Polycyclic aromatic hydrocarbons (PAHs) in atmospheric particulate matter have adverse effects on human health, yet total PAH concentrations should overestimate the toxicity compared to the bioavailable amount of PAHs. To explore this hypothesis, we measured PAHs oral bioavailability in vitro in particulate matter with aerodynamic diameter lower than 10 µm (PM10) using a test that mimics the human digestive system. This assay combines the use of simulated gastrointestinal fluids and a dialysis membrane to simulate intestinal absorption. Results show that oral PAH bioavailability was below 5%, with fluorene, anthracene, acenaphthene and phenanthrene as the most bioavailable PAHs. Data suggest no carcinogenic risk of oral bioavailable PM10-bound PAHs following a health risk assessment via inhalation-ingestion by using benzo(a)pyrene-equivalent carcinogenic concentration and hazard indexes. To our best knowledge, this is the first research study of in vitro oral bioavailability estimation of PM10-associated PAHs.