Recent discoveries of various forms of carbon nanostructure have stimulated research on their applications and hold promise for applications in medicine and other related engineering areas. Although carbon nanotubes (CNTs) are already being produced on a massive scale, few studies have been performed which test the potential harmful effects of this new technology. The authors used a three-dimensional in vitro model of the human airway using a coculture of normal human bronchial epithelial cells and normal human fibroblasts for the health risk assessment of CNTs on the human respiratory systems. The authors used aqueous single-walled carbon nanotube (SWCNT) solution. The average length and diameter of nanotube ropes were about 500 nm and less than 10 nm, respectively. The authors measured the production of nitric oxide (NO) as an inflammatory marker and mitochondrial activity using MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay as a cytotoxic response of the cell layers following exposure of different concentration of aqueous SWCNT solution. The results indicated that NO production was dramatically increased and cell viability was decreased following exposure of different concentrations of SWCNTs. Transepithelial electrical resistance (TER) across the coculture layers was measured to observe the changes in airway physiological function following exposure of different concentrations of SWCNTs. TER value was dramatically decreased following exposure of 20% SWCNT (8 microg/ml). In this study, the authors presented viable alternatives to in vivo tests to evaluate the toxicity of engineered SWCNTs. Cytotoxic/inflammatory responses and barrier function of the human lung layers following exposure of SWCNTs were observed using in vitro coculture system of airway.