Hentunen T A, Cunningham N S, Vuolteenaho O, Reddi A H, Väänänen H K
Department of Anatomy, University of Oulu, Finland.
Bone Miner. 1994 Jun;25(3):183-98. doi: 10.1016/s0169-6009(08)80238-3.
An activity that recruits osteoclasts has been identified and partially characterized from bone matrix. Bone-derived osteoclast recruiting activity (BORA) was co-purified with osteogenin, a bone inductive protein. Osteogenin was extracted from bovine bone with 6 M urea and purified by chromatography on hydroxyapatite, heparin-Sepharose and Sephacryl S-200 gel filtration. The biologically active osteoclast formation-stimulating material was further purified by C18 reverse phase HPLC. BORA is obviously distinct from osteogenin and transforming growth factor beta (TGF-beta), since further purified osteogenin and pure TGF-beta did not stimulate the formation of osteoclast-like cells. BORA (0.1-10 micrograms/ml) stimulated the formation of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells (MNC) in a dose-dependent manner. These multinucleated cells resorbed bone when cultured on bovine bone slices. The effect of BORA is primarily directed to differentiate osteoclast precursors, since it did not stimulate osteoclast function in in vitro resorption assay where disaggregated rat osteoclasts were cultured on bovine bone slices. However, after 24 h preincubation with 50 nM PTH in the mouse calvaria assay, BORA at 10 micrograms/ml significantly stimulated bone resorption.
一种可募集破骨细胞的活性物质已从骨基质中被鉴定出来，并对其进行了部分特性分析。骨源性破骨细胞募集活性物质（BORA）与骨生成素（一种骨诱导蛋白）共同纯化得到。骨生成素用6M尿素从牛骨中提取，然后通过羟基磷灰石、肝素-琼脂糖和Sephacryl S-200凝胶过滤色谱法进行纯化。具有生物活性的破骨细胞形成刺激物质通过C18反相高效液相色谱法进一步纯化。BORA明显不同于骨生成素和转化生长因子β（TGF-β），因为进一步纯化的骨生成素和纯TGF-β不会刺激破骨样细胞的形成。BORA（0.1-10微克/毫升）以剂量依赖的方式刺激抗酒石酸酸性磷酸酶（TRAP）阳性多核细胞（MNC）的形成。当这些多核细胞在牛骨切片上培养时会吸收骨质。BORA的作用主要是促使破骨细胞前体分化，因为在体外吸收试验中，当将分离的大鼠破骨细胞培养在牛骨切片上时，它不会刺激破骨细胞的功能。然而，在小鼠颅骨试验中用50 nM甲状旁腺激素（PTH）预孵育24小时后，10微克/毫升的BORA能显著刺激骨吸收。
