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.
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References
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.
Bauer TW, Muschler GF. Bone graft materials: an overview of the basic science. Clin Orthop Relat Res. 2000;371:10–27.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Jahangir AA, Nunley RM, Mehta S, Sharan A; Fellows at WHP. Bone-graft substitutes in orthopaedic surgery. AAOS Now. 2008;2:1–5.
Karageorgiou V, Kaplan D. Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials. 2005;26:5474–5491.
Laurencin CT, Khan Y, El-Amin SF. Bone graft substitutes. Expert Rev Med Devices. 2006;3:49–57.
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.
LeGeros RZ. Calcium phosphate-based osteoinductive materials. Chem Rev. 2008;108:4742–4753.
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.
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.
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.
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.
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.
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).
Perry CR. Bone repair techniques, bone graft, and bone graft substitutes. Clin Orthop Relat Res. 1999;360:71–86.
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.
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.
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.
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.
Seeherman H, Wozney J, Li R. Bone morphogenetic protein delivery systems. Spine (Phila Pa 1976). 2002;27(16 suppl 1):S16–23.
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.
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.
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.
Tuli SM, Gupta KB. Bridging of large chronic osteoperiosteal gaps by allogeneic decalcified bone matrix implants in rabbits. J Trauma. 1981;21:894–898.
Urist MR. Bone: formation by autoinduction. Science. 1965;150:893–899.
Vaidya R. Transforaminal interbody fusion and the “off label” use of recombinant human bone morphogenetic protein-2. Spine J. 2009;9:667–669.
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.
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.
Williams DF. On the nature of biomaterials. Biomaterials. 2009;30:5897–5909.
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.
Yannas IV, Burke JF. Design of an artificial skin: I. Basic design principles. J Biomed Mater Res. 1980;14:65–81.
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.
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.
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.
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.
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.
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.
Acknowledgments
We thank the following members of the RCSI Tissue Engineering Research Group for their input with this project: John M. O’Byrne FRCSI, Cappagh National Orthopaedic Hospital and Royal College of Surgeons in Ireland; Keith A. Synnott FRCSI, Mater Misericordiae University Hospital and University College Dublin; and Hester McAllister MVB and Lynne Hughes MVB, University Veterinary Hospital and University College Dublin.
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One of the authors certifies that he (FGL), or a member of his immediate family, has or may receive payments or benefits, during the study period, an amount of USD (USD 10,000–USD 100,000), from the clinical research PhD fellowship program of the Health Research Board, Irish Government Department of Health, Dublin, Ireland. Authors John P Gleeson and Fergal J O’Brien hold intellectual property with a commercial product related to the collagen-hydroxyapatite scaffold used in this study.
All ICMJE Conflict of Interest Forms for authors and Clinical Orthopaedics and Related Research editors and board members are on file with the publication and can be viewed on request.
Clinical Orthopaedics and Related Research neither advocates nor endorses the use of any treatment, drug, or device. Readers are encouraged to always seek additional information, including FDA-approval status, of any drug or device prior to clinical use.
Each author certifies that his or her institution approved the animal protocol for this investigation and that all investigations were conducted in conformity with ethical principles of research.
This work was performed at the Department of Anatomy, Royal College of Surgeons in Ireland, Dublin, Ireland; and the Trinity Centre for Bioengineering, Department of Mechanical Engineering, Trinity College, Dublin, Ireland. Three-dimensional micro-CT reconstruction and analysis were done with assistance from Scanco Medical, Wangen-Brüttisellen, Switzerland.
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Lyons, F.G., Gleeson, J.P., Partap, S. et al. Novel Microhydroxyapatite Particles in a Collagen Scaffold: A Bioactive Bone Void Filler?. Clin Orthop Relat Res 472, 1318–1328 (2014). https://doi.org/10.1007/s11999-013-3438-0
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DOI: https://doi.org/10.1007/s11999-013-3438-0