Zhao Yong-Jie, Yin Gang, Liu Bin, Deng Xiao-Qiang, Cao Hai-Yan, Liu Ying
Binzhou Medical University Hospital, Binzhou, Shandong Province, China.
Xing'an League People's Hospital, Ulanhot, Inner Mongolia, China.
Cell Tissue Bank. 2024 Jun;25(2):697-703. doi: 10.1007/s10561-024-10132-5. Epub 2024 Mar 15.
Demineralized bone matrix (DBM) has been regarded as an ideal bone substitute as a native carrier of bone morphogenetic proteins (BMPs) and other growth factors. However, the osteoinductive properties diverse in different DBM products. We speculate that the harvest origin further contributing to variability of BMPs contents in DBM products besides the process technology. In the study, the cortical bone of femur, tibia, humerus, and ulna from a signal donor were prepared and followed demineralizd into DBM products. Proteins in bone martix were extracted using guanidine-HCl and collagenase, respectively, and BMP-2 content was detected by sandwich enzyme-linked immunosorbent assay (ELISA). Variability of BMP-2 content was found in 4 different DBM products. By guanidine-HCl extraction, the average concentration in DBMs harvested from ulna, humerus, tibia, and femur were 0.613 ± 0.053, 0.848 ± 0.051, 3.293 ± 0.268, and 21.763 ± 0.344, respectively (p < 0.05), while using collagenase, the levels were 0.089 ± 0.004, 0.097 ± 0.004, 0.330 ± 0.012, and 1.562 ± 0.008, respectively (p < 0.05). In general, the content of BMP-2 in long bones of Lower limb was higher than that in long bones of upper limb, and GuHCl had remarkably superior extracted efficiency for BMP-2 compared to collagenase. The results suggest that the origin of cortical bones harvested to fabricate DBM products contribute to the variability of native BMP-2 content, while the protein extracted method only changes the measured values of BMP-2.
脱矿骨基质（DBM）作为骨形态发生蛋白（BMPs）和其他生长因子的天然载体，一直被视为理想的骨替代物。然而，不同DBM产品的骨诱导特性各不相同。我们推测，除了加工工艺外，收获来源进一步导致了DBM产品中BMPs含量的变异性。在本研究中，取自单一供体的股骨、胫骨、肱骨和尺骨的皮质骨被制备出来，随后脱矿制成DBM产品。分别使用盐酸胍和胶原酶提取骨基质中的蛋白质，并通过夹心酶联免疫吸附测定（ELISA）检测BMP-2含量。在4种不同的DBM产品中发现了BMP-2含量的变异性。通过盐酸胍提取，从尺骨、肱骨、胫骨和股骨收获的DBM中的平均浓度分别为0.613±0.053、0.848±0.051、3.293±0.268和21.763±0.344（p<0.05），而使用胶原酶时，水平分别为0.089±0.004、0.097±0.004、0.330±0.012和1.562±0.008（p<0.05）。一般来说，下肢长骨中BMP-2的含量高于上肢长骨，并且与胶原酶相比，盐酸胍对BMP-2的提取效率明显更高。结果表明，用于制造DBM产品的皮质骨来源导致了天然BMP-2含量的变异性，而蛋白质提取方法仅改变了BMP-2的测量值。
