White Kristopher, Chalaby Rabab, Lowe Gina, Berlin Jacob, Glackin Carlotta, Olabisi Ronke
Department of Biomedical Engineering, The Henry Samueli School of Engineering, University of California-Irvine, Irvine, CA 92697, USA.
Department of Materials Engineering, University of Technology, Baghdad 10066, Iraq.
Polymers (Basel). 2021 Jul 11;13(14):2274. doi: 10.3390/polym13142274.
We describe a method to assess mineralization by osteoblasts within microspheres using calcein. Fluorescence imaging of calcein bound to the calcium in hydroxyapatite permits assessment of the mineralized portion of the extracellular matrix. Colorimetric imaging of Alizarin Red S complexed with calcium also gives measures of mineralization, and in tissue cultures calcein and Alizarin Red S have been shown to bind to the same regions of mineral deposits. We show that when the mineralization takes place within hydrogel microspheres, Alizarin Red S does not stain mineral deposits as consistently as calcein. As tissue engineers increasingly encapsulate osteoprogenitors within hydrogel scaffolds, calcein staining may prove a more reliable method to assess this mineralization.
我们描述了一种使用钙黄绿素评估微球内成骨细胞矿化的方法。与羟基磷灰石中的钙结合的钙黄绿素的荧光成像可用于评估细胞外基质的矿化部分。与钙络合的茜素红S的比色成像也可用于测量矿化情况，并且在组织培养中，已证明钙黄绿素和茜素红S会结合到矿化沉积物的相同区域。我们发现，当矿化发生在水凝胶微球内时，茜素红S对矿化沉积物的染色不如钙黄绿素稳定。随着组织工程学越来越多地将骨祖细胞封装在水凝胶支架中，钙黄绿素染色可能被证明是评估这种矿化的更可靠方法。
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