Quantitative temporal interrogation in 3D of bioengineered human cartilage using multimodal label-free imaging.
Quantitative temporal interrogation in 3D of bioengineered human cartilage using multimodal label-free imaging.
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DOI：
10.1039/c8ib00050f
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发表时间：
2018-10
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影响因子：
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通讯作者：
Catarina Costa Moura;S. Lanham;Tual Monfort;K. Bourdakos;Rahul S. Tare;R. Oreffo;S. Mahajan
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文献类型：
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作者：
Catarina Costa Moura;S. Lanham;Tual Monfort;K. Bourdakos;Rahul S. Tare;R. Oreffo;S. Mahajan
骨骼干细胞的独特性质在骨骼再生策略的开发中引起了人们的广泛关注。然而，仍然存在一个关键的未满足的需求，即开发定量工具来阐明骨骼细胞发育并使用3D非破坏性技术监测再生组织的形成。无标记的方法，如相干反斯托克斯拉曼散射（汽车），二次谐波产生（SHG）和双光子激发自发荧光（TPEAF）显微镜是微创，非破坏性的，并提出了新的强大的替代传统的成像技术。在这里，我们报告了一个单一的多模式系统中的软骨形成的人类骨骼细胞的时间评估这些技术的组合。使用3D非破坏性平台进行生物工程软骨的评估，其中新参数测量每个细胞的胶原蛋白量，胶原蛋白纤维结构和软骨细胞分布。这种3D无标记时间量化为实时跟踪骨骼细胞发育铺平了道路，并为组织工程和再生医学应用提供了范式转变。
The unique properties of skeletal stem cells have attracted significant attention in the development of strategies for skeletal regeneration. However, there remains a crucial unmet need to develop quantitative tools to elucidate skeletal cell development and monitor the formation of regenerated tissues using non-destructive techniques in 3D. Label-free methods such as coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG) and two-photon excited auto-fluorescence (TPEAF) microscopy are minimally invasive, non-destructive, and present new powerful alternatives to conventional imaging techniques. Here we report a combination of these techniques in a single multimodal system for the temporal assessment of cartilage formation by human skeletal cells. The evaluation of bioengineered cartilage, with a new parameter measuring the amount of collagen per cell, collagen fibre structure and chondrocyte distribution, was performed using the 3D non-destructive platform. Such 3D label-free temporal quantification paves the way for tracking skeletal cell development in real-time and offers a paradigm shift in tissue engineering and regenerative medicine applications.