The effects of acute hydrogen peroxide exposure on respiratory cilia motility and viability

COVID-19 has seen the propagation of alternative remedies to treat respiratory disease, such as nebulization of hydrogen peroxide (H₂O₂). As H₂O₂ has known cytotoxicity, it was hypothesised that H₂O₂ inhalation would negatively impact respiratory cilia function. To test this hypothesis, mouse tracheal samples were incubated with different H₂O₂ concentrations (0.1-1%) then cilia motility, cilia generated flow, and cell death was assessed 0-120 min following H₂O₂ treatment. 0.1-0.2% H₂O₂ caused immediate depression of cilia motility and complete cessation of cilia generated flow. Higher H₂O₂ concentrations (≥0.5%) caused immediate complete cessation of cilia motility and cilia generated flow. Cilia motility and flow was restored 30 min after 0.1% H₂O₂ treatment. Cilia motility and flow remained depressed 120 min after 0.2-0.5% H₂O₂ treatment. No recovery was seen 120 min after treatment with ≥1% H₂O₂. Live/dead staining revealed that H₂O₂ treatment caused preferential cell death of ciliated respiratory epithelia over non-ciliated epithelia, with 1% H₂O₂ causing 35.3 ± 7.0% of the ciliated epithelia cells to die 120 min following initial treatment. This study shows that H₂O₂ treatment significantly impacts respiratory cilia motility and cilia generated flow, characterised by a significant impairment in cilia motility even at low concentrations, the complete cessation of cilia motility at higher doses, and a significant cytotoxic effect on ciliated respiratory epithelial cells by promoting cell death. While this data needs further study using in vivo models, it suggests that extreme care should be taken when considering treating respiratory diseases with nebulised H₂O₂.