FTIR SPECTROSCOPY STUDY ON THE IMPACT OF LOW POWER NEAR-INFRARED LASER ON THE SECONDARY STRUCTURE OF HUMAN HEMOGLOBIN (IN-VITRO)

M. Bysam, A. Sedky, W. Elshemey, H. Mohammed
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Abstract

The low-level laser therapy was used in various biomedical applications for a long time such as improving the rheological properties of blood, wound healing, vascular restenosis, tissue repair. however, its safety has not been well investigated. The present in-vitro study aims to illustrate the influence of low power laser irradiation on the secondary structure of human Haemoglobin (Hb). Fifteen samples of healthy fresh blood have been divided into five groups (3 samples/group) and irradiated using low power infrared laser (wavelength 808 nm, power 4 mw and power density 0.64 mW/cm 2 ) at different irradiation durations (5, 10, 20, 30 min) and zero time for control samples. We used FTIR Spectroscopy followed by some mathematical techniques such as band-narrowing (second-derivative method), spectral subtraction, curve fitting, and integration processes to quantitatively calculate the different contribution ratios of the secondary structure components of irradiated and non-irradiated human hemoglobin samples. It was found no significant change in the contents of the α-helix structure at (5,10,20 min) groups. However, the thirty minutes group (30min) show a significant reduction in the content of α -helix accompanied by a significant increase in the random coil and β-turn structures with (p < 0.05). The decreasing of the alpha-helix structure is explained in terms of photo-inductive degradation to the unordered structure of random coils and β-turns. Extra care has to be taken while the clinical application that includes the intravenous laser irradiation of blood to avoid the protein photodamage caused by the low power laser at long expoture time.
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低功率近红外激光对人血红蛋白二级结构影响的Ftir光谱研究(体外)
长期以来,低水平激光治疗在改善血液流变学特性、伤口愈合、血管再狭窄、组织修复等生物医学领域有着广泛的应用。然而,它的安全性还没有得到很好的调查。本体外研究旨在阐明低功率激光照射对人血红蛋白二级结构的影响。将15份健康新鲜血液样本分为5组(3份/组),分别在不同照射时间(5、10、20、30 min)和对照样品的零照射时间下,采用低功率红外激光(波长808 nm,功率4 mw,功率密度0.64 mw / cm2)照射。采用FTIR光谱法,结合带窄(二阶导数法)、谱差法、曲线拟合和积分等数学方法,定量计算辐照和未辐照人血红蛋白样品二级结构组分的不同贡献比。α-螺旋结构的含量在(5、10、20 min)处理后无明显变化。30min组α -螺旋结构含量显著降低,随机螺旋结构和β-转结构显著增加(p < 0.05)。α -螺旋结构的减少可以用随机线圈和β-匝的无序结构的光诱导降解来解释。临床应用包括静脉内激光照射血液时,要特别注意避免低功率激光长时间照射造成的蛋白质光损伤。
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