title: Polybrene: Observations on cochlear hair cell necrosis and minimal lentiviral transduction of cochlear hair cells.
abstract: Polybrene is widely used to enhance viral transduction; however, little is known about the utility thereof, in enhancing lentiviral transduction of cochlear cells. In the present study, we examined the cytotoxic effects of polybrene, and the further effects thereof, on lentiviral transduction of cochlear cells, especially sensory hair cells. Cochlear basilar membranes of newborn rats were cultured and treated with 0.1-10 μg/mL polybrene for 24h to explore the potential development of ototoxicity. PI staining and TUNEL detection were used to evaluate necrosis or apoptosis of hair cell. Various doses of lentivirus-GFP were added to cochlear organotypic cultures with safe concentrations of polybrene, incubated for 24h, and cultured (in the absence of the virus and polybrene) for a further 48 h. Transduction efficiencies were evaluated. The results showed that polybrene at 0.1 μg/mL was safe to cochlear cells, and 0.5-10 μg/mL concentration induced hair cell necrosis in a dose-dependent manner. However, supporting cells were not damaged. Lentiviral vectors transduced into cochlear cells and 0.1 μg/mL polybrene enhanced transduction efficiency. However, hair cells were hardly transduced with lentiviral vectors either alone or in the presence of 0.1 μg/mL polybrene. The use of polybrene to aid lentiviral transduction of cochlear hair cells requires further attention.
authors.affiliation: Department of Otorhinolaryngology, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, China.
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Han
authors.affiliation: Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, Otolaryngology Institute of Shanghai Jiao Tong University, Shanghai 200233, China.
authors.forename: Dongzhen
authors.initials: D
authors.lastname: Yu
authors.affiliation: Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, Otolaryngology Institute of Shanghai Jiao Tong University, Shanghai 200233, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Song
authors.affiliation: Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, Otolaryngology Institute of Shanghai Jiao Tong University, Shanghai 200233, China.
authors.forename: Jiping
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Otorhinolaryngology, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, China. Electronic address: dongpin64@aliyun.com.
authors.forename: Pin
authors.initials: P
authors.lastname: Dong
authors.affiliation: Department of Otorhinolaryngology, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, China. Electronic address: entdoctorhe@163.com.
authors.forename: Jingchun
authors.initials: J
authors.lastname: He
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006198:Hair Cells, Auditory / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006583:Hexadimethrine Bromide / Q000633:toxicity*; D016086:Lentivirus / Q000235:genetics*; D009336:Necrosis; D017207:Rats, Sprague-Dawley; D046509:Tissue Culture Techniques; D014161:Transduction, Genetic
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D006583:Hexadimethrine Bromide
keywords: Cochlear; Gene transduction; Lentivirus; Ototoxicity; Polybrene