T increase in LDH release from L-2 cells compared to the
T increase in LDH release from L-2 cells compared to the negative control (p < 0.05). Nanoparticle carbon black also resulted in a significant increase in LDH release (p < 0.05) at a particle concentration of 2 mg/ml. High doses of both types of fine particles were also observed to induce similar increases in LDH release but this was not statistically significant. However, maximal LDH release induced by particle treatments was noted to be lower than that observed following cell lysis with Triton X-100 (positive control) possibly due to high particle concentrations interfering with the assay. The negative control in these experiments (untreated cells) showed minimal LDH release. Low doses of particulates (31?00 ) did not appear to show any significant size, composition or surface area-related differences in response.Concentration and time course studies with zymosan activated serum (ZAS) and J774.2 macrophages Figure 2(a) shows the effects of varying concentrations of ZAS on macrophage migration following a 6 hour incuba-tion period. The graph shows that the absorbance increases as the concentration of ZAS increases from 2 to 20 indicating increased numbers of macrophages migrating into or through the polycarbonate filter. Significant increases in macrophage migration were observed at both the 10 and 20 dilutions of ZAS. This was compared to the negative control of foetal bovine serum (FBS) incubated with sterile saline, which showed no significant increases in macrophage migration. Figure 2(b) shows a time course study of macrophage migration towards 10 ZAS. The results indicate that the ZAS induced significant increases (p < 0.001) in macrophage migration at both PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28914615 the 3 hour and 6 hour time points. The highest level of macrophage migration was found at the PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28045099 6 hour time point where migration was found to be 8-fold greater than that of the negative control. The negative control (FBS incubated with sterile saline) did not AMN107 web display any potential for macrophage migration and this may serve as an indicator that random macrophage migration, chemokinesis, was minimal.Page 5 of(page number not for citation purposes)Particle and Fibre Toxicology 2005, 2:http://www.particleandfibretoxicology.com/content/2/1/The other negative control, TNF alone, at comparable concentrations to those used for cell treatments, induced modest macrophage migration.Macrophage migration stimulated by conditioned medium from L-2 Cells exposed to fine and nanoparticle carbon black and TiO2 When compared to the negative control (conditioned medium obtained from untreated cells), nanoparticle carbon black treatment of L-2 cells generated a conditioned medium that induced a significant increase (p < 0.01) in macrophage migration (Figure 4). In comparison, conditioned medium from L-2 cells treated with fine TiO2, CB and nanoparticle TiO2 did not induce any significant increases in macrophage migration. It should also be noted that both fine and nanoparticle carbon black treatment of L-2 cells did induce significant increases in macrophage migration when compared to another negative control; medium incubated with the particles alone (p < 0.01 and p < 0.001 for fine and nanoparticle carbon black respectively). SDS-PAGE electrophoresis of conditioned medium from L2 cells treated with a sub-toxic dose of nanoparticle carbon black A non-reducing SDS-PAGE gel was used to separate the protein components of the conditioned medium obtained from L-2 cells treated for 24 hours with e.
