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首页> 外文期刊>European Biophysics Journal >Investigation of changes in optical attenuation of bone and neuronal cells in organ culture or three-dimensional constructs in vitro with optical coherence tomography: relevance to cytochrome oxidase monitoring
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Investigation of changes in optical attenuation of bone and neuronal cells in organ culture or three-dimensional constructs in vitro with optical coherence tomography: relevance to cytochrome oxidase monitoring

机译：光学相干层析成像技术研究器官培养或三维构建物中骨骼和神经元细胞的光衰减变化：与细胞色素氧化酶监测的相关性

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摘要

Changes in optical attenuation, relevant to cytochrome oxidase, of the rat bone periosteal tissue in explanted culture and human neuronal cells in three-dimensional agarose constructs have been monitored by the use of optical coherence tomography (OCT), with potential applications in tissue engineering and diagnosis. A superluminescent diode (SLD) with a peak emission wavelength (λ=820 nm) that is the near-infrared absorption band of the oxidized form of CytOx was employed. The attenuation coefficient was obtained from the depth-resolved reflectance profiles of liquid phantoms (naphthol green B with intralipid), explant culture (periosteum of calvaria from rats) and cells in 3D agarose constructs. The absorption coefficient of naphthol green B can be accurately quantified by the linear relationship between attenuation coefficients and the concentration. The difference in the attenuation coefficient of astrocytoma cells in agarose before and after reduction of CytOx is 0.26±0.10 mm−1 (n=9), whereas no attenuation is observed with the agarose control. Reduction of the enzyme in periosteal tissue leads to a change in attenuation coefficient of 0.43±0.24 mm−1 (n=7). For comparison, using a biochemical assay, the absorption coefficient of the oxidized-reduced form of CytOx is measured at approximately 8.3±1.5×10−3 mm−1 (n=4) and 8.7±2.5×10−3 mm−1 (n=4) at 820 nm for astrocytoma cells and rat periosteum, respectively. The lower value of CytOx concentration using biochemical versus OCT measurements may result from shifts in the scattering profile and the amplifying influences of multiple heme-based oxidases, indicating that conventional OCT is not specific enough to monitor redox changes in cytochrome oxidase. However, qualitative shifts in oxidation state are apparent using the technique. Our results suggest the potential application of OCT in providing high-resolution tomographic imaging of tissues in organ culture and cells grown in three-dimensional constructs in vitro.
机译：通过使用光学相干断层扫描（OCT）监测了外植培养物中大鼠骨骨膜组织和三维琼脂糖构建物中人神经元细胞的与细胞色素氧化酶相关的光衰减变化，其在组织工程和生物工程中的潜在应用诊断。使用具有作为氧化形式的CytOx的近红外吸收带的峰值发射波长（λ＝ 820nm）的超发光二极管（SLD）。衰减系数是从液体体模（带有脂质内的萘酚绿B），外植体培养物（来自大鼠的颅骨骨膜）和3D琼脂糖构建体中的细胞的深度分辨反射曲线中获得的。萘酚绿B的吸收系数可以通过衰减系数与浓度之间的线性关系来准确地定量。 CytOx还原前后琼脂糖中星形细胞瘤细胞的衰减系数差异为0.26±0.10 mm-1 （n = 9），而在琼脂糖对照组中未观察到衰减。骨膜组织中酶的减少导致衰减系数的变化为0.43±0.24 mm-1 （n = 7）。为了进行比较，使用生化分析，测得的CytOx氧化还原形式的吸收系数约为8.3±1.5×10-3 mm-1 （n = 4）和8.7±2.5星形胶质细胞瘤细胞和大鼠骨膜分别在820 nm处×10−3 mm-1 （n = 4）。使用生化测量与OCT测量得出的CytOx浓度值较低，可能是由于散射曲线的变化以及多种基于血红素的氧化酶的放大影响所致，这表明常规的OCT不够特异性，无法监测细胞色素氧化酶的氧化还原变化。但是，使用该技术可以明显看出氧化态的质变。我们的结果表明，OCT在为器官培养物中的组织和体外三维构建体中生长的细胞提供高分辨率层析成像方面的潜在应用。

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  • 来源
    《European Biophysics Journal》 |2003年第4期|355-362|共8页
  • 作者

    Xiangqun Xu; Ruikang K Wang; Alicia El Haj;

  • 作者单位

    Centre for Science Technology in Medicine School of Medicine Keele University/North Staffordshire Hospital Trust;

    Centre for Science Technology in Medicine School of Medicine Keele University/North Staffordshire Hospital TrustInstitute of Bioscience and Technology Cranfield University;

    Centre for Science Technology in Medicine School of Medicine Keele University/North Staffordshire Hospital Trust;

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  • 正文语种 eng
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  • 关键词

    Absorption; Cytochrome oxidase; Optical coherence tomography; Scattering; Tissue and cells;

    机译：吸收;细胞色素氧化酶;光学相干层析成像;散射;组织和细胞;

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