亲骨荧光素间接标记软骨内成骨的早期血管新生

摘要

背景:虽然当前有多种方法可以观察和检测软骨内成骨早期血管新生,但是均存在一定的不足.目的:探讨四环素和茜素络合酮间接标记软骨内成骨过程中新生血管的可能性.方法:制备新西兰兔双侧桡骨骨缺损模型并植入β-磷酸三钙材料,分别于术后第1天和第15天注射四环素和茜素络合酮,第28天取材.部分标本在墨汁灌注后行硬组织切片荧光/光学显微镜观察,部分标本经脱钙后行免疫组化染色观察,比较软骨内成骨过程中亲骨荧光素标记管腔结构与免疫组化、墨汁灌注标记血管结构的一致性.结果与结论:①亲骨荧光素标记的管腔结构经免疫组化染色证实为CD34阳性的血管结构;②在荧光显微镜下,亲骨荧光素标记的血管形态与墨汁灌注的血管形态一致;荧光素标记后经墨汁灌注的血管在荧光显微镜下可见荧光管腔中有黑色墨汁走行;③此外,亲骨荧光素标记的管腔结构颜色绚丽、三维结构更加生动,可通过不同颜色的荧光显示不同时期的血管新生和演变过程,具有独特的优势,可用于软骨内成骨早期血管发生的形态学检测.%BACKGROUND: There are various methods to observe and detect early angiogenesis in the process of entochondrostosis, but each holds certain deficiencies.OBJECTIVE: To explore the possibility of tetracycline and alizarin complexone as an indirect marker of angiogenesis in the process of entochondrostosis.METHODS: New Zealand white rabbit models of bilateral radial bone defects were prepared, followed by β-tricalcium phosphate implantation, and then given the injection of tetracycline and alizarin complexone at 1 and 15 days,respectively. Samples were collected at 28 days, some of which were observed using fluorescence/light microscope after ink perfusion and hard tissue slicing, and the others were decalcified and observed using immunohistochemistry. The uniformity between lumen structures labeled with bone affinity fluorescein and vascular structures marked by immunohistochemistry and ink perfusion was compared.RESULTS AND CONCLUSION: The lumen structure labeled with bone affinity fluorescein was confirmed to be a CD34 positive vascular structure. Under the fluorescence microscope, the bone affinity fluorescein labeled vascular morphology was consistent with ink perfusion-labeled, and black ink lines could be observed in the lumen structures labeled with bone affinity fluorescein after ink perfusion. In addition, the color of the lumen labeled with fluorescein was more gorgeous,three-dimensional structure more vivid, and the vascular evolution process distinguished more easily by different fluorescein colors, exhibiting unique advantages. Therefore, it is available to detect the early angiogenesis in the process of entochondrostosis.