Safety assessments of tissue irradiation with near infrared fs pulses for multi-photon imaging applications

用于多光子成像应用的近红外飞秒脉冲组织照射的安全评估

基本信息

批准号：
10363397
负责人：
IRENE GEORGAKOUDI
金额：
$ 7.8万
依托单位：
TUFTS UNIVERSITY MEDFORD
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10363397
关键词：
3-Dimensional Address Adoption Animals Apoptosis Architecture Area Biopsy Brain Cardiovascular Diseases Cells Cervical Cervix Uteri Characteristics Contrast Media DNA Damage Databases Dependence Development Diagnosis Diagnostic Disease Dose Early Diagnosis Engineering Ensure Epithelial Evaluation Eye Fibrosis Freezing Genomics Goals Hamsters Health Histology Human Human Engineering Image Immunofluorescence Immunologic In Vitro Label Lasers Light Lighting Malignant Neoplasms Medical center Metabolic Methods Modality Modeling Monitor Morphology Motivation Nature Obesity Optics Oral Organ Photons Physiologic pulse Proteomics Protocols documentation Reporting Reproducibility Resolution Safety Sampling Skin Source Specimen Spottings Squamous Epithelium Stains System Technology Testing Tissue Sample Tissue imaging Tissues Translations Vagina accurate diagnosis base clinical translation comparative density experimental study exposed human population human imaging human tissue imaging approach imaging modality imaging system immune function improved in vivo in vivo imaging insight interest irradiation keratinocyte monolayer multiphoton imaging novel novel therapeutics oral cavity epithelium patient safety safety assessment therapy development translational barrier treatment response wound healing

项目摘要

Multiphoton imaging is emerging as a modality with transformative potential for human diagnostic applications, because it can provide morphological and functional tissue information with micron scale resolution, while obviating the need for a biopsy and exogenous contrast agents. However, these imaging systems rely on novel lasers that deliver light to tissue in the form of highly intense, ultrashort pulses, whose interactions with tissues other than the eye and skin are not well-characterized. The lack of established databases that define maximal permissible exposure (MPE) thresholds and a set of methods and protocols that should be used to demonstrate patient safety for a specific set of irradiation conditions presents a critical barrier to the clinical translation of these imaging methods. Our goal is to establish MPE thresholds for NIR, fs pulsed illumination of non-keratinized human squamous oral and cervical epithelia. We will rely initially on engineered human oral and vaginal- ectocervical epithelial tissues that are highly reproducible and commercially available, yet mimic important morphological and functional human tissue characteristics. Using these tissues, we will examine the impact of illumination wavelength, pulse duration, repetition rate, average power, photon density, and total light dose on tissue morphology assessed via histology and proliferation, DNA damage, and apoptosis examined via immunofluorescence. We will also quantify optical metabolic activity using label-free images acquired during tissue illumination with the parameters of interest. We will identify a subset of illumination parameters that yield no, minimal, or moderate damage and we will test these conditions on human cervical tissues (freshly excised and frozen/thawed), human keratinocyte monolayers, and hamster oral epithelia (in vivo, freshly excised and frozen/thawed). Thus, we expect to identify a protocol for the types of specimens that are likely to yield damage assessments that are most relevant to in vivo human squamous epithelial tissues and the combination of a range of light delivery parameters that are not expected to result in any functional damage. We also anticipate that label-free multi-photon imaging-based characterization of tissue morphology and metabolic function will serve as an independent highly sensitive tissue damage indicator. Our findings will be directly relevant to the safe implementation of multi-photon imaging of the human cervix in vivo; however, the approach and MPE conditions we establish will be straightforward to extend to other tissue types and enable translation of this powerful modality to in vivo human imaging.

多光子成像正在成为一种具有人类诊断应用的变革潜力的方式，因为它可以以微米尺度分辨率提供形态学和功能性组织信息，而消除了需要进行活检和外源性对比剂的需求。但是，这些成像系统依赖于新颖激光器以高度强烈的超短脉冲的形式向组织传递光，其与组织的相互作用除了眼睛和皮肤外，没有很好的表征。缺乏定义最大的既定数据库允许的暴露（MPE）阈值以及一组应用于证明的方法和协议特定辐射条件集的患者安全构成了这些临床翻译的关键障碍成像方法。我们的目标是为NIR建立MPE阈值人类鳞状口腔和宫颈上皮。我们最初将依靠设计的人类口腔和阴道 - 高度可重现且可商购的外宫皮上皮组织，但很重要形态学和功能性人体组织特征。使用这些组织，我们将检查照明波长，脉搏持续时间，重复速率，平均功率，光子密度和总光剂量通过组织学和增殖，DNA损伤和凋亡评估的组织形态学免疫荧光。我们还将使用在此期间获得的无标签图像来量化光学代谢活动带有感兴趣的参数的组织照明。我们将确定产生的照明参数的子集不，最小或中度损害，我们将在人宫颈组织上测试这些条件（新鲜切除和冷冻/解冻），人角质形成细胞单层和仓鼠口腔上皮（体内，新鲜切除和冷冻/解冻）。因此，我们希望确定可能造成损坏的标本类型的协议与体内人类鳞状上皮组织最相关的评估和一个范围的组合预计不会导致任何功能损坏的光输送参数。我们也期待基于无标记的多光子成像的组织形态和代谢功能的表征将服务作为独立的高度敏感的组织损伤指示器。我们的发现将与保险箱直接相关体内人宫颈多光子成像的实施；但是，方法和MPE条件我们确定将直接扩展到其他组织类型，并能够翻译这种强大的方式进行体内人类成像。