ArticleChemical & biomedical imaging2026
Multimodal Biophysical Analysis of Morpho-Phenotypic and Morpho-Functional Dynamics in Normal and Cancer Spheroids within Engineered Microenvironments.
Article in Chemical & biomedical imaging, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Three-dimensional spheroid models provide physiologically relevant systems to investigate cancer progression and stem cell dynamics. However, most existing literature focuses primarily on morphological or biochemical characterization, limiting a comprehensive understanding of the coupled biophysical, morphological, and molecular dynamics of spheroids within engineered microenvironments. Addressing this gap, the present study develops a multimodal biophysical framework to comprehensively assess the morpho-phenotypic, metabolic, and electrochemical characteristics of spheroids. Here, we report a comparative analysis of spheroids derived from mesenchymal stem cells (MSC), a mixed population of cancer stem cells and cancer cells (A549), and differentiated cancer cells (MCF-7), cultured in an ECM-mimetic alginate-gelatin hydrogel. Relevant morpho-phenotypic parameters were analyzed at 3, 7, and 14 days from confocal images and cytoskeletal mapping, revealing distinct trends in size, circularity, solidity, and entropy. All spheroids exhibited time-dependent growth (40-60 to 120-240 μm), with cancer spheroids (A549 and MCF-7) showing significantly greater volumetric expansion. Day 7 emerges as a critical transition point marked by an alteration in the morphological trend. A549 spheroids displayed higher entropy, reflecting increased heterogeneity. Autofluorescence spectroscopy revealed distinct redox dynamics of spheroids from different origins. Cancer spheroids showed enhanced NADH, FAD, and porphyrin signatures reflecting redox alterations and Warburg-like effects, wherein MSC spheroids showed a controlled shift toward oxidative phosphorylation. The impedance spectroscopy further distinguished spheroids through capacitance-resistivity relationships. Notably, A549 spheroids exhibit dynamic transitions between stem-like and malignant states, consistent with their intrinsic heterogeneity. Overall, this integrated approach correlates growth, redox status, and dielectric behavior, offering a translational platform for drug screening, disease modeling, and regenerative medicine.
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