In vitro prediction of inflammatory lung effects of well-dispersed nanomaterials is challenging. Here, the in vitro effects of four colloidal amorphous SiO₂ nanomaterials that differed only by their primary particle size (9, 15, 30, and 55 nm) were analyzed using the rat NR8383 alveolar macrophage (AM) assay. Data were compared to effects of single doses of 15 nm and 55 nm SiO₂ intratracheally instilled in rat lungs. In vitro, all four elicited the release of concentration-dependent lactate dehydrogenase, β-glucuronidase, and tumor necrosis factor alpha, and the two smaller materials also released H₂O₂. All effects were size-dependent. Since the colloidal SiO₂ remained well-dispersed in serum-free in vitro conditions, effective particle concentrations reaching the cells were estimated using different models. Evaluating the effective concentration-based in vitro effects using the Decision-making framework for the grouping and testing of nanomaterials, all four nanomaterials were assigned as "active." This assignment and the size dependency of effects were consistent with the outcomes of intratracheal instillation studies and available short-term rat inhalation data for 15 nm SiO₂. The study confirms the applicability of the NR8383 AM assay to assessing colloidal SiO₂ but underlines the need to estimate and consider the effective concentration of such well-dispersed test materials.
Keywords: 3R method; TNFα; alveolar macrophage; dosimetry; in vitro cytotoxicity; intratracheal instillation; nanomaterial grouping; regulatory hazard assessment; short-term inhalation study (STIS).