光学相干层析术高分辨率活体成像泼尼松龙诱导的骨质疏松斑马鱼模型
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摘要
本研究利用光学相干层析术OCT对泼尼松龙诱导的斑马鱼骨质疏松模型进行活体成像,并结合电镜能谱技术定量分析斑马鱼模型骨质的钙磷元素含量及分布情况,共同探讨OCT方法在基于斑马鱼模型开展的骨质疏松研究中的使用价值。选取40条3月龄野生型斑马鱼暴露于50μmol/L泼尼松龙溶液和含0. 5%DMSO的溶液中(对照组),28. 5℃下培养,分别于第5、10、20天取出浸药组和对照组进行OCT活体成像,比较两者光散射特征。在每个时间点的成像之后,将浸药组的5条斑马鱼处死,然后取颅骨进行元素含量电镜扫描能谱分析。本研究利用50μmol/L泼尼松龙溶液培养斑马鱼至第20天,成功构建了斑马鱼骨质疏松模型。与对照组相比,模型组活体OCT成像显示骨组织光散射减弱,光子量明显减少,呈不均匀分布。能谱元素检查结果说明颅骨内所含钙、磷比例明显下降,证实骨质疏松发生,骨量减少。OCT成像方法在对斑马鱼骨质疏松模型进行活体、实时、无创等研究方面具有重要价值,本试验也为骨质疏松疾病的研究和药物筛选等方面提供了新的有效的方法。
This study aim to explore the application value of OCT( optical coherence tomography) in the study of osteoporosis model of zebrafish. In vivo evaluation of prednisolone-induced osteoporosis of the adult zebrafish was conducted with optical coherence tomography,and the components of the model's bone were measured by scanning electron microscopy. A total of 40 adult wild type zebrafish( 90 day) were selected for this study. Out of 40 fish,30 were exposed to50 μmol/L prednisolone solution,and 10 were set up with 0. 5% DMSO medium( ascertained as controls). The zebrafish were kept in our flowthrough aquaria at a temperature of 28 ℃. The imaging of the bone structure using high resolution optical coherence tomography were performed at days 0,7,14 or 21 after the prednisolone were induced,comparing their light scattering characteristics. At each time point after OCT imaging experiment,the skull of zebrafish was collected for elemental content analysis by electron microscopy. The osteoporosis model using zebrafish induced by 50μmol/L prednisolone solution was successfully developed for 20 days. Compared with the control group,the living OCT imaging in the model group showed that the light scattering of the skull tissue was weakened and the photon quantity was significantly reduced,showing an uneven distribution. The results of energy spectrum element examination demonstrated that,the proportion of calcium and phosphorus in bone decreased significantly,indicating osteoporosis and bone loss.The results demonstrated that OCT is an excellent technology in evaluating in the study of the osteoporosis model of zebrafish in vivo,real-time and non-invasive. This experiment also provides a new and effective method for the study of osteoporosis disease and drug discovey.
This study aim to explore the application value of OCT( optical coherence tomography) in the study of osteoporosis model of zebrafish. In vivo evaluation of prednisolone-induced osteoporosis of the adult zebrafish was conducted with optical coherence tomography,and the components of the model's bone were measured by scanning electron microscopy. A total of 40 adult wild type zebrafish( 90 day) were selected for this study. Out of 40 fish,30 were exposed to50 μmol/L prednisolone solution,and 10 were set up with 0. 5% DMSO medium( ascertained as controls). The zebrafish were kept in our flowthrough aquaria at a temperature of 28 ℃. The imaging of the bone structure using high resolution optical coherence tomography were performed at days 0,7,14 or 21 after the prednisolone were induced,comparing their light scattering characteristics. At each time point after OCT imaging experiment,the skull of zebrafish was collected for elemental content analysis by electron microscopy. The osteoporosis model using zebrafish induced by 50μmol/L prednisolone solution was successfully developed for 20 days. Compared with the control group,the living OCT imaging in the model group showed that the light scattering of the skull tissue was weakened and the photon quantity was significantly reduced,showing an uneven distribution. The results of energy spectrum element examination demonstrated that,the proportion of calcium and phosphorus in bone decreased significantly,indicating osteoporosis and bone loss.The results demonstrated that OCT is an excellent technology in evaluating in the study of the osteoporosis model of zebrafish in vivo,real-time and non-invasive. This experiment also provides a new and effective method for the study of osteoporosis disease and drug discovey.
引文
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[15]JIA Y,BAILEY S T,HWANG T S,et al.Quantitative optical coherence tomography angiography of vascular abnormalities in the living human eye[J].Proceedings of the National Academy of Sciences,2015,112(18):E2395-E2402.
[16]ZHANG J,GE W,YUAN Z.In vivo three-dimensional characterization of the adult zebrafish brain using a 1325 nm spectraldomain optical coherence tomography system with the 27 frame/s video rate[J].Biomedical Optics Express,2015,6(10):3932-3940.
[17]ZHANG J,ZHANG Z,GE W,et al.Long-term in vivo monitoring of injury induced brain regeneration of the adult zebrafish by using spectral domain optical coherence tomography[J].Chinese Optics Letters,2016,14(8):081702-081705.
[2]SICCARDI A J R,PADGETT-VASQUEZ S,GARRIS H W,et al.Dietary strontium increases bone mineral density in intact zebrafish(Danio rerio):a potential model system for bone research[J].Zebrafish,2010,7(3):267-273.
[3]DE VRIEZE E,VAN KESSEL M,PETERS H M,et al.Prednisolone induces osteoporosis-like phenotype in regenerating zebrafish scales[J].Osteoporosis International,2014,25(2):567-578.
[4]SANTORIELLO C,ZON L I.Hooked!Modeling human disease in zebrafish[J].The Journal of Clinical Investigation,2012,122(7):2337-2343.
[5]STWWART A M,BRAUBACH O,SPITSBERGEN J,et al.Zebrafish models for translational neuroscience research:from tank to bedside[J].Trends in Neurosciences,2014,37(5):264-278.
[6]BANDMANN O,BURTON E A.Genetic zebrafish models of neurodegenerative diseases[J].Neurobiology of Disease,2010,40(1):58-65.
[7]GERURTZEN K,VERNET A,FREIDIN A,et al.Immune suppressive and bone inhibitory effects of prednisolone in growing and regenerating zebrafish tissues[J].Journal of Bone and Mineral Research,2017,32(12):2476-2488.
[8]景莉君,王长梅,韦英杰,等.接骨汤抗泼尼松龙诱导斑马鱼骨丢失的作用研究[J].世界科学技术-中医药现代化,2014,16(5):1071-1075.JING Lijun,WANG Changmei,WEI Yingjie,et al.Effect of jiegu decoction on prednisolone-induced zebrafish bone loss[J].Modernization of Traditional Chinese Medicine and Materia Materia-World Science and Technology,2014,16(5):1071-1075.
[9]杨国柱,段涛,卢丽,等.桃红四物汤对泼尼松诱导斑马鱼骨质疏松的治疗作用[J].中国骨质疏松杂志,2017,23(8):1007-1010.YANG Guozhu,DUAN Tao,LU Li,et al.Therapeutic effect of Taohong Siwu Decoction on prednisone induced osteoporosis on zebrafish[J].Chinese Journal of Osteoporosis,2017,23(8):1007-1021.
[10]韦英杰,王长梅,蔡雪婷,等.地塞米松影响骨骼发育的斑马鱼模型的建立[J].药学学报,2013,48(2):255-260.WEI Yingjie,WANG Changmei,CAI Xueting,et al.Establishment of zebrafish osteopenia model induced by dexamethasone[J].Acta Pharmacica Sinica,2013,48(2):255-260.
[11]VAKOC B J,FUKUMURA D,JAIN R K,et al.Cancer imaging by optical coherence tomography:perclinical progress and clinical potential[J].Nature Reviews Cancer,2012,12(5):363-368.
[12]姜宇,姚建铨,王瑞康,等.光学相干层析系统的建立与研究[J].光学仪器,2003,25(2):33-37.JIANG Yu,YAO Jianquan,WANG Ruikang,et al.Establishment and research of optical coherence tomography[J].Optical Instruments,2003,25(2):33-37.
[13]李伟军,方玉宏,董葵,等.去角质化的光学相干成像检测[J].激光生物学报,2017,26(6):508-511.LI Weijun,FANG Yuhong,DONG Kui,et al.Optical coherence tomography imaging of exfoliation[J].Acta Laser Biology Sinica,2017,26(6):508-511.
[14]CHAMIE D,WANG Z,BEZERRA H,et al.Optical coherence tomography and fibrous cap characterization[J].Current Cardiovascular Imaging Reports,2011,4(4):276-283.
[15]JIA Y,BAILEY S T,HWANG T S,et al.Quantitative optical coherence tomography angiography of vascular abnormalities in the living human eye[J].Proceedings of the National Academy of Sciences,2015,112(18):E2395-E2402.
[16]ZHANG J,GE W,YUAN Z.In vivo three-dimensional characterization of the adult zebrafish brain using a 1325 nm spectraldomain optical coherence tomography system with the 27 frame/s video rate[J].Biomedical Optics Express,2015,6(10):3932-3940.
[17]ZHANG J,ZHANG Z,GE W,et al.Long-term in vivo monitoring of injury induced brain regeneration of the adult zebrafish by using spectral domain optical coherence tomography[J].Chinese Optics Letters,2016,14(8):081702-081705.