Skip to main content

Advertisement

Log in

Highly osteogenic PDL stem cell clones specifically express elevated levels of ICAM1, ITGB1 and TERT

  • Original Research
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

Cells derived from the periodontal ligament (PDL) have previously been reported to have stem cell-like characteristics (PDL stem cells; PDLSCs) and play an important part in bone engineering, including that of alveolar bone. However, these populations have been heterogeneous, and thus far no specific marker has yet been established from adult human stem cells derived from PDL tissue. We have previously isolated highly purified single cell-derived PDLSC clones and delineated their phenotypic and functional characteristics. In this report, we further obtained three homogeneous and distinct PDLSC clones demonstrating low, moderate and high mineralized matrix forming ability-namely PC12, PC4 and PC3, respectively, and the expression of mesenchymal stem cell pathway-specific genes in these clones was investigated. PCR array revealed that the expression of intercellular adhesion molecule 1 (ICAM1), integrin beta 1 (ITGB1) and telomerase reverse transcriptase (TERT) was associated with highly osteogenic PDLSC clones, as determined by the expression of key osteoblastic markers and their ability to form alizarin red S positive mineralized matrix in vitro. The present results suggest that these three mesenchymal stem cell-associated markers could potentially be used to isolate PDLSCs with high osteogenic capability for engineering new bone.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Anderson HC (2003) Matrix vesicles and calcification. Curr Rheumatol Rep 5:222–226

    Article  Google Scholar 

  • Barnes GL, Javed A, Waller SM, Kamal MH, Hebert KE, Hassan MQ, Bellahcene A, Van Wijnen AJ, Young MF, Lian JB, Stein GS, Gerstenfeld LC (2003) Osteoblast-related transcription factors Runx2 (Cbfa1/AML3) and MSX2 mediate the expression of bone sialoprotein in human metastatic breast cancer cells. Cancer Res 63:2631–2637

    CAS  Google Scholar 

  • Bartold PM, McCulloch CA, Narayanan AS, Pitaru S (2000) Tissue engineering: a new paradigm for periodontal regeneration based on molecular and cell biology. Periodontol 24:253–269

    Article  CAS  Google Scholar 

  • Bodnar AG, Ouellette M, Frolkis M, Holt SE, Chiu CP, Morin GB, Harley CB, Shay JW, Lichtsteiner S, Wright WE (1998) Extension of life-span by introduction of telomerase into normal human cells. Science 279:349–352

    Article  CAS  Google Scholar 

  • Calderwood DA (2004) Integrin activation. J Cell Sci 117:657–666

    Article  CAS  Google Scholar 

  • Cernuda-Morollon E, Ridley AJ (2006) Rho GTPases and leukocyte adhesion receptor expression and function in endothelial cells. Circ Res 98:757–767

    Article  CAS  Google Scholar 

  • Chaudhary LR, Hofmeister AM, Hruska KA (2004) Differential growth factor control of bone formation through osteoprogenitor differentiation. Bone 34:402–411

    Article  CAS  Google Scholar 

  • Chen XD, Allen MR, Bloomfield S, Xu T, Young M (2003) Biglycan-deficient mice have delayed osteogenesis after marrow ablation. Calcif Tissue Int 72:577–582

    Article  CAS  Google Scholar 

  • Chen XD, Fisher LW, Robey PG, Young MF (2004) The small leucine-rich proteoglycan biglycan modulates BMP-4-induced osteoblast differentiation. FASEB J 18:948–958

    Article  CAS  Google Scholar 

  • Chen SC, Marino V, Gronthos S, Bartold PM (2006) Location of putative stem cells in human periodontal ligament. J Periodontal Res 41:547–553

    Article  CAS  Google Scholar 

  • Czyz J, Wobus A (2001) Embryonic stem cell differentiation: the role of extracellular factors. Differentiation 68:167–174

    Article  CAS  Google Scholar 

  • Digirolamo CM, Stokes D, Colter D, Phinney DG, Class R, Prockop DJ (1999) Propagation and senescence of human marrow stromal cells in culture: a simple colony-forming assay identifies samples with the greatest potential to propagate and differentiate. Br J Haematol 107:275–281

    Article  CAS  Google Scholar 

  • Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsenty G (1997) Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89:747–754

    Article  CAS  Google Scholar 

  • Fu X, Han B, Cai S, Lei Y, Sun T, Sheng Z (2009) Migration of bone marrow-derived mesenchymal stem cells induced by tumor necrosis factor-alpha and its possible role in wound healing. Wound Repair Regen 17:185–191

    Article  Google Scholar 

  • Gay IC, Chen S, MacDougall M (2007) Isolation and characterization of multipotent human periodontal ligament stem cells. Orthod Craniofac Res 10:149–160

    Article  CAS  Google Scholar 

  • Gortz B, Hayer S, Redlich K, Zwerina J, Tohidast-Akrad M, Tuerk B, Hartmann C, Kollias G, Steiner G, Smolen JS, Schett G (2004) Arthritis induces lymphocytic bone marrow inflammation and endosteal bone formation. J Bone Miner Res 19:990–998

    Article  CAS  Google Scholar 

  • Gronthos S, Graves SE, Ohta S, Simmons PJ (1994) The STRO-1+ fraction of adult human bone marrow contains the osteogenic precursors. Blood 84:4164–4173

    CAS  Google Scholar 

  • Gronthos S, Mrozik K, Shi S, Bartold PM (2006) Ovine periodontal ligament stem cells: isolation, characterization, and differentiation potential. Calcif Tissue Int 79:310–317

    Article  CAS  Google Scholar 

  • Ivanovski S, Gronthos S, Shi S, Bartold PM (2006) Stem cells in the periodontal ligament. Oral Dis 12:358–363

    Article  CAS  Google Scholar 

  • Kemoun P, Laurencin-Dalicieux S, Rue J, Vaysse F, Romeas A, Arzate H, Conte-Auriol F, Farges JC, Salles JP, Brunel G (2007) Localization of STRO-1, BMP-2/-3/-7, BMP receptors and phosphorylated Smad-1 during the formation of mouse periodontium. Tissue Cell 39:257–266

    Article  CAS  Google Scholar 

  • Kim YJ, Lee MH, Wozney JM, Cho JY, Ryoo HM (2004) Bone morphogenetic protein-2-induced alkaline phosphatase expression is stimulated by Dlx5 and repressed by Msx2. J Biol Chem 279:50773–50780

    Article  CAS  Google Scholar 

  • Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao YH, Inada M, Sato M, Okamoto R, Kitamura Y, Yoshiki S, Kishimoto T (1997) Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell 89:755–764

    Article  CAS  Google Scholar 

  • Kramer PR, Nares S, Kramer SF, Grogan D, Kaiser M (2004) Mesenchymal stem cells acquire characteristics of cells in the periodontal ligament in vitro. J Dent Res 83:27–34

    Article  CAS  Google Scholar 

  • Kuroki T, Shingu M, Koshihara Y, Nobunaga M (1994) Effects of cytokines on alkaline phosphatase and osteocalcin production, calcification and calcium release by human osteoblastic cells. Br J Rheumatol 33:224–230

    Article  CAS  Google Scholar 

  • Kuru L, Parkar MH, Griffiths GS, Olsen I (2001) Flow cytometry analysis of guided tissue regeneration-associated human periodontal cells. J Periodontol 72:1016–1024

    Article  CAS  Google Scholar 

  • Langer R, Vacanti JP (1993) Tissue engineering. Science 260:920–926

    Article  CAS  Google Scholar 

  • Lekic P, Rojas J, Birek C, Tenenbaum H, McCulloch CA (2001) Phenotypic comparison of periodontal ligament cells in vivo and in vitro. J Periodontal Res 36:71–79

    Article  CAS  Google Scholar 

  • Liddington RC, Ginsberg MH (2002) Integrin activation takes shape. J Cell Biol 158:833–839

    Article  CAS  Google Scholar 

  • Long MW, Robinson JA, Ashcraft EA, Mann KG (1995) Regulation of human bone marrow-derived osteoprogenitor cells by osteogenic growth factors. J Clin Invest 95:881–887

    Article  CAS  Google Scholar 

  • Manolagas SC (2000) Birth and death of bone cells: basic regulatory mechanisms and implications for the pathogenesis and treatment of osteoporosis. Endocr Rev 21:115–137

    Article  CAS  Google Scholar 

  • McCulloch CA, Bordin S (1991) Role of fibroblast subpopulations in periodontal physiology and pathology. J Periodontal Res 26:144–154

    Article  CAS  Google Scholar 

  • Melcher AH (1976) On the repair potential of periodontal tissues. J Periodontol 47:256–260

    Article  CAS  Google Scholar 

  • Murakami Y, Kojima T, Nagasawa T, Kobayashi H, Ishikawa I (2003) Novel isolation of alkaline phosphatase-positive subpopulation from periodontal ligament fibroblasts. J Periodontol 74:780–786

    Article  CAS  Google Scholar 

  • Olsen I, Abraham D, Shelton I, Bou-Gharios G, Muir H, Winchester B (1988) Cell contact induces the synthesis of a lysosomal enzyme precursor in lymphocytes and its direct transfer to fibroblasts. Biochim Biophys Acta 968:312–322

    Article  CAS  Google Scholar 

  • Parisuthiman D, Mochida Y, Duarte WR, Yamauchi M (2005) Biglycan modulates osteoblast differentiation and matrix mineralization. J Bone Miner Res 20:1878–1886

    Article  CAS  Google Scholar 

  • Pischon N, Darbois LM, Palamakumbura AH, Kessler E, Trackman PC (2004) Regulation of collagen deposition and lysyl oxidase by tumor necrosis factor-a in osteoblasts. J Biol Chem 279:30060–30065

    Article  CAS  Google Scholar 

  • Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147

    Article  CAS  Google Scholar 

  • Regazzoni C, Winterhalter KH, Rohrer L (2001) Type I collagen induces expression of bone morphogenetic protein receptor type II. Biochem Biophys Res Commun 283:316–322

    Article  CAS  Google Scholar 

  • Saho T, Kishida T, Hirano H, Hashikawa T, Shimabukuro Y, Murakami S (2003) Induction of CD13 on T-lymphocytes by adhesive interaction with gingival fibroblasts. J Dent Res 82:893–898

    Article  CAS  Google Scholar 

  • Sekiya I, Larson BL, Smith JR, Pochampally R, Cui JG, Prockop DJ (2002) Expansion of human adult stem cells from bone marrow stroma: conditions that maximize the yields of early progenitors and evaluate their quality. Stem Cells 20:530–541

    Article  Google Scholar 

  • Seo BM, Miura M, Gronthos S, Bartold PM, Batouli S, Young M, Brahim J, Robey PG, Wang CY, Shi S (2004) Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet 364:149–155

    Article  CAS  Google Scholar 

  • Shimono M, Ishikawa T, Ishikawa H, Matsuzaki H, Hashimoto S, Muramatsu T, Shima K, Matsuzaka K, Inoue T (2003) Regulatory mechanisms of periodontal regeneration. Microsc Res Tech 60:491–502

    Article  CAS  Google Scholar 

  • Shirakabe K, Terasawa K, Miyama K, Shibuya H, Nishida E (2001) Regulation of the activity of the transcription factor Runx2 by two homeobox proteins, Msx2 and Dlx5. Genes Cells 6:851–856

    Article  CAS  Google Scholar 

  • Sierra J, Villagra A, Paredes R, Cruzat F, Gutierrez S, Javed A, Arriagada G, Olate J, Imschenetzky M, Van Wijnen AJ, Lian JB, Stein GS, Stein JL, Montecino M (2003) Regulation of the bone-specific osteocalcin gene by p300 requires Runx2/Cbfa1 and the vitamin D3 receptor but not p300 intrinsic histone acetyltransferase activity. Mol Cell Biol 23:3339–3351

    Article  CAS  Google Scholar 

  • Simmons PJ, Torok-Storb B (1991) Identification of stromal cell precursors in human bone marrow by a novel monoclonal antibody, STRO-1. Blood 78:55–62

    CAS  Google Scholar 

  • Singhatanadgit W, Donos N, Olsen I (2009) Isolation and characterisation of stem cell clones from adult human ligament. Tissue Eng Part A 15:2625–2636

    Article  CAS  Google Scholar 

  • Takeuchi Y, Nakayama K, Matsumoto T (1996) Differentiation and cell surface expression of transforming growth factor-beta receptors are regulated by interaction with matrix collagen in murine osteoblastic cells. J Biol Chem 271:3938–3944

    Article  CAS  Google Scholar 

  • Takuwa Y, Ohse C, Wang EA, Wozney JM, Yamashita K (1991) Bone morphogenetic protein-2 stimulates alkaline phosphatase activity and collagen synthesis in cultured osteoblastic cells, MC3T3–E1. Biochem Biophys Res Commun 174:96–101

    Article  CAS  Google Scholar 

  • Tanaka Y, Morimoto I, Nakano Y, Okada Y, Hirota S, Nomura S, Nakamura T, Eto S (1995) Osteoblasts are regulated by the cellular adhesion through ICAM-1 and VCAM-1. J Bone Miner Res 10:1462–1469

    Article  CAS  Google Scholar 

  • ten Dijke P, Fu J, Schaap P, Roelen AJ (2003) Signal transduction of bone morphogenetic proteins in osteoblast differentiation. J Bone Joint Surg 85:34–38

    Google Scholar 

  • Turowski P, Adamson P, Greenwood J (2005) Pharmacological targeting of ICAM-1 signaling in brain endothelial cells: potential for treating neuroinflammation. Cell Mol Neurobiol 25:153–170

    Article  CAS  Google Scholar 

  • Viereck V, Siggelkow H, Tauber S, Raddatz D, Schutze N, Hufner M (2002) Differential regulation of Cbfa1/Runx2 and osteocalcin gene expression by vitamin-D3, dexamethasone, and local growth factors in primary human osteoblasts. J Cell Biochem 86:348–356

    Article  CAS  Google Scholar 

  • Young MF, Bi Y, Ameye L, Chen XD (2002) Biglycan knockout mice: new models for musculoskeletal diseases. Glycoconj J 19:257–262

    Article  CAS  Google Scholar 

  • Zakian VA (1995) Telomeres: beginning to understand the end. Science 270:1601–1607

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We acknowledge the financial support from Institute of Dentistry and Thammasat University, Thailand.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Weerachai Singhatanadgit.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sununliganon, L., Singhatanadgit, W. Highly osteogenic PDL stem cell clones specifically express elevated levels of ICAM1, ITGB1 and TERT. Cytotechnology 64, 53–63 (2012). https://doi.org/10.1007/s10616-011-9390-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-011-9390-5

Keywords

Navigation