Cell and Tissue Research

, Volume 321, Issue 3, pp 465–471 | Cite as

Expression and mechanical modulation of matrix metalloproteinase-1 and -2 genes in facial and cranial sutures

  • Rasha Al-Mubarak
  • Adriana Da Silveira
  • Jeremy J. Mao
Regular Article


Craniofacial sutures create a soft tissue interface between various calvarial and facial bones. Facial and cranial sutures show differences in their surrounding anatomical structures and local mechanical strain environments. Despite previous attempts to identify the expression of matrix metalloproteinase genes (MMPs) in cranial sutures, little is known regarding whether facial and cranial sutures differ in MMP expression. We have investigated the expression of MMP-1 and MMP-2 in the premaxillomaxillary suture (PMS; facial suture) and the frontoparietal suture (FPS; cranial suture) in 32-day-old rats with or without the application of cyclic loading. Expression of MMP-1 and MMP-2 was detected by the reverse transcription/polymerase chain reaction technique. At 32 days of postnatal development (n=6), both MMP-1 and MMP-2 were reproducibly expressed in the facial PMS, in comparison with negligible MMP-1 and MMP-2 expression in the cranial FPS. In six age- and sex-matched control rats, cyclic loading at 4 Hz and 1000 mN was applied to the maxilla for two 20-min episodes within a 12-h interval. In some (but not all) cases, cyclic loading induced marked expression of MMP-1 and MMP-2 in the PMS and FPS in comparison with corresponding non-loaded controls. These data confirm our previous finding that short doses of cyclic loading upregulate MMP-2 expression in craniofacial sutures and suggest the possibility that facial and cranial sutures differ in matrix degradation rates during postnatal development.


Matrix metalloproteinase Bone Osteoblasts Mechanical strain Craniofacial sutures Gene expression Rat (Sprague Dawley, male) 



The experimental work described in this manuscript is taken from the Master of Science thesis research by Rasha Al-Mubarak. We thank Drs. Carla Evans and Philip Patston for serving on the thesis committee and for providing valuable suggestions on the thesis project. We are indebted to Aurora Lopez for general technical assistance. We thank two anonymous reviewers whose insightful comments helped improve the quality of the manuscript.


  1. Barille S, Collette M, Thabard W, Bleunven C, Bataille R, Amiot M (2000) Soluable IL-6R (alpha) upregulates IL-6, MMP-1 and MMP-2 secretion in bone marrow stromal cells. Cytokine 12:1426–1429CrossRefPubMedGoogle Scholar
  2. Chin JR, Werb Z (1997) Matrix metalloproteinases regulate morphogenesis, migration and remodeling of epithelium, tongue skeletal muscle and cartilage in the mandibular arch. Development 124:1519–1530PubMedGoogle Scholar
  3. Cohen MM Jr (2000) Merging the old skeletal biology with the new. II. Molecular aspects of bone formation and bone growth. J Craniofac Genet Dev Biol 20:94–106PubMedGoogle Scholar
  4. Collins JM, Ramamoorthy K, Da Silveira A, Patston PA, Mao JJ (2005) Microstrain in intramembranous bones induces altered gene expression of MMP1 and MMP2 in the rat. J Biomech 38:485–492CrossRefPubMedGoogle Scholar
  5. Delaisse J, Engsig M, Everts V, Ovejero M, Ferreras L, Lund L, Vu T, Werb Z, Winding B, Lochter A, Karsdal M, Troen T, Kirkergaard T, Lenhard T, Heegaard A, Neff L, Baron R, Foged T (2000) Proteinases in bone resorption: obvious and less obvious roles. Clin Chim Acta 291:223–234CrossRefPubMedGoogle Scholar
  6. Duncan RL, Turner CH (1995) Mechanotransduction and the functional response of bone to mechanical strain. Calcif Tissue Int 57:344–358CrossRefPubMedGoogle Scholar
  7. Green DD, Hembry RM, Atkinson SJ, Reynolds JJ, Meikle MC (1990) Immunolocalization of collagenase and tissue inhibitor of metalloproteinases (TIMP) in mechanically deformed fibrous joints. Am J Orthod 97:281–288Google Scholar
  8. Greenwald JA, Mehrara BJ, Spector JA, Warren SM, Crisera FE, Fagenholz PJ, Bouletreau PJ, Longaker MT (2000) Regional differentiation of cranial suture-associated dura mater in vivo and in vitro: implications for suture fusion and patency. J Bone Miner Res 15:2413–2430PubMedGoogle Scholar
  9. Gross J, Lapiere CM (1962) Collagenolytic activity in amphibian tissues: a tissue culture assay. Proc Natl Acad Sci USA 54:1197–1204Google Scholar
  10. Ignelzi MA Jr, Wang W, Young AT (2003) Fibroblast growth factors lead to increased Msx2 expression and fusion in calvarial sutures. J Bone Miner Res 18:751–759PubMedGoogle Scholar
  11. Joronen K, Salminen H, Glumoff V, Savontaus M, Vuorio E (2000) Temporospatial expression of tissue inhibitors of matrix metalloproteinases-1, -2 and -3 during development, growth and aging of the mouse skeleton. Histochem Cell Biol 114:157–165PubMedGoogle Scholar
  12. Kokich VG (1976) Age changes in human frontozygomatic sutur. Am J Orthod 69:411–430CrossRefPubMedGoogle Scholar
  13. Kopher RA, Mao JJ (2003) Suture growth modulated by the oscillatory component of micromechanical strain. J Bone Miner Res 18:521–528PubMedGoogle Scholar
  14. Kopher RA, Nudera JA, Wang X, O’Grady K, Mao JJ (2003) Exogenous static and cyclic forces induce characteristic bone strain profiles of rabbit facial sutures. Ann Biomed Eng 31:1125–1131CrossRefPubMedGoogle Scholar
  15. Kusano K, Miyaura C, Inada M, Tamura T, Ito A, Nagase H, Kamoi K, Suda T (1998) Regulation of matrix metalloproteinases (MMP-2, -3, -9, and -13) by interleukin-1 and interleukin-6 in mouse calvaria: association of MMP induction with bone resorption. Endocrinology 139:1338–1345CrossRefGoogle Scholar
  16. Longaker MT (2001) Role of TGF-beta signaling in the regulation of programmed cranial suture fusion. J Craniofac Surg 12:389–390CrossRefPubMedGoogle Scholar
  17. Mao JJ (2002) Mechanobiology of craniofacial sutures. J Dent Res 81:810–816PubMedGoogle Scholar
  18. Mao JJ, H-D Nah (2004) Growth and development: hereditary and mechanical modulations. Am J Orthod 125:676–689Google Scholar
  19. Mao JJ, Wang X, Kopher RA (2003a) Biomechanics of craniofacial sutures: orthopedic implications. Angle Orthod 73:128–135PubMedGoogle Scholar
  20. Mao JJ, Wang X, Mooney MP, Kopher RA, Nudera JA (2003b) Strain induced osteogenesis of the craniofacial suture upon controlled delivery of low-frequency cyclic forces. Front Biosci 8:A10–A17PubMedGoogle Scholar
  21. Matrisian LM (1992) The matrix-degrading metalloproteinases. BioEssays 7:455–463CrossRefGoogle Scholar
  22. Mehrara BJ, Mackool RJ, McCarthy JG, Gittes GK, Longaker MT (1998) Immunolocalization of basic fibroblast growth factor and fibroblast growth factor receptors 1 and 2 in rat cranial sutures. J Craniofac Surg 6:358–363Google Scholar
  23. Meikle MC, Sellers A, Reynolds JJ (1980) Effect of tensile mechanical stress on the synthesis of metalloproteinases by rabbit cranial sutures in vitro. Calcif Tissue Int 30:77–82PubMedGoogle Scholar
  24. Meikle MC, Heath JK, Reynolds JJ (1984) The use of in vitro models for investigating the response of fibrous joints to tensile mechanical stress. Am J Orthod 85:141–153CrossRefPubMedGoogle Scholar
  25. Miyawaki S, Forbes DP (1987) The morphologic and biochemical effects of tensile force application to the interparietal suture of the Sprague–Dawley rat. Am J Orthod 92:123–133Google Scholar
  26. Mooney MP, Burrows AM, Smith TD, Losken HW, Opperman LA, Dechant J, Kreithen AM, Kapueu R, Cooper GM, Ogle RC, Siegel MI (2001) Correction of coronal suture synostosis using suture and dura mater allograft rabbits with familial craniosynostosis. Cleft Palate Craniofac J 38:206–225CrossRefPubMedGoogle Scholar
  27. Mosley JR (2000) Osteoporosis and bone functional adaptation: mechanobiological regulation of bone architecture in growing and adult bone, a review. J Rehabil Res Dev 37:189–199PubMedGoogle Scholar
  28. Mosley JR, Lanyon LE (1998) Strain rate as a controlling influence on adaptive modeling in response to dynamic loading of the ulna in growing male rats. Bone 23:313–318CrossRefPubMedGoogle Scholar
  29. Mosley JR, March BM, Lynch J, Lanyon LE (1997) Strain magnitude related changes in whole bone architecture in growing rats. Bone 20:191–198CrossRefPubMedGoogle Scholar
  30. Moss ML (1954) Growth of the calvaria in the rat. Am J Anat 94:333–361CrossRefPubMedGoogle Scholar
  31. Nacamuli RP, Fong KD, Warren SM, Fang TD, Song HM, Helms JA, Longaker MT (2003) Markers of osteoblast differentiation in fusing and nonfusing cranial sutures. Plast Reconstr Surg 112:1328–1335CrossRefPubMedGoogle Scholar
  32. Nacamuli RP, Song HM, Fang TD, Fong KD, Mathy JA, Shi YY, Salim A, Longaker MT (2004) Quantitative transcriptional analysis of fusing and nonfusing cranial suture complexes in mice. Plast Reconstr Surg 114:1818–1825CrossRefPubMedGoogle Scholar
  33. Oberheim MC, Mao JJ (2002) Bone strain patterns of the zygomatic complex in response to simulated orthopedic forces. J Dent Res 81:608–612PubMedGoogle Scholar
  34. Opperman LA (2000) Cranial sutures as intramembranous bone growth sites. Dev Dyn 219:472–485CrossRefPubMedGoogle Scholar
  35. Opperman LA, Passarelli RW, Morgan EP, Reintjes M, Ogle RC (1995) Cranial sutures require interactions with the dura mater to resist osseous obliteration in vitro. J Bone Miner Res 10:1978–1987PubMedGoogle Scholar
  36. Opperman LA, Moursi AM, Sayne JR, Wintergerst AM (2002) Transforming growth factor-beta 3(Tgf-beta3) in a collagen gel delays fusion of the rat posterior interfrontal suture in vivo. Anat Rec 267:120–130CrossRefPubMedGoogle Scholar
  37. Rafferty KL, Herring SW (1999) Craniofacial sutures: morphology, growth, and in vivo masticatory strains. J Morphol 242:167–179CrossRefPubMedGoogle Scholar
  38. Rubin CT, Lanyon LE (1984) Osteoregulatory nature of mechanical stimuli: function as determinant for adaptive remodeling in bone. J Orthop Res 5:300–310CrossRefGoogle Scholar
  39. Rubin CT, Lanyon LE (1985) Regulation of bone mass by mechanical strain magnitude. Calcif Tissue Int 37:411–417PubMedGoogle Scholar
  40. Shingleton WD, Hodges DJ, Bzrick P, Cawston TE (1996) Collagenase: a key enzyme in collagen turnover. Biochem Cell Biol 74:759–775PubMedGoogle Scholar
  41. Spector J, Mehrara B, Greenwald J, Saadeh P, Steinbrech D, Smith L, Longaker M (2000) A molecular analysis of the isolated rat posterior frontal and sagittal sutures: differences in gene expression. Plast Reconstr Surg 106:852–861CrossRefPubMedGoogle Scholar
  42. Takahara M, Naruse T, Takagi M, Orui H, Ogino T (2004) Matrix metalloproteinase-9 expression, tartrate-resistant acid phosphatase activity, and DNA fragmentation in vascular and cellular invasion into cartilage preceding primary endochondral ossification in long bones. J Orthop Res 22:1050–1057CrossRefPubMedGoogle Scholar
  43. Turner CH, Forwood MR, Rho JY, Yoshikawa T (1994) Mechanical loading thresholds for lamellar and woven bone formation. J Bone Mer Res 9:87–97Google Scholar
  44. Warren SM, Longaker MT (2001) The pathogenesis of craniosynostosis in the fetus Yonsei. Med J 42:646–659Google Scholar
  45. Weijs WA, De Jongh HJ (1977) Strain in mandibular alveolar bone during mastication in the rabbit. Arch Oral Biol 22:667–675CrossRefPubMedGoogle Scholar
  46. Wilkie AOM, Morriss-Kay GM (2001) Genetics of craniofacial development and malformation. Nature 2:458–468Google Scholar
  47. Yen EHK, Yue CS, Suga DM (1989) Effect of force level on synthesis of type III and type I collagen in mouse interparietal suture. J Dent Res 68:1746–1751PubMedGoogle Scholar
  48. Zhou Z, Apte SS, Soininen R, Cao R, Baaklini GY, Rauser RW, Wang J, Cao Y, Tryggvason K (2000) Impaired endochondral ossification and angiogenesis in mice deficient in membrane-type matrix metalloproteinase I. Proc Natl Acad Sci USA 97:4052–4057CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Rasha Al-Mubarak
    • 1
  • Adriana Da Silveira
    • 1
  • Jeremy J. Mao
    • 1
  1. 1.Tissue Engineering Laboratory, Rm 237University of Illinois at Chicago, MC 841ChicagoUSA

Personalised recommendations