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Optimization of the CUPRAC-CAT assay BoxBehnken Design (BBD) was applied as a function of response surface methodology (RSM) to achieve optimal conditions of the CUPRAC-CAT method

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Additionally, multi-parameter screening based on cell phenotypes can be performed using AI image analysis systems to assist high-content imaging (HCI), simultaneously quantifying -SMA expression, cell migration (Transwell assays), and collagen contraction (3D collagen gel models), automatically identifying myofibroblast morphology changes (such as pseudopod retraction), significantly enhancing the efficiency of screening in drug development (Weisbart et al., 2024

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