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双光子激发的 NAD(P)H 和 FAD 代谢活性的荧光寿命成像在骨植入物存活诊断中的应用

Two-Photon-Excited FLIM of NAD(P)H and FAD-Metabolic Activity of Fibroblasts for the Diagnostics of Osteoimplant Survival.

机构信息

Laboratory of Laser Molecular Imaging and Machine Learning, Tomsk State University, Lenin Ave. 36, 634050 Tomsk, Russia.

Independent Researcher, 10178 Berlin, Germany.

出版信息

Int J Mol Sci. 2024 Feb 13;25(4):2257. doi: 10.3390/ijms25042257.

DOI:10.3390/ijms25042257
PMID:38396933
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10889693/
Abstract

Bioinert materials such as the zirconium dioxide and aluminum oxide are widely used in surgery and dentistry due to the absence of cytotoxicity of the materials in relation to the surrounding cells of the body. However, little attention has been paid to the study of metabolic processes occurring at the implant-cell interface. The metabolic activity of mouse 3T3 fibroblasts incubated on yttrium-stabilized zirconium ceramics cured with aluminum oxide (ATZ) and stabilized zirconium ceramics (Y-TZP) was analyzed based on the ratio of the free/bound forms of cofactors NAD(P)H and FAD obtained using two-photon microscopy. The results show that fibroblasts incubated on ceramics demonstrate a shift towards the free form of NAD(P)H, which is observed during the glycolysis process, which, according to our assumptions, is related to the porosity of the surface of ceramic structures. Consequently, despite the high viability and good proliferation of fibroblasts assessed using an MTT test and a scanning electron microscope, the cells are in a state of hypoxia during incubation on ceramic structures. The FLIM results obtained in this work can be used as additional information for scientists who are interested in manufacturing osteoimplants.

摘要

生物惰性材料，如二氧化锆和氧化铝，由于材料对身体周围细胞无细胞毒性，因此被广泛应用于外科和牙科。然而，人们对发生在植入物-细胞界面的代谢过程的研究关注甚少。本研究基于双光子显微镜获得的辅助因子 NAD(P)H 和 FAD 的游离/结合形式的比率，分析了在经过氧化铝（ATZ）和稳定氧化锆（Y-TZP）处理的掺钇氧化锆陶瓷上孵育的小鼠 3T3 成纤维细胞的代谢活性。结果表明，在陶瓷上孵育的成纤维细胞向 NAD(P)H 的游离形式转移，这在糖酵解过程中观察到，根据我们的假设，这与陶瓷结构表面的多孔性有关。因此，尽管使用 MTT 试验和扫描电子显微镜评估的成纤维细胞活力高且增殖良好，但在陶瓷结构上孵育时，细胞处于缺氧状态。本工作中获得的 FLIM 结果可作为对制造骨植入物感兴趣的科学家的附加信息。

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