ABSTRACT Optoporation enables high‐precision delivery of biomolecules for treating diseases through cell therapy, gene editing, and personalized therapeutics. Nanoparticle‐based optoporation offers high transfection efficiency by minimizing cell damage, thus enhancing the overall cellular health. This study demonstrates the versatility of using Titanium nitride nanoparticles (TiN NPs)‐mediated optoporation for the delivery of small to large cargos, achieving high transfection efficiency alongside cell viability. TiN NPs adsorbed on the cell membrane, when irradiated with a laser fluence of 12.8 mJ/cm 2 at an 850 nm wavelength, generate plasmonic bubbles, facilitating the delivery of cargo into the cells. By using this platform, wide range of cargos/biomolecules such as propidium iodide (PI) dye (668.4 Da), dextran (3 kDa), small interfering RNA (siRNA) (13.3 kDa), enhanced green fluorescence protein (EGFP) expression plasmid DNA (229.4 kDa), and β‐ galactosidase enzyme (465 kDa) are delivered into diverse mammalian cell lines (L929, MG‐63, and N2a) with high transfection efficiency and cell viability. The functionality of the transfected β‐galactosidase enzyme is assessed by measuring its enzymatic activity, verifying the effectiveness of the transfection process. For small PI molecules, MG‐63 cells demonstrated a delivery efficiency of 98% with a cell viability of 99%. In contrast, for oversized cargo (enzyme, 465 kDa), the transfection efficiency and cell viability achieved were 97% and 99%, respectively. Furthermore, human mesenchymal stem cells (hMSCs) are transfected with the EGFP plasmid and β‐galactosidase enzyme, demonstrating a transfection efficiency of 98% and cell viability of 99%. To study the cytotoxicity of TiN NPs, an MTT assay, Kaplan‐Meier survival analysis, and behavioral assessment of the in vivo zebrafish model are conducted. The plots suggest that TiN NPs do not pose neurotoxic effects. Thus, this platform has demonstrated inherent potential for cell reprogramming and applications in medicine, molecular biology, and cellular biology.
Balasubramaniam et al. (2026) studied this question.