Cochlear Transfection Gene Guinea Pigs Mediates Atoh1-EGFP Based Hyaluronic Acid Modified Polyethyleneimine Nanoparticles

Abstract

To modify polyethyleneimine (PEI) nanoparticles using hyaluronic acid (HA) to prepare a novel nonviral vector and use it to coat Atoh1-EGFP plasmid to detect its translocation in living guinea pig cochlea dyeing efficiency. Atoh1-EGFP plasmid was extracted and characterized using a Zetasizer particle size analyzer. HA/PEI/DNA complexion was characterized and introduced into the round window membrane. EGFP green fluorescence carried in the Atoh1 plasmid was observed by confocal microscopy. The transfection results were verified by Western blot and reverse transcription polymerase chain reaction (RT-PCR) from the perspective of protein and nucleic acid to verify its expression results. In this study, HA-modified PEI nanoparticles are negatively-charged nanoscale gene carrier complexes. After the Atoh1-EGFP plasmid was introduced into the cochlea, the results of confocal microscopy showed that the inner and outer hair cells of the basement membrane could be detected in green fluorescent protein. The transfection efficiency of basement membrane is as high as 81.7±4.71%, while the transversion is 33.8±9.02%. Western Blot and RT-PCR also confirmed that the Atoh1 gene can be successfully transfected on the basement membrane. The gene transfection of cochlea may be achieved by HA-modified PEI nanoparticle gene vector with no obvious toxicity to basement membrane cells. It is also an ideal inner-end gene transfection vector owing to its simple synthesis method and low cost.