Quantification of the collagenolytic activity of isolated osteoclasts by enzyme‐linked immunosorbent assay
Quantification of the collagenolytic activity of isolated osteoclasts by enzyme‐linked immunosorbent assay
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通过酶联免疫吸附测定定量分离破骨细胞的溶胶原活性
DOI：
10.1002/jbmr.5650110212
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发表时间：
1996
影响因子：
6.2
通讯作者：
M. Bonde
中科院分区：
文献类型：
--
作者：
N. Foged;J. Delaissé;P. Hou;Henriette Lou;Takuya Sato;B. Winding;M. Bonde
骨吸收坑的几何定义和测量困难是骨或牙本质基质上培养的孤立破骨细胞进行骨吸收定量分析的主要问题。在本研究中，我们开发了一种用于体外骨吸收定量的酶联免疫吸附试验（ELISA），该试验可特异性定量通过骨或牙本质切片上培养的骨细胞的吸收作用释放到培养基中的I型胶原片段。在牛、大象和人基质上以不同密度接种并培养不同时期的分离兔破骨细胞，观察到吸收陷窝的形成和抗原性胶原片段的释放之间始终存在高度相关性。在进一步支持胶原溶解作用的骨形成性质中，当在作用非常不同但典型的小凹形成抑制剂存在下培养兔骨细胞时，也观察到小凹形成和胶原溶解之间的高度一致性，即，碳酸酐酶抑制剂乙酰唑胺、半胱氨酸蛋白酶抑制剂环氧琥珀酰-L-亮氨酰氨基-（4-胍基）丁烷（E-64）、磷脂酰肌醇3-激酶抑制剂渥曼青霉素和双膦酸盐伊班膦酸盐（BM 21.0955）。总之，ELISA代表了一种简单、精确和客观的方法，通过定量分离的破骨细胞的胶原溶解活性来动态监测体外骨吸收。
Difficulties in the geometrical definition and measurement of resorption pits is a major problem for the quantitative analysis of bone resorption by isolated osteoclasts cultured on bone or dentin substrates. In this study we developed an enzyme‐linked immunosorbent assay (ELISA) for quantification of bone resorption in vitro, which specifically quantifies type I collagen fragments released into the culture medium by the resorptive action of bone cells cultured on slices of bone or dentin. A consistently high correlation between the formation of resorption pits and the release of antigenic collagen fragments was observed for isolated rabbit osteoclasts seeded at various densities and cultured for various periods on bovine, elephant, and human substrates. In a further support of the osteoclastic nature of the collagenolytic effects, a high consistency between pit formation and collagenolysis was also observed when the rabbit bone cells were cultured in the presence of very differently acting but typical inhibitors of pit formation, i.e., the carbonic anhydrase inhibitor acetazolamide, the cysteine proteinase inhibitor epoxysuccinyl‐L‐leucylamido‐(4‐guanodino)butane (E‐64), the phosphatidyl‐inositol 3‐kinase inhibitor wortmannin, and the bisphosphonate ibandronate (BM 21.0955). In conclusion, the ELISA represents a simple, precise, and objective way to dynamically monitor bone resorption in vitro through quantification of the collagenolytic activity of isolated osteoclasts.
影响因子：
15.9
作者：
CARANO, A;TEITELBAUM, SL;BLAIR, HC
通讯作者：
BLAIR, HC