Satomura K, Hiraiwa K, Nagayama M
First Department of Oral and Maxillofacial Surgery, School of Dentistry, Tokushima University, Japan.
Bone Miner. 1991 Jul;14(1):41-54. doi: 10.1016/0169-6009(91)90101-5.
Rat bone marrow stromal cells were cultured in the presence of 10 nM dexamethasone and various concentrations of beta-glycerophosphate. At day 12-15, some nodules consisting of polygonal cells were formed in all culture conditions, and these nodules were mineralized 2-3 days later. beta-Glycerophosphate significantly enhanced nodule formation at concentrations of not less than 5 mM. The mineralized nodules formed in the absence of beta-glycerophosphate were examined using phase-contrast microscopy, undemineralized and demineralized tissue histology, histochemistry for alkaline phosphatase, immunohistochemistry for type I, II, and III collagen, energy dispersive X-ray microanalysis, electron diffraction, and Fourier transform infrared spectroscopy (FT-IR). Mineralized nodules had histological characteristic similar to bone. Cells associated with nodules exhibited high alkaline phosphatase activity, and extracellular matrix of the nodules predominantly consisted of type I collagen. X-Ray microanalysis showed the presence of Ca and P in the mineralized area, and electron diffraction pattern showed the mineral to have apatite crystal structure. Moreover FT-IR indicated that the mineral was a mixture of hydroxyapatite and carbonateapatite. From these observations, it is concluded that the mineralized nodules formed in our culture system are truly bone-like.
将大鼠骨髓基质细胞在含有10 nM地塞米松和不同浓度β-甘油磷酸的条件下进行培养。在第12 - 15天，所有培养条件下均形成了一些由多边形细胞组成的结节，这些结节在2 - 3天后矿化。β-甘油磷酸在浓度不低于5 mM时显著增强结节形成。对在无β-甘油磷酸条件下形成的矿化结节进行了相差显微镜检查、未脱矿和脱矿组织组织学检查、碱性磷酸酶组织化学检查、I型、II型和III型胶原免疫组织化学检查、能量色散X射线微分析、电子衍射和傅里叶变换红外光谱（FT - IR）分析。矿化结节具有与骨相似的组织学特征。与结节相关的细胞表现出高碱性磷酸酶活性，结节的细胞外基质主要由I型胶原组成。X射线微分析显示矿化区域存在钙和磷，电子衍射图谱显示矿物质具有磷灰石晶体结构。此外，FT - IR表明该矿物质是羟基磷灰石和碳酸磷灰石的混合物。从这些观察结果可以得出结论，我们培养系统中形成的矿化结节确实类似骨。
