Akademik Veri Yönetim Sistemi
34th Annual Conference of the European Society for Biomaterials (ESB), Turin, İtalya, 7 - 11 Eylül 2025, ss.2535, (Özet Bildiri)
Introduction
Protein-based scaffolds such as GelMA are widely used in 3D drug metabolism models. While RGD-mediated integrin signaling is well known to support adhesion and proliferation, its possible influence on drug-metabolizing enzyme expression and metabolite profiles remains largely unexplored. Studies suggest that cellular bioelectric states—such as membrane potential and ion flux—can affect gene expression and metabolic state (1, 2). Based on this, we propose that scaffold degradation products may indirectly shape drug metabolism through integrin-associated modulation of cellular electrical behavior.
Materials and Methods
To test this, Caco-2 cells will be cultured in 2D and 3D GelMA scaffolds. Real-time impedance measurements (EIS, Gamry 1010E), calcium imaging, and gene/protein analysis will be used to assess bioelectric changes and associated shifts in the expression of drug-metabolizing enzymes, particularly cytochrome P450 3A4 (CYP3A4). HPLC will be used to analyze the metabolite profiles of a CYP3A4 probe substrate. Liver microsomal assays with and without synthetic degradation peptides will serve as acellular controls, limited to phase I metabolic reactions. Electrode-integrated 3D printed wells will be designed based on prior methods (3).
Results
We anticipate that 3D scaffold degradation will result in measurable bioelectric changes—such as alterations in impedance or membrane voltage—which may correlate with changes in cytochrome P450 enzyme expression. These changes are expected to influence not only the overall quantity of drug metabolites but also their qualitative composition. Specifically, 3D conditions may lead to the generation of a broader or altered range of metabolic products, potentially including distinct stereoisomers. Since certain stereoisomers can differ in biological activity or toxicity, this aspect of metabolic variability is particularly relevant. Importantly, GelMA degradation is expected to release RGD-containing peptides, which may engage integrin receptors and initiate downstream signaling cascades such as PI3K/AKT and MAPK/ERK. These pathways are known to influence both ion channel regulation and gene expression, including that of CYP enzymes (4). Such integrin-mediated signaling could therefore modulate the bioelectric state of the cell and contribute to functional reprogramming of its metabolic profile. Comparisons with conventional 2D cultures will help identify whether the effects are specific to the 3D microenvironment, while acellular microsomal assays—limited to phase I reactions—will clarify whether observed changes result from direct enzyme modulation or cell-dependent responses to scaffold-derived biochemical cues.
Discussion
This work proposes a mechanistic framework connecting degradable scaffold signals to drug metabolism outcomes in 3D cancer models. By incorporating both electrochemical and molecular readouts, it addresses a gap in understanding how ECM-derived peptides, such as RGD motifs, may affect cellular metabolism through integrin-mediated signaling and bioelectric modulation. While experimental validation is underway, this approach may offer a new dimension to biomaterial design in pharmacological testing, emphasizing the functional consequences of scaffold–cell interaction beyond structural support.
Conclusions
This study outlines a framework for investigating how ECM-derived biochemical and biophysical cues may impact drug metabolism in vitro. By examining a possible link between scaffold degradation and metabolic variability, we aim to contribute to the development of more robust and predictive 3D culture systems for pharmacological applications.
References
[1] Levin M. et al., 2012, doi: 10.1146/annurev-bioeng-071811-150114
[2] Sheth, M., & Esfandiari, L. 2022, doi: 10.3389/fonc.2022.846917
[3] Paivana G. et al., 2019, doi: 10.3390/bios9040136
[4] Becchetti, A. et al., 2019, doi: 10.1016/j.tcb.2018.12.005