Clinical Orthopaedics and Related Research®

, Volume 472, Issue 4, pp 1318–1328

Novel Microhydroxyapatite Particles in a Collagen Scaffold: A Bioactive Bone Void Filler?

  • Frank G. Lyons
  • John P. Gleeson
  • Sonia Partap
  • Karen Coghlan
  • Fergal J. O’Brien
Basic Research

Abstract

Background

Treatment of segmental bone loss remains a major challenge in orthopaedic surgery. Traditional techniques (eg, autograft) and newer techniques (eg, recombinant human bone morphogenetic protein-2 [rhBMP-2]) have well-established performance limitations and safety concerns respectively. Consequently there is an unmet need for osteoinductive bone graft substitutes that may eliminate or reduce the use of rhBMP-2.

Questions/purposes

Using an established rabbit radius osteotomy defect model with positive (autogenous bone graft) and negative (empty sham) control groups, we asked: (1) whether a collagen-glycosaminoglycan scaffold alone can heal the defect, (2) whether the addition of hydroxyapatite particles to the collagen scaffold promote faster healing, and (3) whether the collagen-glycosaminoglycan and collagen-hydroxyapatite scaffolds are able to promote faster healing (by carrying a low dose rhBMP-2).

Methods

A 15-mm transosseous radius defect in 4-month-old skeletally mature New Zealand White rabbits were treated with either collagen-hydroxyapatite or collagen-glycosaminoglycan scaffolds with and without rhBMP-2. Autogenous bone graft served as a positive control. Time-series radiographs at four intervals and postmortem micro-CT and histological analysis at 16 weeks were performed. Qualitative histological analysis of postmortem explants, and qualitative and volumetric 3-D analysis of standard radiographs and micro-CT scans enabled direct comparison of healing between test groups.

Results

Six weeks after implantation the collagen-glycosaminoglycan group had callus occupying greater than ½ the defect, whereas the sham (empty) control defect was still empty and the autogenous bone graft defect was completely filled with unremodeled bone. At 6 weeks, the collagen-hydroxyapatite scaffold groups showed greater defect filling with dense callus compared with the collagen-glycosaminoglycan controls. At 16 weeks, the autogenous bone graft groups showed evidence of early-stage medullary canal formation beginning at the proximal and distal defect borders. The collagen-glycosaminoglycan and collagen-glycosaminoglycan-rhBMP-2 groups had nearly complete medullary canal formation and anatomic healing at 16 weeks. However, collagen-hydroxyapatite-rhBMP-2 scaffolds showed the best levels of healing, exhibiting a dense callus which completely filled the defect.

Conclusions

The collagen-hydroxyapatite scaffold showed comparable healing to the current gold standard of autogenous bone graft. It also performed comparably to collagen-glycosaminoglycan-rhBMP-2, a representative commercial device in current clinical use, but without the cost and safety concerns.

Clinical Relevance

The collagen-glycosaminoglycan scaffold may be suitable for a low load-bearing defect. The collagen-hydroxyapatite scaffold may be suitable for a load-bearing defect. The rhBMP-2 containing collagen-glycosaminoglycan and collagen-hydroxyapatite scaffolds may be suitable for established nonunion defects.

References

  1. 1.
    Alam MI, Asahina I, Ohmamiuda K, Takahashi K, Yokota S, Enomoto S. Evaluation of ceramics composed of different hydroxyapatite to tricalcium phosphate ratios as carriers for rhBMP-2. Biomaterials. 2001;22:1643–1651.PubMedCrossRefGoogle Scholar
  2. 2.
    Bauer TW, Muschler GF. Bone graft materials: an overview of the basic science. Clin Orthop Relat Res. 2000;371:10–27.PubMedCrossRefGoogle Scholar
  3. 3.
    Boden SD, Zdeblick TA, Sandhu HS, Heim SE. The use of rhBMP-2 in interbody fusion cages: definitive evidence of osteoinduction in humans: a preliminary report. Spine (Phila Pa 1976). 2000;25:376–381.CrossRefGoogle Scholar
  4. 4.
    Curtin CM, Cunniffe GM, Lyons FG, Bessho K, Dickson GR, Duffy GP, O’Brien FJ. Innovative collagen nano-hydroxyapatite scaffolds offer a highly efficient non-viral gene delivery platform for stem cell-mediated bone formation. Adv Mater. 2012;24:749–754.PubMedCrossRefGoogle Scholar
  5. 5.
    Department of Health and Human Services, US. Agency for Healthcare Research and Quality report. Bone Morphogenetic Protein: The State of the Evidence for On-Label and Off-Label Use. Available at: http://www.ahrq.gov/clinic/ta/comments/boneprotein/. Accessed 05/12/2011.
  6. 6.
    FDA Centers for Devices and Radiological Health. Corrspondence to RW Treharne and Premarket Approval Application (PMS) Supplement. Re: P000058. InFUSE™ Bone Graft/LT-CAGE™ Lumbar Tapered Fusion Device. Available at: http://www.accessdata.fda.gov/cdrh_docs/pdf/P000058a.pdf. Accessed May 12, 2011.
  7. 7.
    FDA Centers for Devices and Radiological Health. Summary of Safety and Effectiveness Data. Available at: http://www.accessdata.fda.gov/cdrh_docs/pdf/P000058b.pdf. Accessed May 12, 2011.
  8. 8.
    FDA Centers for Devices and Radiological Health. Correspondence to AJ LaForte and Conditions of Approval for an HDE for OP-1™. Available at: http://www.accessdata.fda.gov/cdrh_docs/pdf/h010002a.pdf. Accessed May 12, 2011.
  9. 9.
    FDA. Centers for Devices and Radiological Health. Correspondence to D Ellis and Premarket Approval for INFUSE® bone graft. Available at: http://www.accessdata.fda.gov/cdrh_docs/pdf/p000054a.pdf. Accessed May 12, 2011.
  10. 10.
    Garrett MP, Kakarla UK, Porter RW, Sonntag VK. Formation of painful seroma and edema after the use of recombinant human bone morphogenetic protein-2 in posterolateral lumbar spine fusions. Neurosurgery. 2010;66:1044–1049; discussion 1049.PubMedCrossRefGoogle Scholar
  11. 11.
    Gleeson JP, Plunkett NA, O’Brien FJ. Addition of hydroxyapatite improves stiffness, interconnectivity and osteogenic potential of a highly porous collagen-based scaffold for bone tissue regeneration. Eur Cell Mater. 2010;20:218–230.PubMedGoogle Scholar
  12. 12.
    Haugh MG, Jaasma MJ, O’Brien FJ. The effect of dehydrothermal treatment on the mechanical and structural properties of collagen-GAG scaffolds. J Biomed Mater Res A. 2009;89:363–369.PubMedCrossRefGoogle Scholar
  13. 13.
    Hedberg EL, Kroese-Deutman HC, Shih CK, Crowther RS, Carney DH, Mikos AG, Jansen JA. In vivo degradation of porous poly(propylene fumarate)/poly(DL-lactic-co-glycolic acid) composite scaffolds. Biomaterials. 2005;26:4616–4623.PubMedCrossRefGoogle Scholar
  14. 14.
    Jahangir AA, Nunley RM, Mehta S, Sharan A; Fellows at WHP. Bone-graft substitutes in orthopaedic surgery. AAOS Now. 2008;2:1–5.Google Scholar
  15. 15.
    Karageorgiou V, Kaplan D. Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials. 2005;26:5474–5491.PubMedCrossRefGoogle Scholar
  16. 16.
    Laurencin CT, Khan Y, El-Amin SF. Bone graft substitutes. Expert Rev Med Devices. 2006;3:49–57.PubMedCrossRefGoogle Scholar
  17. 17.
    Le Nihouannen D, Daculsi G, Saffarzadeh A, Gauthier O, Delplace S, Pilet P, Layrolle P. Ectopic bone formation by microporous calcium phosphate ceramic particles in sheep muscles. Bone. 2005;36:1086–1093.PubMedCrossRefGoogle Scholar
  18. 18.
    LeGeros RZ. Calcium phosphate-based osteoinductive materials. Chem Rev. 2008;108:4742–4753.PubMedCrossRefGoogle Scholar
  19. 19.
    Lyons FG, Al-Munajjed AA, Kieran SM, Toner ME, Murphy CM, Duffy GP, O’Brien FJ. The healing of bony defects by cell-free collagen-based scaffolds compared to stem cell-seeded tissue engineered constructs. Biomaterials. 2010;31:9232–9243.PubMedCrossRefGoogle Scholar
  20. 20.
    McKay B, Sandhu HS. Use of recombinant human bone morphogenetic protein-2 in spinal fusion applications. Spine (Phila Pa 1976). 2002;27(16 suppl 1):S66–85.CrossRefGoogle Scholar
  21. 21.
    Murphy CM, Haugh MG, O’Brien FJ. The effect of mean pore size on cell attachment, proliferation and migration in collagen-glycosaminoglycan scaffolds for bone tissue engineering. Biomaterials. 2010;31:461–466.PubMedCrossRefGoogle Scholar
  22. 22.
    O’Brien FJ, Harley BA, Yannas IV, Gibson L. Influence of freezing rate on pore structure in freeze-dried collagen-GAG scaffolds. Biomaterials. 2004;25:1077–1086.PubMedCrossRefGoogle Scholar
  23. 23.
    O’Brien FJ, Harley BA, Yannas IV, Gibson LJ. The effect of pore size on cell adhesion in collagen-GAG scaffolds. Biomaterials. 2005;26:433–441.PubMedCrossRefGoogle Scholar
  24. 24.
    O’Brien FJ, Plunkett NA, Gleeson JP, inventors: A collagen/hydroxyapatite composite scaffold, and process for the production thereof. US patent US8435552 (B2)-2013-05-07; European patent EP2117617 (B1)-2013-08-1; Australia patent AU2008212526 (A1).Google Scholar
  25. 25.
    Perry CR. Bone repair techniques, bone graft, and bone graft substitutes. Clin Orthop Relat Res. 1999;360:71–86.PubMedCrossRefGoogle Scholar
  26. 26.
    Ratko TA, Belinson SE, Samson DJ, Bonnell C, Ziegler KM, Aronson N. Bone morphogenetic protein: the state of the evidence of on-label and off-label use. Technology Assessment Report. Available at: http://www.cms.gov/DeterminationProcess/downloads/id75ta.pdf?bcsi_scan_9CF786ACA806128D=0&bcsi_scan_filename=id75ta.pdf. Accessed May 12, 2011.
  27. 27.
    Roy TD, Simon JL, Ricci JL, Rekow ED, Thompson VP, Parsons JR. Performance of degradable composite bone repair products made via three-dimensional fabrication techniques. J Biomed Mater Res A. 2003;66:283–291.PubMedCrossRefGoogle Scholar
  28. 28.
    Schuessele A, Mayr H, Tessmar J, Goepferich A. Enhanced bone morphogenetic protein-2 performance on hydroxyapatite ceramic surfaces. J Biomed Mater Res A. 2009;90:959–971.PubMedCrossRefGoogle Scholar
  29. 29.
    Sciadini MF, Johnson KD. Evaluation of recombinant human bone morphogenetic protein-2 as a bone-graft substitute in a canine segmental defect model. J Orthop Res. 2000;18:289–302.PubMedCrossRefGoogle Scholar
  30. 30.
    Seeherman H, Wozney J, Li R. Bone morphogenetic protein delivery systems. Spine (Phila Pa 1976). 2002;27(16 suppl 1):S16–23.CrossRefGoogle Scholar
  31. 31.
    Shields LB, Raque GH, Glassman SD, Campbell M, Vitaz T, Harpring J, Shields CB. Adverse effects associated with high-dose recombinant human bone morphogenetic protein-2 use in anterior cervical spine fusion. Spine (Phila Pa 1976). 2006;31:542–547.CrossRefGoogle Scholar
  32. 32.
    Smucker JD, Petersen EB, Fredericks DC. Assessment of MASTERGRAFT PUTTY as a graft extender in a rabbit posterolateral fusion model. Spine (Phila Pa 1976). 2012;37:1017–1021.CrossRefGoogle Scholar
  33. 33.
    Tierney CM, Haugh MG, Liedl J, Mulcahy F, Hayes B, O’Brien FJ. The effects of collagen concentration and crosslink density on the biological, structural and mechanical properties of collagen-GAG scaffolds for bone tissue engineering. J Mech Behav Biomed Mater. 2009;2:202–209.PubMedCrossRefGoogle Scholar
  34. 34.
    Tuli SM, Gupta KB. Bridging of large chronic osteoperiosteal gaps by allogeneic decalcified bone matrix implants in rabbits. J Trauma. 1981;21:894–898.PubMedCrossRefGoogle Scholar
  35. 35.
    Urist MR. Bone: formation by autoinduction. Science. 1965;150:893–899.PubMedCrossRefGoogle Scholar
  36. 36.
    Vaidya R. Transforaminal interbody fusion and the “off label” use of recombinant human bone morphogenetic protein-2. Spine J. 2009;9:667–669.PubMedCrossRefGoogle Scholar
  37. 37.
    Vines JB, Lim DJ, Anderson JM, Jun HW. Hydroxyapatite nanoparticle reinforced peptide amphiphile nanomatrix enhances the osteogenic differentiation of mesenchymal stem cells by compositional ratios. Acta Biomater. 2012;8:4053–4063.PubMedCentralPubMedCrossRefGoogle Scholar
  38. 38.
    Wakimoto M, Ueno T, Hirata A, Iida S, Aghaloo T, Moy PK. Histologic evaluation of human alveolar sockets treated with an artificial bone substitute material. J Craniofac Surg. 2011;22:490–493.PubMedCrossRefGoogle Scholar
  39. 39.
    Williams DF. On the nature of biomaterials. Biomaterials. 2009;30:5897–5909.PubMedCrossRefGoogle Scholar
  40. 40.
    Wong DA, Kumar A, Jatana S, Ghiselli G, Wong K. Neurologic impairment from ectopic bone in the lumbar canal: a potential complication of off-label PLIF/TLIF use of bone morphogenetic protein-2 (BMP-2). Spine J. 2008;8:1011–1018.PubMedCrossRefGoogle Scholar
  41. 41.
    Yannas IV, Burke JF. Design of an artificial skin: I. Basic design principles. J Biomed Mater Res. 1980;14:65–81.PubMedCrossRefGoogle Scholar
  42. 42.
    Yannas IV, Burke JF, Gordon PL, Huang C, Rubenstein RH. Design of an artificial skin: II. Control of chemical composition. J Biomed Mater Res. 1980;14:107–132.PubMedCrossRefGoogle Scholar
  43. 43.
    Yannas IV, Lee E, Orgill DP, Skrabut EM, Murphy GF. Synthesis and characterization of a model extracellular matrix that induces partial regeneration of adult mammalian skin. Proc Natl Acad Sci U S A. 1989;86:933–937.PubMedCentralPubMedCrossRefGoogle Scholar
  44. 44.
    Yasko AW, Lane JM, Fellinger EJ, Rosen V, Wozney JM, Wang EA. The healing of segmental bone defects, induced by recombinant human bone morphogenetic protein (rhBMP-2): a radiographic, histological, and biomechanical study in rats. J Bone Joint Surg Am. 1992;74:659–670.PubMedGoogle Scholar
  45. 45.
    Yuan H, De Bruijn JD, Zhang X, Van Blitterswijk CA, De Groot K. Use of an osteoinductive biomaterial as a bone morphogenetic protein carrier. J Mater Sci Mater Med. 2001;12:761–766.PubMedCrossRefGoogle Scholar
  46. 46.
    Yuan H, Fernandes H, Habibovic P, de Boer J, Barradas A, de Ruiter A, Walsh W, van Blitterswijk C, de Bruijn J. Osteoinductive ceramics as a synthetic alternative to autologous bone grafting. Proc Natl Acad Sci U S A. 2010;107:13614–13619.PubMedCentralPubMedCrossRefGoogle Scholar
  47. 47.
    Yuan H, Van Den Doel M, Li S, Van Blitterswijk CA, De Groot K, De Bruijn JD. A comparison of the osteoinductive potential of two calcium phosphate ceramics implanted intramuscularly in goats. J Mater Sci Mater Med. 2002;13:1271–1275.PubMedCrossRefGoogle Scholar

Copyright information

© The Association of Bone and Joint Surgeons® 2013

Authors and Affiliations

  • Frank G. Lyons
    • 1
    • 2
    • 3
    • 4
  • John P. Gleeson
    • 1
    • 2
  • Sonia Partap
    • 1
    • 2
  • Karen Coghlan
    • 1
    • 2
    • 5
  • Fergal J. O’Brien
    • 1
    • 2
    • 5
  1. 1.Tissue Engineering Research Group, Department of AnatomyRoyal College of Surgeons in IrelandDublin 2Ireland
  2. 2.Trinity Centre for BioengineeringTrinity College DublinDublin 2Ireland
  3. 3.Cappagh National Orthopaedic HospitalDublinIreland
  4. 4.Mater Misericordiae University HospitalDublinIreland
  5. 5.Advanced Materials and Bioengineering Research (AMBER) CentreRoyal College of Surgeons in Ireland, Trinity College DublinDublinIreland

Personalised recommendations