Anthocyanin Encapsulated Nanoparticles as a Pulmonary Delivery System

Figure 7

The protective effect of carboxymethyl chitosan (CMC) nanoparticles (E-CMC and HB-CMC) and an equivalent dose of free HB-anthocyanin (HB) against NNKOAc-induced DNA damage in BEAS-2B cells. The protective effect was measured by alkaline and neutral comet assays. BEAS-2B cells were treated with 2 mg/mL E-CMC, HB-CMC (comprised 4 μg/mL C3G equivalent anthocyanin), and 4 μg/mL HB-anthocyanin for 3 h separately, following exposure to 100 μM NNKOAc for another 3 h. (a) Alkaline comet mages represent both single- and double-strand breaks. The HB-CMC nanoparticles significantly reduced NNKOAc-induced tail length (b), tail DNA percentage (c), and tail moment (d) as quantified by alkaline comet assay. (e) Neutral comet images represent the DNA double-strand breaks. The HB-CMC nanoparticles did not attenuate the NNKOAc-induced tail length (B), tail DNA percentage (c), and tail moment (d) as quantified by neutral comet assay. Nuclear DNA comets were stained with SYBR Green I dye, and images were taken by fluorescence microscope. The scale =100 μm. The comet tail length, percent DNA in tail, and tail moment (tail moment = measure of tail length × measure of DNA in the tail) were quantified in (30–100 nuclei from each treatment) using Fiji ImageJ and Open Comet Version 1.3 software. The results represent three independent experiments (). One-way analysis of variance was performed () with Tukey’s pairwise comparison (at ) for mean comparison. and indicate statistical differences at and 0.001, respectively, with . NS: results do not significantly different.

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