Fig 1.
A) Schematic diagram of the blast wave generator (front view). The dimension is in millimetres (mm). B) The blast wave generator includes pressure transducers at different positions. A cracking pressure transducer in the pressure reservoir, two overpressure sensors on the left and right sides of the rat and a transducer for recording the breathing frequency.
Fig 2.
Histopathology of rat blast injury.
Rats were exposed to the pressure wave at 3.5 cm from the nozzle. Animals were sacrificed either 10 min A), 3 hours B) or 24 hours C) after blast exposure, non-traumatized animals served as sham controls E) + D). Visualized at an original magnification of 40-fold A)–C) + E) and 10-fold in D).
Fig 3.
Blast injury-related protein accumulation in BALFs.
Animals were sacrificed 10 min (n = 5), 1.5 h (n = 3), 3 h (n = 3), 6 h (n = 3), 12 h (n = 4), 24 h (n = 3) or 96 hours (n = 3) after blast exposure, non-traumatized animals served as sham controls (n = 9). Protein concentration was determined in the BALF. Data were analysed by one-way ANOVA and Dunnett’s multiple comparison test: **p <0.01 vs. control. Data represent means ± SEM, number of individual experiments (n) in parentheses.
Fig 4.
Blast injury-related pulmonary haemorrhage.
The animals were sacrificed 10 min (n = 5), 1.5 h (n = 3), 3 h (n = 3), 6 h (n = 3), 12 h (n = 3), 24 h (n = 3) or 96 hours (n = 3) after blast exposure, non-traumatized animals served as sham controls (n = 8). RBCs were determined in BALFs. Data were analysed by one-way ANOVA and Dunnett’s multiple comparison test: *p <0.05 and **p <0.01 vs. control. Data are means ± SEM, number of experiments (n) in parentheses.
Fig 5.
LDH release after thoracic trauma.
The animals were sacrificed 10 min (n = 5), 1.5 h (n = 3), 3 h (n = 3), 6 h (n = 3), 12 h (n = 4), 24 h (n = 3) or 96 hours (n = 3) after blast exposure, non-traumatized animals served as sham controls (n = 8). LDH activity was determined in BALFs. Data were analysed by one-way ANOVA and Dunnett’s multiple comparison test: *p <0.05 and **p <0.01 vs. control. Data are means ± SEM, number of experiments (n).
Fig 6.
Thorax trauma-induced weight gain.
Lung wet weight and dry weight were determined at different time-points after pressure wave exposure. The wet to dry weight ratios were calculated. Data were analysed by one-way ANOVA and Bonferroni’s multiple comparison test: **p <0.01 1.5 h vs. sham control and 6 h after blast, respectively.
Fig 7.
Prostanoid release in the early phase after thoracic trauma.
The animals were sacrificed 10 min (n = 6), 1.5 h (n = 3), 3 h (n = 3), 6 h (n = 3), 12 h (n = 4), 24 h (n = 3) or 96 hours (n = 3) after blast exposure, non-traumatized animals served as sham controls (n = 9). A) TxB2 release and B) 6-keto PGF1α were determined in BALFs. Data were analysed by one-way ANOVA and Tukey’s multiple comparison test. *** p <0.001: 10min vs. all other groups. Data represent means ± SEM, number of experiments (n) in parentheses.
Fig 8.
Time course of TNF and IL-6 release after thoracic trauma.
The animals were sacrificed 10 min (n = 6), 1.5 h (n = 3), 3 h (n = 3), 6 h (n = 3), 12 h (n = 4), 24 h (n = 3) or 96 hours (n = 3) after blast exposure, non-traumatized animals served as sham controls (n = 9). A) TNF and B) IL-6 release were determined in BALFs. Data were statistically analysed by one-way ANOVA and Dunnett’s multiple comparison test: *p <0.05 and ** p <0.01 vs. sham control. Data represent means ± SEM, number of experiments (n) in parentheses.
Fig 9.
Decrease in BAL IL-10 after thoracic trauma.
The animals were sacrificed 10 min (n = 6), 1.5 h (n = 3), 3 h (n = 3), 6 h (n = 3), 12 h (n = 4), 24 h (n = 3) or 96 hours (n = 3) after blast exposure, non-traumatized animals served as sham controls (n = 9). IL-10 release was assessed in BALFs. Data were statistically analysed by one-way ANOVA and Dunnett’s multiple comparison test: *p<0.05 and **p<0.01 vs. sham control. Data are means ± SEM, number of experiments (n) in parentheses.
Fig 10.
Time course of sTNFR p55 and p75 BAL concentrations in A) as well as the p55/ p75 ratio in B). The animals were sacrificed 10 min (n = 6), 1.5 h (n = 3), 3 h (n = 3), 6 h (n = 3), 12 h (n = 4), 24 h (n = 3) or 96 hours (n = 3) after blast exposure, non-traumatized animals served as sham controls (n = 9). Levels of sTNFR p55 and p75 were assessed in BALFs. Data were analysed by one-way ANOVA and Dunnett’s multiple comparison test: *p <0.05 and **p <0.01 vs. sham control. Unpaired t-test was performed to compare differences in p55 and p75 levels at different time-points: **p = 0.0074 for p75 vs. p55 and *p = 0.04 for p75 versus p55 at 24 hours and 96 hours after blast, respectively. Data are means ± SEM, number of experiments (n) in parentheses. Linear regression between p55 and p75 release revealing a Pearson r of r = 0.96; p <0.0001.
Fig 11.
Time-course of neutrophil (PMN) infiltration into the alveoli after thoracic trauma.
The animals were sacrificed 10 min (n = 6), 1.5 h (n = 3), 3 h (n = 3), 6 h (n = 3), 12 h (n = 4), 24 h (n = 3) or 96 hours (n = 3) after blast exposure, non-traumatized animals served as sham controls (n = 9). Statistical analysis was performed by one-way ANOVA and Dunnett’s multiple comparison test: * p <0.05 vs. sham control. Data are expressed as mean ± SEM, number of experiments (n).
Fig 12.
Cytospin pictures after blast exposure.
A) sham control, B) 24 hours after blast exposure. AM: alveolar macrophages; Neu: neutrophils; Lym: lymphocytes; RBC: red blood cells.
Table 1.
Summary of time-dependent BAL findings compared to non-traumatized sham controls.
Fig 13.
CINC-3 measurement in the BALF of traumatized rat lungs at different time-points after blast exposure: 10 min (n = 6), 1.5 h (n = 3), 6 h (n = 3), 12 h (n = 4), 24 h (n = 3), 24 h (n = 3) and 96 h (n = 3).
Non-traumatized animals served as sham controls (n = 9). Statistical analysis was performed by one-way ANOVA (p = 0.0014) and Dunnett’s multiple comparison test: **p <0.01 vs. sham control. Data are expressed as mean ± SEM, number of experiments (n).