Jones Jake D, Ramser Hallie E, Woessner Alan E, Quinn Kyle P
Department of Biomedical Engineering, University of Arkansas, 123 John A. White Jr. Engineering Hall, Fayetteville, AR, 72701, USA.
Commun Biol. 2018 Nov 19;1:198. doi: 10.1038/s42003-018-0206-4. eCollection 2018.
Chronic wounds are difficult to diagnose and characterize due to a lack of quantitative biomarkers. Label-free multiphoton microscopy has emerged as a useful imaging modality capable of quantifying changes in cellular metabolism using an optical redox ratio of FAD/(NADH+FAD) autofluorescence. However, the utility of an optical redox ratio for long-term in vivo monitoring of tissue metabolism has not been robustly evaluated. In this study, we demonstrate how multiphoton microscopy can be used to monitor changes in the metabolism of individual full-thickness skin wounds in vivo. 3D optical redox ratio maps and NADH fluorescence lifetime images identify differences between diabetic and control mice during the re-epithelialization of wounds. These metabolic changes are associated with a transient increase in keratinocyte proliferation at the wound edge. Our study demonstrates that high-resolution, non-invasive autofluorescence imaging can be performed in vivo and that optical redox ratios can serve as quantitative optical biomarkers of impaired wound healing.
由于缺乏定量生物标志物，慢性伤口难以诊断和表征。无标记多光子显微镜已成为一种有用的成像方式，能够利用FAD/（NADH+FAD）自发荧光的光学氧化还原比来量化细胞代谢的变化。然而，光学氧化还原比在组织代谢长期体内监测中的效用尚未得到充分评估。在本研究中，我们展示了多光子显微镜如何用于体内监测单个全层皮肤伤口的代谢变化。3D光学氧化还原比图和NADH荧光寿命图像识别出糖尿病小鼠和对照小鼠在伤口再上皮化过程中的差异。这些代谢变化与伤口边缘角质形成细胞增殖的短暂增加有关。我们的研究表明，可以在体内进行高分辨率、非侵入性自发荧光成像，并且光学氧化还原比可以作为伤口愈合受损的定量光学生物标志物。
