Researcher(s)
- Ryan Svenson, Chemical Engineering, University of Delaware
Faculty Mentor(s)
- Catherine Fromen, CBE, University of Delaware
- April Kloxin, CBE, MSEG, University of Delaware
Abstract
The extracellular matrix (ECM) is a highly dynamic and complex network of biological macromolecules that provides essential structural and functional support to tissues and organs. The ECM regulates key cellular functions, including adhesion, migration, differentiation, and signaling. To better replicate native cellular environments in vitro, short amino acid sequences derived from ECM proteins—such as RGD (from fibronectin), IKVAV (from laminin), GFOGER, and YIGSR (both from collagen)—have been developed. These bioactive peptides can mimic ECM functions and serve as tools for studying cell–matrix interactions. This project investigates how ECM-derived peptides influence macrophage polarization, particularly the differentiation of macrophages into pro-inflammatory (M1) or anti-inflammatory (M2) phenotypes.
Using solid-phase peptide synthesis, peptides were synthesized, cleaved, purified via high-performance liquid chromatography, and verified by mass spectrometry to confirm proper synthesis. Preliminary experimentation showed that RGD sequences showed the most promise in producing an effect on macrophages in 2D and 3D models, so the project focused on synthesizing RGDS, a scrambled control (RGSD), and KaGWGRGDS. RGDS was dosed to THP-1-derived macrophages in a 2D environment to evaluate its effect on polarization. Preliminary results revealed limited and inconsistent effects of RGDS on macrophage polarization in 2D environments, suggesting that soluble dosing alone may not effectively engage cell–matrix signaling pathways. Macrophage polarization was verified through flow cytometry and ELISA protocols to analyze surface markers and protein secretion, respectively. Moving forward, research will prioritize incorporating ECM-derived peptides into 3D hydrogels to better mimic physiological conditions. Additionally, the study will expand to evaluate other ECM-derived sequences such as IKVAV, GFOGER, and YIGSR to more comprehensively assess how specific peptide cues influence macrophage phenotype and function.