In vivo label-free characterization of aged skin to predict delayed wound healing

老化皮肤的体内无标记表征以预测伤口愈合延迟

基本信息

批准号：
9922191
负责人：
Kyle Patrick Quinn
金额：
$ 34.71万
依托单位：
UNIVERSITY OF ARKANSAS AT FAYETTEVILLE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9922191
关键词：
Advanced Development Affect Age Aging American Biochemical Biological Assay Biological Markers Cell Proliferation Cell physiology Cells Cellular Structures Characteristics Clinic Clinical Clinical Management Clinical assessments Collagen Contrast Media Dependence Deposition Dermal Development Diabetes Mellitus Diabetic Foot Ulcer Dyes Educational workshop Elastin Elderly Excision Extracellular Matrix Fibroblasts Fluorescence Foundations Functional disorder Generations Goals Health Histology Hyperglycemia Image Immunohistochemistry Impaired healing Impaired wound healing Individual Infection Injury Keratin Label Life Expectancy Measures Mechanics Metabolic Metabolic dysfunction Metabolism Methods Mitochondria Modeling Monitor Mus NADH Optics Outcome Oxidation-Reduction Predisposition Process Regression Analysis Research Skin Skin Aging Skin wound healing Source Stains Streptozocin Structure Techniques Technology Testing Three-Dimensional Imaging Time Tissues Ulcer Weight-Bearing state age effect age related aged aging population base candidate marker cell motility chronic wound cofactor comorbidity crosslink diabetic experimental study extracellular fluorescence lifetime imaging healing imaging modality in vivo innovation keratinocyte mitochondrial dysfunction mouse model multi-scale modeling multiphoton imaging multiphoton microscopy non-diabetic non-healing wounds non-invasive monitor optical imaging pentosidine preclinical development preclinical study prevent primary outcome product development prognostic response second harmonic skin regeneration skin ulcer two-photon wound wound care wound healing wound treatment

项目摘要

Project Summary: Age-related delays in wound healing have been attributed to altered dermal microstructure, mitochondrial dysfunction, and reduced cellular proliferation. However, an inability to non-destructively measure these characteristics has limited their use in guiding wound care and product development. The long-term goal of this project is to establish non-invasive, real-time, quantitative optical biomarkers to predict age-related delays in skin wound healing. The specific objective of this proposal is to utilize label-free multiphoton microscopy techniques, including two-photon excited fluorescence, fluorescence lifetime imaging, and second harmonic generation to identify and develop metabolic and microstructural biomarkers of aged skin that are associated with delayed closure and susceptibility to mechanical re-injury. Our central hypothesis is that the natural fluorescence of cells and their surrounding matrix can be quantified to provide sensitive biomarkers of age- related wound healing impairment. To test this hypothesis and evaluate the effect of aging, controlled pre- clinical studies will be performed using mice with ages ranging from 12.5-75% of their life expectancy, independent of common chronic wound comorbidities such as diabetes mellitus. In Aim 1, we will non- invasively monitor wound healing dynamics in individual live mice and identify differences in wound metabolism during the healing process using the natural fluorescence of NADH and FAD. In Aim 2, we quantify extracellular matrix organization and composition using multiphoton microscopy to predict wound strength through image-based multiscale modeling. In Aim 3, we will evaluate the effect of diabetes mellitus on the age-related optical biomarkers of wound healing developed in Aims 1 and 2. Quantitative comparisons of cellular and extracellular biomarkers will be made across different ages and validated against histology and immunohistochemistry. The expected outcome of this project is a set of candidate biomarkers for use in guiding therapy that are capable of discriminating age-related alterations and common comorbidities known to impair healing. With multiphoton microscopy technology already making its way into the clinic, the imaging methods developed here have the potential for immediate impacts in the clinical assessment and management of wounds.

项目概要：与年龄相关的伤口愈合延迟归因于真皮微观结构、线粒体的改变功能障碍，细胞增殖减少。然而，无法无损地测量这些特性限制了它们在指导伤口护理和产品开发中的使用。长期目标是该项目旨在建立非侵入性、实时、定量的光学生物标志物来预测与年龄相关的延迟在皮肤伤口愈合中。该提案的具体目标是利用无标记多光子显微镜技术，包括双光子激发荧光、荧光寿命成像和二次谐波一代来识别和开发与老化皮肤相关的代谢和微观结构生物标志物延迟闭合且易受机械性再次损伤。我们的中心假设是自然可以量化细胞及其周围基质的荧光，以提供敏感的年龄生物标志物相关的伤口愈合障碍。为了检验这一假设并评估衰老的影响，控制预临床研究将使用年龄为预期寿命 12.5-75% 的小鼠进行，独立于常见的慢性伤口合并症，例如糖尿病。在目标 1 中，我们将不有创地监测个体活小鼠的伤口愈合动态并识别伤口代谢的差异在愈合过程中使用 NADH 和 FAD 的天然荧光。在目标 2 中，我们量化使用多光子显微镜预测伤口强度的细胞外基质组织和成分通过基于图像的多尺度建模。在目标 3 中，我们将评估糖尿病对目标 1 和 2 中开发的与年龄相关的伤口愈合光学生物标志物。将在不同年龄阶段制作细胞和细胞外生物标志物，并根据组织学和免疫组织化学。该项目的预期成果是一组候选生物标志物，用于指导治疗能够区分年龄相关的改变和已知的常见合并症损害愈合。随着多光子显微镜技术已经进入临床，成像这里开发的方法有可能对临床评估和管理产生直接影响的伤口。