The complexing of cationic copolymer MPC30-DEA70 with TGF-β1 antisense oligodeoxynucleotide and transfection into cardiomyocytes in vitro.

Although gene therapy is an attractive option for the treatment of cardiovascular diseases, the ideal gene delivery systems are still under investigation and must meet the following criteria: safety, adequate gene transfer efficiency, and stable expression of the transgene for a duration appropriate for treating the disease. In this study, we developed a cationic phosphorylcholine-containing diblock copolymer, namely MPC₃₀-DEA₇₀, as carrier systems to deliver a chemically synthesized transforming growth factor-beta 1(TGF-β1) antisense oligonucleotide (AS-ODN) into cardiomyocytes (CMs) to observe the cell transfection efficiency of MPC₃₀-DEA₇₀ and the inhibition effect on the expression of TGF-β1. MPC₃₀-DEA₇₀/TGF-β1 AS-ODN complexes were formed through complexation between copolymer MPC₃₀-DEA₇₀ (N) and AS-ODN (P) at different N/P ratios and were characterized by DNA electrophoresis. Notably, the cytotoxicity and cell growth inhibition assay showed that the MPC₃₀-DEA₇₀ had low cytotoxicity to CMs within the effective transfection dosage range (<20 μL/mL). CLSM/TEM images displayed that most of the AS-ODN molecules engulfed by cells were located around the cell nuclei, and a few entered into the cell nuclei without harming the organelles in the cell. Transfection studies from CMs indicated a steady increase of transfection efficiency with increasing N/P ratios. The expression levels of TGF-β1 mRNA and protein in CMs were significantly inhibited at high N/P ratios. This study shows that MPC₃₀-DEA₇₀ can function as an effective transgenic vector into CMs and that TGF-β1 AS-ODN delivered by MPC₃₀-DEA₇₀ can silence the expression of the TGF-β1 gene efficiently and specifically and thereafter antagonize TGF-β1-mediated biological function in cardiomyocytes. [ABSTRACT FROM AUTHOR]