Raman spectroscopy demonstrates prolonged alteration of bone chemical composition following extremity localized irradiation.
Raman spectroscopy demonstrates prolonged alteration of bone chemical composition following extremity localized irradiation.
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DOI：
10.1016/j.bone.2013.08.014
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发表时间：
2013-11
期刊：
影响因子：
4.1
通讯作者：
Morris, Michael D.
中科院分区：
文献类型：
--
作者：
Gong, Bo;Oest, Megan E.;Mann, Kenneth A.;Damron, Timothy A.;Morris, Michael D.
对附肢骨骼进行放射治疗可能会导致发生灾难性骨折、骨愈合延迟或不愈合的风险增加，并且随后可能需要多次手术和截肢。生物力学研究表明，受辐照的骨骼更脆，但其原因尚不清楚，也不能用骨骼结构或数量的变化来解释，这表明受辐照的骨骼材料特性发生了重大变化。拉曼光谱提供了一种评估骨骼矿物质和基质成分化学性质的方法，这有助于解释辐射后脆化。在这项研究中，我们使用局部照射的小鼠胫骨模型并进行拉曼光谱来检验以下假设：照射后骨化学的变化与骨质量的降低一致，并且在照射后长期持续存在。 12周龄雌性BALB/F小鼠接受单侧局部后肢照射，每天4次，每次5Gy（4×5Gy），总计20Gy，并在照射后1、4、8、12和26周处死（n=6/组）。取出经过照射的（右）和未经照射的对侧对照（左）胫骨，并通过近端皮质骨表面的非偏振和偏振拉曼光谱进行评估。使用的拉曼参数包括矿物质/基质比率、矿物质结晶度、碳酸盐/磷酸盐比率、胶原交联比率和去极化比率。照射后 1 周，胶原交联率显着增加，基质去极化率明显降低，并且这种情况持续了 26 周。除第 8 周和第 26 周外，所有时间点的矿物质去极化率均观察到类似的显着下降。与对照组相比，照射后 4 周，矿物质/基质比率显着增加，矿物质结晶度增加，碳酸盐/磷酸盐比率降低。然而，在照射后 12 周，这些参数向相反方向移动，导致与对照相比，矿物质/基质比率显着降低，结晶度降低，碳酸盐/磷酸盐比率增加。第 26 周时，矿物质/基质、结晶度和碳酸盐/磷酸盐比率恢复正常。在该小鼠模型中，拉曼光谱报告了辐射引起的骨矿物质和胶原蛋白交联异常，这些异常最早在辐射后 1 周就显现出来，并持续 26 周。这张照片是一张严重受损的照片，之后有人试图进行重建。我们假设，由辐射损伤胶原蛋白形成的病理性交联在改变的重塑过程中吸收不良，因此在有缺陷的支架上形成新组织，导致骨脆性增加。
Radiotherapy to the appendicular skeleton can cause an increased risk of developing catastrophic fractures with delayed bone healing or non-union, and may subsequently require multiple procedures and amputation. Biomechanical studies suggest that irradiated bone is more brittle, but the cause is unclear and cannot be explained by changes to bone structure or quantity, suggesting that there are crucial changes in irradiated bone material properties. Raman spectroscopy provides a means to assess the chemical properties of the mineral and matrix constituents of bone, which could help explain post-radiation embrittlement. In this study we use a murine tibial model with focal irradiation and perform Raman spectroscopy to test the hypothesis that changes in bone chemistry following irradiation is consistent with reduced bone quality and persists in the long term after irradiation. Female BALB/F mice aged 12 weeks were subjected to unilateral, localized hindlimb irradiation in 4 daily 5 Gy fractions (4×5Gy) totaling 20 Gy, and were euthanized at 1, 4, 8, 12, and 26 weeks post-irradiation (n=6/group). The irradiated (right) and non-irradiated contralateral control (left) tibiae were explanted and assessed by non-polarized and polarized Raman spectroscopy over the proximal cortical bone surface. Raman parameters used included the mineral/matrix ratio, mineral crystallinity, carbonate/phosphate ratio, collagen cross-link ratio, and depolarization ratio. Significantly increased collagen cross-link ratio and decreased depolarization ratio of matrix were evident at 1 week after irradiation and this persisted through 26 weeks. A similar significant decrease was observed for depolarization ratio of mineral at all time points except 8 and 26 weeks. At 4 weeks after irradiation there was a significantly increased mineral/matrix ratio, increased mineral crystallinity, and decreased carbonate/phosphate ratio compared to controls. However, at 12 weeks after irradiation these parameters had moved in the opposite direction, resulting in a significantly decreased mineral/matrix ratio, decreased crystallinity and increased carbonate/phosphate ratio compared to controls. At 26 weeks, mineral/matrix, crystallinity and carbonate/phosphate ratios had returned to normal. In this mouse model, Raman spectroscopy reports both bone mineral and collagen cross-link radiation-induced abnormalities that are evident as early as one week after irradiation and persist for 26 weeks. The picture is one of extensive damage, after which there is an attempt at remodeling. We hypothesize that pathological cross-links formed by radiation damage to collagen are poorly resorbed during the altered remodeling process, so that new tissue is formed on a defective scaffold, resulting in increased bone brittleness.
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