European Spine Journal

, Volume 21, Issue 9, pp 1740–1747 | Cite as

Effects of a perfusion bioreactor activated novel bone substitute in spine fusion in sheep

  • Jesper Roed Sørensen
  • Kariatta Ester Koroma
  • Ming Ding
  • David Wendt
  • Stig Jespersen
  • Maria Vinther Juhl
  • Naseem Theilgaard
  • Ivan Martin
  • Søren Overgaard
Original Article

Abstract

Purpose

To evaluate the effect of a large perfusion-bioreactor cell-activated bone substitute, on a two-level large posterolateral spine fusion sheep model.

Methods

A 50 mm long porous biphasic-calcium–phosphate bone substitute reinforced with poly(d,l-lactide) and, activated with bone marrow derived mononuclear-cells (BMNC) was used. Eighteen sheep were divided into two groups and one group (n = 9) had BMNC-activated bone substitutes and cell-free substitutes implanted. The second group (n = 9) had autograft supplemented with BMNC and regular autograft implanted. The implant material was alternated between spine level L2–L3 and L4–L5 in both groups. MicroCT was used to compare the spine fusion efficacy and bone structure of the two groups as well as the implanted bone substitutes and non-implanted substitutes.

Results

After 4½ months six sheep survived in both groups and we found five spine levels were fused when using activated bone substitute compared to three levels with cell-free bone substitute (p = 0.25). Five sheep fused at both levels in the autograft group. A significant increased bone density (p < 0.05) and anisotropy (p < 0.05) was found in the group of activated bone substitutes compared to cell-free bone substitute and no difference existed on the other parameters. The implanted bone substitutes had a significant higher bone density and trabecular thickness than non-implanted bone substitutes, thus indicating that the PLA reinforced BCP had osteoconductive properties (p < 0.05). No effect of the supplemented BMNC to autograft was observed. The autograft group had a significant higher bone density, trabecular thickness and degree of anisotropy than the implanted bone substitutes (p < 0.05), but a lower connectivity density existed (p < 0.05). This indicates that though the activated substitute might have a similar fusion efficacy to autograft, the fusion bridge is not of equal substance.

Conclusion

We found that bioreactor-generated cell-based bone substitutes seemed superior in fusion ability when compared to cell-free bone substitute and comparable to autograft in fusion ability, but not in bone structure. This combined with the favorable biocompatible abilities and strength comparable to human cancellous bone indicates that it might be a suitable bone substitute in spine fusion procedures.

Keywords

Bone graft substitute Poly(d,l-lactide) enhanced hydroxyappatite/β-tricalciumphosphate Perfusion bioreactor Posterolateral spine fusion Microarchitecture 

References

  1. 1.
    Gazdag AR, Lane JM, Glaser D, Forster RA (1995) Alternatives to autogenous bone graft: efficacy and indications. J Am Acad Orthop Surg 3:1–8PubMedGoogle Scholar
  2. 2.
    Heary RF, Schlenk RP, Sacchieri TA, Barone D, Brotea C (2002) Persistent iliac crest donor site pain: independent outcome assessment. Neurosurgery 50:510–516PubMedGoogle Scholar
  3. 3.
    Banwart JC, Asher MA, Hassanein RS (1995) Iliac crest bone graft harvest donor site morbidity. A statistical evaluation. Spine (Phila Pa 1976) 20:1055–1060CrossRefGoogle Scholar
  4. 4.
    France JC, Yaszemski MJ, Lauerman WC, Cain JE, Glover JM, Lawson KJ et al (1999) A randomized prospective study of posterolateral lumbar fusion. Outcomes with and without pedicle screw instrumentation. Spine (Phila Pa 1976) 24:553–560CrossRefGoogle Scholar
  5. 5.
    Glassman SD, Howard JM, Sweet A, Carreon LY (2010) Complications and concerns with osteobiologics for spine fusion in clinical practice. Spine (Phila Pa 1976) 35:1621–1628CrossRefGoogle Scholar
  6. 6.
    Rihn JA, Kirkpatrick K, Albert TJ (2010) Graft options in posterolateral and posterior interbody lumbar fusion. Spine (Phila Pa 1976) 35:1629–1639CrossRefGoogle Scholar
  7. 7.
    Miyazaki M, Tsumura H, Wang JC, Alanay A (2009) An update on bone substitutes for spinal fusion. Eur Spine J 18:783–799PubMedCrossRefGoogle Scholar
  8. 8.
    Kim SS, Sun PM, Jeon O, Yong CC, Kim BS (2006) Poly(lactide-co-glycolide)/hydroxyapatite composite scaffolds for bone tissue engineering. Biomaterials 27:1399–1409PubMedCrossRefGoogle Scholar
  9. 9.
    Eshraghi S, Das S (2010) Mechanical and microstructural properties of polycaprolactone scaffolds with one-dimensional, two-dimensional, and three-dimensional orthogonally oriented porous architectures produced by selective laser sintering. Acta Biomater 6:2467–2476PubMedCrossRefGoogle Scholar
  10. 10.
    Wang Y, Uemura T, Dong J, Kojima H, Tanaka J, Tateishi T (2003) Application of perfusion culture system improves in vitro and in vivo osteogenesis of bone marrow-derived osteoblastic cells in porous ceramic materials. Tissue Eng 9:1205–1214PubMedCrossRefGoogle Scholar
  11. 11.
    Wendt D, Stroebel S, Jakob M, John GT, Martin I (2006) Uniform tissues engineered by seeding and culturing cells in 3D scaffolds under perfusion at defined oxygen tensions. Biorheology 43:481–488PubMedGoogle Scholar
  12. 12.
    Wendt D, Marsano A, Jakob M, Heberer M, Martin I (2003) Oscillating perfusion of cell suspensions through three-dimensional scaffolds enhances cell seeding efficiency and uniformity. Biotechnol Bioeng 84:205–214PubMedCrossRefGoogle Scholar
  13. 13.
    Volkmer E, Drosse I, Otto S, Stangelmayer A, Stengele M, Kallukalam BC et al (2008) Hypoxia in static and dynamic 3D culture systems for tissue engineering of bone. Tissue Eng 14:1331–1340CrossRefGoogle Scholar
  14. 14.
    Ding M, Dalstra M, Danielsen CC, Kabel J, Hvid I, Linde F (1997) Age variations in the properties of human tibial trabecular bone. J Bone Joint Surg Br 79:995–1002PubMedCrossRefGoogle Scholar
  15. 15.
    Ding M, Hvid I (2000) Quantification of age-related changes in the structure model type and trabecular thickness of human tibial cancellous bone. Bone 26:291–295PubMedCrossRefGoogle Scholar
  16. 16.
    Fields AJ, Eswaran SK, Jekir MG, Keaveny TM (2009) Role of trabecular microarchitecture in whole-vertebral body biomechanical behavior. J Bone Miner Res 24:1523–1530PubMedCrossRefGoogle Scholar
  17. 17.
    Henriksen SS, Ding M, Vinther JM, Theilgaard N, Overgaard S (2011) Mechanical strength of ceramic scaffolds reinforced with biopolymers is comparable to that of human bone. J Mater Sci- Mater Med 22:1111–1118PubMedCrossRefGoogle Scholar
  18. 18.
    Braccini A, Wendt D, Jaquiery C, Jakob M, Heberer M, Kenins L et al (2005) Three-dimensional perfusion culture of human bone marrow cells and generation of osteoinductive grafts. Stem Cells 23:1066–1072PubMedCrossRefGoogle Scholar
  19. 19.
    Scaglione S, Braccini A, Wendt D, Jaquiery C, Beltrame F, Quarto R et al (2006) Engineering of osteoinductive grafts by isolation and expansion of ovine bone marrow stromal cells directly on 3D ceramic scaffolds. Biotechnol Bioeng 93:181–187PubMedCrossRefGoogle Scholar
  20. 20.
    Braccini A, Wendt D, Farhadi J, Schaeren S, Heberer M, Martin I (2007) The osteogenicity of implanted engineered bone constructs is related to the density of clonogenic bone marrow stromal cells. J Tissue Eng Regen Med 1:60–65PubMedCrossRefGoogle Scholar
  21. 21.
    Ding M, Odgaard A, Hvid I (1999) Accuracy of cancellous bone volume fraction measured by micro-CT scanning. J Biomech 32:323–326PubMedCrossRefGoogle Scholar
  22. 22.
    Hildebrand T, Rüegsegger P (1997) A new method for the model-independent assessment of thickness in three-dimensional images. J Microsc 185:67–75CrossRefGoogle Scholar
  23. 23.
    Hildebrand T, Ruegsegger P (1997) Quantification of bone microarchitecture with the structure model index. Comput Methods Biomech Biomed Eng 1:15–23CrossRefGoogle Scholar
  24. 24.
    Hildebrand T, Laib A, Muller R, Dequeker J, Ruegsegger P (1999) Direct three-dimensional morphometric analysis of human cancellous bone: microstructural data from spine, femur, iliac crest, and calcaneus. J Bone Miner Res 14:1167–1174PubMedCrossRefGoogle Scholar
  25. 25.
    Odgaard A (1997) Three-dimensional methods for quantification of cancellous bone architecture. Bone 20:315–328PubMedCrossRefGoogle Scholar
  26. 26.
    Wang L, Hu YY, Wang Z, Li X, Li DC, Lu BH et al (2009) Flow perfusion culture of human fetal bone cells in large beta-tricalcium phosphate scaffold with controlled architecture. J Biomed Mater Res A 91:102–113PubMedGoogle Scholar
  27. 27.
    Olivier V, Hivart P, Descamps M, Hardouin P (2007) In vitro culture of large bone substitutes in a new bioreactor: importance of the flow direction. Biomed Mater 2:174–180PubMedCrossRefGoogle Scholar
  28. 28.
    Ding M, Odgaard A, Danielsen CC, Hvid I (2002) Mutual associations among microstructural, physical and mechanical properties of human cancellous bone. J Bone Joint Surg Br 84:900–907PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Jesper Roed Sørensen
    • 1
  • Kariatta Ester Koroma
    • 1
  • Ming Ding
    • 1
  • David Wendt
    • 3
  • Stig Jespersen
    • 1
  • Maria Vinther Juhl
    • 2
  • Naseem Theilgaard
    • 2
  • Ivan Martin
    • 3
  • Søren Overgaard
    • 1
  1. 1.Department of Orthopaedics and Traumatology, Odense University Hospital, Institute of Clinical ResearchUniversity of Southern DenmarkOdense CDenmark
  2. 2.Centre for Plastic TechnologyDanish Technological InstituteTaastrupDenmark
  3. 3.Departments of Surgery and of BiomedicineUniversity Hospital BaselBaselSwitzerland

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