<p>Tissue engineering seeks to develop biomimetic substitutes for damaged tissues using natural or synthetic materials functionalized with cells or biologically active compounds. Understanding how cell-scaffold interactions influence cellular behavior is critical for optimizing tissue engineering strategies. Polylactic acid (PLA), a biodegradable and biocompatible polymer approved by the U.S. FDA, is commonly used for scaffolds fabrication. Mesenchymal stromal cells (MSCs) are widely utilized in regenerative medicine due to their capacity to differentiate into mesodermal lineages and their secretomes, which exhibit robust paracrine activity. To date, few studies have investigated how scaffold characteristics, such as materials and/or architecture, modulate secretome composition using integrated multi-omics approaches. An untargeted metabolomics workflow combined with label-free proteomics was employed to analyze ASC-derived secretomes, obtained from adipose tissue-derived MSCs (ASCs) cultured in PLA-discs and PLA-scaffolds, as well as on conventional 2D polystyrene (PS) culture surfaces. A similar proteomic distribution was observed when secretomes from PLA-discs and PLA-scaffolds were compared. On the other hand, significant changes in the proteomic profile were observed between PLA-discs and 2D-PS secretomes. Proteins belonging to carbohydrate metabolism, cell motility, vasculature development, and oxidative stress response, among others, were increased in PLA-discs. The metabolomic profile shows significant differences, with metabolites related to glucose metabolism, such as pyruvate and lactic acid, increased in PLA secretomes. Our multi-omic approach demonstrates that the PLA constructs introduced here can modulate the secretome of ASCs, inducing significant changes in its composition, highlighting the influence of culture format on the secretory capacity of ASCs.</p>