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Application of Raman Micro-spectroscopic and Fourier transformed Infrared Micro-spectroscopic Techniques for Clinical Test.

拉曼显微光谱和傅里叶变换红外显微光谱技术在临床测试中的应用。

基本信息

批准号：
14370793
负责人：
MIYOSHI Norio
金额：
$ 7.36万
依托单位：
University of Fukui (2004-2005)福井医科大学 (2002-2003)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2005
项目状态：
已结题

项目摘要

(1) FT-IR microscopic imaging of frosen-sectioning experimental tumor tissue :Our purpose is to develop infrared (IR) microspectroscopy as a new optical diagnostic tool to support conventional lightscopic techniques in investigating the viability of carcinoma tissues and to develop its use in the evaluation of the early effects of anticancer therapy by monitoring the IR spectra in the necrotic area.We ammased for 4 weeks after the isotransplantation of mouse squamoouscell carcinoma into the thigh of mice. The borders of the necrotic area of frozen tissue specimens were investigated by Fourier-transform IR microspectroscopy and conventional histological staining.In the results, a significantly higher accumulation of cholesterol was observed in the necrotic tissue of a carcinoma. The mechanism of this phenomenon is hitherto unrecognized. We proposed that the accumulated cholesterol may lie extracellularly as a result of the ruptured plasma and internal membranes after the swelling of the … More necrotic cells brought on by hypoxia. The analysis of the secondary structure of protein revealed that the amount of β-sheet increased significantly in striking contrast to the decreasing amounts of α-helix in a necrotic area of a carcinoma. It is plausible that this structural conversion of protein was because of the liquid-autooxidation products, such as cholesterol oxide but not cholesterol itself, which processes cell toxicity and could be generated in a necrotic area.(2) Raman microscopic imaging of hard tissues (Calcified Cartilage) :The aim of the present study was to investigate the quantity and quality of hydroxyapatite (HA) in the zone of calcified cartilage of articular cartilage and underlying subchondral bone by Fourier transform infrared microscopy (FTIRM).The mineral:matrix ratio was determined by integrating areas of υ1, υ3 bands at 900-1200cm^<-1> to Amide I band at 1585-1725cm^<-1>. The CO_3:PO_4 ratio was determined by integrating areas of υ2 carbonate band at 850-900cm^<-1> to υ1. υ3 bands. In the control, the mean mineral:matrix ratio of the medial side was significantly lower than the lateral side at the zone of the calcified cartilage and subchondral bone. The mean CO_3:PO_4 ratio was significantly higher in the medial side than the lateral one at the zone of the calcified cartilage and subchondral bone. In contrast, the results obtained from the model showed no significant differences between the two sides. Our results showed diminished deposition and increased maturation of HA in control. These findings may correlate with the pathological process of knee OA, and indicate that FTIRM is a useful tool for determining the variability of HA in OA. Less

(1)实验肿瘤组织切片的FT-IR显微成像：我们的目的是开发红外(IR)显微光谱作为一种新的光学诊断工具，以支持传统的光学技术来研究肿瘤组织的生存能力，并通过监测坏死区的红外光谱来开发其在评估抗癌治疗早期效果方面的应用。用傅立叶变换红外光谱分析和常规组织学染色研究了冷冻组织标本坏死区的边界，结果发现，在肿瘤的坏死组织中，胆固醇积聚显著增加。到目前为止，人们还没有认识到这种现象的机制。我们推测，在…肿胀后，累积的胆固醇可能存在于细胞外，这是由于血浆和内膜破裂的结果。缺氧导致更多的坏死细胞。对蛋白质二级结构的分析表明，在肿瘤坏死区，β-折叠的数量明显增加，而α-螺旋的数量却减少了。(2)硬组织(钙化软骨)的拉曼显微成像：本研究的目的是用傅立叶变换红外光谱仪研究关节软骨钙化软骨区和软骨下骨中羟基磷灰石(HA)的数量和质量。通过积分υ1，υ3条带900-1200 cm；酰胺I带在1585-1725 cm^-1；CO_3：PO_4的比值是通过将υ_2碳酸盐谱带850-900 cm~(-1)的面积积分到υ_1、υ_3谱带的面积来确定的。对照组钙化软骨和软骨下骨带内侧的平均矿物质/基质比明显低于外侧。钙化软骨和软骨下骨带内侧的平均CO_3/PO_4值显著高于外侧。相比之下，从模型得到的结果显示双方没有显著差异。我们的结果显示对照组透明质酸沉积减少，成熟度增加。这些发现可能与膝骨性关节炎的病理过程有关，提示FTIRM是一种有用的工具，可以用来确定膝骨性关节炎患者透明质酸的变异性。较少