Figure 1. Effects of silicone breast implant infused media on adverse outcomes in C elegans:(A) Brood size was estimated after 48-hour exposure to SBI-infused media. The number of progeny was classified into bins, where there was expected to be more than 50 viable offspring in control conditions (bin 3, green bars). If 10-50 progeny were produced, this was shown with blue bars (bin 2). Orange bars (bin 1) indicate up to 10 viable offspring and red bars are indicative of no viable progeny (bin 0). No acute toxicity of the parents was observed in any condition. For each condition, n>26 from 2 independent biological replicates and the percentage of worms in each bin is shown. A
chi2 test for homogeneity resulted in a p=0.01. (B) The health of the offspring was assessed by observing their mobility. A healthy worm displays a rapid head-to-tail body bend and, this is observed in 100% of the offspring (bin 3, green bars) or in less than 50% of the offspring (bin 2, blue bars). Abnormal mobility was noted when the worms had an uncoordinated thrashing motion which is indicative of neurotoxicity, shown by orange bars (bin 1) and dead nematodes are indicated with red bars (bin 0, there were none in this category). For each condition, n>26 from 2 independent biological replicates and the percentage of worms in each bin is shown. Statistical analysis (
chi2 test) gives p<0.0001. (C) Modified Oil Red O (MORO) staining of C. elegans indicates that silicones are located in cryosections of 48-hour exposed animals. (i) Three sections of C. elegans exposed to control conditions. Some small areas of positive MORO stain are indicated by arrows. (ii) Animals exposed to 17-day SBI-infused S-complete show a clear and distinctive MORO stain, that is positive for silicone, highlighted with an arrow. Scale bar is 50µm. A table to show the number of samples which had MORO stain present (detected) or not (undetected). In the control worms, the detected MORO stain is at such low levels this is considered background. (D) Nematodes are placed on silicone breast implant infused NGM as L4s and incubated at 20oC for 48 hours. The cytoplasmic expression of DAF-16::GFP (light grey bars) is a result of the absence of stress. As the conditions become more stressful, the expression of the reporter becomes more nuclear (black bars). Intermediate localisation (dark grey bars) indicates some animals are stressed as their DAF-16::GFP can be found in both the nucleus and cytoplasm in the same worm. For each condition, n>30 in 1 biological replicate. The percentage of worms in each category is shown with p<0.001 indicated by a
chi2 test for homogeneity.