Assessing Prussian Blue Nanoparticle Labeling of Mesenchymal Stem Cells: In‑Vitro Effects on Viability, Proliferation, Migration, Differentiation, Cytoskeleton, and Protein Expression

Abstract

Mesenchymal stem cells (MSCs) are widely applied in regenerative medicine, yet their post‑transplant fate remains poorly understood. Magnetic resonance imaging (MRI) offers a non‑invasive means to track transplanted MSCs, and Prussian blue nanoparticles (PBNPs) have emerged as a promising MRI contrast agent. In this study, we synthesized PBNPs and evaluated their influence on mouse MSCs (C3H10T1/2) in vitro. Real‑time cell analysis (RTCA) revealed that PBNPs did not alter cell viability, proliferation, or migration. Oil Red O and Alizarin Red staining confirmed preserved adipogenic and osteogenic differentiation. Phalloidin staining and scanning electron microscopy (SEM) showed intact cytoskeletal architecture. Western blotting indicated unchanged expression of β‑catenin and vimentin. In vitro MRI of PBNP‑labeled cell pellets displayed clear T2 signal attenuation, even after 14 days of osteogenic induction. Collectively, PBNPs can effectively label MSCs without compromising their biological properties, supporting their use for cell tracking in regenerative therapies.

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