Bone Formation in a Scaffold Composed of Cylindrical Hydroxyapatite and Tryptophan- or Lysine-Coated Sponge in Vivo


Because of the three-dimensional structure of bone or hard tissue such as a tooth, a scaffold is necessary for its regeneration by cellular engineering. Commonly, for in vivo examination, hydroxyapatite (HA) has been used as such a scaffold. Cylindrical HA with a hollow center, which included a columnar formalin-treated polyvinyl alcohol sponge, was used in this examination as a scaffold. The sponge had been coated with L-tryptophan or L-lysine before insertion into the hollow center of the HA. Rat bone marrow cells (rBMCs) derived from the femur were seeded in the sponge before insertion into the hollow center of HA. The number of rBMCs seeded in each sponge was 1.5 × 106. These scaffolds were implanted subcutaneously into the backs of Fischer 344 rats for 6 weeks. In the amino-acid-coated sponge in HA, osteogenesis was found histologically. An osteocalcin level of approximately 10 ug was measured in the scaffolds with L-tryptophan-coated formalized poly-vinyl alcohol sponge containing rBMCs, 4 ug on average in the scaffolds with L-lysine-coated sponge containing the cells and about 2 ug in each scaffold with uncoated sponge containing the cells. The structure of the scaffolds used in this study was thought to be suitable for osteogenesis by rBMCs. It was concluded that tryptophan, as a factor for bone formation by stem cells, functioned by promoting cell adhesion and the differentiation of stem cells into osteoblasts.

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Yoshikawa, M. , Kakigi, H. , Maeda, H. , Nishikawa, I. , Ikenaga, H. , Inamoto, T. and Tsuji, N. (2015) Bone Formation in a Scaffold Composed of Cylindrical Hydroxyapatite and Tryptophan- or Lysine-Coated Sponge in Vivo. Journal of Biomedical Science and Engineering, 8, 389-398. doi: 10.4236/jbise.2015.86037.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Shim, I.K., Jung, M.R., Kim, K.H., Seol, Y.J., Park, Y.J., Park, W.H. and Lee, S.J. (2010) Novel Three-Dimensional Scaf-folds of Poly(L-Lactic Acid) Microfibers Using Electrospinning and Mechanical Expansion: Fabrication and Bone Regeneration. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 95, 150-160.
[2] Yoshikawa, M., Tshji, N., Shimomura, Y., Hayashi, H. and Ohgushi, H. (2008) Osteogenesis Depending on Geometry of Porous Hydroxyapatite Scaffolds. Calcified Tissue International, 83, 139-145.
[3] Yoshikawa, M., Tshji, N., Toda, T. and Ohgushi, H. (2007) Osteogenic Effect of Hyaluronic Acid Sodium Salt in the Pores of a Hydroxyapatite Scaffold. Materials Science and Engineering: C, 27, 220-226.
[4] Masica D.L., Gray, J.J. and Shaw, W.J. (2011) Partial High-Resolution Structure of Phosphorylated and Nonphosphorylated Leucine-Rich Amelogenin Protein Adsorbed to Hydroxyapatite. The Journal of Physical Chemistry C: Nanomaterials and Interfaces, 115, 13775-13785.
[5] Yabuuchi, T., Yoshikawa, M., Kakigi, H. and Hayashi, H. (2014) Hybrid Scaffolds Composed of Amino-Acid Coated Sponge and Hydroxyapatite for Hard Tissue Formation by Bone Marrow Cells. Journal of Biomedical Science and Engineering, 7, 316-329.
[6] Sibilia, V., Pagani, F., Lattuada, N., Greco, A. and Guidobono, F. (2009) Linking Chronic Tryptophan Deficiency with Impaired Bone Metabolism and Reduced Bone Accrual in Growing Rats. Journal of Cellular Biochemistry, 107, 890-898.
[7] Torricelli, P., Fini, M., Giavaresi, G. and Giardino, R. (2001) Bone Tissue Cultures: An in Vitro Model for the Evaluation of Bone Defect Healing after L-Arginine and L-Lysine Administration. Artificial Cells, Blood Substitutes and Biotechnology, 29, 325-334.
[8] Visser, J.J. and Hoekman, K. (1994) Arginine Supplementation in the Prevention and Treatment of Osteoporosis. Medical Hypotheses, 43, 339-342.
[9] Ida-Yonemochi, H., Nakatomi, M., Harada, H., Takata, H., Baba, O. and Ohshima, H. (2011) Glucose Uptake Mediated by Glucose Transporter 1 Is Essential for Early Tooth Morphogenesis and Size Determination of Murine Molars. Developmental Biology, 363, 52-61.
[10] Traphagen, S. and Yelick, P.C. (2009) Reclaiming a Natural Beauty: Whole-Organ Engineering with Natural Extra-Cellular Materials. Regenerative Medicine, 4, 747-758.
[11] Komine, A., Suenaga, M., Nakao, K., Tsuji T. and Tomooka, Y. (2007) Tooth Regeneration from Newly Established Cell Lines from a Molar Tooth Germ Epithelium. Biochemical and Biophysical Research Communications, 355, 758- 763.
[12] Ohazama, A. and Sharpe, P.T. (2004) TNF Signalling in Tooth Development. Current Opinion in Genetics & Development, 14, 513-519.
[13] Tucker, A. and Sharpe, P. (2004) The Cutting-Edge of Mammalian Development: How the Embryo Makes Teeth. Nature Reviews Genetics, 5, 499-508.
[14] Desai, N., Ludgin, J., Goldberg, J. and Falcone, T. (2013) Development of a Xeno-Free Non-Contact Co-Culture System for Derivation and Maintenance of Embryonic Stem Cells Using a Novel Human Endometrial Cell Line. Journal of Assisted Reproduction and Genetics, 30, 609-615.
[15] Antebi, B., Cheng, X., Harris, J.N., Gower, L.B., Chen, X.D. and Ling, J. (2013) Biomimetic Collagen-Hydroxyapatite Composite Fabricated via a Novel Perfusion-Flow Mineralization Technique. Tissue Engineering Part C: Methods, 19, 487-496.
[16] Sopyana, I., Mel, M., Ramesh, S. and Khalid, K.A. (2007) Porous Hydroxyapatite for Artificial Bone Applications. Science and Technology of Advanced Materials, 8, 116-123.
[17] Beyth, S., Schroeder, J. and Liebergall, M. (2011) Stem Cells in Bone Diseases: Current Clinical Practice. British Medical Bulletin, 99, 199-210.
[18] Sakai, K., Yamamoto, A., Matsubara, K., Nakamura, S., Naruse, M., Yamagata, M., Sakamoto, K., Tauchi, R., Wakao, N., Imagama, S., Hibi, H., Kadomatsu, K., Ishiguro, N. and Ueda, M. (2012) Human Dental Pulp-Derived Stem Cells Promote Locomotor Recovery after Complete Transection of the Rat Spinal Cord by Multiple Neuro-Regenerative Mechanisms. The Journal of Clinical Investigation, 122, 80-90.
[19] Ganz, J., Arie, I., Ben-Zur, T., Dadon-Nachum, M., Pour, S., Araidy, S., Pitaru, S. and Offen, D. (2014) Astrocyte-Like Cells Derived from Human Oral Mucosa Stem Cells Provide Neuroprotection in Vitro and in Vivo. Stem Cells Translational Medicine, 3, 375-386.
[20] Zhang, Q.Z., Nguyen, A.L., Yu, W.H. and Le, A.D. (2012) Human Oral Mucosa and Gingiva: A Unique Reservoir for Mesenchymal Stem Cells. Journal of Dental Research, 91, 1011-1018.
[21] Yoshikawa, M., Shimomura, Y., Kakigi, H., Tsuji, N., Yabuuchi, T. and Hayashi, H. (2012) Effect of L-Lysine in Culture Medium on Nodule Formation by Bone Marrow Cells. Journal of Biomedical Science and Engineering, 5, 587-592.
[22] 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.
[23] Yoshikawa, M., Tsuji, N., Kakigi, H., Yabuuchi, T., Shimomura, Y., Hayashi, H. and Ohgushi, H. (2010) Dextran Coating on and among Fibers of Polymer Sponge Scaffold for Osteogenesis by Bone Marrow Cells in Vivo. Journal of Biomedical Science and Engineering, 3, 751-757.
[24] Mauney, J.R., Blumberg, J., Pirun, M., Volloch, V., Vunjak-Novakovic, G. and Kaplan, D.L. (2004) Osteogenic Differentiation of Human Bone Marrow Stromal Cells on Partially Demineralized Bone Scaffolds in Vitro. Tissue Engineering, 10, 81-92.
[25] Togami, W., Sei, A., Okada, T., Taniwaki, T., Fujimoto, T., Tahata, S., Nagamura, K., Nakanishi, Y. and Mizuta, H. (2015) Effects of the Water-Holding Capability of Polyvinyl Formal Sponges on Osteogenic Ability in in Vivo Experiments. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 103, 188-194.
[26] Gustafsson, Y., Haag, J., Jungebluth, P., Lundin, V., Lim, M.L., Baiguera, S., Ajalloueian, F., Del Gaudio, C., Bianco, A., Moll, G., Sjoqvist, S., Lemon, G., Teixeira, A.I. and Macchiarini, P. (2012) Viability and Proliferation of Rat MSCs on Adhesion Protein-Modified PET and PU Scaffolds. Biomaterials, 33, 8094-8103.
[27] Kaper, T., Looger, L.L., Takanaga, H., Platten, M., Steinman, L. and Frommer, W.B. (2007) Nanosensor Detection of an Immunoregulatory Tryptophan Influx/Kynurenine Efflux Cycle. PLoS Biology, 5, e257.
[28] Sibilia, V., Pagani, F., Dieci, E., Mrak, E., Marchese, M., Zarattini, G. and Guidobono, F. (2013) Dietary Tryptophan Manipulation Reveals a Central Role for Serotonin in the Anabolic Response of Appendicular Skeleton to Physical Activity in Rats. Endocrine, 44, 790-802.
[29] Ducy, P. and Karsenty, G. (2010) The Two Faces of Serotonin in Bone Biology. The Journal of Cell Biology, 191, 7-13.
[30] Conconi, M.T., Tommasini, M., Muratori, E. and Parnigotto, P.P. (2001) Essential Amino Acids Increase the Growth and Alkaline Phosphatase Activity in Osteoblasts Cultured in Vitro. II Farmaco, 56, 755-761.

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