脱矿骨基质骨诱导活性的定量敏感体外测定法。

Quantitative and sensitive in vitro assay for osteoinductive activity of demineralized bone matrix.

作者信息

Han Bo, Tang Baowei, Nimni Marcel E

机构信息

Tissue Engineering Laboratory, Department of Surgery, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA.

出版信息

J Orthop Res. 2003 Jul;21(4):648-54. doi: 10.1016/S0736-0266(03)00005-6.

DOI:10.1016/S0736-0266(03)00005-6

PMID:12798064

Abstract

A sensitive, rapid, reliable and quantitative method to check the bone forming potential of demineralized bone matrix (DBM) has been developed. The osteoinductivity of the bone morphogenetic proteins (BMPs), present in DBM, can be measured in vitro using a pluripotent myoblast C2C12 cell line. Alkaline phosphatase activity induced by co-incubation of DBM with C2C12 cells was dose-responsive and corresponds to the amount of active BMPs in DBM. Bone forming potential was simultaneously tested in vivo by implanting DBM intra-muscularly in nude rats. ALP activity induced in C2C12 cells, correlated with bone formation in vivo (r=0.88), determined by alkaline phosphatase activity, mineralization density and histomorphology of the DBM explants. Results from DBM batches, originating from five established Bone Banks, showed good consistency between in vitro and in vivo assays. However, DBM activity varied widely from bank to bank as well as from batch to batch within the same bank.

摘要

已开发出一种灵敏、快速、可靠且定量的方法来检测脱矿骨基质（DBM）的骨形成潜力。DBM中存在的骨形态发生蛋白（BMP）的骨诱导活性可在体外使用多能成肌细胞C2C12细胞系进行测量。DBM与C2C12细胞共同孵育诱导的碱性磷酸酶活性呈剂量依赖性，且与DBM中活性BMP的量相对应。通过将DBM肌内植入裸鼠体内，同时在体内测试骨形成潜力。C2C12细胞中诱导的碱性磷酸酶活性与体内骨形成相关（r = 0.88），体内骨形成通过DBM外植体的碱性磷酸酶活性、矿化密度和组织形态学来确定。来自五个成熟骨库的DBM批次的结果表明，体外和体内试验之间具有良好的一致性。然而，不同骨库之间以及同一骨库内不同批次之间的DBM活性差异很大。