{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T13:00:30Z","timestamp":1790600430064,"version":"4.1.0"},"reference-count":60,"publisher":"American Chemical Society (ACS)","issue":"9","license":[{"start":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T00:00:00Z","timestamp":1777420800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001843","name":"Science and Engineering Research Board","doi-asserted-by":"publisher","award":["PAIR\/2025\/000021"],"award-info":[{"award-number":["PAIR\/2025\/000021"]}],"id":[{"id":"10.13039\/501100001843","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001843","name":"Science and Engineering Research Board","doi-asserted-by":"publisher","award":["WEA\/2023\/000015"],"award-info":[{"award-number":["WEA\/2023\/000015"]}],"id":[{"id":"10.13039\/501100001843","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>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\u201360 to 120\u2013240 \u03bcm), 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\u2013resistivity 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.<\/jats:p>","DOI":"10.1021\/cbmi.5c00255","type":"journal-article","created":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T11:44:11Z","timestamp":1777463051000},"page":"2010-2022","source":"Crossref","is-referenced-by-count":0,"title":["Multimodal Biophysical\nAnalysis of Morpho-Phenotypic\nand Morpho-Functional Dynamics in Normal and Cancer Spheroids within\nEngineered Microenvironments"],"prefix":"10.1021","volume":"4","author":[{"given":"Srijani","family":"Dey","sequence":"first","affiliation":[{"name":"Indian Institute of Engineering Science and Technology , , Shibpur, ,","place":["Howrah, India, 711103"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shreya","family":"Adhikary","sequence":"additional","affiliation":[{"name":"Indian Institute of Engineering Science and Technology , , Shibpur, ,","place":["Howrah, India, 711103"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Priyanka","family":"Das","sequence":"additional","affiliation":[{"name":"National Institute of Pharmaceutical Education and Research , , ,","place":["Kolkata, India, 700054"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7404-4917","authenticated-orcid":true,"given":"Pallab","family":"Datta","sequence":"additional","affiliation":[{"name":"National Institute of Pharmaceutical Education and Research , , ,","place":["Kolkata, India, 700054"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Subhadip","family":"Chakraborty","sequence":"additional","affiliation":[{"name":"Institute of Engineering and Management , Management House, D-1, Sector-V, Saltlake Electronics Complex , , ,","place":["Kolkata, West Bengal, India, 700091"]},{"name":"New Town University , , Area, Plot no. III, B\/5, New Town Rd, Action Area III, Newtown , , ,","place":["Kolkata, West Bengal, India, 700160"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9710-9448","authenticated-orcid":true,"given":"Ananya","family":"Barui","sequence":"additional","affiliation":[{"name":"Indian Institute of Engineering Science and Technology , , Shibpur, ,","place":["Howrah, India, 711103"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,4,29]]},"reference":[{"issue":"19","key":"2026092808072747400_cit1","doi-asserted-by":"publisher","first-page":"4846","DOI":"10.3390\/cancers15194846","article-title":"Three-dimensional in\nvitro tumor spheroid models for evaluation of anticancer therapy:\nrecent 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Biotechnol."},{"key":"2026092808072747400_cit33","author":"Adhikary","journal-title":"Label Free in Situ Characterization of 3d Tumor Spheroids\nby Microfluidic-Based\nBioimpedance Spectroscopy"},{"key":"2026092808072747400_cit34","doi-asserted-by":"publisher","DOI":"10.1039\/D5LC00540J","article-title":"High-throughput impedance\nmonitoring in 3D tumor cultures: a multiplex, microfluidic-free platform\nfor drug screening","author":"Marino","year":"2025","journal-title":"Lab Chip"},{"issue":"8","key":"2026092808072747400_cit35","doi-asserted-by":"publisher","first-page":"1028","DOI":"10.1021\/acssensors.6b00272","article-title":"Electrical impedance spectroscopy for microtissue spheroid analysis\nin hanging-drop networks","volume":"1","author":"Schmid","year":"2016","journal-title":"ACS sensors"},{"key":"2026092808072747400_cit36","doi-asserted-by":"publisher","first-page":"605","DOI":"10.3389\/fphys.2017.00605","article-title":"Comparative analysis\nof 3D bladder tumor spheroids obtained by forced floating and hanging\ndrop methods for drug screening","volume":"8","author":"Amaral","year":"2017","journal-title":"Front. Physiol."},{"issue":"17","key":"2026092808072747400_cit37","doi-asserted-by":"publisher","first-page":"4189","DOI":"10.1039\/C8AN00729B","article-title":"Electrical impedance\ntomography for real-time and label-free cellular viability assays\nof 3D tumour spheroids","volume":"143","author":"Wu","year":"2018","journal-title":"Analyst"},{"issue":"1","key":"2026092808072747400_cit38","doi-asserted-by":"publisher","first-page":"455","DOI":"10.1186\/s13104-025-07524-5","article-title":"Impact of different\n3D culture environments on spheroid\nmorphology, drug response, and invasion in pancreatic cancer cells","volume":"18","author":"Karakas","year":"2025","journal-title":"BMC Res. Notes"},{"issue":"2","key":"2026092808072747400_cit39","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1007\/s44164-024-00074-3","article-title":"Scaffold-free development\nof multicellular tumor spheroids with spatial characterization of\nstructure and metabolic radial profiles","volume":"3","author":"Bess","year":"2024","journal-title":"In\nvitro models"},{"issue":"3","key":"2026092808072747400_cit40","doi-asserted-by":"publisher","first-page":"140","DOI":"10.1089\/adt.2018.895","article-title":"Quantitative\nsize-based\nanalysis of tumor spheroids and responses to therapeutics","volume":"17","author":"Thakuri","year":"2019","journal-title":"ASSAY and Drug Development Technologies"},{"issue":"4","key":"2026092808072747400_cit41","doi-asserted-by":"publisher","first-page":"2000810","DOI":"10.1002\/adhm.202000810","article-title":"Cancer Cell Invasion of Mammary Organoids\nwith Basal-In Phenotype","volume":"10","author":"Parigoris","year":"2021","journal-title":"Adv. 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App."},{"key":"2026092808072747400_cit43","doi-asserted-by":"publisher","first-page":"106897","DOI":"10.1016\/j.compbiomed.2023.106897","article-title":"A mechanobiological model for tumor spheroid evolution\nwith application to glioblastoma: a continuum multiphysics approach","volume":"159","author":"Carrasco-Mantis","year":"2023","journal-title":"Computers in Biology and Medicine"},{"issue":"127","key":"2026092808072747400_cit44","doi-asserted-by":"publisher","first-page":"20160851","DOI":"10.1098\/rsif.2016.0851","article-title":"Measurement\nof oxygen tension within\nmesenchymal stem cell spheroids","volume":"14","author":"Murphy","year":"2017","journal-title":"J. R. 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Ther."},{"issue":"18","key":"2026092808072747400_cit56","doi-asserted-by":"publisher","first-page":"5184","DOI":"10.1158\/0008-5472.CAN-14-0663","article-title":"Quantitative\noptical\nimaging of primary tumor organoid metabolism predicts drug response\nin breast cancer","volume":"74","author":"Walsh","year":"2014","journal-title":"Cancer research"},{"key":"2026092808072747400_cit57","doi-asserted-by":"publisher","first-page":"510","DOI":"10.1016\/j.freeradbiomed.2019.09.004","article-title":"Mitochondrial fragmentation, elevated mitochondrial\nsuperoxide and respiratory supercomplexes disassembly is connected\nwith the tamoxifen-resistant phenotype of breast cancer cells","volume":"143","author":"Tomkova\u0301","year":"2019","journal-title":"Free Radical Biol. 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