Assessment of glycosaminoglycan content in bone using Raman spectroscopy.
Assessment of glycosaminoglycan content in bone using Raman spectroscopy.
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DOI：
10.1016/j.bone.2023.116751
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发表时间：
2023-06
期刊：
影响因子：
4.1
通讯作者：
Wang, Xiaodu
中科院分区：
文献类型：
--
作者：
Heath, Savannah;Han, Yan;Hua, Rui;Roy, Anuradha;Jiang, Jean;Nyman, Jeffry S.;Wang, Xiaodu
糖胺聚糖（GAG）负责保持骨组织韧性以及调节细胞外基质中的胶原形成和矿化。然而，目前用于表征骨中GAG的方法是破坏性的，因此无法捕获实验组之间GAG的原位变化或差异。作为替代方案，拉曼光谱是一种非破坏性方法，可以检测GAG和其他骨成分的同时变化。在本研究中，我们假设硫酸化GAG的两个最显著的拉曼峰（约1066 cm−1和约1378 cm−1）可用于检测骨中GAG含量的差异。为了检验这一假设，使用了三种实验模型：体外模型（从人尸体骨中酶促去除GAG）、体内小鼠模型（双糖链聚糖KO与WT）和离体老化模型（比较年轻和老年供体的尸体骨样品）。将所有的拉曼测量结果与阿尔新蓝测量结果进行比较，以确认拉曼光谱法在检测骨中GAG变化中的有效性。不管不同的模型，发现骨的拉曼光谱中的~ 1378 cm −1峰在相对于磷酸盐相（~ 960 cm −1）归一化时对骨中GAG含量的变化唯一敏感;即，1378 cm −1/960 cm −1（峰强度比）或1370- 1385 cm −1/930- 980 cm −1（积分峰面积比）。相比之下，1070 cm −1峰（包括GAG的另一个主峰（1066 cm −1））似乎由于碳酸盐（CO 3）在相似峰范围内的同时变化而无法检测骨中GAG的变化。本研究验证了拉曼光谱检测骨基质中GAG水平原位治疗、基因型和年龄相关变化的能力。
Glycosaminoglycans (GAGs) are responsible for preserving bone tissue toughness as well as regulating collagen formation and mineralization in the extracellular matrix. However, current methods for characterization of GAGs in bone are destructive, thus unable to capture in situ changes or differences in GAGs between experimental groups. As an alternative, Raman spectroscopy is a non-destructive method and can detect concurrent changes in GAGs and other bone constituents. In this study, we hypothesized that the two most prominent Raman peaks of sulfated GAGs (at ~1066 cm−1 and at ~1378 cm−1) could be used to detect differences in GAGs content of bone. To test this hypothesis, three experimental models were utilized: an in vitro model (enzymatic removal of GAGs from human cadaver bone), an in vivo mouse model (biglycan KO vs. WT), and an ex vivo aging model (comparing cadaveric bone samples from young and old donors). All Raman measurements were compared to Alcian blue measurements to confirm the validity of Raman spectroscopy in detecting GAGs changes in bone. Irrespective of different models, it was found that the ~1378cm−1 peak in Raman spectra of bone was uniquely sensitive to changes of GAGs content in bone when normalized with respect to the phosphate phase (~960cm−1); i.e., 1378cm−1/960cm−1 (peak intensity ratio) or 1370-1385cm−1/930-980cm−1 (integrated peak area ratio). In contrast, the 1070cm−1 peak, which includes another major peak of GAGs (1066cm−1), seemed to be compromised to detect changes of GAGs in bone due to concurrent changes of carbonate (CO3) in the similar peak range. This study validates the ability of Raman spectroscopy to detect in situ treatment-, genotype-, and age-related changes in GAG levels of bone matrix.
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影响因子：
6.2
作者：
Chen, XD;Shi, ST;Young, MF
通讯作者：
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影响因子：
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作者：
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