Modification of primary amines to higher order amines reduces in vivo hematological and immunotoxicity of cationic nanocarriers through TLR4 and complement pathways

Randall Toy; Pallab Pradhan; Vijayeetha Ramesh; Nelson C. Di Paolo; Blake Lash; Jiaying Liu; Emmeline L. Blanchard; Philip J. Santangelo; Dmitry M. Shayakhmetov; Krishnendu Roy

The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/647305 doi: bioRxiv preprint The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/647305 doi: bioRxiv preprint 30 and 150 minutes after bPEI nanoparticle injection (n=4-5). Outliers were identified and removed using the Grubbs test (p=0.1). Error bars represent SD of the mean. Statistical differences in experiments between two groups were determined using a 2-tailed, unpaired Student's t-test assuming unequal variance. Statistical differences in experiments between more than two groups were determined using one-way ANOVA followed by Tukey's test. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/647305 doi: bioRxiv preprint bPEI nanoparticles in RAW 264.7 macrophages using a flow cytometry-based proximity ligation assay (n=3-4). Outliers were identified and removed using Grubbs' test (p=0.1). (B-G) Mouse BMDCs were treated with unmodified bPEI nanoparticles (bPEI-TPP), IAA-modified bPEI nanoparticles (bPEI-IAA-TPP), unmodified chitosan nanoparticles (Ch-TPP), or IAA-modified chitosan nanoparticles (Ch-IAA-TPP) for 24h. Cell supernatants were collected and IL-6, IL-1 and TNF-were measured (n=5). Error bars represent SD of the mean. Statistical differences in experiments between two groups were determined using a 2-tailed, unpaired Student's t-test assuming unequal variance. Statistical differences in experiments between more than two groups were determined using one-way ANOVA followed by Tukey's test. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/647305 doi: bioRxiv preprint