Abstract
Photodynamic therapy (PDT) is a treatment which uses light-activated compounds to produce reactive oxygen species, leading to membrane damage and cell death. Multicellular cancer spheroids are a preferable alternative for PDT evaluation in comparison to monolayer cell cultures due to their ability to better mimic in vivo avascular tumour characteristics such as hypoxia and cell-cell interactions, low cost, and ease of production. However, inconsistent growth kinetics and drug responsiveness causes poor experimental reproducibility and limits their usefulness. Herein, we used image analysis to establish a link between human melanoma C8161 spheroid morphology and drug responsiveness. Spheroids were pre-selected based on sphericity, area, and diameter, reducing variation in experimental groups before treatment. Spheroid morphology after PDT was analyzed using AnaSP and ReViSP, MATLAB-based open-source software, obtaining nine different parameters. Spheroids displayed a linear response between biological assays and morphology, with area (R2 = 0.7219) and volume (R2 = 0.6138) showing the best fit. Sphericity, convexity, and solidity were confirmed as poor standalone indicators of spheroid viability. Our results indicate spheroid morphometric parameters can be used to accurately screen inefficient treatment combinations of novel compounds.
Highlights
Photodynamic therapy (PDT) is an FDA-approved cancer treatment which produces reactive oxygen species through the specific excitation of a photosensitizer (PS) at a given wavelength
Our results show the potential of image-based analysis for 3D cell culture models to complement in vitro data and highlight key areas of opportunity for their use in drug screening
This work presented an approach for using spheroid morphometric parameters for screening drug response to various treatment combinations
Summary
Photodynamic therapy (PDT) is an FDA-approved cancer treatment which produces reactive oxygen species through the specific excitation of a photosensitizer (PS) at a given wavelength. It is multifactorial, requiring the evaluation of parameters like irradiation wavelength, light dose, and drug concentration (Cho et al, 2013). 3D cell culture models replicate in vivo conditions such as hypoxia, dormancy, and cell-cell interactions more accurately than 2D models. Multicellular cancer spheroids (MCTS) can replicate in vivo conditions such as hypoxia, dormancy, and cell-cell interactions more accurately, making them an ideal model for evaluating PDT parameters (Imamura et al, 2015)
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