JBiSE  Vol.7 No.6 , May 2014
Hybrid Scaffolds Composed of Amino-Acid Coated Sponge and Hydroxyapatite for Hard Tissue Formation by Bone Marrow Cells
ABSTRACT
A formalin-treated polyvinyl-alcohol (PVF) sponge is convenient as a scaffold because its configuration is easily modified. However, coating the sponge with an adhesive chemical agent is necessary to attach bone marrow cells (BMCs) to the sponge structure. Moreover, it was considered that a hybrid scaffold composed of a sponge and enveloped cylindrical porous hydroxyapatite (HA) would be convenient. In this study, the effect of leucine (Leu) coating on a PVF sponge was examined for osteogenesis on an HA/PVF hybrid scaffold by rat BMCs (rBMCs). In an in vivo assessment, the sponge immersed in Leu solution (10 mg/ml) was inserted into the hollow center of cylindrical HA. The sponge received 1.5 × 106 rBMCs obtained from male Fischer 344 rats. The hybrid scaffolds were then implanted subcutaneously of syngeneic rats for 6 weeks. In vitro assessment of Leu to hard tissue formation with coating on the well or addition in rBMC culture medium was also performed in a 6-well plate for 2 weeks. In vivo examinations showed the excellent effect of Leu coating on PVF sponge. Leu-coated PVF sponge in the scaffolds showed marked new bone formation in the pores by histological examination. Leu-coated PVF sponge showed a high quantity of osteocalcine (OC). HA might prevent the release of rBMCs from PVF as a barrier. In in vitro examinations, the quantity of OC in rBMC culture with and without the addition of Leu in culture medium showed no significant difference. However, addition of Leu showed significant ALP activity level in culture medium. Leu coating in culture plate wells showed no influence on the quantity of OC. It was concluded from the results that Leu might prevent the emigration of rBMCs to the outside of the scaffold and promote the differentiation of cells to osteoblasts in the scaffold.

Cite this paper
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. doi: 10.4236/jbise.2014.76034.
References
[1]   Wen, Y., Wang, F., Zhang, W., Li, Y., Yu, M., Nan, X., Chen, L., Yue, W., Xu, X. and Pei, X. (2012) Application of Induced Pluripotent Stem Cells in Generation of a Tissue-Engineered Tooth-Like Structure. Tissue Engineering: Part A, 18, 1677-1685.
http://dx.doi.org/10.1089/ten.tea.2011.0220

[2]   Mallon, B.S., Hamilton, R.S., Kozhich, O.A., Johnson, K.R., Fann, Y.C., Rao, M.S. and Robey, P.G. (2014) Comparison of the Molecular Profiles of Human Embryonic and Induced Pluripotent Stem Cells of Isogenic Origin. Stem Cell Research, 12, 376-386.
http://dx.doi.org/10.1016/j.scr.2013.11.010

[3]   Mastrogiacomo, M., Muraglia, A., Komlev, V., Peyrin, F., Rustichelli, F., Crovace, A. and Cancedda. R. (2005) Tissue Engineering of Bone: Search for a Better Scaffold. Orthodontics & Craniofacial Research, 8, 277-284.
http://dx.doi.org/10.1111/j.1601-6343.2005.00350.x

[4]   Ohgushi, H., Kotobuki, N., Funaoka, H., Machida, H., Hirose, M., Tanaka, Y. and Takakura, Y. (2005) Tissue Engineered Ceramic Artificial Joint—Ex Vivo Osteogenic Differentiation of Patient Mesenchymal Cells on Total Ankle Joints for Treatment of Osteoarthritis. Biomaterials, 26, 4654-4661.
http://dx.doi.org/10.1016/j.biomaterials.2004.11.055

[5]   Okamoto, M., Dohi, Y., Ohgushi, H., Shimaoka, H., Ikeuchi, M., Matsushima, A., Yonemasu, K. and Hosoi, H. (2006) Influence of the Porosity of Hydroxyapatite Ceramics on in Vitro and in Vivo Bone Formation by Cultured Rat Bone Marrow Stromal Cells. Journal of Materials Science Materials in Medicine, 17, 327-336.
http://dx.doi.org/10.1007/s10856-006-8232-z

[6]   Appleford, M.R., Oh, S., Oh, N. and Ong, J.L. (2009) In Vivo Study on Hydroxyapatite Scaffolds with Trabecular Architecture for Bone Repair. Journal of Biomedical Materials Research Part A, 89A, 1019-1027.
http://dx.doi.org/10.1002/jbm.a.32049

[7]   Herath, H.M., Di Silvio, L. and Evans, J.R. (2010) Biological Evaluation of Solid Free Formed, Hard Tissue Scaffolds for Orthopedic Applications. Journal of Applied Biomaterials & Functional Materials, 8, 89-96.

[8]   Yoshikawa, M., Tsuji, 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.
http://dx.doi.org/10.1016/j.msec.2006.05.014

[9]   Jeong, W.K., Oh, S.H., Lee, J.H. and Im, G.I. (2008) Repair of Osteochondral Defects with a Construct of Esenchymal Stem Cells and a Polydioxanone/Poly (Vinyl Alcohol) Scaffold. Biotechnology and Applied Biochemistry, 49, 155-164.
http://dx.doi.org/10.1042/BA20070149

[10]   Charlton, D.C., Peterson, M.G., Spiller, K., Lowman, A., Torzilli, P.A. and Maher, S.A. (2008) Semi-Degradable Scaffold for Articular Cartilage Replacement. Tissue Engineering: Part A, 14, 207-213.
http://dx.doi.org/10.1089/ten.a.2006.0344

[11]   Thomas, L.V., Arun, U., Remya, S. and Nair, P.D. (2009) A Biodegradable and Biocompatible PVA-Citric Acid Polyester with Potential Applications as Matrix for Vascular Tissue Engineering. Journal Materials Science: Materials in Medicine, 20, S259-S269.
http://dx.doi.org/10.1007/s10856-008-3599-7

[12]   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 & Engineering, 3, 751-757.
http://dx.doi.org/10.4236/jbise.2010.38100

[13]   Lemus, D. (1995) Contributions of Heterospecific Tissue Recombinations to Odontogenesis. The International Journal of Developmental Biology, 39, 291-297.

[14]   Smith, A.J. and Lesot, H. (2001) Induction and Regulation of Crown Dentinogenesis: Embryonic Events as a Template for Dental Tissue Repair? Critical Reviews in Oral Biology & Medicine, 12, 425-437.
http://dx.doi.org/10.1177/10454411010120050501

[15]   Gronthos, S., Brahim, J., Li, W., Fisher, L.W., Cherman, N., Boyde, A., DenBesten, P., Robey, P.G. and Shi, S. (2002) Stem Cell Properties of Human Dental Pulp Stem Cells. Journal of Dental Research, 81, 531-535.
http://dx.doi.org/10.1177/154405910208100806

[16]   Jo, Y.Y., Lee, H.J., Kook, S.Y., Choung, H.W., Park, J.Y., Chung, J.H., Choung, Y.H., Kim, E.S., Yang, H.C. and Choung, P.H. (2007) Isolation and Characterization of Postnatal Stem Cells from Human Dental Tissues. Tissue Engineering, 13, 767-773.
http://dx.doi.org/10.1089/ten.2006.0192

[17]   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.
http://dx.doi.org/10.1177/0022034509340867

[18]   Balic, A., Aguila, H.L., Caimano, M.J., Francone, V.P. and Mina, M. (2010) Characterization of Stem and Progenitor Cells in the Dental Pulp of Erupted and Unerupted Murine Molars. Bone, 46, 1639-1651.
http://dx.doi.org/10.1016/j.bone.2010.02.019

[19]   Duailibi, M.T., Duailibi, S.E., Young, C.S., Bartlett, J.D., Vacanti, J.P. and Yelick, P.C. (2004) Bioengineered Teeth from Cultured Rat Tooth Bud Cells. Journal of Dental Research, 83, 523-528.
http://dx.doi.org/10.1177/154405910408300703

[20]   Lin, N.H., Gronthos, S. and Bartold, P.M. (2008) Stem Cells and Periodontal Regeneration. Australian Dental Journal, 53, 108-121.
http://dx.doi.org/10.1111/j.1834-7819.2008.00019.x

[21]   Inoue, K., Ohgushi, H., Yoshikawa, T., Okumura, M., Sempuku, T., Tamai, S. and Dohi, Y. (1997) The Effect of Aging on Bone Formation in Porous Hydroxyapatite: Biochemical and Histological Analysis. Journal of Bone and Mineral Research, 12, 989-994.
http://dx.doi.org/10.1359/jbmr.1997.12.6.989

[22]   Toquet, J., Rohanizadeh, R., Guicheux, J., Couillaud, S., Passuti, N., Daculsi, G. and Heymann, D. (1999) Osteogenic Potential in Vitro of Human Bone Marrow Cells Cultured on Macroporous Biphasic Calcium Phosphate Ceramic. Journal of Biomedical Materials Research, 44, 98-108.
http://dx.doi.org/10.1002/(SICI)1097-4636(199901)44:1<98::AID-JBM11>3.0.CO;2-P

[23]   Livingston, T., Ducheyne, P. and Garino, J. (2002) In Vivo Evaluation of a Bioactive Scaffold for Bone Tissue Engineering. Journal of Biomedical Materials Research, 62, 1-13.
http://dx.doi.org/10.1002/jbm.10157

[24]   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.
http://dx.doi.org/10.1007/s00223-008-9157-y

[25]   Young, C.S., Terada, S., Vacanti, J.P., Honda, M., Bartlett, J.D. and Yelick, P.C. (2002) Tissue Engineering of Complex Tooth Structures on Biodegradable Polymer Scaffolds. Journal of Dental Research, 81, 695-700.
http://dx.doi.org/10.1177/154405910208101008

[26]   Chang, Y.S., Oka, M., Kobayashi, M., Gu, H.O., Li, Z.L., Nakamura, T. and Ikada, Y. (1996) Significance of Interstitial Bone Ingrowth under Load-Bearing Conditions: A Comparison between Solid and Porous Implant Materials. Biomaterials, 17, 1141-1148.
http://dx.doi.org/10.1016/0142-9612(96)85917-5

[27]   Gimble, J.M., Zvonic, S., Floyd, Z.E., Kassem, M. and Nuttall, M.E. (2006) Playing with Bone and Fat. Journal of Cellular Biochemistry, 98, 251-266.
http://dx.doi.org/10.1002/jcb.20777

[28]   Maniatopoulos, C., Sodek, J. and Melcher, A.H. (1988) Bone Formation in Vitro by Stromal Cells Obtained from Bone Marrow of Young Adult Rats. Cell and Tissue Research, 254, 317-330.
http://dx.doi.org/10.1007/BF00225804

[29]   Yoshikawa, T., Ohgushi, H. and Tamai, S. (1996) Immediate Bone Forming Capability of Prefabricated Osteogenic Hydroxyapatite. Journal of Biomedical Materials Research, 32, 481-492.
http://dx.doi.org/10.1002/(SICI)1097-4636(199611)32:3<481::AID-JBM23>3.0.CO;2-I

[30]   Yoshikawa, T., Ohgushi, H., Dohi, Y. and Davies, J.E. (1997) Viable Bone Formation in Porous Hydroxyl Apatite: Marrow Cell-Derived in Vitro Bone on the Surface of Ceramics. Bio-Medical Materials and Engineering, 7, 49-58.

[31]   Jemvall, J. and Thesleff, I. (2000) Reiterative Signaling and Patterning during Mammalian Tooth Morphogenesis. Mechanisms of Development, 92, 19-29.
http://dx.doi.org/10.1016/S0925-4773(99)00322-6

[32]   Koussoulakou, D.S., Margaritis, L.H. and Koussoulakos, S.L. (2009) A Curriculum Vitae of Teeth: Evolution, Generation, Regeneration. International Journal of Biological Sciences, 5, 226-243.
http://dx.doi.org/10.7150/ijbs.5.226

[33]   Tucker, A. and Sharpe, P. (2004) The Cutting-Edge of Mammalian Development; How the Embryo Makes Teeth. Nature Reviews Genetics, 5, 499-508.
http://dx.doi.org/10.1038/nrg1380

[34]   Zhang, W., Walboomers, X.F., Van Kuppevelt, T.H., Daamen, W.F., Van Damme, P.A., Bian, Z. and Jansen, J.A. (2008) In Vivo Evaluation of Human Dental Pulp Stem Cells Differentiated towards Multiple Lineages. Journal of Tissue Engineering and Regenerative Medicine, 2, 117-125.
http://dx.doi.org/10.1002/term.71

[35]   Yen, A.H. and Sharpe, P.T. (2006) Regeneration of Teeth Using Stem Cell-Based Tissue Engineering. Expert Opinion on Biological Therapy, 6, 9-16.
http://dx.doi.org/10.1517/14712598.6.1.9

[36]   Morsczeck, C., Schmalz, G., Reichert, T.E., Völlner, F., Galler, K. and Driemel, O. (2008) Somatic Stem Cells for Regenerative Dentistry. Clinical Oral Investigations, 12, 113-118.
http://dx.doi.org/10.1007/s00784-007-0170-8

[37]   Nosrat, A., Li, K.L., Vir, K., Hicks, M.L. and Fouad, A.F. (2013) Is Pulp Regeneration Necessary for Root Maturation? Journal of Endodontics, 39, 1291-1295.
http://dx.doi.org/10.1016/j.joen.2013.06.019

[38]   Steindorff, M.M., Lehl, H., Winkel, A. and Stiesch, M. (2014) Innovative Approaches to Regenerate Teeth by Tissue Engineering. Archives of Oral Biology, 59, 158-166.
http://dx.doi.org/10.1016/j.archoralbio.2013.11.005

[39]   Tan, X.W., Perera, A.P., Tan, A., Tan, D., Khor, K.A., Beuerman, R.W. and Mehta, J.S. (2011) Comparison of Candidate Materials for a Synthetic Osteo-Odonto Keratoprosthesis Device. Investigative Ophthalmology & Visual Science, 52, 21-29.
http://dx.doi.org/10.1167/iovs.10-6186

[40]   Zhang, W., Walboomers, X.F., van Osch, G.J., van den Dolder, J. and Jansen, J.A. (2008) Hard Tissue Formation in a Porous HA/TCP Ceramic Scaffold Loaded with Stromal Cells Derived from Dental Pulp and Bone Marrow. Tissue Engineering Part A, 14, 285-294.
http://dx.doi.org/10.1089/tea.2007.0146

[41]   Yoshikawa, M., Tsuji, N., Shimomura, Y., Hayashi, H. and Ohgushi, H. (2007) Effects of Laminin for Osteogenesis in Porous Hydroxyapatite. Macromolecular Symposia, 253, 172-178.
http://dx.doi.org/10.1002/masy.200750724

[42]   Mastrangelo, F., Nargi, E., Carone, L., Dolci, M., Caciagli, F., Ciccarelli, R., De Lutiis, M.A., Karapanou, V., Shaik, B.Y., Conti, P. and Teté, S.J. (2008) Tridimensional Response of Human Dental Follicular Stem Cells onto a Synthetic Hydroxyapatite Scaffold. Journal of Health Science, 54, 154-161.
http://dx.doi.org/10.1248/jhs.54.154

[43]   Hayakawa, S., Li, Y., Tsuru, K., Osaka, A., Fujii, E. and Kawabata, K. (2009) Preparation of Nanometer-Scale Rod Array of Hydroxyapatite Crystal. Acta Biomaterialia, 5, 2152-2160.
http://dx.doi.org/10.1016/j.actbio.2009.02.018

[44]   Bi, Y., Stuelten, C.H., Kilts, T., Wadhwa, S., Iozzo, R.V., Robey, P.G., Chen, X.D. and Young, M.F. (2005) Extracellular Matrix Proteoglycans Control the Fate of Bone Marrow Stromal Cells. The Journal of Biological Chemistry, 280, 30481-30489.
http://dx.doi.org/10.1074/jbc.M500573200

[45]   Warotayanont, R., Zhu, D., Snead, M.L. and Zhou, Y. (2008) Leucine-Rich Amelogenin Peptide Induces Osteogenesis in Mouse Embryonic Stem Cells. Biochemical and Biophysical Research Communications, 367, 1-6.
http://dx.doi.org/10.1016/j.bbrc.2007.12.048

[46]   Wen, X., Cawthorn, W.P., MacDougald, O.A., Stupp, S.I., Snead, M.L. and Zhou, Y. (2011) The Influence of Leucine-Rich Amelogenin Peptide on MSC Fate by Inducing Wnt10b Expression. Biomaterials, 32, 6478-6486.
http://dx.doi.org/10.1016/j.biomaterials.2011.05.045

[47]   Chen, C.H., Yeh, M.L., Geyer, M., Wang, G.J., Huang, M.H., Heggeness, M.H., Höök, M. and Luo, Z.P. (2006) Inter-actions between Collagen IX and Biglycan Measured by Atomic Force Microscopy. Biochemical and Biophysical Research Communications, 339, 204-208.
http://dx.doi.org/10.1016/j.bbrc.2005.10.205

[48]   Huttunen, M.M., Pekkinen, M., Ahlström, M.E. and Lamberg-Allardt, C.J. (2007) Effects of Bioactive Peptides Iso-leucine-Proline-Proline (IPP), Valine-Proline-Proline (VPP) and Leucine-Lysine-Proline (LKP) on Gene Expression of Osteoblasts Differentiated from Human Mesenchymal Stem Cells. British Journal of Nutrition, 98, 780-788.
http://dx.doi.org/10.1017/S0007114507744434

[49]   Zhang, W., Yang, N. and Shi, X.M. (2008) Regulation of Mesenchymal Stem Cell Osteogenic Differentiation by Glucocorticoid-Induced Leucine Zipper (GILZ). The Journal of Biological Chemistry, 283, 4723-4729.
http://dx.doi.org/10.1074/jbc.M704147200

[50]   Waddington, R.J., Roberts, H.C., Sugars, R.V. and Schönherr, E. (2003) Differential Roles for Small Leucine-Rich Proteoglycans in Bone Formation. European Cells & Materials, 6, 12-21.

[51]   Chen, X.D., Allen, M.R., Bloomfield, S., Xu, T. and Young, M. (2003) Biglycan-Deficient Mice Have Delayed Osteo-genesis after Marrow Ablation. Calcified Tissue International, 72, 577-582.
http://dx.doi.org/10.1007/s00223-002-1101-y

[52]   Bi, Y., Nielsen, K.L., Kilts, T.M., Yoon, A., Karsdal, M.A., Wimer, H.F., Greenfield, E.M., Heegaard, A.M. and Young, M.F. (2006) Biglycan Deficiency Increases Osteoclast Differentiation and Activity Due to Defective Osteo-blasts. Bone, 38, 778-786.
http://dx.doi.org/10.1016/j.bone.2005.11.005

[53]   Wallace, J.M., Rajachar, R.M., Chen, X.D., Shi, S., Allen, M.R., Bloomfield, S.A., Les, C.M., Robey, P.G, Young, M.F. and Kohn, D.H. (2006) The Mechanical Phenotype of Biglycan-Deficient Mice Is Bone- and Gender-Specific. Bone, 39, 106-116.
http://dx.doi.org/10.1016/j.bone.2005.12.081

[54]   Cool, S.M. and Nurcombe, V. (2005) Substrate Induction of Osteogenesis from Marrow-Derived Mesenchymal Pre-cursors. Stem Cells and Development, 14, 632-642.
http://dx.doi.org/10.1089/scd.2005.14.632.

[55]   Costa-Pinto, A.R., Correlo, V.M., Sol, P.C., Bhattacharya, M., Charbord, P., Delorme, B., Reis, R.L. and Neves, N.M. (2009) Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells Seeded on Melt Based Chitosan Scaffolds for Bone Tissue Engineering Applications. Biomacromolecules, 10, 2067-2073.
http://dx.doi.org/10.1021/bm9000102.

[56]   Zajaczkowski, M.B., Cukierman, E., Galbraith, C.G. and Yamada, K.M. (2003) Cell-Matrix Adhesions on Poly (Vinyl Alcohol) Hydrogels. Tissue Engineering, 9, 525-533.
http://dx.doi.org/10.1089/107632703322066705

[57]   Tsuji, Y., Yoshimura, N., Aoki, H., Sharov, A.A., Ko, M.S., Motohashi, T. and Kunisada, T. (2008) Maintenance of Undifferentiated Mouse Embryonic Stem Cells in Suspension by the Serum- and Feeder-Free Defined Culture Condition. Developmental Dynamics, 237, 2129-2138.
http://dx.doi.org/10.1002/dvdy.21617

 
 
Top