것으로 나타났다. 결론적으로, 클로로필린 처치는 치주인대 세포의 생활력 유지에 도움이 되는 것으로 사료된다.

키워드

참고문헌

  1. Andreasen JO, Hjorting-Hansen E, Replantation of teeth: I. Radiographic and clinical study of 110 human teeth replanted after accidental loss. Acta Odontol Scand 1966;24:263-85 https://doi.org/10.3109/00016356609028222
  2. Andreasen JO, Hjorting-Hansen E, Replantation of teeth: .II. Histological study of 22 replanted anterior teeth in humans. Acta Odontol Scand 1966;24:287-306 https://doi.org/10.3109/00016356609028223
  3. Blomlof L. Milk and saliva as possible storage media for traumatically exarticulated teeth prior to replantation. Swed Dent J 1981; (suppl, 8):5-23
  4. Krasner P, Rankow HJ. New philosophy for treatment of avulsed teeth. Oral Surg Oral Med Oral Pathol 1995;79:616-23 https://doi.org/10.1016/S1079-2104(05)80105-2
  5. Layug ML, Barrett EJ, Kenny DJ, Interim storage of avulsed permanent teeth. J Can Dent Assoc 1998 May;64(5):357-63, 365-9
  6. Nordenvall KJ. Milk as storage medium for exarticulated teeth: report of a case. ASDC J Dent Child 1992;59:150-5
  7. Ashkenazi M, Sarnat H, Keila S. In vitro viability. mitogenicity and clonogenic capacity of periodontal ligament cells after storage in six different media. Endod Dent Traumatol 1999;15:149-156 https://doi.org/10.1111/j.1600-9657.1999.tb00793.x
  8. Marino TG. West LA, Liewehr FR et al. Determination of periodontal ligament cell viability in long shelf-life milk. J Endodon 2000;26:699-702 https://doi.org/10.1097/00004770-200012000-00005
  9. Harkacz O, Cames D, Walker W. Determination of periodontal ligament cell viability in the oral rehydration fluid Gatorade and milks of varying fat content. J Endodon 1997;23:687-90 https://doi.org/10.1016/S0099-2399(97)80402-5
  10. American Association of Endodontists. Treatment of the avulsed tooth. Recommended guidelines of the American Association of Endodontists. Endodontic Today 1994
  11. Olson BD, Mailhot JM, Anderson RW et al. Comparison of various transport media on human periodontal ligament cell viability. J Endodon 1997;23:676-79 https://doi.org/10.1016/S0099-2399(97)80399-8
  12. Todo S, Nery J, Yanaga K, Podesta I, Gordon RD, Starzl TE. Extended preservation of human liver grafts with UW solution. JAMA 1989;261:711-4 https://doi.org/10.1001/jama.261.5.711
  13. Hiltz J. Trope M. Vitality of human lip fibroblasts in milk, Hank's balanced salt solution and ViaSpan storage media. Endod Dent Traumatol 1991;7:69-72 https://doi.org/10.1111/j.1600-9657.1991.tb00187.x
  14. Hupp JG. Trope M, Mesaros SV, Aukhil I. Tritiated thymidine uptake in periodontal ligament cells of dog's teeth stored in various media for extended periods. Endod Dent Traumatol 1997;13:223-7 https://doi.org/10.1111/j.1600-9657.1997.tb00044.x
  15. Ashkenazi M, Marouni M, Sarnat H. In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in four media at room temperature. Endod Dent Traumatol 2000;16:63-70 https://doi.org/10.1034/j.1600-9657.2000.016002063.x
  16. Fischer S, Maclean AA, Liu M, Cardella JA, Slutsky AS, Suga M, Moreira JF. Keshavjee S. Dynamic changes in apoptotic and necrotic cell death correlate with severity of ischsmia-reperfusion injury in lung transplantation. Am J Respir Crit Care Med 2000;162(5):1932-9 https://doi.org/10.1164/ajrccm.162.5.9910064
  17. Sitasaward SL, Patole MS, Effect of cryopreservation on lipid peroxidation in chicken cornea. Indian J Exp BioI 1998;36(10):1025
  18. Hyon SH, Kim DH. Long-term preservation of rat pancreatic islets under physiological conditions. J Biotechnol 2001;23:85(3):241-6
  19. Kamat JP, Boloor KK, Devasagayam TP. Chlorophyllin as an effective antioxidant against membrane damage in vitro and ex vivo. Biochim Biophys Acta 2000 Sep 27;1487(2-3):113-27 https://doi.org/10.1016/S1388-1981(00)00088-3
  20. Chung WY, Lee JM, Park MY, Yook JI, Kim J, Chung AS, Surh YJ, Park KK. Inhibitory effects of chlorophyllin on 7,12-dimethylbenz[a]anthracene-induced bacterial mutagenesis and mouse skin carcinogenesis. Cancer Lett 1999 Oct 18;145(1-2):57-64. https://doi.org/10.1016/S0304-3835(99)00229-3
  21. Kim J, Yook JI, Park KK, Jung SY, Hong JC, Kim KJ, Kim JA, Chung WY. Anti-promotion effect of chiorophyllin in DMBA-TPA-induced mouse skin carcinogenesis. Anticancer Res. 2000 May-Jun;20(3A):1493-8
  22. Dashwood RH: Chlorophylls as anticarcinogens (Review). In J Oncol 1997;10:721-27
  23. Kumar SS. Devasagayam TP. Bhushan B, Verma NC. Scavenging of reactive species by chlorophyllin: an ESR study. Free Radic Res 2001 Nov;35(5):563-74 https://doi.org/10.1080/10715760100301571
  24. Gomez LA, Alekseev AE, Aleksandrova LA, Brady PA, Terzic A. Use of the MTT assay in adult ventricular cardiomyocytes to assess viability:
  25. Anderson HJ, Jong GD, Vincent I, Roberge M. Short note. Flow cytometry of mitotic cells. Exp Cell Res 1998;238:498-502 https://doi.org/10.1006/excr.1997.3862
  26. Kuru L, Parkar MH, Griffiths GS, Newman HN, Olsen I. Flow cytometry analysis of gingival and periodontal ligament cells. J Dent Res 1998 April;77(4):555-564 https://doi.org/10.1177/00220345980770040801
  27. Schwartz O. Cryopreservation as long-term storage of teeth for transplantation or replantation. Int J Oral Maxillofac Surg 1986;15:30-32 https://doi.org/10.1016/S0300-9785(86)80008-4
  28. Andreasen JO. Atlas of replantation and transplantation of teeth. Philadelphia: WB Saunders, 1992;242-56
  29. Blomlof L. Otteskog P. Viability of human periodontal ligament cells after storage in milk or saliva. Scand J Dent Res 1980;88:436-40
  30. Pettiette M, Hupp J, Mesaros S, Trope M. Periodontal healing of extracted dog's teeth air-dried for extended periods and soaked in various media. Endod Dent Traumatol 1997;13:113-8 https://doi.org/10.1111/j.1600-9657.1997.tb00023.x
  31. Huang SC. Remeikis NA. Daniel JC. Effects of longterm exposure of human periodontal ligament cells to milk and other solutions. J Endodon 1996;22:30-33 https://doi.org/10.1016/S0099-2399(96)80233-0
  32. Somerrman MJ, Archer SY, Imm GR, Foster RA. A comparative study of human ligament cells and gingival fibroblasts in vitro. J Dent Res 1988 January;67(1) :66-70 https://doi.org/10.1177/00220345880670011301
  33. Lekic PC, Kenny DJ, Moe HK, Barrett EJ. McCulloch AG, Relationship of the clonogenic capacity to plating efficiency and dye staining of human periodontal ligament cells: implication for tooth replantation. J Periodont Res 1996;31:294-300 https://doi.org/10.1111/j.1600-0765.1996.tb00496.x
  34. D' Alessandro AM, Kalayoglu M, Sollinger HW, Pirsch JD, Southard JH. Belzer FO. Current status of organ preservation with University of Wisconsin solution. Arch Pathol Lab Med. 1991;115(3):306-310
  35. Halberstadt M, Athmann S, Winter R, Hagenah M. Impact of transportation on short-term preserved corneas preserved in Optisol-GS, Likorol, Likorol-DX. and MK-medium. Cornea 2000;19(6):788-791 https://doi.org/10.1097/00003226-200011000-00006
  36. Santanam N, Ramachandran S, Parthasarathy S. Oxygen radicals. antioxidants. and lipid peroxidation. Semin Reprod Endocrinol 1998;16(4):275-80 https://doi.org/10.1055/s-2007-1016287
  37. Halliwell B, Chirico S. Lipid peroxidation: its mechanism. measurement, and significance. Am J Clin Nutr 1993;57(suppl):715S-25S https://doi.org/10.1093/ajcn/57.5.715S
  38. Halliwell B, Murcia A, Chirico S, Aruoma OI. Free radicals and antioxidants in food and in vivo: what they do and how they work. Critical reviews in food science and nutrition. 1995;35(1&2):7-20 https://doi.org/10.1080/10408399509527682
  39. Tewari S. Gupta V. Bhattacharya S. Comparative study of antioxidant potential of tea with and without additives. Indian J Physiol Pharmacol 2000 Apr;44(2): 215-9
  40. Kamat JP. Boloor KK, Devasagayam TP. Chlorophyllin as an effective antioxidant against membrane damage n vitro and ex vivo. Biochim Biophy Acta 2000 Sep 27;1487(2-3):113-27 https://doi.org/10.1016/S1388-1981(00)00088-3