Integrative Infochemistry of Polyhydroxyalkanoate Films: Multimodal Data, Topological Tuning, and Cell-Surface...

This study investigates seeding and cultivation of C2C12 myoblast cells on poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P3HBV) films synthesized at four polymer concentrations, yielding films with progressively increasing lateral thickness and diverse surface topographies. Cell attachment was evaluated after 24 h, alongside correlations between surface topography and cellular behavior. Surface characteristics across films at each thickness were quantified using atomic force microscopy (AFM), scanning electron microscopy (SEM), advanced topological data analysis (TDA), and threshold relief analysis. C2C12 myoblast growth was imaged via fluorescence microscopy, with quantitative metrics extracted using the Cellpose Plus toolbox. Statistical modeling revealed correlations between surface features and cell morphology. SEM imaging further corroborated pore characteristics against cell growth metrics. Results demonstrate a linear relationship between surface topography and cellular responses, underscoring the critical role of substrate morphology in modulating cell behavior, where films synthesized at high polymer concentrations (5-7% w/v) exhibit enhanced cell attachment. The 85:15 P3HBV copolymer was selected for its favorable thermomechanical properties, high biocompatibility, and cost-effectiveness for large-scale biosynthesis. Our work presents a reproducible data-driven pipeline for analyzing cell-surface interactions, combining TDA and image segmentation metrics to explore the behavior of surface morphology under varying material conditions.