The transfection efficiency of newly developed calcium phosphate nanoparticles in reprogramming of fibroblast cells

Abstract

Induced pluripotent stem cells (iPSCs) represent a groundbreaking advancement in stem cell research, offering new avenues for regenerative therapy and disease modeling. This study explores the application of calcium phosphate (CaP) nanoparticles in gene transfer studies involving iPSCs. By meticulously analyzing the characteristics of CaP nanoparticles-such as mean particle size and zeta potential-using dynamic light scattering (DLS), the research provides insights into their potential application in iPSC-related research. In addition, in vitro assessments were conducted on L929 mouse fibroblast cells to evaluate the biocompatibility of CaP nanoparticles, further demonstrating their potential utility. The results from the MTT assay indicate no significant toxic effects across various concentrations (ranging from 5 to 25 mu g/mu L), highlighting their safety profile and supporting their use in iPSC-related studies. Additionally, the CaP nanoparticle group exhibited lower total oxidant levels, suggesting potential antioxidant properties or the ability to mitigate oxidative stress. This reduction in oxidant levels may contribute to maintaining cell health and normal cellular functions, complemented by higher total antioxidant levels observed in the CaP group. Moreover, increased levels of cyclin D1 in the CaP group indicate enhanced cellular activity in proliferation and division processes, particularly significant during the G1 phase of the cell cycle. In summary, calcium phosphate nanoparticles play a multifaceted role in iPSC-related research, showcasing antioxidant properties, supporting cell proliferation, and potentially enhancing cell survival by inhibiting apoptosis. This research underscores their efficacy as non-viral carriers for genetic studies and sheds light on their application in gene transfer experiments. Successful transfection of L929 cells with a selected plasmid encoding Oct4-Sox2-Klf4-Myc-GFP demonstrates the effectiveness of CaP nanoparticles in delivering genetic material into cells. With a transfection efficiency persisting at 85% (+/- 3.4%) over a 72-h observation period, coupled with significant expression levels of key genes OCT-4 and SSEA-4 detected via flow cytometry (88%), CaP nanoparticles emerge as a promising tool for facilitating cellular reprogramming.