대한소아치과학회지 (Journal of the korean academy of Pediatric Dentistry)

  • 제45권2호
  • /
  • Pages.242-249
  • /
  • 2018
  • /
  • 1226-8496(pISSN)
  • /
  • 2288-3819(eISSN)
대한소아치과학회 (Korean Academy of Pediatric Dentistry)
권호 표지
DOI QR코드

DOI QR Code

과잉치 치수 세포와 치주인대 세포의 유전자 발현 비교

Comparison of Gene Expression from Supernumerary Dental Pulp and Periodontal Ligament Stem Cells

  • 이상은 (단국대학교 치과대학 소아치과학교실) ;
  • 김종빈 (단국대학교 치과대학 소아치과학교실) ;
  • 김종수 (단국대학교 치과대학 소아치과학교실)
  • Lee, Sangeun (Department of Pediatric Dentistry, School of Dentistry, Dankook University) ;
  • Kim, Jongbin (Department of Pediatric Dentistry, School of Dentistry, Dankook University) ;
  • Kim, Jongsoo (Department of Pediatric Dentistry, School of Dentistry, Dankook University)
  • 투고 : 2018.01.16
  • 심사 : 2018.03.13
  • 발행 : 2018.05.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

이 연구의 목적은 정량적 실시간 중합효소 연쇄 반응법을 이용하여 발거된 과잉치 치수 및 치주인대 줄기세포의 유전자 발현을 비교하는 것이다. 전신병력이 없는 만 6세 남아 2명의 상악 전치부에 매복된 과잉치를 발거하였다. 같은 날 치수 및 치주인대 세포를 채취하였고, 3계대까지 계대배양하였다. 분석을 위해 상아질모세포 특이 유전자인 Alkaline phosphatase (ALP), Dentin Matrix Protein 1 (DMP-1), Dentin sialophosphoprotein (DSPP), Osteocalcin (OCN) 그리고 Osteonectin (ONT)을 사용했고, glyceraldehyde 3-phosphate dehydrogenase (GAPDH)을 대조군으로 설정했다. 과잉치 치수 세포에서는 ONT, OCN, ALP, DMP-1, DSPP 순서로 발현량이 많았고, 치주인대 세포에서는 DMP-1과 DSPP의 순서만 바뀌었다. 치주인대 세포보다 치수 세포에서 모든 유전자의 발현량이 많았다. 이러한 상아질모세포의 특성을 고려해 보았을 때, 다른 조직으로의 분화 가능성이 있는 과잉치 줄기세포는 유용한 공여부로서 그 잠재력이 있음을 알 수 있었다.

The purpose of this study is to compare the properties of dental pulp and periodontal ligament stem cells from extracted supernumerary teeth by quantitative real-time PCR. Impacted supernumerary teeth in the maxillary anterior region were extracted. Dental pulp and periodontal ligament cells were collected from extracted supernumerary teeth on the same day. After isolation and culture of cells, compare characterization of them by using qRT-PCR. Primer sequences for odontoblasts are ONT, ALP, OCN, DMP-1 and DSPP. On dental pulp group, ONT has the largest quantity of gene expression, followed by OCN, ALP, DMP-1 and DSPP. On periodontal ligament group, ONT has the largest quantity of gene expression, followed by OCN, ALP, DSPP and DMP-1. Analysis of quantitative gene expression data using relative quantification showed that the expression of all genes decreased in periodontal ligament cells. Dental pulp and periodontal ligament stem cells from supernumerary teeth have the properties of odontoblasts. Considering that properties, supernumerary teeth were considered a useful donor site of dental pulp and periodontal ligament stem cells.

키워드

참고문헌

  1. Irma Thesleff, Pekka Nieminent : Tooth morphogenesis and cell differentiation. Current Opinion in Cell Biology, 8:844-850, 1996. https://doi.org/10.1016/S0955-0674(96)80086-X
  2. Ruch JV : Odontoblast Differentiation and the Formation of the Odontoblast Layer. J Dent Res, 64:489-498, 1985. https://doi.org/10.1177/002203458506400402
  3. Couve E, Osorio R, Schmachtenberg O : The Amazing Odontoblast: Activity, Autophagy, and Aging. J Dent Res, 92:765-772, 2013. https://doi.org/10.1177/0022034513495874
  4. Sasaki T, Garant PR : Structure and Organization of Odontoblasts. Anat Rec, 245:235-249, 1996. https://doi.org/10.1002/(SICI)1097-0185(199606)245:2<235::AID-AR10>3.0.CO;2-Q
  5. Papagerakis P, Berdal A, Macdougall M, et al. : Investigation of Osteocalcin, Osteonectin, and Dentin Sialophosphoprotein in Developing Human Teeth. Bone, 30:377-385, 2002. https://doi.org/10.1016/S8756-3282(01)00683-4
  6. Asaumi JI, Shibata Y, Kishi K, et al. : Radiographic examination of mesiodens and their associated complications. Dentomaxillofac Radiol, 33:125-127, 2004. https://doi.org/10.1259/dmfr/68039278
  7. Omer RS, Anthonappa RP, King NM : Determination of the optimum time for surgical removal of unerupted anterior supernumerary teeth. Pediatr Dent, 32:14-20, 2010.
  8. Huang GT, Gronthos S, Shi S : Mesenchymal Stem Cells Derived from Dental Tissues vs. Those from Other Sources: Their Biology and Role in Regenerative Medicine. J Dent Res, 88:792-806, 2009. https://doi.org/10.1177/0022034509340867
  9. Ulmer FL, Winkel A, Kohorst P, Stiesch M : Stem Cells - Prospects in Dentistry. Schweiz Monatsschr Zahnmed, 120:860-883, 2010.
  10. Gronthos S, Brahim J, Gehron Robey P, et al. : Stem Cell Properties of Human Dental Pulp Stem Cells. J Dent Res, 81:531-535, 2002. https://doi.org/10.1177/154405910208100806
  11. Seo BM, Miura M, Shi S, et al. : Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet, 364:149-155, 2004. https://doi.org/10.1016/S0140-6736(04)16627-0
  12. Song JS, Kim SH, Jung HS, et al. : Characterization of Stem Cells Obtained from the Dental Pulp and Periodontal Ligament of Deciduous Teeth. Tissue Eng Regen Med, 7:575-582, 2010.
  13. Livak KJ, Schmittgen TD : Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the $2-{\Delta}{\Delta}CT$ Method. Methods, 25:402-408, 2001. https://doi.org/10.1006/meth.2001.1262
  14. Gadkar VJ, Filion M : New Developments in Quantitative Real-time Polymerase Chain Reaction Technology. Curr Issues Mol Biol, 16:1-6, 2014.
  15. Shih WY, Hsieh CY, Tsai TP : Clinical evaluation of the timing of mesiodens removal. J Chin Med Assoc, 79:345-350, 2016. https://doi.org/10.1016/j.jcma.2015.10.013
  16. Russell KA, Folwarczna MA : Mesiodens - Diagnosis and Management of a Common Supernumerary Tooth. J Can Dent Assoc, 69:362-366, 2003.
  17. Arana-Chavez VE, Massa LF : Odontoblasts: the cells forming and maintaining dentine. Int J Biochem Cell Biol, 36:1367-1373, 2004. https://doi.org/10.1016/j.biocel.2004.01.006
  18. Couve E : Ultrastructural changes during the life cycle of human odontoblasts. Arch Oral Biol, 31:643-651, 1986. https://doi.org/10.1016/0003-9969(86)90093-2
  19. Goldberg M, Kulkarni AB, Young M, Boskey A : Dentin: Structure, Composition and Mineralization. Front Biosci, 3:711-735, 2011.
  20. Begue-Kirn C, Smith AJ, Lesot H, et al. : Comparative analysis of TGFBs, BMPs, IGF1, msxs, fibronectin, osteonectin and bone sialoprotein gene expression during normal and in vitro-induced odontoblast differentiation. Int J Dev Biol, 38:405-420, 1994.
  21. Termine JD, Kleinman HK, Martin GR, et al. : Osteonectin, A Bone-Specific Protein Linking Mineral to Collagen. Cell, 26:99-105, 1981. https://doi.org/10.1016/0092-8674(81)90037-4
  22. Abd-Elmeguid A, Abdeldayem M, Yu DC, et al. : Osteocalcin Expression in Pulp Inflammation. J Endod, 39:865-872, 2013. https://doi.org/10.1016/j.joen.2012.12.035
  23. Beertsen W, Van den Bos T : Alkaline Phosphatase Induces the Deposition of Calcified Layers in Relation to Dentin: An in vitro Study to Mimic the Formation of Afibrillar Acellular Cementum. J Dens Res, 70:176-181, 1991. https://doi.org/10.1177/00220345910700030401
  24. Shiba H, Nakanishi K, Kurihara H, et al. : Proliferative Ability and Alkaline Phosphatase Activity with In Vivo Cellular Aging in Human Pulp Cells. J Endod, 29:9-11, 2003. https://doi.org/10.1097/00004770-200301000-00003
  25. Park JH, Song JS, Lee JH, et al. : Characteristics of Stem Cells Derived from the Periodontal Ligament of Supernumerary Teeth. Tissue Eng Regen Med, 8:123-131, 2011.
  26. Jacob A, Zhang Y, George A : Transcriptional regulation of dentin matrix protein 1 (DMP1) in odontoblasts and osteoblasts. Connect Tissue Res, 55:107-112, 2014. https://doi.org/10.3109/03008207.2014.923850
  27. Alessandra T, Viviana S, Alessandra RJ, et al. : Distribution of DMP-1 and DSPP in human carious dentin: an immunohistochemical analysis. Italian J Anat Embryol, 116:192, 2011.
  28. Suzuki S, Haruyama N, Nishimura F, Kulkarni AB : Dentin sialophosphoprotein and dentin matrix protein-1: Two highly phosphorylated proteins in mineralized tissues. Arc Oral Biol, 57:1165-1175, 2012. https://doi.org/10.1016/j.archoralbio.2012.03.005
  29. Kamata N, Fujimoto R, Yasumoto S, et al. : Immortalization of human dental papilla, dental pulp, periodontal ligament cells and gingival fibroblasts by telomerase reverse transcriptase. J Oral Pathol Med, 33:417-423, 2004. https://doi.org/10.1111/j.1600-0714.2004.00228.x
  30. Hyoun S, Park SH, Choi GW : The Comparison of Gene Expression from Human Dental Pulp Cells and Periodontal Ligament cells. Restor Dent Endod, 34:430-441, 2009.