Calcified microspheres as biological entities and their isolation from bone
Calcified microspheres as biological entities and their isolation from bone
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DOI：
10.1023/a:1003707909842
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发表时间：
1999-07-01
期刊：
影响因子：
--
通讯作者：
Carter, DH
中科院分区：
文献类型：
--
作者：
Aaron, JE;Oliver, B;Carter, DH
钙化的微球，直径约1 μ m，出现在骨形成的部位，在那里它们投资于胶原基质，变得汇合，然后消失。骨盐、脂质、纤维连接蛋白和酸性磷酸酶在骨软化、酸蚀和正常人骨中的无机相的颗粒组织化学染色证实了颗粒边界不会随着压实而消失，而仅仅是变形。它们作为离散对象的持久性通过应用从未成熟小鼠颅骨和成熟牛股骨的胶原基质中分离它们的方法得到证实。已有五种方法用于提取它们，包括(i)生化，（ii）化学，（iii）机械，（iv）热源和(v)生物分离。在光学显微镜下，所有分离物均由相似的离散物体和桥接组件组成，其双折射随处理而变化。脱钙后，它们的有机“幽灵”仍然存在。每个分离的微球都有一个复杂的亚结构，由非胶原钙化的细丝团围绕着一个密度较小的中心。丝状物直径为5nm，周期性为5nm，丝状物间有规则的细连接。结论是，微球是独立的，复杂的，普遍的和核心的遏制（即包装）磷酸钙在骨。它们的提取将使进一步的分析成为可能。
Calcified microspheres, about 1 mu m in diameter, appear at sites of bone formation where they invest the collagenous matrix, become confluent and disappear. Evidence that the particle boundaries are not lost with compaction but merely deformed is supported in section by the granular histochemical staining of the inorganic phase for bone salt, lipid, fibronectin and acid phosphatase in osteomalacic, acid-etched and normal human bone. Their persistence as discrete objects is confirmed by the application of methods for their isolation from the collagenous matrix of immature mouse calvarium and mature bovine femur. Five methods have been used to extract them and include (i) biochemical, (ii) chemical, (iii) mechanical, (iv) pyrogenous and (v) biological separation. Under the optical microscope, all isolates consisted of similar discrete objects and bridged assemblies, whose birefringence varied with treatment. After decalcification, their organic 'ghosts' remained. Each isolated microsphere had a complex substructure of clusters of non-collagenous calcified filaments surrounding a less dense centre. The filaments were 5 nm in diameter with a 5 nm periodicity and regular fine interfilamentous connections. It is concluded that the microspheres are independent, complex, pervasive and central to the containment (i.e. packaging) of calcium phosphate in bone. Their extraction will enable further analysis.