Flexivent device

WT and Adam9^−/− mice were anesthetized with i.p. injections of ketamine (100 mg/kg) xylazine (10 mg/kg), and acepromazine (3 mg/kg) and 30–100 mU of bleomycin in 30 μl of endotoxin-free saline or 30 μl of saline alone was delivered to WT vs. Adam9^−/− mice by the IT route. In one cohort of mice, weight loss and mortality was monitored over 21 days. Other cohorts of mice were euthanized at intervals over 10 days and BAL was performed. Absolute numbers of PMN, macrophages, monocytes, and lymphocytes were counted in BAL samples 3, 7, and 10 days after instilling bleomycin.
In other cohorts of mice, respiratory mechanics (lung elastance, lung resistance, and quasi-static compliance) were measured on anesthetized and tracheostomized mice using a digitally-controlled mechanical ventilator (FlexiVent device; SCIREQ, Inc., Montreal, Quebec, Canada) 7 days after instilling 100 mU of bleomycin or saline, exactly as described previously (41 (link)). Wet-to-dry lung weight ratios were also measured after euthanizing the animals.
Collagen deposition in the lung was measured 21 days after delivering bleomycin or saline by measuring hydroxyproline levels in hydrolysates of left lungs and staining formalin-fixed right lung sections with Masson’s trichrome stain, as described previously (42 (link)).