Anthocyanin Encapsulated Nanoparticles as a Pulmonary Delivery System

Figure 5

The effect of carboxymethyl chitosan (CMC) nanoparticles (E-CMC), HB encapsulated with CMC (HB-CMC), and an equivalent dose of free HB-anthocyanin (HB) against NNKOAc-induced oxidative stress in BEAS-2B cells. 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) Intracellular reactive oxygen species (ROS) levels were assessed by measuring the dichlorofluorescein status in treated cells by flow cytometry. The CMC nanoparticles reduced (graphs skewed towards left) the NNKOAc-induced ROS generation (skewed towards the right). (b) The expression of p-Nrf2 and antioxidant enzymes, determined by western blot. (c–g) The bar graphs present the quantitative data of the DCFDA flow cytometry data and the expression of p-Nrf2, superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx). The results represent three independent experiments. One-way analysis of variance was performed () with Bonferroni pairwise comparison (at ) for mean comparison. , , and indicate statistical differences at , 0.01, and 0.001, respectively, with . NS: results do not significantly different; E-CMC: anthocyanin-free carboxymethyl chitosan nanoparticles; HB-CMC: haskap berry anthocyanin encapsulated nanoparticles; HB: haskap berry anthocyanin; DCFDA: 2,7-dichlorofluorescein diacetate; C3G: cyanidin-3-O-glucoside.

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