Effects of laminin on hard tissue formation by bone marrow cells in vivo and in vitro


The effect of laminin on hard tissue formation using rat bone marrow cells was assessed. Rat bone marrow cells were obtained from femora of 6-week-old male Fischer 344 rats. In this in vivo examination, porous cylindrical hydroxyapatite scaffolds with a hollow center were immersed in 100 mg/ml laminin solution and air-dried. Rat bone marrow cells in 200 ml culture medium at 1 × 106 cells/ml were seeded in the scaffolds. The scaffolds were implanted into the dorsal subcutis of 7-week-old male Fischer 344 rats for 6 weeks. The scaffolds were then removed and examined histologically. For in vitro examinations, 1 × 105 rat bone marrow cells in 2 ml culture medium were then cultured with the addition of dexamethasone and laminin. Rat bone marrow cells were also cultured in laminin-coated culture plates. In vitro examinations showed the effectiveness of laminin for hard tissue formation from the results of biochemical and immunochemical analysis. From the in vivo examination, laminin coating of the scaffolds induced hard tissue in the pores with the cells. It is concluded that laminin is useful for bone formation, as in an in vitro culture study using bone marrow cells, in hydroxyapatite scaffolds in vivo.

Share and Cite:

Yoshikawa, M. , Kakigi, H. , Yabuuchi, T. and Hayashi, H. (2014) Effects of laminin on hard tissue formation by bone marrow cells in vivo and in vitro. Journal of Biomedical Science and Engineering, 7, 15-23. doi: 10.4236/jbise.2014.71003.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Pittenger, M.F. and Martin, B.J. (2004) Mesenchymal stem cells and their potential as cardiac therapeutics. Circulation Research, 95, 9-20.
[2] Barry, F.P. and Murphy, J.M. (2004) Mesenchymal stem cells: Clinical applications and biological characterization. The International Journal of Biochemistry & Cell Biology, 36, 568-584.
[3] Petite, H., Viateau, V., Bensaid, W., Meunier, A., De Pollak, C., Bourguignon, M., Oudina, K., Sedel, L. and Guillemin, G. (2000) Tissue-engineered bone regeneration. Nature Biotechnology, 18, 959-963.
[4] Caplan, A.I. and Bruder, S.P. (2001) Mesenchymal stem cells: Building blocks for molecular medicine in the 21st century. Trends in Molecular Medicine, 7, 259-264.
[5] Derubeis, A.R. and Cancedda, R. (2004) Bone marrow stromal cells (BMSCs) in bone engineering: Limitations and recent advances. Annals of Biomedical Engineering, 32, 160-165.
[6] Yoshikawa, M., Tsuji, N., Shimomura, Y., Hayashi, H. and Ohgushi, H. (2008) Osteogenesis depending on geometry of porous hydroxyapatite scaffolds. Calcified Tissue International, 83, 139-145.
[7] Akahane, M., Ohgushi, H., Yoshikawa, T., Sempuku, T., Tamai, S., Tabata, S. and Dohi, Y. (1999) Osteogenic phenotype expression of allogeneic rat marrow cells in porous hydroxyapatite ceramics. Journal of Bone and Mineral Research, 14, 561-568.
[8] Huang, G.T., Gronthos, S. and Shi, S. (2009) Mesenchymal stem cells derived from dental tissues vs. those from other sources: Their biology and role in regenerative medicine. Journal of Dental Research, 88, 792-806.
[9] Sengupta, S., Park, S.H., Patel, A., Carn, J., Lee, K. and Kaplan, D.L. (2010) Hypoxia and amino acid supplementation synergistically promote the osteogenesis of human mesenchymal stem cells on silk protein scaffolds. Tissue Engineering Part A, 16, 3623-3634.
[10] Huang, C.H., Tseng, W.Y., Yao, C.C., Jeng, J.H., Young, T.H. and Chem, Y.J. (2010) Glucosamine promotes osteogenic differentiation of dental pulp stem cells through modulating the level of the transforming growth factor-?type I receptor. Journal of Cellular Physiology, 225, 140-151. http://dx.doi.org/10.1002/jcp.22206
[11] Bohl, K.S., Shon, J., Rutherford, B. and Mooney, D.J. (1998) Role of synthetic extracellular matrix in development of engineered dental pulp. Journal of Biomaterials Science, Polymer Edition, 9, 749-764.
[12] Srisuwan, T., Tilkorn, D.J., Al-Benna, S., Vashi, A., Penington, A., Messer, H.H., Abberton, K.M. and Thompson, E.W. (2012) Survival of rat functional dental pulp cells in vascularized tissue engineering chambers. Tissue and Cell, 44, 111-121.
[13] Leye Benoist, F., Gaye Ndiaye, F., Kane, A.W., Benoist, H.M. and Farge, P. (2012) Evaluation of mineral trioxide aggregate (MTA) versus calcium hydroxide cement (Dycal?) in the formation of a dentine bridge: A randomised controlled trial. International Dental Journal, 62, 33-39.
[14] Yoshikawa, M., Tsuji, N. and Toda, T. (2004) Hard tissue formation by cultured dental pulp cells and bone marrow cells. Journal of Osaka Dental University, 38, 119-125.
[15] Santiago, J.A., Pogemiller, R. and Ogle, B.M. (2009) Heterogeneous differentiation of human mesenchymal stem cells in response to extended culture in extracellular matrices. Tissue Engineering Part A, 15, 3911-3922.
[16] Burridge, K., Fath, K., Kelly, T., Nuckolls, G. and Turner, C. (1988) Focal adhesions: Transmembrane junctions between the extracellular matrix and the cytoskeleton. Annual Review of Cell Biology, 4, 487-525.
[17] Gu, Y.C., Kortesmaa, J., Tryggvason, K., Persson, J., Ekblom, P., Jacobsen, S.E. and Ekblom, M. (2003) Laminin isoform-specific promotion of adhesion and migration of human bone marrow progenitor cells. Blood, 101, 877-885. http://dx.doi.org/10.1182/blood-2002-03-0796
[18] D’Ippolito, G., Schiller, P.C., Ricordi, C., Roos, B.A. and Howard, G.A. (1999) Age-related osteogenic potential of mesenchymal stromal cells from human vertebral bone marrow. Journal of Bone and Mineral Research, 14, 1115-1122.
[19] Ohgushi, H., Dohi, Y., Yoshikawa, T., Tamai, S., Tabata, S., Okunaga, K. and Shibuya, T. (1996) Osteogenic differentiation of cultured marrow stromal stem cells on the surface of bioactive glass ceramics. Journal of Biomedical Materials Research, 32, 341-348.
[20] Klees, R.F., Salasznyk, R.M., Kingsley, K., Williams, W.A., Boskey, A. and Plopper, G.E. (2005) Laminin-5 induces osteogenic gene expression in human mesenchymal stem cells through an ERK-dependent pathway. Molecular Biology of the Cell, 16, 881-890.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.