Analysis of the role of ECM in modulating electroporation efficiency in 3D HeLa cultures

Electrochemotherapy relies on transient membrane electroporation to enhance chemotherapeutic drug uptake,; yet its in vitro efficacy in three-dimensional tumor models is critically governed by the biochemical and structural features of the extracellular matrix. Here, we dissect the role of scaffold extracellular matrix composition in modulating electroporation thresholds in three-dimensional (3D) HeLa cultures. Two hyaluronic acid-based scaffolds were designated: (i) hyaluronic acid-based scaffolds, modified with 5% w/w of self-assembling EAbuK peptide incorporating the IKVAV cell-adhesive motif and (ii) hyaluronic acid-based scaffolds added with 5% weigh/weight of exogenous calf Type I collagen. The control is the scaffold hyaluronic acid-only, without functionalization. The data obtained with 3D experiments were compared with 2D experimental results. These findings highlight the influence of scaffold-induced endogenous extracellular matrix remodeling on electroporation efficiency and underscore the importance of extracellular matrix architecture in refining 3D electrochemotherapy models for improved therapeutic applications.