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

• 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
• 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

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.

项目摘要： 与年龄相关的伤口愈合延迟归因于皮肤微结构改变，线粒体 功能障碍和降低的细胞增殖。但是，无力测量这些 特征限制了他们在指导伤口护理和产品开发中的使用。长期目标 该项目是建立非侵入性，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。该提案的具体目的是利用无标签的多光子显微镜 技术，包括两光激发荧光，荧光寿命成像和第二次谐波 识别和发展与年龄皮肤的代谢和微结构生物标志物 闭合延迟和对机械重新伤害的敏感性。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，以提供年龄的敏感生物标志物 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，受控的 临床研究将使用年龄范围为12.5-75％的预期寿命的小鼠进行 独立于常见的慢性伤口合并症，例如糖尿病。在AIM 1中，我们将 侵入性监测单个活小鼠的伤口愈合动力学并确定伤口代谢的差异 在使用NADH和FAD的自然荧光的愈合过程中。在AIM 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组成 通过基于图像的多尺度建模。在AIM 3中，我们将评估糖尿病对 在目标1和2中发展了与年龄相关的伤口愈合的光学生物标志物。定量比较 细胞和细胞外生物标志物将在不同年龄段制造，并针对组织学和 免疫组织化学。该项目的预期结果是一组候选生物标志物用于 能够区分与年龄相关的改变和已知的共同合并症的指导疗法 损害康复。随着多光子显微镜技术已经进入诊所，成像 这里开发的方法有可能立即影响临床评估和管理 伤口。