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A simple cell motility assay demonstrates differential motility of human periodontal ligament fibroblasts, gingival fibroblasts, and pre-osteoblasts

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Abstract

During periodontal regeneration, multiple cell types can invade the wound site, thereby leading to repair. Cell motility requires interactions mediated by integrin receptors for the extracellular matrix (ECM), which might be useful in guiding specific cell populations into the periodontal defect. Our data demonstrate that fibroblasts exhibit differential motility when grown on ECM proteins. Specifically, gingival fibroblasts are twice as motile as periodontal ligament fibroblasts, whereas osteoblasts are essentially non-motile. Collagens promote the greatest motility of gingival fibroblasts in the following order: collagen III>collagen V>collagen I. Differences in motility do not correlate with cell proliferation or integrin expression. Osteoblasts display greater attachment to collagens than does either fibroblast population, but lower motility. Gingival fibroblast motility on collagen I is generally mediated by α2 integrins, whereas motility on collagen III involves α1 integrins. Other integrins (α10 or α11) may also contribute to gingival fibroblast motility. Thus, ECM proteins do indeed differentially promote the cell motility of periodontal cells. Because of their greater motility, gingival fibroblasts have more of a potential to invade periodontal wound sites and to contribute to regeneration. This finding may explain the formation of disorganized connective tissue masses rather than the occurrence of the true regeneration of the periodontium.

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References

  • Akizuki T, Oda S, Komaki M, Tsuchioka H, Kawakatsu N, Kikuchi A, Yamato M, Okano T, Ishikawa I (2005) Application of periodontal ligament cell sheet for periodontal regeneration: a pilot study in beagle dogs. J Periodontal Res 40:245–251

    Article  PubMed  Google Scholar 

  • Alliot-Licht B, De Lange GL, Gregoire M (1997) Effects of hydroxyapatite particles on periodontal ligament fibroblast-like cell behavior. J Periodontol 68:158–165

    PubMed  CAS  Google Scholar 

  • Badylak SF (2002) The extracellular matrix as a scaffold for tissue reconstruction. Semin Cell Dev Biol 13:377–383

    Article  PubMed  CAS  Google Scholar 

  • Bartold PM, Walsh LJ, Narayanan AS (2000) Molecular and cell biology of the gingiva. Periodontol 2000 24:28–55

    Article  PubMed  CAS  Google Scholar 

  • Bengtsson T, Aszodi A, Nicolae C, Hunziker EB, Lundgren-Akerlund E, Fassler R (2005) Loss of alpha10beta1 integrin expression leads to moderate dysfunction of growth plate chondrocytes. J Cell Sci 118:929–936

    Article  PubMed  CAS  Google Scholar 

  • Bernick S, Paule W, Ertl D, Nishimoto SK, Nimni ME (1989) Cellular events associated with the induction of bone by demineralized bone. J Orthop Res 7:1–11

    Article  PubMed  CAS  Google Scholar 

  • Bland YS, Ashhurst DE (1996) Changes in the distribution of fibrillar collagens in the collateral and cruciate ligaments of the rabbit knee joint during fetal and postnatal development. Histochem J 28:325–334

    Article  PubMed  CAS  Google Scholar 

  • Bumann A, Carvalho RS, Schwarzer CL, Yen EH (1997) Collagen synthesis from human PDL cells following orthodontic tooth movement. Eur J Orthod 19:29–37

    Article  PubMed  CAS  Google Scholar 

  • Butler WT, Birkedal-Hansen H, Beegle WF, Taylor RE, Chung E (1975) Proteins of the periodontium. Identification of collagens with the [alpha1(I)]2alpha2 and [alpha1(III)]3 structures in bovine periodontal ligament. J Biol Chem 250:8907–8912

    PubMed  CAS  Google Scholar 

  • Camper L, Hellman U, Lundgren-Akerlund E (1998) Isolation, cloning, and sequence analysis of the integrin subunit alpha10, a beta1-associated collagen binding integrin expressed on chondrocytes. J Biol Chem 273:20383–20389

    Article  PubMed  CAS  Google Scholar 

  • Camper L, Holmvall K, Wangnerud C, Aszodi A, Lundgren-Akerlund E (2001) Distribution of the collagen-binding integrin alpha10beta1 during mouse development. Cell Tissue Res 306:107–116

    Article  PubMed  CAS  Google Scholar 

  • Caton J, Greenstein G, Zappa U (1992) Guided tissue regeneration using Vicryl periodontal mesh. Compendium 13:202–208

    PubMed  CAS  Google Scholar 

  • Elices MJ, Hemler ME (1989) The human integrin VLA-2 is a collagen receptor on some cells and a collagen/laminin receptor on others. Proc Natl Acad Sci USA 86:9906–9910

    Article  PubMed  CAS  Google Scholar 

  • Fernyhough W, Page RC (1983) Attachment, growth and synthesis by human gingival fibroblasts on demineralized or fibronectin-treated normal and diseased tooth roots. J Periodontol 54:133–140

    PubMed  CAS  Google Scholar 

  • Gaunt WA, Osborn JW, TenCate AR (1971) The Periodontal Ligament. Advanced Dental Histology. Williams and Wilins, Baltimore

    Google Scholar 

  • Giannopoulou C, Cimasoni G (1996) Functional characteristics of gingival and periodontal ligament fibroblasts. J Dent Res 75:895–902

    PubMed  CAS  Google Scholar 

  • Groeneveld MC, Everts V, Beertsen W (1995) Alkaline phosphatase activity in the periodontal ligament and gingiva of the rat molar: its relation to cementum formation. J Dent Res 74:1374–1381

    PubMed  CAS  Google Scholar 

  • Hallman M, Lundgren S, Sennerby L (2001) Histologic analysis of clinical biopsies taken 6 months and 3 years after maxillary sinus floor augmentation with 80% bovine hydroxyapatite and 20% autogenous bone mixed with fibrin glue. Clin Implant Dent Relat Res 3:87–96

    PubMed  CAS  Google Scholar 

  • Harris SA, Enger RJ, Riggs BL, Spelsberg TC (1995) Development and characterization of a conditionally immortalized human fetal osteoblastic cell line. J Bone Miner Res 10:178–186

    Article  PubMed  CAS  Google Scholar 

  • Hasegawa M, Yamato M, Kikuchi A, Okano T, Ishikawa I (2005) Human periodontal ligament cell sheets can regenerate periodontal ligament tissue in an athymic rat model. Tissue Eng 11:469–478

    Article  PubMed  CAS  Google Scholar 

  • Hay ED (1981) Extracellular matrix. J Cell Biol 91:205s–223s

    Article  PubMed  CAS  Google Scholar 

  • Hou LT, Liu CM, Wong MY (1995) Expression of matrix proteins in cloned fibroblasts derived from periodontal tissue under different cell growth densities. Proc Natl Sci Counc Repub China [B] 19:176–184

    CAS  Google Scholar 

  • Hou LT, Yaeger JA (1993) Cloning and characterization of human gingival and periodontal ligament fibroblasts. J Periodontol 64:1209–1218

    PubMed  CAS  Google Scholar 

  • Kerkvliet EH, Jansen IC, Schoenmaker T, Beertsen W, Everts V (2003) Collagen type I, III and V differently modulate synthesis and activation of matrix metalloproteinases by cultured rabbit periosteal fibroblasts. Matrix Biol 22:217–227

    Article  PubMed  CAS  Google Scholar 

  • Kim JK, Xu Y, Xu X, Keene DR, Gurusiddappa S, Liang X, Wary KK, Hook M (2005) A novel binding site in collagen type III for integrins alpha1beta1 and alpha2beta1. J Biol Chem 280:32512–32520

    Article  PubMed  CAS  Google Scholar 

  • Klekotka PA, Santoro SA, Wang H, Zutter MM (2001) Specific residues within the alpha 2 integrin subunit cytoplasmic domain regulate migration and cell cycle progression via distinct MAPK pathways. J Biol Chem 276:32353–32361

    Article  PubMed  CAS  Google Scholar 

  • Kreis S, Schonfeld HJ, Melchior C, Steiner B, Kieffer N (2005) The intermediate filament protein vimentin binds specifically to a recombinant integrin alpha2/beta1 cytoplasmic tail complex and co-localizes with native alpha2/beta1 in endothelial cell focal adhesions. Exp Cell Res 305:110–121

    Article  PubMed  CAS  Google Scholar 

  • Kuru L, Griffiths GS, Petrie A, Olsen I (1999) Alkaline phosphatase activity is upregulated in regenerating human periodontal cells. J Periodontal Res 34:123–127

    Article  PubMed  CAS  Google Scholar 

  • Lallier T, Bronner-Fraser M (1992) Alpha 1 beta 1 integrin on neural crest cells recognizes some laminin substrata in a Ca(2+)-independent manner. J Cell Biol 119:1335–1345

    Article  PubMed  CAS  Google Scholar 

  • Lallier TE, Spencer A, Fowler MM (2005) Transcript profiling of periodontal fibroblasts and osteoblasts. J Periodontol 76:1044–1055

    Article  PubMed  CAS  Google Scholar 

  • Lekic P, Sodek J, McCulloch CA (1996) Osteopontin and bone sialoprotein expression in regenerating rat periodontal ligament and alveolar bone. Anat Rec 244:50–58

    Article  PubMed  CAS  Google Scholar 

  • Lochter A, Navre M, Werb Z, Bissell MJ (1999) Alpha1 and alpha2 integrins mediate invasive activity of mouse mammary carcinoma cells through regulation of stromelysin-1 expression. Mol Biol Cell 10:271–282

    PubMed  CAS  Google Scholar 

  • Martin M, Gantes B, Garrett S, Egelberg J (1988) Treatment of periodontal furcation defects. I. Review of the literature and description of a regenerative surgical technique. J Clin Periodontol 15:227–231

    Article  PubMed  CAS  Google Scholar 

  • Nanci A, Bosshardt DD (2006) Structure of periodontal tissues in health and disease. Periodontol 2000 40:11–28

    Article  PubMed  Google Scholar 

  • Narayanan AS, Page RC (1983a) Biosynthesis and regulation of type V collagen in diploid human fibroblasts. J Biol Chem 258:11694–11699

    PubMed  CAS  Google Scholar 

  • Narayanan AS, Page RC (1983b) Connective tissues of the periodontium: a summary of current work. Coll Relat Res 3:33–64

    PubMed  CAS  Google Scholar 

  • Nguyen L, Lekic P, McCulloch CA (1997) Collagen implants do not preserve periodontal ligament homeostasis in periodontal wounds. J Periodontal Res 32:419–429

    Article  PubMed  CAS  Google Scholar 

  • Nishimura F, Terranova VP (1996) Comparative study of the chemotactic responses of periodontal ligament cells and gingival fibroblasts to polypeptide growth factors. J Dent Res 75:986–992

    PubMed  CAS  Google Scholar 

  • Niyibizi C, Kavalkovich K, Yamaji T, Woo SL (2000) Type V collagen is increased during rabbit medial collateral ligament healing. Knee Surg Sports Traumatol Arthrosc 8:281–285

    Article  PubMed  CAS  Google Scholar 

  • Ohshima M, Kuwata F, Otsuka K, Saito R, Sato K, Shioji S, Suzuki K (1988) Alkaline phosphatase activities of cultured human periodontal ligament cells. J Nihon Univ Sch Dent 30:208–217

    PubMed  CAS  Google Scholar 

  • Palaiologou AA, Yukna RA, Moses R, Lallier TE (2001) Gingival, dermal, and periodontal ligament fibroblasts express different extracellular matrix receptors. J Periodontol 72:798–807

    Article  PubMed  CAS  Google Scholar 

  • Popova SN, Rodriguez-Sanchez B, Liden A, Betsholtz C, Van Den Bos T, Gullberg D (2004) The mesenchymal alpha11beta1 integrin attenuates PDGF-BB-stimulated chemotaxis of embryonic fibroblasts on collagens. Dev Biol 270:427–442

    Article  PubMed  CAS  Google Scholar 

  • Rao LG, Wang HM, Kalliecharan R, Heersche JN, Sodek J (1979) Specific immunohistochemical localization of type I collagen in porcine periodontal tissues using the peroxidase-labelled antibody technique. Histochem J 11:73–82

    Article  PubMed  CAS  Google Scholar 

  • Reddi AH (1981) Cell biology and biochemistry of endochondral bone development. Coll Relat Res 1:209–226

    PubMed  CAS  Google Scholar 

  • Riikonen T, Vihinen P, Potila M, Rettig W, Heino J (1995) Antibody against human alpha 1 beta 1 integrin inhibits HeLa cell adhesion to laminin and to type I, IV, and V collagens. Biochem Biophys Res Commun 209:205–212

    Article  PubMed  CAS  Google Scholar 

  • Robinson KA, Li J, Mathison M, Redkar A, Cui J, Chronos NA, Matheny RG, Badylak SF (2005) Extracellular matrix scaffold for cardiac repair. Circulation 112:I135–I143

    Article  PubMed  Google Scholar 

  • Romanos GE, Schroter-Kermani C, Hinz N, Wachtel HC, Bernimoulin JP (1991) Immunohistochemical localization of collagenous components in healthy periodontal tissues of the rat and marmoset (Callithrix jacchus). II. Distribution of collagen types IV, V and VI. J Periodontal Res 26:323–332

    Article  PubMed  CAS  Google Scholar 

  • Saelman EU, Nieuwenhuis HK, Hese KM, de Groot PG, Heijnen HF, Sage EH, Williams S, McKeown L, Gralnick HR, Sixma JJ (1994) Platelet adhesion to collagen types I through VIII under conditions of stasis and flow is mediated by GPIa/IIa (alpha 2 beta 1-integrin). Blood 83:1244–1250

    PubMed  CAS  Google Scholar 

  • Schon M, Schon MP, Kuhrober A, Schirmbeck R, Kaufmannn R, Klein CE (1996) Expression of the human alpha2 integrin subunit in mouse melanoma cells confers the ability to undergo collagen-directed adhesion, migration and matrix reorganization. J Invest Dermatol 106:1175–1181

    Article  PubMed  CAS  Google Scholar 

  • Senger DR, Claffey KP, Benes JE, Perruzzi CA, Sergiou AP, Detmar M (1997) Angiogenesis promoted by vascular endothelial growth factor: regulation through alpha1beta1 and alpha2beta1 integrins. Proc Natl Acad Sci USA 94:13612–13617

    Article  PubMed  CAS  Google Scholar 

  • Shimomura T, Jia F, Niyibizi C, Woo SL (2003) Antisense oligonucleotides reduce synthesis of procollagen alpha1 (V) chain in human patellar tendon fibroblasts: potential application in healing ligaments and tendons. Connect Tissue Res 44:167–172

    PubMed  CAS  Google Scholar 

  • Somerman MJ, Archer SY, Imm GR, Foster RA (1988) A comparative study of human periodontal ligament cells and gingival fibroblasts in vitro. J Dent Res 67:66–70

    PubMed  CAS  Google Scholar 

  • Steffensen B, Duong AH, Milam SB, Potempa CL, Winborn WB, Magnuson VL, Chen D, Zardeneta G, Klebe RJ (1992) Immunohistological localization of cell adhesion proteins and integrins in the periodontium. J Periodontol 63:584–592

    PubMed  CAS  Google Scholar 

  • Strayhorn CL, Garrett JS, Dunn RL, Benedict JJ, Somerman MJ (1999) Growth factors regulate expression of osteoblast-associated genes. J Periodontol 70:1345–1354

    Article  PubMed  CAS  Google Scholar 

  • Takada Y, Wayner EA, Carter WG, Hemler ME (1988) Extracellular matrix receptors, ECMRII and ECMRI, for collagen and fibronectin correspond to VLA-2 and VLA-3 in the VLA family of heterodimers. J Cell Biochem 37:385–393

    Article  PubMed  CAS  Google Scholar 

  • Takeshita A, Niu ZG, Hanazawa S, Takara I, Higuchi H, Katayama I, Kitano S (1992) Effect of interleukin-1 beta on gene expressions and functions of fibroblastic cells derived from human periodontal ligament. J Periodontal Res 27:250–255

    Article  PubMed  CAS  Google Scholar 

  • Takiguchi T, Kobayashi M, Nagashima C, Yamaguchi A, Nishihara T, Hasegawa K (1999) Effect of prostaglandin E2 on recombinant human bone morphogenetic protein-2-stimulated osteoblastic differentiation in human periodontal ligament cells. J Periodontal Res 34:431–436

    Article  PubMed  CAS  Google Scholar 

  • Tardieu-Moreau C, Pourreau-Schneider N, Colomb E, Kopp F, Martin PM, Franquin JC (1992) Effects of culture passages on collagen immunostaining in human dental pulp and gingival fibroblasts. J Biol Buccale 20:169–174

    PubMed  CAS  Google Scholar 

  • Tenenbaum HC (1990) Cellular origins and theories of differentiation of bone-forming cells. In: Hall BK (ed) Bone, vol 1. The osteoblast and osteocyte. Telford, New Jersey, pp 41–69

    Google Scholar 

  • Tiger CF, Fougerousse F, Grundstrom G, Velling T, Gullberg D (2001) Alpha11beta1 integrin is a receptor for interstitial collagens involved in cell migration and collagen reorganization on mesenchymal nonmuscle cells. Dev Biol 237:116–129

    Article  PubMed  CAS  Google Scholar 

  • Tung PS, Domenicucci C, Wasi S, Sodek J (1985) Specific immunohistochemical localization of osteonectin and collagen types I and III in fetal and adult porcine dental tissues. J Histochem Cytochem 33:531–540

    PubMed  CAS  Google Scholar 

  • Urabe M, Hosokawa R, Chiba D, Sato Y, Akagawa Y (2000) Morphogenetic behavior of periodontium on inorganic implant materials: an experimental study of canines. J Biomed Mater Res 49:17–24

    Article  PubMed  CAS  Google Scholar 

  • Urist MR, DeLange RJ, Finerman GA (1983) Bone cell differentiation and growth factors. Science 220:680–686

    Article  PubMed  CAS  Google Scholar 

  • Velling T, Risteli J, Wennerberg K, Mosher DF, Johansson S (2002) Polymerization of type I and III collagens is dependent on fibronectin and enhanced by integrins alpha 11beta 1 and alpha 2beta 1. J Biol Chem 277:37377–37381

    Article  PubMed  CAS  Google Scholar 

  • Vihinen P, Riikonen T, Laine A, Heino J (1996) Integrin alpha 2 beta 1 in tumorigenic human osteosarcoma cell lines regulates cell adhesion, migration, and invasion by interaction with type I collagen. Cell Growth Differ 7:439–447

    PubMed  CAS  Google Scholar 

  • Wang HM, Nanda V, Rao LG, Melcher AH, Heersche JN, Sodek J (1980) Specific immunohistochemical localization of type III collagen in porcine periodontal tissues using the peroxidase-antiperoxidase method. J Histochem Cytochem 28:1215–1223

    PubMed  CAS  Google Scholar 

  • Werfully S, Areibi G, Toner M, Bergquist J, Walker J, Renvert S, Claffey N (2002) Tensile strength, histological and immunohistochemical observations of periodontal wound healing in the dog. J Periodontal Res 37:366–374

    Article  PubMed  CAS  Google Scholar 

  • Wikesjo UM, Nilveus RE, Selvig KA (1992) Significance of early healing events on periodontal repair: a review. J Periodontol 63:158–165

    PubMed  CAS  Google Scholar 

  • Yamaguchi M, Shimizu N, Shibata Y, Abiko Y (1996) Effects of different magnitudes of tension-force on alkaline phosphatase activity in periodontal ligament cells. J Dent Res 75:889–894

    Article  PubMed  CAS  Google Scholar 

  • Zantop T, Gilbert TW, Yoder MC, Badylak SF (2006) Extracellular matrix scaffolds are repopulated by bone marrow-derived cells in a mouse model of Achilles tendon reconstruction. J Orthop Res 24:1299–1309

    Article  PubMed  Google Scholar 

  • Zhou H, Kramer RH (2005) Integrin engagement differentially modulates epithelial cell motility by RhoA/ROCK and PAK1. J Biol Chem 280:10624–10635

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Cell Biology and Anatomy, Center of Excellence in Oral and Craniofacial Biology, Louisiana State University Health Science Center, School of Dentistry, 1100 Florida Avenue, New Orleans, LA, 70119, USA

    Thomas E. Lallier, Jackie Sonnier & Amber Spencer

  2. Department of Endodontics, Louisiana State University Health Science Center, School of Dentistry, 1100 Florida Avenue, New Orleans, LA, 70119, USA

    Quinton W. Miner Jr

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  1. Thomas E. Lallier
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  2. Quinton W. Miner Jr
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  3. Jackie Sonnier
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  4. Amber Spencer
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Correspondence to Thomas E. Lallier.

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This research was supported by the Louisiana Board of Regents through the Millennium Trust Health Excellence Fund, HEF-(2000-05)-04.

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Lallier, T.E., Miner, Q.W., Sonnier, J. et al. A simple cell motility assay demonstrates differential motility of human periodontal ligament fibroblasts, gingival fibroblasts, and pre-osteoblasts. Cell Tissue Res 328, 339–354 (2007). https://doi.org/10.1007/s00441-006-0372-4

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