Quantitative Biophotonics for Tissue Characterization and Function

用于组织表征和功能的定量生物光子学

• 批准号: 10007486
• 负责人: Amir H Gandjbakhche
• 金额: $ 75.31万
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10007486
• 关键词: Abdomen; Affinity; Algorithms; Anatomy; Animals; Anisotropy; Anterior; Asphyxia; Asses; Behavioral; Biological Phenomena; Biophotonics; Blood; Blood Vessels; Blood Volume; Cells; Cellular Metabolic Process; Cerebral Palsy; Cervical Ripening; Cervix Uteri; Cheek structure; Clinical; Clinical Endocrinology; Clinical Trials; Collaborations; Colposcopes; Computer Simulation; Computer software; Contralateral; Contrast Media; Country; Cushing Syndrome; Data; Data Analyses; Dermis; Development; Devices; Diagnosis; Diagnostic Procedure; Diffuse; Disease remission; Early Diagnosis; Early Intervention; Elements; Endocrinology; Ensure; Environment; Epidermal Growth Factor; Epidermis; Extracellular Matrix; Face; Fatty acid glycerol esters; Fetal Growth Retardation; Fetal health; Florida; Fluorescence; Fourier Analysis; Geometry; Goals; Hand; Hela Cells; Hemoglobin; Human; Hydrocortisone; Hypoxia; Image; Image Analysis; In Vitro; International; Journals; Kaposi Sarcoma; Knowledge; Label; Lasers; Lesion; Light; Link; Lipids; Location; Longitudinal Studies; Malignant - descriptor; Malignant Neoplasms; Maternal Health; Maternal-fetal medicine; Measurement; Measures; Melanins; Membrane; Methodology; Methods; Modeling; Monitor; Monte Carlo Method; Morbidity - disease rate; National Institute of Child Health and Human Development; Near-Infrared Spectroscopy; Normal tissue morphology; Operative Surgical Procedures; Optical Coherence Tomography; Optics; Oxygen; Oxyhemoglobin; Patients; Perinatology; Pharmacotherapy; Physiological; Pilot Projects; Pituitary-dependent Cushing' s disease; Placenta; Plasma; Point-of-Care Systems; Population Control; Postoperative Period; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Pregnancy Outcome; Premature Birth; Prevalence; Property; Publishing; Research; Research Personnel; Resolution; Scientist; Signal Transduction; Site; Skin; Source; Spectrum Analysis; Standardization; Structure; System; Techniques; Testing; Thermography; Thick; Third Pregnancy Trimester; Time; Tissue imaging; Tissues; Transillumination; Treatment Protocols; Treatment outcome; Ultrasonography; United States National Institutes of Health; Universities; Uterus; Variant; Walking; Water; Western Blotting; Wireless Technology; absorption; attenuation; base; bench to bedside; chromophore; cobaltous chloride; comparative; computerized data processing; density; deoxyhemoglobin; design; detector; early detection biomarkers; experimental study; fetus hypoxia; flexibility; fluorescence lifetime imaging; follow-up; graphical user interface; healthy pregnancy; high risk; hypoxia inducible factor 1; imaging system; improved; in vivo; indexing; instrumentation; interest; light intensity; light scattering; light weight; novel; novel diagnostics; pressure; prototype; receptor; response; spectrograph; spectroscopic imaging; theories; tomography; tool; treatment strategy; tumor; user-friendly

Monitoring the placental oxygenation is critical to ensure a healthy pregnancy outcome. Abnormalities in placental oxygenation has been associated with preeclampsia and intrauterine growth restriction, fetal hypoxia, asphyxia, and cerebral palsy. Quantifying placental oxygenation can help early detection of such complications. This project is driven by the lack of patient-friendly devices to measure the oxygenation of the anterior placenta. A wearable, wireless system that can monitor the placental oxygenation dynamically are realized using Functional Near Infrared Spectroscopy (fNIRS). In parallel to the in-vivo studies, we are also interested in investigating placenta at cellular level to control the oxygen levels and understand its effect on cell metabolism. We intend to 1) find the baseline for the normal pregnancies and standardize the oxygenation data across pregnancies, 2) correlate the oxygenation data with the pregnancy outcomes and 3) study placental cell metabolism in-vitro at physiologically relevant variations of oxygen level using a novel biophotonic methodology, Dynamic Full Field Optical Coherence Tomography (DFFOCT). For the in-vivo study, we designed a wearable, wireless, non-invasive and inexpensive device to measure the anterior placental oxygenation in a subject-friendly environment. This prototype is light-weight and compact that can be placed at different abdominal locations for efficient and localized measurement of oxygenation. The NIRS device uses near infrared light with two different wavelengths of 760 nm and 840 nm that are sensitive to changes in oxy-hemoglobin and deoxy-hemoglobin. Our system consists of two light detectors and six LED sources to probe different tissue depth and distinguish the oxygenation between maternal layers and placental tissue. Flexible geometry of the optical probe of the device ensures effective optical contact with the skin without exerting excess pressure. The efficiency of the device was examined for different melanin volume fractions, fat thicknesses and uterus thicknesses to separate the placental oxygenation from the maternal layers contamination. Since the light passes through several tissue compartments, we have developed the multi-layer model based on Monte Carlo simulation that includes optical properties (scattering and absorption) of skin (dermis and epidermis), fat, uterus and placental tissue. For precise calculation of the oxygenation, prior knowledge of the optical properties of the given tissues such as scattering (how much tissue scatters the light) and absorption (how well tissue absorbs the light) is needed. We use a dual wavelength LED source with photodiode array unit built in-house to find the optical properties of the placenta. The attenuation coefficient (as a function of scattering and absorption coefficient) is calculated based on the reflection curve (light intensity as a function of source-detector distance) from placental tissues with and without blood. Monte Carlo simulations along with our multi-layer model helped us build a system that considers skin melanin content, fat and uterus thickness into the calculation of oxygenation index. In collaboration with Maternal-Fetal Medicine, Imaging, and Behavioral Development Affinity Group (Dr. Roberto Romero) at NICHD, Wayne State University, and USUHS we have the opportunity to test our device through pilot studies. We have started our first pilot study with Wayne State University, where we measure the oxygenation of the placenta during the last trimester in normal pregnancies to find a baseline of placental oxygenation. Meanwhile, we refine our data analysis software by incorporating anatomical data from subjects. Ultra-sound imaging gives us the fat and uterus thicknesses we need for the analysis. We hope to detect pregnancy complications in their earlier stages to improve both maternal and fetal health. Our initial in-vitro experiments on HeLa cells confirmed the ability of DFFOCT to detect the changes in intra-cellular activity as a function of ambient oxygenation. We use our segmentation algorithm to identify the DFFOCT signal from the cells grown on porous membrane and use Fourier analysis to understand the dynamic activity occurring within the cells. We compared the signal from cells grown under hypoxia and physoxia (physiological oxygenation) and observed unique differences in the Fourier spectra at these conditions. Control experiments with Cobalt chloride were used to mimic hypoxia. We also ran Western blot to ensure the effects of hypoxia on expression of Hif-1 protein in the cells. We plan to do further experiments to understand the link between oxygenation and the DFFOCT signal. This will help us in using DFFOCT as a tool to asses activity of placental cells under altered oxygenation. Facial plethora is one of the earliest described clinical features of Cushings syndrome (CS). In collaboration with SEG at NICHD, we have quantified changes of facial plethora in CS as an early assessment of cure. Non-invasive multi-spectral near-infrared imaging was performed on the right cheek of the patients before and after surgery. Patients were defined as cured by post-operative measurements of plasma cortisol less than 3 (mcg/dl), and/or adrenocortical insufficiency for which they received replacement. Results indicate that a decrease in facial plethora after surgery, as evidenced by decrease in blood volume fraction, is correlated with cure of CS. The first set of results were published in the journal of clinical endocrinology & metabolism. We also showed that water content fraction could be used as a new biomarker of early cure in patients with CS. We ran our methodology for first (3-6 months after surgery, N=22) and second (6 months after the first follow up, N=10) post-surgery follow up, where all subjects are identified clinically in remission state. As a next step for our bench to bedside goal, we have developed a new hand-held multispectral camera to be used a point-of-care system. The device uses a high-resolution CMOS camera with on-chip filters. Images with resolution of 256X256 pixels are acquired simultaneously at eight different near-infrared wavelengths (700-980 nm). We have also developed a user-friendly graphical interface for data processing in Matlab. Assessment of tumor development in patients can facilitate treatment strategies and early intervention. In our recent published study, we designed time-resolved fluorescence lifetime imaging to distinguish bound Human Epidermal Growth Factor 2 (HER2) specific affibody probes to HER2 receptors in live animals. Our results show that changes in fluorescence lifetime of the bound contrast agent can be used to rapidly assess the high to mid-level expression of HER2 expressing tumors in-vivo. In another study, we aimed to use the PReterm IMaging system based on colposcope to characterize uterine cervix structure in a longitudinal study of low- and high-risk (prior preterm birth (PTB) or a sonographic short cervix) patients. Polarization imaging is an effective tool to measure optical anisotropy in birefringent materials, such as the cervix's extracellular matrix and to predict cervical ripening and potentially to diagnose pre-term birth. We developed a handheld colposcope device for active polarization imaging of the cervix. Through our under-review collaboration with Wayne State Universitys Perinatology Research Branch and Florida International University we will test our system in a control population and those with PTB prevalence.

监测胎盘氧合对于确保健康的妊娠结局至关重要。胎盘氧合的异常与子痫前期和子宫内生长限制，胎儿缺氧，窒息和脑瘫有关。量化胎盘氧合可以有助于尽早发现此类并发症。该项目是由于缺乏患者友好型设备来驱动的，无法测量前胎盘的氧合。可穿戴的无线系统，可以使用功能性近红外光谱（FNIRS）来动态监测胎盘氧合。与体内研究并行，我们也有兴趣研究细胞水平的胎盘以控制氧气水平并了解其对细胞代谢的影响。我们打算1）找到正常妊娠的基线，并标准化妊娠的氧合数据，2）将氧合数据与妊娠结局相关联和3）3）3）研究胎盘细胞代谢在生理上相关的氧气变化的胎盘细胞代谢，使用新型的生物体性方法，使用新型的生物体性方法学，动态完整现场辅导（动态完整的均匀绘画）（Dffffoct）（Dffffoct）。 在体内研究中，我们设计了一种可穿戴，无线，无创和廉价的设备，以测量受试者友好环境中的前胎盘氧合。该原型是轻巧和紧凑的，可以放置在不同的腹部位置，以进行有效和局部的氧合测量。 NIRS设备在红外光中使用了两个不同的波长为760 nm和840 nm的波长，这些波长对氧气血红蛋白和脱氧 - 血红蛋白的变化敏感。我们的系统由两个光检测器和六个LED来源组成，可探测不同的组织深度，并区分孕妇层和胎盘组织之间的氧合。设备的光学探针的柔性几何形状可确保与皮肤的有效光接触，而不会施加过多的压力。检查了设备的效率不同的黑色素体积分数，脂肪厚度和子宫厚度，以将胎盘氧合与母体层污染分开。由于光经过了几个组织室，因此我们基于蒙特卡洛模拟开发了多层模型，其中包括皮肤（真皮和表皮），脂肪，子宫和胎盘组织的光学特性（散射和吸收）。为了精确计算氧合，对给定组织的光学特性（例如散射（散射多少组织散射）和吸收（组织吸收光线充足）的先验知识。我们使用带有光电二极管阵列单元内置的双波长LED源来查找胎盘的光学特性。根据反射曲线（光强度作为源 - 探测器距离的函数）计算衰减系数（作为散射和吸收系数的函数）是从有或没有血液的胎盘组织中计算出来的。蒙特卡洛模拟以及我们的多层模型帮助我们建立了一个将皮肤黑色素含量，脂肪和子宫厚度考虑到氧合指数计算中的系统。 在NICHD，Wayne State University和USUHS的母亲 - 狂热医学，成像和行为发展亲和力小组（Roberto Romero博士）中，我们有机会通过试点研究来测试我们的设备。我们已经在韦恩州立大学开始了第一项试点研究，在正常怀孕的最后三个月期间，我们测量了胎盘的氧合，以找到胎盘氧合的基线。同时，我们通过合并受试者的解剖学数据来完善数据分析软件。超声成像为我们提供了分析所需的脂肪和子宫厚度。我们希望在早期阶段发现妊娠并发症，以改善孕产妇和胎儿健康。 我们在HeLa细胞上进行的初始体外实验证实了DFFOCT检测细胞内活性随环境氧合的函数的能力。我们使用分割算法从多孔膜上生长的细胞中识别DFFOCT信号，并使用傅立叶分析来了解细胞内发生的动态活性。我们比较了在缺氧和植物学下生长的细胞的信号（生理氧合），并观察到在这些条件下傅立叶光谱中的独特差异。用氯化钴进行的对照实验用于模仿缺氧。我们还运行了蛋白质印迹，以确保缺氧对细胞中HIF-1蛋白表达的影响。我们计划进行进一步的实验，以了解氧合和DFFOCT信号之间的联系。这将有助于我们将DFFOCT用作在氧合改变下胎盘细胞活性的工具。 面部太多是库欣综合征（CS）的最早描述的临床特征之一。在与NICHD的SEG合作，我们将CS中面部过多的变化量化为治疗的早期评估。手术前后，在患者的右脸颊上进行了非侵入性多光谱近红外成像。患者被定义为通过小于3（MCG/DL）的血浆皮质醇的术后测量和/或肾上腺皮质不足替代的疗法。结果表明，手术后面部过多的降低，如血容量分数的降低所证明，与CS的治疗相关。第一组结果发表在《临床内分泌与代谢杂志》杂志上。我们还表明，在CS患者中，水含量分数可以用作早期治疗的新生物标志物。我们运行了第一次（手术后3-6个月，n = 22）和第二个随访后的第二个（n = 10之后的6个月）进行手术后的方法，该方法在手术后随访中进行了随访，其中所有受试者均以缓解状态识别。作为我们的长凳到床边目标的下一步，我们开发了一个新的手持式多光谱摄像头，以被使用一个护理点系统。该设备使用带有芯片过滤器的高分辨率CMOS摄像头。分辨率为256x256像素的图像以八个不同的近红外波长（700-980 nm）同时获取。我们还开发了一个用户友好的图形接口，用于MATLAB中的数据处理。 评估患者的肿瘤发展可以促进治疗策略和早期干预。在我们最近发表的研究中，我们设计了时间分辨的荧光寿命成像，以区分生物动物中HER2受体的结合人类表皮生长因子2（HER2）特定的杂物探针。我们的结果表明，结合对比剂的荧光寿命的变化可用于快速评估体体表达HER2表达肿瘤的高度至中级表达。 在另一项研究中，我们旨在使用基于阴道镜的早产系统来表征子宫子宫颈结构，这是对低风险和高风险（先前的早产（PTB）或超声范围）患者的纵向研究。极化成像是测量双重材料（例如子宫颈外基质基质）并预测子宫颈成熟并有可能诊断前末期出生的有效工具。我们开发了一种手持式阴道镜设备，用于子宫颈的主动极化成像。通过与韦恩州立大学围产科研究部和佛罗里达国际大学的审视不足合作，我们将在控制人群中测试我们的系统以及PTB患病率的人。