Assessment of collagen quality associated with non-enzymatic cross-links in human bone using Fourier-transform infrared imaging.
Assessment of collagen quality associated with non-enzymatic cross-links in human bone using Fourier-transform infrared imaging.
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DOI：
10.1016/j.bone.2017.01.015
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发表时间：
2017-04
期刊：
影响因子：
4.1
通讯作者：
Busse B
中科院分区：
文献类型：
--
作者：
Schmidt FN;Zimmermann EA;Campbell GM;Sroga GE;Püschel K;Amling M;Tang SY;Vashishth D;Busse B
衰老和许多疾病，尤其是糖尿病，与骨基质中非酶交联的积累有关。非酶交联，也称为晚期糖基化终末产物 (AGE)，发生在胶原组织水平，与可塑性降低和骨折风险增加相关。在这项研究中，傅里叶变换红外（FTIR）成像用于检测与人骨胶原中非酶交联形成相关的光谱变化。在这里，在一个体外核糖治疗组和另一个体内双膦酸盐治疗组中研究了非酶交联谱。通过 FTIR 成像，通过酰胺 I 峰内 1678/1692 cm−1 子带的面积比（称为非酶交联比 (NE-xLR)）测量与非酶交联相关的胶原蛋白质量的二维 (2D) 空间分布。与对照组相比，肋骨治疗组的 NE-xLR 增加了 35%（p < 0.005），间质骨组织更容易形成非酶交联。超高效液相色谱、荧光显微镜和荧光分析证实了对照组和肋骨组中非酶交联含量与 NE-xLR 比率之间的相关性。二维骨微结构的高分辨率 FTIR 成像显示，组织年龄较高的骨区域（即间质骨）中非酶交联的积累增强。这种非酶交联比 (NE-xLR) 使研究人员不仅可以研究骨骼中 AGE 的总体含量，还可以研究其空间分布，该分布随骨骼老化和糖尿病而变化，并提供骨骼骨折倾向的额外测量。
Aging and many disease conditions, most notably diabetes, are associated with the accumulation of non-enzymatic cross-links in the bone matrix. The non-enzymatic crosslinks, also known as advanced glycation end products (AGEs), occur at the collagen tissue level, where they are associated with reduced plasticity and increased fracture risk. In this study, Fourier-transform infrared (FTIR) imaging was used to detect spectroscopic changes associated with the formation of non-enzymatic cross-links in human bone collagen. Here, the non-enzymatic cross-link profile was investigated in one cohort with an in vitro ribose treatment as well as another cohort with an in vivo bisphosphonate treatment. With FTIR imaging, the two-dimensional (2D) spatial distribution of collagen quality associated with non-enzymatic cross-links was measured through the area ratio of the 1678/1692 cm−1 subbands within the amide I peak, termed the non-enzymatic crosslink-ratio (NE-xLR). The NE-xLR increased by 35% in the ribation treatment group in comparison to controls (p< 0.005), with interstitial bone tissue being more susceptible to the formation of non-enzymatic cross-links. Ultra high performance liquid chromatography, fluorescence microscopy, and fluorometric assay confirm a correlation between the non-enzymatic cross-link content and the NE-xLR ratio in the control and ribated groups. High resolution FTIR imaging of the 2D bone microstructure revealed enhanced accumulation of non-enzymatic cross-links in bone regions with higher tissue age (i.e., interstitial bone). This non-enzymatic cross-link ratio (NE-xLR) enables researchers to study not only the overall content of AGEs in the bone but also its spatial distribution, which varies with skeletal aging and diabetes mellitus and provides an additional measure of bone's propensity to fracture.