Evidence of nanobacterial-like structures in calcified human arteries and cardiac valves
Evidence of nanobacterial-like structures in calcified human arteries and cardiac valves
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DOI：
10.1152/ajpheart.00075.2004
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发表时间：
2004-09-01
影响因子：
4.8
通讯作者：
Lieske, JC
中科院分区：
文献类型：
--
作者：
Miller, VM;Rodgers, G;Lieske, JC
介导血管钙化的机制仍不完全清楚。被认为是一种细菌(纳米细菌)的纳米级物体与钙化的地质标本、人类肾结石和卵巢癌中的沙门氏菌小体有关。实验的目的是评估人类血管组织中是否存在类似的纳米级物体。来自德克萨斯州奥斯汀心脏医院和明尼苏达州罗切斯特市梅奥诊所的钙化性人动脉瘤(n=8)、颈动脉斑块(n=2)、股动脉斑块(n=2)和心脏瓣膜病患者的心脏瓣膜(n=2)和非钙化性动脉瘤(n=2)作为手术废物收集。对每个标本的整体或相邻切片进行电子显微镜检查，进行磷酸钙染色，或用商用抗体(8D10)进行染色。过滤后的动脉瘤匀浆(0.2um)培养，用8D10抗体与PicoGreen染色或与[H-3]尿苷孵育。钙磷染色在所有钙化组织中分布不均。8D10免疫染色在有无磷酸钙的区域也呈不均匀分布。对免疫阳性区域的分析表明，球体的大小从30到100 nm不等，具有钙和磷的光谱模式(高能色散光谱)。从钙化但不是来自非钙化动脉瘤培养的纳米颗粒被DNA特异性染料识别并掺入放射性标记的尿苷，脱钙后，它们在电子显微镜下显示包含细胞壁。因此，从地质标本和人类肾结石中分离出来的纳米级颗粒类似于被描述为纳米细菌的颗粒，可以在钙化的人类心血管组织中可视化并进行培养。
Mechanisms mediating vascular calcification remain incompletely understood. Nanometer scale objects hypothesized to be a type of bacteria ( nanobacteria) are associated with calcified geological specimens, human kidney stones, and psammona bodies in ovarian cancer. Experiments were designed to evaluate human vascular tissue for the presence of similar nanometer-scale objects. Calcified human aneurysms (n = 8), carotid plaques ( n = 2), femoral arterial plaques ( n = 2), and cardiac valves ( n = 2) and noncalcified aneurysms from patients with bicuspid aortic valve disease ( n = 2) were collected as surgical waste from the Heart Hospital of Austin, Austin, Texas, and Mayo Clinic, Rochester, Minnesota. Whole mounts or adjacent sections from each specimen were examined by electron microscopy, stained for calcium phosphate, or stained with a commercially available antibody (8D10). Filtered (0.2 mum) homogenates of aneurysms were cultured and costained with 8D10 antibody followed by PicoGreen to detect DNA or incubated with [H-3] uridine. Staining for calcium phosphate was heterogeneously distributed within all calcified tissues. Immunological staining with 8D10 was also heterogeneously distributed in areas with and without calcium phosphate. Analysis of areas with positive immunostaining identified spheres ranging in size from 30 to 100 nm with a spectral pattern of calcium and phosphorus (high-energy dispersive spectroscopy). Nanosized particles cultured from calcified but not from noncalcified aneurysms were recognized by a DNA-specific dye and incorporated radiolabeled uridine, and, after decalcification, they appeared via electron microscopy to contain cell walls. Therefore, nanometer-scale particles similar to those described as nanobacteria isolated from geological specimens and human kidney stones can be visualized in and cultured from calcified human cardiovascular tissue.