Journal of Periodontal and Implant Science

Korean Academy of Periodontology (대한치주과학회)
권호 표지
DOI QR코드

DOI QR Code

Identification of stemness and differentially expressed genes in human cementum-derived cells

  • Lee, EunHye (Dental Research Institute, Seoul National University) ;
  • Kim, Young-Sung (Department of Periodontics, Asan Medical Center, University of Ulsan College of Medicine) ;
  • Lee, Yong-Moo (Department of Periodontology and Dental Research Institute, Seoul National University School of Dentistry) ;
  • Kim, Won-Kyung (Department of Periodontics, Asan Medical Center, University of Ulsan College of Medicine) ;
  • Lee, Young-Kyoo (Department of Dentistry, Uijeongbu Eulji Medical Center, Eulji University) ;
  • Kim, Su-Hwan (Department of Periodontics, Asan Medical Center, University of Ulsan College of Medicine)
  • Received : 2021.04.23
  • Accepted : 2021.07.09
  • Published : 2021.10.30
Download PDF
⟨ Previous Next ⟩

Abstract

Purpose: Periodontal treatment aims at complete regeneration of the periodontium, and developing strategies for periodontal regeneration requires a deep understanding of the tissues composing the periodontium. In the present study, the stemness characteristics and gene expression profiles of cementum-derived cells (CDCs) were investigated and compared with previously established human stem cells. Candidate marker proteins for CDCs were also explored. Methods: Periodontal ligament stem cells (PDLSCs), pulp stem cells (PULPSCs), and CDCs were isolated and cultured from extracted human mandibular third molars. Human bone marrow stem cells (BMSCs) were used as a positive control. To identify the stemness of CDCs, cell differentiation (osteogenic, adipogenic, and chondrogenic) and surface antigens were evaluated through flow cytometry. The expression of cementum protein 1 (CEMP1) and cementum attachment protein (CAP) was investigated to explore marker proteins for CDCs through reverse-transcription polymerase chain reaction. To compare the gene expression profiles of the 4 cell types, mRNA and miRNA microarray analysis of 10 samples of BMSCs (n=1), PDLSCs (n=3), PULPSCs (n=3), and CDCs (n=3) were performed. Results: The expression of mesenchymal stem cell markers with a concomitant absence of hematopoietic markers was observed in PDLSCs, PULPSCs, CDCs and BMSCs. All 4 cell populations also showed differentiation into osteogenic, adipogenic, and chondrogenic lineages. CEMP1 was strongly expressed in CDCs, while it was weakly detected in the other 3 cell populations. Meanwhile, CAP was not found in any of the 4 cell populations. The mRNA and miRNA microarray analysis showed that 14 mRNA genes and 4 miRNA genes were differentially expressed in CDCs vs. PDLSCs and PULPSCs. Conclusions: Within the limitations of the study, CDCs seem to have stemness and preferentially express CEMP1. Moreover, there were several up- or down-regulated genes in CDCs vs. PDLSCs, PULPSCs, and BMSCs and these genes could be candidate marker proteins of CDCs.

Keywords

Acknowledgement

This study was supported by a grant (2013-541) from the Asan Institute for Life Sciences, Seoul, Korea.

References

  1. Gottlow J, Nyman S, Lindhe J, Karring T, Wennstrom J. New attachment formation in the human periodontium by guided tissue regeneration. Case reports. J Clin Periodontol 1986;13:604-16. https://doi.org/10.1111/j.1600-051X.1986.tb00854.x
  2. Nyman S, Lindhe J, Karring T, Rylander H. New attachment following surgical treatment of human periodontal disease. J Clin Periodontol 1982;9:290-6. https://doi.org/10.1111/j.1600-051X.1982.tb02095.x
  3. Tonetti MS, Cortellini P, Suvan JE, Adriaens P, Baldi C, Dubravec D, et al. Generalizability of the added benefits of guided tissue regeneration in the treatment of deep intrabony defects. Evaluation in a multi-center randomized controlled clinical trial. J Periodontol 1998;69:1183-92. https://doi.org/10.1902/jop.1998.69.11.1183
  4. Tonetti MS, Prato GP, Cortellini P. Factors affecting the healing response of intrabony defects following guided tissue regeneration and access flap surgery. J Clin Periodontol 1996;23:548-56. https://doi.org/10.1111/j.1600-051X.1996.tb01823.x
  5. Park JY, Jeon SH, Choung PH. Efficacy of periodontal stem cell transplantation in the treatment of advanced periodontitis. Cell Transplant 2011;20:271-85. https://doi.org/10.3727/096368910X519292
  6. Liu Y, Zheng Y, Ding G, Fang D, Zhang C, Bartold PM, et al. Periodontal ligament stem cell-mediated treatment for periodontitis in miniature swine. Stem Cells 2008;26:1065-73. https://doi.org/10.1634/stemcells.2007-0734
  7. Park JY, Park CH, Yi T, Kim SN, Iwata T, Yun JH. rhBMP-2 pre-treated human periodontal ligament stem cell sheets regenerate a mineralized layer mimicking dental cementum. Int J Mol Sci 2020;21:3767. https://doi.org/10.3390/ijms21113767
  8. Cho H, Tarafder S, Fogge M, Kao K, Lee CH. Periodontal ligament stem/progenitor cells with protein-releasing scaffolds for cementum formation and integration on dentin surface. Connect Tissue Res 2016;57:488-95. https://doi.org/10.1080/03008207.2016.1191478
  9. Grzesik WJ, Kuzentsov SA, Uzawa K, Mankani M, Robey PG, Yamauchi M. Normal human cementum-derived cells: isolation, clonal expansion, and in vitro and in vivo characterization. J Bone Miner Res 1998;13:1547-54. https://doi.org/10.1359/jbmr.1998.13.10.1547
  10. Liu HW, Yacobi R, Savion N, Narayanan AS, Pitaru S. A collagenous cementum-derived attachment protein is a marker for progenitors of the mineralized tissue-forming cell lineage of the periodontal ligament. J Bone Miner Res 1997;12:1691-9. https://doi.org/10.1359/jbmr.1997.12.10.1691
  11. Alvarez-Perez MA, Narayanan S, Zeichner-David M, Rodriguez Carmona B, Arzate H. Molecular cloning, expression and immunolocalization of a novel human cementum-derived protein (CP-23). Bone 2006;38:409-19. https://doi.org/10.1016/j.bone.2005.09.009
  12. Komaki M, Iwasaki K, Arzate H, Narayanan AS, Izumi Y, Morita I. Cementum protein 1 (CEMP1) induces a cementoblastic phenotype and reduces osteoblastic differentiation in periodontal ligament cells. J Cell Physiol 2012;227:649-57. https://doi.org/10.1002/jcp.22770
  13. Bosshardt DD. Are cementoblasts a subpopulation of osteoblasts or a unique phenotype? J Dent Res 2005;84:390-406. https://doi.org/10.1177/154405910508400501
  14. Foster BL. Methods for studying tooth root cementum by light microscopy. Int J Oral Sci 2012;4:119-28. https://doi.org/10.1038/ijos.2012.57
  15. Matthews BG, Roguljic H, Franceschetti T, Roeder E, Matic I, Vidovic I, et al. Gene-expression analysis of cementoblasts and osteoblasts. J Periodontal Res 2016;51:304-12. https://doi.org/10.1111/jre.12309
  16. Salmon CR, Tomazela DM, Ruiz KG, Foster BL, Paes Leme AF, Sallum EA, et al. Proteomic analysis of human dental cementum and alveolar bone. J Proteomics 2013;91:544-55. https://doi.org/10.1016/j.jprot.2013.08.016
  17. Kitagawa M, Tahara H, Kitagawa S, Oka H, Kudo Y, Sato S, et al. Characterization of established cementoblast-like cell lines from human cementum-lining cells in vitro and in vivo. Bone 2006;39:1035-42. https://doi.org/10.1016/j.bone.2006.05.022
  18. Wu D, Ikezawa K, Parker T, Saito M, Narayanan AS. Characterization of a collagenous cementum-derived attachment protein. J Bone Miner Res 1996;11:686-92. https://doi.org/10.1002/jbmr.5650110517
  19. Kim SH, Kim KH, Seo BM, Koo KT, Kim TI, Seol YJ, et al. Alveolar bone regeneration by transplantation of periodontal ligament stem cells and bone marrow stem cells in a canine peri-implant defect model: a pilot study. J Periodontol 2009;80:1815-23. https://doi.org/10.1902/jop.2009.090249
  20. Gronthos S, Mankani M, Brahim J, Robey PG, Shi S. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci U S A 2000;97:13625-30. https://doi.org/10.1073/pnas.240309797
  21. Seo BM, Miura M, Gronthos S, Bartold PM, Batouli S, Brahim J, et al. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet 2004;364:149-55. https://doi.org/10.1016/S0140-6736(04)16627-0
  22. Estrela C, Alencar AH, Kitten GT, Vencio EF, Gava E. Mesenchymal stem cells in the dental tissues: perspectives for tissue regeneration. Braz Dent J 2011;22:91-8. https://doi.org/10.1590/S0103-64402011000200001
  23. Sonoyama W, Liu Y, Yamaza T, Tuan RS, Wang S, Shi S, et al. Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study. J Endod 2008;34:166-71. https://doi.org/10.1016/j.joen.2007.11.021
  24. Shi S, Robey PG, Gronthos S. Comparison of human dental pulp and bone marrow stromal stem cells by cDNA microarray analysis. Bone 2001;29:532-9. https://doi.org/10.1016/S8756-3282(01)00612-3
  25. Kim SH, Kim YS, Lee SY, Kim KH, Lee YM, Kim WK, et al. Gene expression profile in mesenchymal stem cells derived from dental tissues and bone marrow. J Periodontal Implant Sci 2011;41:192-200. https://doi.org/10.5051/jpis.2011.41.4.192
  26. Kawashima N. Characterisation of dental pulp stem cells: a new horizon for tissue regeneration? Arch Oral Biol 2012;57:1439-58. https://doi.org/10.1016/j.archoralbio.2012.08.010
  27. Taha MF, Hedayati V. Isolation, identification and multipotential differentiation of mouse adipose tissuederived stem cells. Tissue Cell 2010;42:211-6. https://doi.org/10.1016/j.tice.2010.04.003
  28. Tropel P, Noel D, Platet N, Legrand P, Benabid AL, Berger F. Isolation and characterisation of mesenchymal stem cells from adult mouse bone marrow. Exp Cell Res 2004;295:395-406. https://doi.org/10.1016/j.yexcr.2003.12.030
  29. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999;284:143-7. https://doi.org/10.1126/science.284.5411.143
  30. Carnes DL, Maeder CL, Graves DT. Cells with osteoblastic phenotypes can be explanted from human gingiva and periodontal ligament. J Periodontol 1997;68:701-7. https://doi.org/10.1902/jop.1997.68.7.701
  31. Torii D, Konishi K, Watanabe N, Goto S, Tsutsui T. Cementogenic potential of multipotential mesenchymal stem cells purified from the human periodontal ligament. Odontology 2015;103:27-35. https://doi.org/10.1007/s10266-013-0145-y
  32. Hoz L, Romo E, Zeichner-David M, Sanz M, Nunez J, Gaitan L, et al. Cementum protein 1 (CEMP1) induces differentiation by human periodontal ligament cells under three-dimensional culture conditions. Cell Biol Int 2012;36:129-36. https://doi.org/10.1042/CBI20110168
  33. Nunez J, Sanz M, Hoz-Rodriguez L, Zeichner-David M, Arzate H. Human cementoblasts express enamel-associated molecules in vitro and in vivo. J Periodontal Res 2010;45:809-14. https://doi.org/10.1111/j.1600-0765.2010.01291.x
  34. Paula-Silva FW, Ghosh A, Arzate H, Kapila S, da Silva LA, Kapila YL. Calcium hydroxide promotes cementogenesis and induces cementoblastic differentiation of mesenchymal periodontal ligament cells in a CEMP1- and ERK-dependent manner. Calcif Tissue Int 2010;87:144-57. https://doi.org/10.1007/s00223-010-9368-x
  35. Gronthos S, Zannettino AC, Graves SE, Ohta S, Hay SJ, Simmons PJ. Differential cell surface expression of the STRO-1 and alkaline phosphatase antigens on discrete developmental stages in primary cultures of human bone cells. J Bone Miner Res 1999;14:47-56. https://doi.org/10.1359/jbmr.1999.14.1.47
  36. Bronckers AL, Farach-Carson MC, Van Waveren E, Butler WT. Immunolocalization of osteopontin, osteocalcin, and dentin sialoprotein during dental root formation and early cementogenesis in the rat. J Bone Miner Res 1994;9:833-41. https://doi.org/10.1002/jbmr.5650090609
  37. Pitaru S, Narayanan SA, Olson S, Savion N, Hekmati H, Alt I, et al. Specific cementum attachment protein enhances selectively the attachment and migration of periodontal cells to root surfaces. J Periodontal Res 1995;30:360-8. https://doi.org/10.1111/j.1600-0765.1995.tb01288.x
  38. Arzate H, Zeichner-David M, Mercado-Celis G. Cementum proteins: role in cementogenesis, biomineralization, periodontium formation and regeneration. Periodontol 2000 2015;67:211-33. https://doi.org/10.1111/prd.12062
  39. Arzate H, Jimenez-Garcia LF, Alvarez-Perez MA, Landa A, Bar-Kana I, Pitaru S. Immunolocalization of a human cementoblastoma-conditioned medium-derived protein. J Dent Res 2002;81:541-6. https://doi.org/10.1177/154405910208100808