The Effects of Various Extracellular Matrices on Motility of Cultured MC3T3-E1 Cell

다양한 세포외기질이 배양 골아세포의 이동에 미치는 영향

  • Park, Beyoung Yun (Institute for Human Tissue Restoration, Department of Plastic and Reconstructive Surgery, Yonsei University College of Medicine,) ;
  • Seo, Sang Woo (Institute for Human Tissue Restoration, Department of Plastic and Reconstructive Surgery, Yonsei University College of Medicine,) ;
  • Lee, Won Jai (Institute for Human Tissue Restoration, Department of Plastic and Reconstructive Surgery, Yonsei University College of Medicine,) ;
  • Ryu, Chang Woo (Institute for Human Tissue Restoration, Department of Plastic and Reconstructive Surgery, Yonsei University College of Medicine,) ;
  • Rah, Dong Kyun (Institute for Human Tissue Restoration, Department of Plastic and Reconstructive Surgery, Yonsei University College of Medicine,) ;
  • Son, Hyun Joo (Brain Korea 21 Project for Medical Science, Medical Engineering, Yonsei University College of Medicine) ;
  • Park, Jong Chul (Brain Korea 21 Project for Medical Science, Medical Engineering, Yonsei University College of Medicine)
  • 박병윤 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소) ;
  • 서상우 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소) ;
  • 이원재 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소) ;
  • 류창우 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소) ;
  • 나동균 (연세대학교 의과대학 성형외과학교실 인체조직복원연구소) ;
  • 손현주 (연세대학교 의과대학 의학공학교실, BK21 의과학사업단) ;
  • 박종철 (연세대학교 의과대학 의학공학교실, BK21 의과학사업단)
  • Received : 2004.11.09
  • Published : 2005.03.10

Abstract

Chemotactic migration of bone forming cell, osteoblast, is an important event during bone formation, bone remodeling, and fracture healing. Migration of cells is mediated by adhesion receptors, such as integrins, that link the cell to extracellular matrix ligands, type I collagen, fibronectin, laminin and depend on interaction between integrin and extracellular ligand. Our study was designed to investigate the effect of extracellular matrix like fibronectin, laminin, type I collagen on migration of osteoblast. Migration distance and speed of MC3T3-E1 cell on extracellular matrix-coated glass were measured for 24 hours using 0.01% type I collagen, 0.01% fibronectin, 100 microliter/ml laminin. The migration distance and speed of MC3T3-E1 cell was compared using a video-microscopy system. To determine migration speed, cells were viewed with a 4 phase- contrast lens and video recorded. Images were captured using a color CCD camera and saved in 8-bit full-color mode. The migration distance on 0.01% type I collagen or 0.01% fibronectin was longer than that on $100{\mu}l/ml$ laminin-coated glass. The migration speed on fibronectin-coated glass was 68 micrometer/hour which was fastest. The migration speed on type I collagen-coated glass was similar with that on fibronectin-coated glass. The latter two migration speeds were faster than that on no-coated glass. On the other hand, the average migration speed on laminin-coated glass was 37micrometer/hour and not different from that of control group. In conclusion, the extracelluar matrix ligands such as type I collagen and fibronectin seem to play an important role in cell migration. The type I collagen or fibronectin coated scaffold is more effective for migration of osteoblast in tissue engineering process.

Keywords

References

  1. Imai S, Kaksonen M, Raulo E, Kinnunen T, Fages C, Meng X, Lakso M, Rauvala H: Osteoblast recruitment and bone formation enhanced by cell matrix-associated heparinbinding growth-associated molecule(HB-GAM). J Cell Biol 143: 1113, 1998 https://doi.org/10.1083/jcb.143.4.1113
  2. Grzesik WJ, Gehron-Robey P: Bone matrix RGD glycoproteins: immunolocaliztion and interaction with primary osteoblastic bone cells in vitro. J Bone Miner Res 9: 487, 1997
  3. Gehron-Robey P: The biochemistry of bone. Endocrinol Metab Clin North Am 18: 858, 1989
  4. Damsky CH, Werb Z: Signal transduction by integrin receptors for extracellular matrix: cooperative processing of extracellular information. Curr Opin Cell Biol 4: 772, 1992 https://doi.org/10.1016/0955-0674(92)90100-Q
  5. Sastry SK, Horwitz AF: Integrin cytoplasmic domains: mediators of cytoskeletal linkage and extra- and intracellular initiated transmembrane signaling. Curr Opin Cell Biol 5: 819, 1993 https://doi.org/10.1016/0955-0674(93)90031-K
  6. Beck K, Hunter I, Engel J: Structures and function of laminin: anatomy of a multidomain glycoprotein. FASEB J 4: 148, 1990 https://doi.org/10.1096/fasebj.4.2.2404817
  7. Engel J: Laminins and other strange proteins. Biochemistry 31: 10643, 1992 https://doi.org/10.1021/bi00122a014
  8. Timpl R, Brown JC: The laminins. Matrix Biol 14: 275, 1994 https://doi.org/10.1016/0945-053X(94)90192-9
  9. Gover J, Gowen M: Are MG-63 and HOS TE 85 human osteosarcoma cell lines representative models of the osteoblastic phenotype? Bone 15: 585, 1994 https://doi.org/10.1016/8756-3282(94)90305-0
  10. Gronthos S, Stewart K, Graves SE, Hay S, Simmons PJ: Integrin expression and function on human osteoblast-like cells. J Bone Miner Res 12: 1189, 1997 https://doi.org/10.1359/jbmr.1997.12.8.1189
  11. Saito T, Albelda SM, Brighton CT: Identification of integrin receptors on cultured human bone cells. J Orthop Res 12: 384, 1994 https://doi.org/10.1002/jor.1100120311
  12. Lacouture ME, Schaffer IL, Klickstein LB: A comparison of type I collagen, fibronectin, and vitronectin in supporting adhesion of mechanically strained osteoblasts. J Bone Miner Res 17: 481, 2002 https://doi.org/10.1359/jbmr.2002.17.3.481
  13. Roche P, Rousselle P, Lissitzky JC, Delmas PD, Malaval L: Isoform-specific attachment of osteoprogenitors to laminins: mapping to the short arms of laminin-1. Exp Cell Res 250: 465, 1999 https://doi.org/10.1006/excr.1999.4518
  14. Carvalho RS, Kostenuik PJ, Salih E, Bumann A, Gerstenfeld LC: Selective adhesion of osteoblastic cells to different integrin ligands induces osteopontin gene expression. Matrix Biol 22: 241, 2003 https://doi.org/10.1016/S0945-053X(03)00038-6
  15. Stossel TP: On the crawling of animal cells. Science 260: 1086, 1993 https://doi.org/10.1126/science.8493552
  16. McCarthy J, Turley EA: Effects of extracellular matrix components on cell locomotion. Crit. Rev. Oral Biol Med 4: 619, 1993 https://doi.org/10.1177/10454411930040050101
  17. Pavalko FM, Chen NX, Turner CH, Burr DB, Atkinson S, Hsieh YF, Qiu J, Duncan RL: Fluid shear-induced mechanical signaling in MC3T3-E1 osteoblasts requires cytoskeleton-integrin interactions. Am J Physiol 275: 1591, 1998 https://doi.org/10.1152/ajpcell.1998.275.6.C1591
  18. Takeuchi Y, Nakayama K, Matsumoto T: Differentiation and cell surface expression of transforming growth factor-beta receptors are regulated by interaction with matrix collagen in murine osteoblastic cells. J BioI Chem 271: 3938, 1996 https://doi.org/10.1074/jbc.271.7.3938