Structural firefighting exposes firefighters to intense physical and thermal stress, and emissions of health-burden compounds [e.g., particulate matter and polycyclic aromatic hydrocarbons (PAHs)]. The use of a self-contained breathing apparatus (SCBA) is mandatory during structure fires, although in the overhaul phase, firefighters often remove it. The characterization of inhaled PAHs during structure fires remains scarce, particularly for European firefighters. This study assessed, for the first time, the levels of PAH in the air of firefighters outside and inside the SCBA during training firefighting activities and at fire stations on regular working days. The control group included 13 male volunteer firefighters without active participation in firefighting in the previous 7 days, while the exposed group included 31 professional sapper firefighters. Outside PAHs were monitored with a pre-treated polyurethane foam (PUF), placed in the personal protective equipment, while a cotton disk was placed inside the SCBA. PAHs were quantified by high-performance liquid chromatography. Total PAH levels in PUFs of exposed firefighters were higher than PUFs from the control group (11.5–28.0 µg/m³ versus 1.41 µg/m³), and significantly higher than levels detected inside the SCBA (1.96-10.40 ng/m³). Regarding carcinogenic PAHs, levels quantified outside the SCBA were higher for exposed firefighters than the control group (5.80-17.2 µg/m³ versus 0.84 µg/m³), and superior to the inside SCBA’ levels (0.77-0.93 ng/m³) (p<0.005). International occupational guidelines were not exceeded. The inside–outside ratios (<1) suggested that ambient PAHs contributed to the levels quantified inside the SCBA. Extracts collected outside and inside the SCBA caused cytotoxicity in human alveolar and bronchial cell lines. Moreover, the collected samples were applied to an in vitro co-culture air–blood barrier model to assess permeability. Carcinogenic benzo(a)pyrene (100%) permeated the model. Overall, the negative effects of fire emissions on human respiratory cell lines were confirmed, suggesting that fire pollutants contribute to firefighters’ respiratory health burden.