Development of an improved bone washing and demineralisation process to produce large demineralised human cancellous bone sponges
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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.
KeywordsDemineralised bone sponge Vacuum Protein DNA Allograft
We acknowledge support for this work from NHSBT.
Conflict of interest
The authors declare that they have no conflict of interest.
- Bethesda MD (2006) American association of tissue banks. Standards for tissue banking, (11th ed) American association of tissue banksGoogle Scholar
- 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 119–142Google Scholar
- 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]Google Scholar
- 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]Google Scholar
- 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):373–379Google Scholar
- 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]Google Scholar
- Stevens A (1990) Pigments and minerals. In: Bancroft JD, Stevens A (eds) Theory and practice of histological techniques, 3rd edn. Churchill Livingstone, Edinburgh, pp 245–267Google Scholar
- 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, TotowaGoogle Scholar