Institute of Biothermal Science and Technology, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai, China.
Tianjin Key Laboratory of Neurotrauma Repair, Institute of Neurotrauma Repair, Characteristic Medical Center of Chinese People's Armed Police Force, Tianjin, China.
J Biomed Mater Res B Appl Biomater. 2024 Jan;112(1):e35358. doi: 10.1002/jbm.b.35358.
Allogenic demineralized bone matrix (DBM), processed to expose bioactive proteins imbedded by calcium salts, is widely used for bone repair and regeneration as an alternative to the autologous bone graft. However, demineralized bone matrices from tissue banks vary significantly in residual calcium content and osteogenicity for clinical bone regeneration. The present study produced DBM with various residual calcium contents by partial demineralization using ethylenediaminetetraacetic acid disodium (EDTA) and hydrochloric acid. Compositional analysis reveals that, as the percent weight loss of bone materials increases from 0% to 74.9% during demineralization, the residual calcium content of DBM decreases from 24.8% to 0.2% and collagen content increases from 29.7% to 92.6%. Calorimetrical analysis and Fourier transform infrared (FTIR) analysis demonstrated that demineralization to the residual calcium content of <4% enables the complete exposure and/or release of bone collagen fibers and other bioactive molecules. In order to evaluate the relationship between the extent of demineralization and the osteogenicity of DBM, DBM particles were fabricated with the aid of acellular dermal matrix (ADM) microfibers to form flexible foam-like DBM/ADM composites. Proteomic analysis identified various type collagens and bone formation-related bioactive molecules in both ADM and DBM. Using the rat bilateral Φ = 5 mm calvarium defect repair model, the study had shown that the DBM/ADM composite with ~20% DBM residual calcium (e.g., ~40% calcium being removed) maximized the osteogenicity for bone defect repair after 4 and 8 weeks. DBM with ~40% calcium removal had the maximal osteogenicity presumably through the sustained release of bioactive molecules during the process of bone regeneration.
同种异体脱矿骨基质（DBM），经处理后暴露嵌入钙盐的生物活性蛋白，作为自体骨移植物的替代品，广泛用于骨修复和再生。然而，来自组织库的脱矿骨基质在用于临床骨再生时，其残留钙含量和成骨活性差异很大。本研究通过使用乙二胺四乙酸二钠（EDTA）和盐酸进行部分脱矿作用，生产出具有不同残留钙含量的 DBM。组成分析表明，随着脱矿过程中骨材料的失重百分比从 0%增加到 74.9%，DBM 的残留钙含量从 24.8%降至 0.2%，胶原含量从 29.7%增加到 92.6%。量热分析和傅里叶变换红外（FTIR）分析表明，脱矿至残留钙含量<4%可使骨胶原纤维和其他生物活性分子完全暴露和/或释放。为了评估脱矿程度与 DBM 成骨活性之间的关系，借助无细胞真皮基质（ADM）微纤维制备了 DBM 颗粒，形成了具有柔韧性的泡沫状 DBM/ADM 复合材料。蛋白质组学分析鉴定了 ADM 和 DBM 中各种类型的胶原蛋白和与骨形成相关的生物活性分子。使用大鼠双侧Φ=5mm 颅骨缺损修复模型，研究表明，残留钙含量约为 20%（例如，去除约 40%的钙）的 DBM/ADM 复合材料在 4 周和 8 周后对骨缺损修复的成骨活性最大。残留钙含量约为 40%的 DBM 具有最大的成骨活性，可能是通过在骨再生过程中持续释放生物活性分子。
