Abstract
Shaped demineralised bone matrices (DBM) made from cancellous bone have important uses in orthopaedic and dental procedures, where the properties of the material allow its insertion into confined defects, therefore acting as a void filler and scaffold onto which new bone can form. The sponges are often small in size, <1.0 cm3. In this study, we report on an improved bone washing and demineralisation process that allows production of larger DBM sponges (3.375 or 8.0 cm3) from deceased donor bone. These sponges were taken through a series of warm water washes, some with sonication, centrifugation, 100 % ethanol and two decontamination chemical washes and optimally demineralised using 0.5 N hydrochloric acid under vacuum. Demineralisation was confirmed by quantitative measurement of calcium and qualitatively by compression. Protein and DNA removal was also determined. The DBM sponges were freeze dried before terminal sterilisation with a target dose of 25 kGy gamma irradiation whilst frozen. Samples of the sponges were examined histologically for calcium, collagen and the presence of cells. The data indicated lack of cells, absence of bone marrow and a maximum of 1.5 % residual calcium.
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References
Bethesda MD (2006) American association of tissue banks. Standards for tissue banking, (11th ed) American association of tissue banks
Bleazey S, Brigado SA (2012) Reconstruction of complex osteochondral lesions of the Talus with cylindrical sponge allograft and particulate juvenile cartilage graft : provisional results with a short-term follow-up. Foot Ankle Spec 5:300–305
Bradbury P, Gordon KC (1990) Connective tissue and stains. In: Bancroft JD, Stevens A (eds) Theory and practice of histological techniques, 3rd edn. Churchill Livingstone, Edinburgh, pp 119–142
Burkus JK, Gornet MF, Dickman CA, Zdeblick TA (2002) Anterior lumbar interbody fusion using rhBMP-2 with tapered interbody cages. J Spinal Disord Tech 15(5):337–349
Eagle MJ, Rooney P, Kearney JN (2014) Optimized demineralization of human cancellous bone by application of a vacuum. J Biomed Res B [e-pub ahead of print]
Eagle MJ, Man J, Rooney P, Hogg P, Kearney JN (2015) Assessment of an improved bone washing protocol for deceased donor human bone. Cell Tissue Bank 16(1):83–90
Elsalanty ME, Por YC, Genecov DG, Salyer KE, Wang Q, Barcelo CR, Troxler K, Gendler E, Opperman LA (2008) Recombinant human BMP-2 enhances the effects of materials used for reconstruction of large cranial defects. J Oral Maxillofac Surg 66:277–285
Galli MM, Protzman NM, Bleazey ST, Brigado SA (2014) Role of demineralised allograft subchondral bone in the treatment of shoulder lesions of the Talus: clinical results with two-year follow up. J Foot Ankle Surg [e-pub ahead of print]
Girasole G, Muro G, Mintz A, Chertoff J (2013) Transforaminal lumbar interbody fusion rates in patients using a novel titanium implant and demineralised cancellous bone sponge. Int J Spine Surg 7(1):95–100
Gruskin E, Doll BA, Futrell FW, Schmitz JP, Hollinger JO (2012) Demineralized bone matrix in bone repair: history and use. Adv Drug Deliv Rev 64(12):1063–1077
Holt DJ, Grainger DW (2012) Demineralized bone matrix as a vehicle for delivering endogenous and exogenous therapeutics in bone repair. Adv Drug Deliv Rev 64(12):1123–1128
Liu G, Sun J, Li Y, Zhou H, Cui L, Liu W, Cao Y (2008) Evaluation of partially demineralized osteoporotic cancellous bone matrix combined with human bone marrow stromal cells for tissue engineering: an in vitro and in vivo study. Calcif Tissue Int 83:176–185
Mauney JR, Jaquiery C, Volloch V, Heberer M, Martin I, Kaplan D (2005) In vitro and in vivo evaluation of differentially demineralized cancellous bone scaffolds combined with human bone marrow stromal cells for tissue engineering. Biomaterials 26:3173–3185
Mulliken JB, Glowaki J, Kaban LB, Folkman J, Murray JE (1981) Use of demineralised allogeneic bone implants for the correction of maxillocraniofacial deformities. Ann Surg 194:366–372
Ohta H, Wakitani S, Tensho K, Horiuchi H, Wakabayashi S, Saito N, Nakamura Y, Nozaki K, Imai Y, Takaoka K (2005) The effects of heat on the biological activity of recombinant human bone morphogenetic protein-2. J Bone Miner Metab 23:420
Qiu QQ, Connor J (2008) Effects of γ-irradiation, storage and hydration on osteoinductivity of DBM and DBM/AM composite. J Biomed Res A 87(2):373–379
Rhyu KH, Jung MH, Yoo JJ, Lee MC, Seong SC, Kim HJ (2003) In vitro release of vancomycin from vancomycin-loaded blood coated demineralised bone. Int Orthop 27:53–55
Rosenthal R, Folkman J, Glowacki J (1999) Demineralized bone implants for non-union fractures, bone cysts, and fibrous lesions. Clin Orthop Relat Res 364:61–69
Shi T, Niedzinski JR, Samaniego A, Bogdansky S, Atkinson BL (2012) Adipose-derived stem cells combined with a demineralised cancellous bone substrate for bone regeneration. Tissue Eng Part A 18:1313–1321
Smith CA, Richardson SM, Eagle MJ, Rooney P, Board T, Hoyland JA (2014) The use of a novel bone allograft wash process to generate a biocompatible, mechanically stable and osteoinductive biological scaffold for use in tissue engineering. J Tissue Eng Regen Med [e-pub ahead of print]
Stevens A (1990) Pigments and minerals. In: Bancroft JD, Stevens A (eds) Theory and practice of histological techniques, 3rd edn. Churchill Livingstone, Edinburgh, pp 245–267
Wolfinbarger L, Eisenlohr LM, Ruth K (2008) Demineralized bone matrix: maximizing new bone formation for successful bone implantation. In: Pietrzak W (ed) Orthopedic biology and medicine: musculoskeletal tissue regeneration biological materials and methods. Humana Press, Totowa
Zhang N, Zhou M, Zhang Y, Wang X, Ma S, Dong L, Yang T, Ma L, Li B (2014) Porcine bone grafts defatted by lipase: efficacy of defatting and assessment of cytocompatibility. Cell Tissue Bank 15:357–367
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We acknowledge support for this work from NHSBT.
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Eagle, M.J., Rooney, P. & Kearney, J.N. Development of an improved bone washing and demineralisation process to produce large demineralised human cancellous bone sponges. Cell Tissue Bank 16, 569–578 (2015). https://doi.org/10.1007/s10561-015-9504-y
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DOI: https://doi.org/10.1007/s10561-015-9504-y