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A Raman-Derived Biophysical Commitment Index for Predicting Nanosecond-Pulsed-Electric-Field-Induced Neuronal Differentiation in PC-12 Cells

Author(s): Benjamin Scott1
1Institute for Environmental Analysis Greenfield University United States
Benjamin Scott
Institute for Environmental Analysis Greenfield University United States

Abstract

nsPEFs constitute a temporally sharp physical perturbation, independent of sustained chemical input, for probing the impact on intracellular state. In PC-12 cells, nsPEFs induce neuritogenesis, enhanced neurite outgrowth when NGF-treated, increased C–H Raman scattering intensity, cell volume contraction, and intracellular heating. The research question posed here concerns the existence of a relationship between the early nsPEF-induced changes observable by Raman spectroscopy and the subsequent neuronal commitment of the treated cell population. A novel definition of the Biophysical Commitment Index (BCI), based on cytoplasmic crowding effects, cell-area shrinkage, thermal dynamics of the intracellular medium, and nsPEF{}-waveform-related parameters, serves as a physiologically motivated predictor of nsPEF-induced neuronal commitment. Herein, the BCI{} predictor is determined by hierarchical modeling, mediated by Raman-observables and bright-field morphometrics of the individual cells nested within biological replicates, in a way consistent with leave-one-dish-out cross-validation. Contributions of this work include the development of a physically intuitive model relating nsPEF-triggered C–H and O–H vibrations to further neuronal commitment; a validation scheme that accounts for the nested structure of experimental data and preserves the distinction between treatment assignment and early intracellular state; and an experimental design for assessing label-free Raman spectroscopy as a biomarker of drug-free neuronal differentiation.

Keywords: nanosecond pulsed electric fields; Raman microscopy; PC-12 cells; neuronal differentiation; intracellular temperature; molecular crowding; neurite outgrowth; biophysical commitment index
Copyright © 2025 Benjamin Scott. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.