同步辐射微束荧光分析技术在兔髌骨-髌腱界面中钙、锌元素分布的应用

Investigation of calcium and zinc distribution in rabbit patella -patellar tendon interface using synchrotron radiation micro X -ray fluorescence technique

目的利用同步辐射微束荧光分析技术(SR-μXRF)表征兔髌骨-髌腱界面(PPTI)中钙、锌元素的含量及分布特点。方法兔PPTI经切片及抛光处理后,于上海光源行SR-μXRF检测,检测后样本行苏木素-伊红(HE)及樊基森(Van-Gieson)染色进行匹配观察。结果组织学染色显示兔PPTI钙化的纤维软骨层厚度为(163.00±42.00) μm;SR-μXRF检测可见兔PPTI中钙含量最大值93 853 cps(位于钙化纤维软骨层与软骨下骨的交界区)是髌腱的(28.00±9.05)倍;锌含量最大值10 100 cps(位于钙化与未钙化纤维软骨层的交界区)是髌腱的(3.20±0.65)倍;钙含量的最大值是锌的(8.70±3.20)倍;钙、锌含量的最大值所处位置相距(135.00±16.00) μm,与组织学染色结果一致(r^2=0.935)。结论SR-μXRF可超微分辨地表征PPTI中微量元素的含量及分布特征.

Objective The unique distributions and maps of calcium and zinc at the regions of interest in patella-patellar tendon interface (PPTI) were detected by synchrotron radiation micro X-ray fluorescence (SR-μXRF).Methods Healthy New Zealand rabbits were used in this experiment. Five harvested PPTI samples were cut along the sagittal plane of the PPTI by a microtome. Sections of PPTI were then scanned by SR-μXRF at BL15U1 of shanghai synchrotron radiation facility (SSRF) in China. Then the samples were used for histological staining with hematoxylin and eosin (HE) and Van-Gieson.ResultsHistological staining showed that the thickness of calcified fibrocartilage in PPTI was (163.00±42.00) μm;The distributions and characterization of calcium and zinc of PPTI were successfully obtained via the SR-μXRF. The maximum calcium content [93 853 cps, (28.00±9.05) times of that of patellar tendon] was found in the transitional zone of calcified fibrocartilage region and the patella. A significant accumulation of zinc [10 100 cps, (3.20±0.65) times of that of patellar tendon] was exhibited in the transition area of tidemark. The maximum content of zinc was (8.70±3.20) times of that of calcium. The distribution distance between the maximum content of calcium and zinc was (135.00±16.00) μm, which was consistent with histological staining (r2=0.935). Conclusion SR-μXRF served as a high-precision and high-resolution instrument to qualitatively assess the distributions of calcium and zinc in the PPTI, which provides a new insight for the study of bone tendon interface repair.