Background: Poly (β-amino Ester) nanocarriers show promise for gene therapy, but their effectiveness can be limited by the environment within the body. This study aims to understand how common cell culture media components affect optimized PBAE nanocarrier performance in gene delivery.
Methods: Optimized PBAE was synthesized based on Michael addition reaction and characterized by different assays, this study employed techniques like DLS and TEM to characterize PBAE nanocarriers, followed by cellular uptake analysis (flow cytometry and confocal imaging) and evaluation of gene expression under different polymer/DNA ratio ratios and media conditions.
Results: The nanocarriers exhibited size under 200 nm and surface positive charge, with high encapsulation efficiency (up to 95%). Cellular uptake, transfection efficiency, and cytotoxicity were evaluated. Flow cytometry analysis revealed high cellular uptake (over 77% at 1 hour and up to 95% after 3 hours) and good viability. Transfection efficiency reached up to 80% with 2 μg DNA, particularly at weight ratios of 60 and 90.
Conclusion: The study also identified factors affecting transfection efficiency, including serum concentration and antibiotics in the culture medium, highlighting the importance of optimizing these conditions for future applications.
Keywords: Gene delivery; PBAE; extracellular conditions; nanocarriers; polymeric nanoparticles.
Our study focused on a new way to deliver medicine directly to cells using tiny particles called nanocarriers. These tiny particles are made from a special material called PBAE. Our study found that certain things around these nanocarriers, like antibiotics and high levels of a substance in the cell medium, can make it harder for the medicine to get into the cells. We also found that while the PBAE material is safe on its own, it can become more harmful when used in the nanocarriers at high amounts. By figuring out the best conditions for these nanocarriers, we hope to make gene therapy, a treatment for diseases like cancer, work better and be safer.