Cigarette smoke (CS) is a risk factor for cardiovascular and chronic obstructive pulmonary disease diseases. Being a complex aerosol with over 6,000 chemicals, it is difficult to determine the contribution of individual constituents to overall CS toxicity. The biological impact of total particulate matter (TPM) from the reference cigarette 3R4F and prototypic modified risk tobacco products (MRTPs) was investigated in normal human bronchial epithelial (NHBE) and human coronary artery endothelial (HCAEC) cells, two cell types relevant to respiratory and vascular diseases. Using an impedance-based real-time cellular analysis system, we optimized the experimental conditions (e.g., dose-ranges and exposure times) for subsequent high content screening (HCS) analysis. Toxicological HCS assessment was conducted using seven multi-parametric indicators of cellular toxicity. In the case of 3R4F, the results showed a clear dose-response and cytotoxicity at doses of TPM above 100µg/ml. No effects were observed for the TPM from prototypic MRTPs. Furthermore, HCAEC appeared to be more sensitive than NHBE cells in their response to TPM. The combination of impedance-based and HCS technologies provides a robust scientific basis for in vitro systems biology-based risk assessment for MRTPs.