Baugh Lauren M, Liu Zhiyi, Quinn Kyle P, Osseiran Sam, Evans Conor L, Huggins Gordon S, Hinds Philip W, Black Lauren D, Georgakoudi Irene
Department of Biomedical Engineering, Tufts University, Medford, MA, 02155, USA.
Department of Biomedical Engineering, University of Arkansas, Fayetteville, AR, 72701, USA.
Nat Biomed Eng. 2017 Nov;1(11):914-924. doi: 10.1038/s41551-017-0152-3. Epub 2017 Nov 6.
Calcifications occur during the development of healthy bone, and at the onset of calcific aortic-valve disease (CAVD) and many other pathologies. Although the mechanisms regulating early calcium deposition are not fully understood, they may provide targets for new treatments and for early interventions. Here, we show that two-photon excited fluorescence (TPEF) can provide quantitative and sensitive readouts of calcific nodule formation, in particular in the context of CAVD. Specifically, by means of the decomposition of TPEF spectral images from excised human CAVD valves and from rat bone prior to and following demineralization, as well as from calcific nodules formed within engineered gels, we identified an endogenous fluorophore that correlates with the level of mineralization in the samples. We then developed a ratiometric imaging approach that provides a quantitative readout of the presence of mineral deposits in early calcifications. TPEF should enable non-destructive, high-resolution imaging of three-dimensional tissue specimens for the assessment of the presence of calcification.
钙化现象在健康骨骼发育过程中出现，也在钙化性主动脉瓣疾病（CAVD）及许多其他病症发病时出现。尽管调节早期钙沉积的机制尚未完全明确，但这些机制可能为新的治疗方法和早期干预提供靶点。在此，我们表明双光子激发荧光（TPEF）能够对钙化结节形成进行定量且灵敏的检测，尤其是在CAVD的背景下。具体而言，通过对脱矿前后的切除的人类CAVD瓣膜、大鼠骨骼以及工程凝胶中形成的钙化结节的TPEF光谱图像进行分解，我们鉴定出一种内源性荧光团，它与样品中的矿化水平相关。然后我们开发了一种比率成像方法，可对早期钙化中矿物质沉积的存在情况进行定量检测。TPEF应能实现对三维组织标本的无损、高分辨率成像，以评估钙化的存在情况。
