Adkisson H D, Strauss-Schoenberger J, Gillis M, Wilkins R, Jackson M, Hruska K A
Department of Medicine, Barnes-Jewish Hospital/Washington University, St. Louis, Missouri 63110, USA.
J Orthop Res. 2000 May;18(3):503-11. doi: 10.1002/jor.1100180326.
We developed a reproducible, relatively rapid bioassay that quantitatively correlates with the osteoinductive capacity of demineralized bone matrix obtained from human long bones. We have found that Saos human osteosarcoma cells proliferate in response to incubation with demineralized bone matrix and that an index of this proliferative activity correlates with demineralized bone matrix-induced osteogenesis in vivo. The bioassay (Saos cell proliferation) had an interassay coefficient of variation of 23 +/- 2% and an intra-assay coefficient of 11 +/- 1%. Cell proliferation was normalized to a standard sample of demineralized bone matrix with a clinically high osteoinductive capacity, which was assigned a value of one. The Saos cell proliferation for each sample was related to the standard and assigned a value placing it into the low (0.00-0.39), intermediate (0.40-0.69), or high (0.70-1.49) osteoinductive index group. Osteoinduction of human demineralized bone matrix was quantitated by expressing new bone formation as a function of the total bone volume (new bone plus the demineralized bone powder). The demineralized bone matrix was placed in pouches formed in the rectus abdominis muscles of athymic rats, and endochondral bone formation was assessed at 35 days following implantation, when marrow spaces in the ossicles were formed by new bone bridging the spaces between demineralized bone matrix particles. The proliferative index correlated with the area of new bone formation in histological sections of the newly formed ossicles. When the proliferative index (the osteoinductive index) was divided into low, intermediate, and high groups, the correlation between it and new bone formation (osteoinduction) was 0.850 (p < 0.0005) in 25 samples of demineralized bone matrix. There was no overlap in the osteoinduction stimulated between the samples with low and high osteoinductive indices. We conclude that the proliferation assay is useful for the routine screening of bone allograft donors for osteoinductive potential. Furthermore, the two-dimensional area of new bone formation, as it relates to total new bone area, is a quantitative measure of osteoinduction.
我们开发了一种可重复、相对快速的生物测定方法，该方法与从人长骨获得的脱矿骨基质的骨诱导能力具有定量相关性。我们发现，Saos人骨肉瘤细胞在与脱矿骨基质孵育后会增殖，并且这种增殖活性的指标与脱矿骨基质在体内诱导的成骨作用相关。该生物测定法（Saos细胞增殖）的批间变异系数为23±2%，批内变异系数为11±1%。细胞增殖以具有临床高骨诱导能力的脱矿骨基质标准样品进行标准化，该标准样品被赋予值1。每个样品的Saos细胞增殖与标准样品相关，并根据其值将其归入低（0.00 - 0.39）、中（0.40 - 0.69）或高（0.70 - 1.49）骨诱导指数组。通过将新骨形成表示为总骨体积（新骨加脱矿骨粉）的函数来定量人脱矿骨基质的骨诱导作用。将脱矿骨基质置于无胸腺大鼠腹直肌中形成的囊中，并在植入后35天评估软骨内骨形成，此时小骨中的骨髓腔由新骨桥接脱矿骨基质颗粒之间的间隙形成。增殖指数与新形成小骨组织切片中新骨形成的面积相关。当将增殖指数（骨诱导指数）分为低、中、高组时，在25个脱矿骨基质样品中，其与新骨形成（骨诱导）之间的相关性为r = 0.850（p < 0.）。低骨诱导指数和高骨诱导指数的样品之间刺激的骨诱导没有重叠。我们得出结论，增殖测定法可用于常规筛选骨移植供体的骨诱导潜力。此外，与总新骨面积相关的新骨形成的二维面积是骨诱导的定量指标。
