Relationships between trabecular bone remodeling and bone vascularization: A quantitative study
Relationships between trabecular bone remodeling and bone vascularization: A quantitative study
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DOI：
10.1016/s8756-3282(02)00677-4
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发表时间：
2002-04-01
期刊：
影响因子：
4.1
通讯作者：
Lafage-Proust, MH
中科院分区：
文献类型：
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作者：
Barou, O;Mekraldi, S;Lafage-Proust, MH
除了其在骨发育中的众所周知的作用之外，血管化在骨细胞迁移以进行骨重建和转移性肿瘤侵袭中起主要作用。然而，用于识别骨中血管的各种技术从未被测试用于骨小梁血管定量，而骨重建定量参数通常被评估。在这种情况下，我们开发和比较了各种组织学技术，用于可视化大鼠骨血管，以量化它们。首先，通过心内输注测试了几种产品，以使骨血管网不透明。最好的结果是使用印度墨水-1%琼脂糖溶液或印度墨水饱和硫酸钡溶液，然后通过X射线显微放射照相术。第二，我们还进行了组织酶学和免疫组织化学染色，以确定类型或血管。碱性磷酸酶（用于内皮细胞）和三磷酸腺苷酶（ATP酶）染色（用于平滑肌细胞）均不能提供足够低的背景以进行血管识别和定量。对于免疫组织化学，分析了各种特定的血管成分：层粘连蛋白，平滑肌细胞α-肌动蛋白，因子VIII和凝集素Griffonia difolia。抗层粘连蛋白和抗平滑肌细胞α-肌动蛋白抗体给出了最佳的定量结果。第三，在优化这些技术后，我们对两组12只大鼠进行定量骨和血管组织形态计量学，测量骨重建和血管数量和面积参数。两组之间未观察到统计学差异，证实了我们测量的可重复性。血管数量与组织动力学参数之间存在显著相关性;即骨形成率与印度墨水阳性血管面积（p < 0.009，r（2）= 0.54）和α-肌动蛋白阳性血管数量（p < 0.05，r（2）= 0.66）呈正相关。此外，我们报告了可重复的技术，可视化和定量的血管在骨，也允许同时进行传统的骨组织形态计量学。这种方法有助于研究人员更好地了解正常或病理过程中松质骨及其血管化之间的功能和解剖关系。(Bone 30：604-612; 2002）（C）Elsevier Science Inc. All rights reserved.
Beside its well-known role in bone development, vascularization plays a major role in bone cell migration for bone remodeling and metastatic tumor invasion. However, the various techniques used to identify vessels in bone have never been tested for trabecular bone vessel quantification, whereas bone remodeling quantitative parameters are commonly assessed. In this context, we developed and compared various histological techniques used to visualize blood vessels in rat bone in order to quantify them. First, several products were tested by intracardiac infusion to opacify the bone vascular network. The best results were obtained using either an India ink-1% agarose solution or an India ink-saturated barium sulfate solution followed by X-ray microradiography. Second, to identify the types or vessels, we also performed histoenzymology and immunohistochemistry stainings. Neither alkaline phosphatase (for endothelial cells) nor adenosine triphosphatase (ATPase) stainings (for smooth muscle cells) provided a low enough background to allow for vessel identification and quantification. For immunohistochemistry, various specific vessel constituents were analyzed: laminin, smooth muscle cell alpha-actin, factor VIII, and lectin Griffonia simplifolia. Anti-laminin and anti-smooth muscle cell alpha-actin antibodies gave the best results for quantification. Third, after optimization of these techniques, we performed quantitative bone and vessel histomorphometry on two groups of 12 rats each, for which bone remodeling and vessel number and area parameters were measured. No statistical differences were observed between the two groups, confirming the reproducibility of our measurements. A significant relationship was found between vessel number and histodynamic parameters; that is, bone formation rate correlated positively with India ink-positive vessel area (p < 0.009, r(2) = 0.54) and alpha-actin-positive vessel number (p < 0.05, r(2) = 0.66). Furthermore, we report reproducible techniques for visualization and quantification of vessels in bone that also allowed for simultaneous conventional bone histomorphometry. This methodology should help researchers to better understand the functional and anatomical relationship between trabecular bone and its vascularization during normal or pathological processes. (Bone 30:604-612; 2002) (C) 2002 by Elsevier Science Inc. All rights reserved.