Guangzhou Institute of Traumatic Surgery, The Fourth Affiliated Hospital, Jinan University School of Medicine, 396 Tong Fu Zhonglu Road, Hai Zhu District, Guangzhou, 510220, Guangdong, People's Republic of China,
Calcif Tissue Int. 2014 Mar;94(3):338-47. doi: 10.1007/s00223-013-9815-6. Epub 2013 Nov 20.
Bone is a mineralized connective tissue that is continuously and microstructurally remodeled. Altered bone formation and microstructure arise in pathological bone conditions such as osteoporosis, osteonecrosis, fracture repair, and Paget disease of bone. A proper and objective assessment of bone formation and microstructure will provide insight into the understanding of bone pathogenesis and remodeling. Here, new bone formation ex vitro and its microstructure were evaluated in in vivo multiple sequential polychrome-labeled samples using confocal laser scanning microscopy (CLSM), which generated clearer and more reliable images of thick bone sections than conventional fluorescence microscopy (CFM). Intriguingly, fine details of the bone microstructural features, including the mineralization fronts, quiescent versus active osteons, and Volkmann's channel, were elucidated using CLSM, which defines the relationship between morphological changes and function, when combined with differential interference contrast microscopy. Furthermore, CLSM provided objective evaluations of bone formation, such as the ratio of labeled areas of new bone formation in a rabbit model when compared with CFM. Altogether, new bone formation and its microstructure can be evaluated more adequately using a combination of CLSM and DIC microscopies.
骨骼是一种矿化的结缔组织，不断地进行微观结构重塑。在病理性骨条件下，如骨质疏松症、骨坏死、骨折修复和骨 Paget 病，会出现改变的骨形成和微观结构。对骨形成和微观结构进行适当和客观的评估，将深入了解骨发病机制和重塑。在这里，使用共聚焦激光扫描显微镜 (CLSM) 对体内多个连续多色标记样本中的体外新骨形成及其微观结构进行了评估，与传统荧光显微镜 (CFM) 相比，CLSM 生成的厚骨切片图像更清晰、更可靠。有趣的是，使用 CLSM 可以阐明骨微观结构特征的细微细节，包括矿化前沿、静止和活跃的骨单位以及沃克曼氏管，当与微分干涉对比显微镜结合使用时，它可以定义形态变化与功能之间的关系。此外，CLSM 还可以对骨形成进行客观评估，例如与 CFM 相比，在兔模型中评估新骨形成的标记区域比例。总之，使用 CLSM 和 DIC 显微镜的组合可以更充分地评估新骨形成及其微观结构。
