Abstract
Introduction. Interbody fusion requires a structural member to carry load while the autograft or osteoinductive agent stimulates bone formation. In the present study, we evaluated the potential use of extracted nucleated cells from bone marrow mixed in hyaluronic acid gel as an osteoinductive agent, in comparison to Collagraft loaded with nucleated cells or rhBMP-2 in the porous tantalum ring, in an anterior lumbar interbody fusion (ALIF) in pigs.
Methods. Four 3-month-old female Danish landrace pigs were employed in the current study. Bone marrow was collected by means of aspiration, from the medullary cavity of the proximal tibia. The nucleated cells were isolated with a Ficoll step gradient centrifugation. The cell adhered rate after 24 hours of cultivation and ALP activity in the osteogenic medium following 4 days of cultivation were measured. Cell numbers in the porous tantalum discs were assessed by CyQuant measurement, and fluorescent live/dead cell staining in the porous tantalum discs was performed after periods of 24 hours and 7 days of cultivation. The nucleated cells mixed in hyaluronic acid gel were cultivated on slides for 24 hours, 7 days and 21 days. The live/dead cell staining, ALP staining or osteocalcin staining, were performed. A porous tantalum ring was loaded with nucleated cells in hyaluronic acid gel or packed with Collagraft strips also with nucleated cells or rhBMP-2. Immediately after preparation, one of three implants was inserted into L2-3, L4-5 or L6-7 respectively. The pigs were killed 3 months postoperatively. The lumbar spine specimens were prepared for histological evaluation.
Results. The adhered rate, of the nucleated cells, was 2.26 ± 0.56‰. ALP activity was no different in the osteogenic culture compared to DMEM/10%FBS alone. Cell numbers and live/dead cells showed no difference in the porous tantalum discs. Histological appearance showed that nucleated cells mixed with hyaluronic acid gel, had more mature bone in the central hole of the porous tantalum ring, compared to Collagraft strips with nucleated cells or rhBMP-2. Bone volume fraction did not differ within the three porous tantalum rings; however, more marrow space in the central hole of the porous tantalum ring was present when nucleated cells mixed with hyaluronic acid gel (57.4%) compared to Collagraft strips with rhBMP-2 (29.7%).
Conclusion. In the current study, we demonstrate that nucleated cells, which were isolated from bone marrow intraoperatively, could be used to replace autograft if nucleated cells mixed with hyaluronic acid gel or with Collagraft strips packed into the porous tantalum ring in the pig ALIF model.
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14.7. References
Bagley, J., Rosenzweig, M., Marks, D.F., and Pykett, M.J., 1999, Extended culture of multipotent hematopoietic progenitors without cytokine augmentation in a novel three-dimensional device, Exp. Hematol. 27(3):496–504.
Banwart, J.C., Asher, M.A., and Hassanein, R.S., 1995, Iliac crest bone graft harvest donor site morbidity. A statistical evaluation, Spine 20(9):1055–1060.
Bensaid, W., Triffitt, J.T., Blanchat, C., Oudina, K., Sedel, L., and Petite, H., 2003, A biodegradable fibrin scaffold for mesenchymal stem cell transplantation, Biomaterials 24(14):2497–2502.
Burkus, J.K., Transfeldt, E.E., Kitchel, S.H., Watkins, R.G., and Balderston, R.A., 2002, Clinical and radiographic outcomes of anterior lumbar interbody fusion using recombinant human bone morphogenetic protein-2, Spine 27(21):2396–2408.
Christie, M.J., 2002, Clinical applications of Trabecular Metal, Am. J. Orthop. 31(4):219–220.
Collier J.H., Camp J.P., Hudson T.W., and Schmidt C.E., 2000, Synthesis and characterization of polypyrrolehyaluronic acid composite biomaterials for tissue engineering applications, J. Biomed. Mater. Res. 50(4):574–584.
Cornell, C.N., Lane, J.M., Chapman, M., Merkow, R., Seligson, D., Henry, S., Gustilo, R., and Vincent, K., 1991, Multicenter trial of Collagraft as bone graft substitute, J. Orthop. Trauma. 5(1):1–8.
Ehring, B., Biber, K., Upton, T.M., Plosky, D., Pykett, M., and Rosenzweig, M., 2003, Expansion of HPCs from cord blood in a novel 3D matrix, Cytotherapy 5(6):490–499.
Huang, L., Cheng, Y.Y., Koo, P.L., Lee, K.M., Qin, L., Cheng, J.C., and Kumta, S.M., 2003, The effect of hyaluronan on osteoblast proliferation and differentiation in rat calvarial-derived cell cultures, J. Biomed. Mater. Res. A 66(4):880–884.
Li, H., Zou, X., Xue, Q., Egund, N., Lind, M., and Bunger, C., 2004, Anterior lumbar interbody fusion with carbon fiber cage loaded with bioceramics and platelet-rich plasma. An experimental study on pigs, Eur. Spine J. 13(4):354–358.
Muschler, G.F., and Midura, R.J., 2002, Connective tissue progenitors: practical concepts for clinical applications, Clinical Orthop. 395:66–80.
Ringe, J., Kaps, C., Burmester, G.R., and Sittinger, M., 2002, Stem cells for regenerative medicine: Advances in the engineering of tissues and organs, Naturwissenschaften. 89(8):338–351.
Ringe, J., Leinhase, I., Endres, M., Stich, S., Loch, A., and Sittinger, M., 2003, Human periosteal cells and mesenchymal stem cells for tissue engineering: A comparative study, Tissue Eng. 9(4):783–784.
Rosenzweig, M., Pykett, M., Marks, D.F., Johnson, R.P., 1997, Enhanced maintenance and retroviral transduction of primitive hematopoietic progenitor cells using a novel three-dimensional culture system, Gene Ther. 4(9):928–936.
Sidhu, K.S., Prochnow, T.D., Schmitt, P., Fischgrund, J., Weisbrode, S., and Herkowitz, H.N., 2001, Anterior cervical interbody fusion with rhBMP-2 and tantalum in a goat model, Spine J. 1(5):331–340.
Toh, Y.C., Ho, S.T., Zhou, Y., Hutmacher, D.W., and Yu, H., 2005, Application of a polyelectrolyte complex coacervation method to improve seeding efficiency of bone marrow stromal cells in a 3D culture system, Biomaterials 26(19):4149–4160.
Ushida, T., Furukawa, K., Toita, K., and Tateishi, T., 2002, Three-dimensional seeding of chondrocytes encapsulated in collagen gel into PLLA scaffolds, Cell Transplant. 11(5):489–494.
West D.C., Hampson I.N., Arnold F., and Kumar S., 1985, Angiogenesis induced by degradation products of hyaluronic acid, Science 228(4705):1324–1326.
Wigfield, C., Robertson, J., Gill, S., and Nelson, R., 2003, Clinical experience with porous tantalum cervical interbody implants in a prospective randomized controlled trial, Br. J. Neurosurg. 17(5):418–425.
Xue, Q., Li, H., Zou, X., Christensen, F.B., Lind, M., and Bünger, C., 2005, Healing properties of allograft from alendronate-treated animal in lumbar spine interbody cage fusion, Eur. Spine J. 14(3):222–226.
Younger, E.M. and Chapman, M.W.. 1989, Morbidity at bone graft donor sites, Orthop Trauma. 3:192–195.
Zou, X., Li, H., Baatrup, A., Lind, M., and Bunger, C., 2003, Engineering of bone tissue with porcine bone marrow stem cells in three-dimensional trabecular metal: in vitro and in vivo studies, APMIS Suppl. 111(109):127–132.
Zou, X., Li, H., Baatrup, A., Bünger, C., and Lind, M., 2004a, Stimulation of porcine bone marrow stromal cells by hyaluronan, dexamethasone and rhBMP-2, Biomaterials 25(23):5375–5385.
Zou, X., Li, H., Bünger, M., Egund, N., Lind, M., and Bünger, C., 2004b, Bone ingrowth characteristics of porous tantalum and carbon fiber interbody devices: an experimental study in pigs, Spine J. 4(1):99–105.
Zou, X., Li, H., Teng, X., Xue, Q., Egund, N., Lind, M., and Bünger, C., 2005, Pedicle screw fixation enhances Aanterior lumbar interbody fusion with porous tantalum cages: an experimental study in gigs, Spine 30(14):E392–E399.
Zou, X., Xue, Q., Li, H., Bünger, M., Lind, M., and Bünger, C., 2003, Effect of alendronate on bone ingrowth into porous tantalum and carbon fiber interbody devices: An experimental study on spinal fusion in pigs, Acta Orthop Scand. 74(5):596–603.
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Zou, X., Li, H., Zou, L., Mygind, T., Lind, M., Bünger, C. (2006). Porous Tantalum Trabecular Metal Scaffolds in Combination with a Novel Marrow Processing Technique to Replace Autograft. In: Fisher, J.P. (eds) Tissue Engineering. Advances in Experimental Medicine and Biology, vol 585. Springer, Boston, MA. https://doi.org/10.1007/978-0-387-34133-0_14
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DOI: https://doi.org/10.1007/978-0-387-34133-0_14
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