<br>用拉曼光谱评估人类皮质骨糖化介导的变化,Journal of Biophotonics

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用拉曼光谱评估人类皮质骨糖化介导的变化
Journal of Biophotonics ( IF 2.0 ) Pub Date : 2018-05-06 , DOI: 10.1002/jbio.201700352
Mustafa Unal _{1,

2,

3} , Sasidhar Uppuganti _{1,

3} , Calen J Leverant ₄ , Amy Creecy _{1,

3,

5} , Mathilde Granke _{1,

3} , Paul Voziyan ₆ , Jeffry S Nyman _{1,

2,

3,

5}

Affiliation

建立一种无损方法来空间评估晚期糖基化终产物（AGE）是研究 AGE 在骨质量中的机制作用的潜在有用步骤。为了检验骨基质拉曼光谱 (RS) 分析中酰胺 I 的形状随 AGE 积累而变化的假设，我们将配对的尸体皮质骨分别在核糖或葡萄糖溶液和对照溶液中孵育 4 周和 16 周， 37°C。 RS 使用 20X 物镜和 830 nm 激光采集每根骨头 10 个光谱，对 AGE 积累敏感（通过戊糖苷和荧光 AGE 的生化测量证实）。使用 0.1 M 核糖、0.5 M 核糖或 0.5 M 葡萄糖进行糖化时，Hyp/Pro 比率增加。当使用二阶导数光谱或差异光谱的局部最大值直接计算时，糖化还降低了酰胺 I 子峰比率 (cm ^-1 ) 1668/1638 和 1668/1610，尽管处理方法（例如，平均光谱与单个光谱）最大限度地减少噪音影响核糖孵化骨骼差异的检测。然而，当使用间接子带拟合计算时，糖化并不影响这些子峰比率，包括基质成熟度比率（1668/1690）。酰胺 I 亚峰比率可能反映了胶原蛋白 I 结构的变化。

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Assessing glycation‐mediated changes in human cortical bone with Raman spectroscopy

Establishing a non‐destructive method for spatially assessing advanced glycation end‐products (AGEs) is a potentially useful step toward investigating the mechanistic role of AGEs in bone quality. To test the hypothesis that the shape of the amide I in the Raman spectroscopy (RS) analysis of bone matrix changes upon AGE accumulation, we incubated paired cadaveric cortical bone in ribose or glucose solutions and in control solutions for 4 and 16 weeks, respectively, at 37°C. Acquiring 10 spectra per bone with a 20X objective and a 830 nm laser, RS was sensitive to AGE accumulation (confirmed by biochemical measurements of pentosidine and fluorescent AGEs). Hyp/Pro ratio increased upon glycation using either 0.1 M ribose, 0.5 M ribose or 0.5 M glucose. Glycation also decreased the amide I sub‐peak ratios (cm⁻¹) 1668/1638 and 1668/1610 when directly calculated using either second derivative spectrum or local maxima of difference spectrum, though the processing method (eg, averaged spectrum vs individual spectra) to minimize noise influenced detection of differences for the ribose‐incubated bones. Glycation however did not affect these sub‐peak ratios including the matrix maturity ratio (1668/1690) when calculated using indirect sub‐band fitting. The amide I sub‐peak ratios likely reflected changes in the collagen I structure.

更新日期：2018-05-06

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