EHC-93, EHC-98, EHC-2000, SRM-1648, SRM-1649 or TiO2 (?), Tukey test, p?0.05, Dose main effect, p?0.001, Doses 0 or 10 vs. SigmaPlot, version 12.5 (Systat Software, Inc., San Jose, CA, USA). Hierarchical clustering of the pattern of cytokine secretion by A549 and J774 in response to particle exposure were conducted using the GenePattern webtool (http://www.broadinstitute.org/cancer/software/genepattern) , and visualized as heatmaps using Java TreeView plugin version 1.16.r2 (http://jtreeview.sourceforge.net) . Linear regression between corresponding individual or combined potency estimates in vitro and in vivo was conducted using Sigmaplot v12.5 and depicted using Microsoft Excel 2010 (Microsoft Corp., Redmond, WA, USA). The strength of the relationship between every two variables was described by a correlation coefficient R and the significance of the hypothesis test by the p-value of 0.05 (two-tailed test), or one-tailed test, where applicable (i.e. consistent directionality of the variables). The correlations presented are performed between in vitro and in vivo matched endpoints across all particles, based on individual particle potencies (Table?1) or combined potency estimates (average of endpoints) for toxicity, inflammation, or integrated Ziprasidone D8 inflammation plus toxicity (Tables?2, ?,3,3, and ?and5)5) all eight particles tested in vitro (EHC-93, EHC-98, EHC-2000, SRM-1648, SRM-1649, DWR1, TiO2, SiO2) or for five particles tested in vivo (EHC-2000, SRM-1649, DWR1, TiO2, SiO2) for Table?5. Table 1 Pearson correlations for cytotoxic potency and cytokine inductions in cell lines exposed to particles (2-tailed) (2-tailed) (2-tailed) (2-tailed) not detectable In vitro integrated particle potency To summarize all cytotoxic particle effects in Ziprasidone D8 vitro across cell types, biological reactivity R CELLS was derived by averaging the absolute values of the cytotoxic potencies, V of the particles in A549 and J774A.1 cells and the AhR activity in H1L1.1c2 cells, assuming biological reactivity of the particles as any deviation from baseline. From the R CELLS estimate, SRM-1648 and SRM-1649 particles had the highest overall cell potency in contrast to DWR1 and CRI particles which showed the lowest values (Fig.?5a). Open in a separate window Fig. 5 Biological reactivity estimate, R CELLS was derived by averaging the cytotoxic potencies of the particles in A549 and J774A.1 cells, as well as the AhR activity in H1L1.1c2 cells (a). A combined inflammatory estimate, I-V CELLS, was obtained for each particle by averaging particle inflammatory potency estimates adjusted for cell viability of J774A.1 and A549 cells. The I-V LO represents the lower estimate (IL-10 and IL-6 potency subtracted from average potency), while I-V HI is the higher estimate (IL-10 potency subtracted, IL-6 added to average potency) (b). An integrated potency estimate of the particles, I CELLS was determined by averaging the biological reactivity Ziprasidone D8 (cytotoxic potency) and inflammatory estimates for each particle. I LO represents the lower estimate (IL-10 and IL-6 potency subtracted from average potency). I HI Gja7 represents the higher estimate (IL-10 potency subtracted, IL-6 added to average potency) (c) Similarly, an inflammatory potency estimate I-V CELLS, was obtained for each particle by averaging the cell viability-adjusted inflammatory potency values Ziprasidone D8 of the particles across both cell lines for all cytokines detected, for each inflammatory scenario (Fig.?5b). The estimates impacted the magnitude of particle potency ranking, but the ranking was comparable between the different scenarios, where the urban particles, except EHC-93 were more potent than mineral particles and DWR1. Lastly, an overall integrated potency estimate, I CELLS of the particles was calculated by averaging the biological reactivity, R CELLS with inflammatory indices, I-V CELLSfor each particle (Fig.?5c). The I CELLS profile was similar to the profile of the inflammatory indices I-V CELLS. Toxicity of particles in vivo BALB/c mice were exposed for 24?h to a subset of particles selected based on contrasting in vitro potency (EHC-2000, SRM-1649, CRI, TiO2 and DWR1) by intratracheal instillation (IT). Ziprasidone D8 Acute pulmonary toxicity was observed in mice exposed to the high dose (250?g) of the particles, as shown by a general increase.