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.

Nanomaterials for Inner Ear Diseases: Challenges, Limitations and Opportunities

“…No cytotoxicity was observed even at 4 mg/mL, but the vector significantly prolonged drug release and action time. Li et al [ 71 ] prepared a novel non-viral vector (hyaluronic acid-modified polyethylene imine nanoparticles) for cochlear gene transfection, and the transfection efficiency of the basal membrane was up to 81.7 ± 4.71%, with no obvious toxicity to the basal membrane cells. Xu et al [ 72 ] used superparamagnetic iron oxide nanoparticles to label mesenchymal stem cells to evaluate the therapeutic effect of sensory neural hearing loss and found that the nanoparticle labeling efficiency was up to 95% and can also supplement iron.…”

Novel Nanoencapsulation Technology and its Potential Role in Bile Acid‐Based Targeted Gene Delivery to the Inner Ear

“…Both positive and negatively charged nanoparticles have been explored for therapy delivery into the inner ear. [ 116,141 ] Positively charged nanoparticles have been reported to show improved permeation at the expense of increased toxicity. Here, positively charged nanoparticles were hypothesized to bind to the negatively charged RWM and result in improved permeation.…”

Low-Molecular-Weight Hyaluronic Acid Contributes to Noise-Induced Cochlear Inflammation

“…In recent years, hyaluronic acid (HA) has often been used as an important adjuvant agent in the management of different inner ear problems, for example, in cochlear implant electrode insertion or gene delivery [Abi Zeid Daou and Bassim, 2020;Li et al, 2020]. Our previous study demonstrated that increased HA was involved in the toxicity of silver nanoparticles to the kidney and cochlea [Feng et al, 2015].…”