PDF(Pubmed)
Abstract：
Dystrophic muscle is characterized by necrosis/regeneration cycles, inflammation, and fibro-adipogenic development. Conventional histological stainings provide essential topographical data of this remodeling but may be limited to discriminate closely related pathophysiological contexts. They fail to mention microarchitecture changes linked to the nature and spatial distribution of tissue compartment components. We investigated whether label-free tissue autofluorescence revealed by Synchrotron deep ultraviolet (DUV) radiation could serve as an additional tool for monitoring dystrophic muscle remodeling. Using widefield microscopy with specific emission fluorescence filters and microspectroscopy defined by high spectral resolution, we analyzed samples from healthy dogs and two groups of dystrophic dogs: naïve (severely affected) and MuStem cell-transplanted (clinically stabilized) animals. Multivariate statistical analysis and machine learning approaches demonstrated that autofluorescence emitted at 420-480 nm by the Biceps femoris muscle effectively discriminates between healthy, dystrophic, and transplanted dog samples. Microspectroscopy showed that dystrophic dog muscle displays higher and lower autofluorescence due to collagen cross-linking and NADH respectively than that of healthy and transplanted dogs, defining biomarkers to evaluate the impact of cell transplantation. Our findings demonstrate that DUV radiation is a sensitive, label-free method to assess the histopathological status of dystrophic muscle using small amounts of tissue, with potential applications in regenerative medicine.
摘要：
营养不良肌肉的特征是坏死/再生周期，炎症,和纤维脂肪发育。常规的组织学染色提供了这种重塑的基本地形数据，但可能仅限于区分密切相关的病理生理学背景。他们没有提到与组织区室成分的性质和空间分布有关的微结构变化。我们研究了同步加速器深紫外线（DUV）辐射显示的无标记组织自发荧光是否可以作为监测营养不良性肌肉重塑的附加工具。使用具有特定发射荧光滤光片的宽视场显微镜和由高光谱分辨率定义的显微光谱学，我们分析了来自健康狗和两组营养不良狗的样本:未治疗(受严重影响)和MuStem细胞移植(临床稳定)动物。多变量统计分析和机器学习方法表明，由股二头肌在420-480nm发出的自发荧光有效地区分健康，营养不良,和移植的狗样本。显微光谱显示，营养不良犬的肌肉由于胶原蛋白交联和NADH而分别显示出比健康犬和移植犬更高和更低的自发荧光。定义生物标志物以评估细胞移植的影响。我们的研究结果表明，DUV辐射是一种敏感的，使用少量组织评估营养不良肌肉的组织病理学状态的无标签方法，在再生医学中具有潜在的应用。
