Eagle Mark J, Rooney Paul, Kearney John N
NHSBT, Tissue Services R&D, 14 Estuary Banks, Speke, Liverpool, L24 8RB, UK.
Cell Tissue Bank. 2015 Dec;16(4):569-78. doi: 10.1007/s10561-015-9504-y. Epub 2015 Mar 4.
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 cm(3). 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 cm(3)) 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.
由松质骨制成的成型脱矿骨基质（DBM）在骨科和牙科手术中具有重要用途，该材料的特性使其能够插入狭窄的缺损处，从而充当空隙填充物和新骨可以在其上形成的支架。这些海绵通常尺寸较小，<1.0 cm³。在本研究中，我们报告了一种改进的骨清洗和脱矿过程，该过程允许从已故捐赠者的骨中生产更大的DBM海绵（3.375或8.0 cm³）。这些海绵经过一系列温水洗涤，有些还进行了超声处理、离心、100%乙醇处理以及两次去污化学洗涤，并在真空下使用0.5 N盐酸进行最佳脱矿处理。通过钙的定量测量和压缩定性来确认脱矿。还测定了蛋白质和DNA的去除情况。DBM海绵在最终灭菌前进行冷冻干燥，目标剂量为25 kGy伽马射线辐照，同时保持冷冻状态。对海绵样本进行组织学检查，以检测钙、胶原蛋白和细胞的存在情况。数据表明没有细胞、没有骨髓，且残留钙最多为1.5%。
