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Cultured meat production requires scaffolds that mimic the body's natural environment, such as hydrogel scaffolds, which guide the development of satellite muscle cells into new muscle fibers. In native muscle, the extracellular matrix (ECM) transmits physical and chemical signals to cells that regulate cell growth and differentiation. During myogenesis, the cells sense the stiffness of the environment, which influences their progression; therefore, replicating that stiffness in the hydrogel is key to achieving efficient muscle development. This study presents the synthesis and characterization of thiol-functionalized silica nanoparticles (fSNPs) in terms of size, morphology, and surface chemistry using dynamic light scattering, scanning and transmission electron microscopy, and Ellman's reagent to quantify thiol density. These fSNPs were incorporated into a polyethylene glycol-norbornene (PEG-N) hydrogel, crosslinked via thiol:ene click chemistry and designed to degrade enzymatically. Hydrogel stiffness was evaluated through bulk rheology in both dry and swollen states to assess the impact of nanoparticle incorporation. The incorporation of fSNPs is shown to increase hydrogel stiffness in a tunable manner. A live/dead assay was conducted with human mesenchymal stem cells (hMSCs) to ensure cell viability after 24 hours, with no observed cell death. Preliminary results indicate that stiffness increases with fSNP incorporation. This approach provides a versatile and reproducible platform for the design of tunable structural supports, with potential applications in cellular agriculture, tissue engineering and development of advanced biomimetic systems. Keywords: Nanoparticles; Cultured Meat; Hydrogel Scaffolds; Thiol-Functionalized

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Purdue University / 2025

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