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<p>Three-dimensional (3D) culture systems are becoming common in tissue regeneration research, for better mimic in vivo&#xa0;conditions. However, non-destructive evaluation of 3D cultures remains challenging. Electrochemical impedance spectroscopy (EIS) could represent an alternative and automated testing for <i>in vitro</i> research. In this study, hydrogels consisting of 1%alginate/2%agarose were combined with gingival (GT-MSCs) and dental pulp (DP-MSCs) stem cells. Cells were cultured in hydrogels, assessed via microscopy, resazurin assay, and Calcein-AM staining, and compared to EIS measurements. EIS was measured (10<sup>6</sup>–10<sup>−2</sup>&#xa0;Hz) to optimize viability sensing. Hydrogels supported viability, with EIS revealing distinct dielectric behaviors: DP-MSCs showed slow impedance rise during the whole testing (1.4 × 10<sup>5</sup> Ω·cm<sup>2</sup> at 10<sup>–2</sup>&#xa0;Hz at 168&#xa0;h), indicating low cell growth, while GT-MSCs exhibited higher impedance (1.6 × 10<sup>4</sup> Ω·cm<sup>2</sup> at 120&#xa0;h), reflecting higher viability. EIS proved to be a valuable for culture monitoring, distinguishing cell-type-specific responses and validating hydrogel efficacy in supporting cell growth.</p> Graphical abstract <p></p>

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Non-destructive evaluation of dental stem cell proliferation in 3D hydrogel cultures via electrochemical impedance

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年1月5日

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  • Patricia González-Alva,
  • Miguel Angel Hernández-Gallegos,
  • Guadalupe Rosario Fajardo-Orduña,
  • Juan José Montesinos

摘要

Three-dimensional (3D) culture systems are becoming common in tissue regeneration research, for better mimic in vivo conditions. However, non-destructive evaluation of 3D cultures remains challenging. Electrochemical impedance spectroscopy (EIS) could represent an alternative and automated testing for in vitro research. In this study, hydrogels consisting of 1%alginate/2%agarose were combined with gingival (GT-MSCs) and dental pulp (DP-MSCs) stem cells. Cells were cultured in hydrogels, assessed via microscopy, resazurin assay, and Calcein-AM staining, and compared to EIS measurements. EIS was measured (106–10−2 Hz) to optimize viability sensing. Hydrogels supported viability, with EIS revealing distinct dielectric behaviors: DP-MSCs showed slow impedance rise during the whole testing (1.4 × 105 Ω·cm2 at 10–2 Hz at 168 h), indicating low cell growth, while GT-MSCs exhibited higher impedance (1.6 × 104 Ω·cm2 at 120 h), reflecting higher viability. EIS proved to be a valuable for culture monitoring, distinguishing cell-type-specific responses and validating hydrogel efficacy in supporting cell growth.

Graphical abstract