Solid Lipid Nanoformulation of Berberine Attenuates Doxorubicin Triggered in vitro Inflammation in H9c2 Rat Cardiomyocytes

Abstract

Aim: To evaluate berberine solid lipid nanoparticles' efficacy against doxorubicin-induced cardiotoxicity. Background: Berberine (Ber) is cardioprotective, but its oral bioavailability is low and its effect in chemotherapy-induced cardiotoxicity has not been studied. Objective: Solid lipid nanoparticles (SLNs) of berberine chloride were prepared, characterized and evaluated in vitro against Doxorubicin induced cardiomyocyte injury. Methodology: Berberine loaded SLNs (Ber-SLNs) were synthesized using water-in-oil microemulsion technique with tripalmitin, Tween 80 and poloxamer 407. Ber-SLNs were evaluated for preventive effect against toxicity of Doxorubicin in H9c2 cells. The culture was pre-treated (24 h) with Ber (10 µM) and Ber-SLNs (1 and 10 µM) and exposed to 1 µM of Doxorubicin (Dox) was added for 3 h. The cell viability (LDH (Lactate dehydrogenase) assay and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)), levels of Creatine kinase-MB (CK-MB), Nitrite, MDA (Malondialdehyde), ROS (Reactive oxygen species) generation and apoptotic DNA (Deoxyribonucleic acid) content were assessed. Results: Ber-SLNs had a mean particle size of 13.12±1.188 nm, zeta potential of -1.05 ± 0.08 mV, poly-dispersity index (PDI) of 0.317 ± 0.05 and entrapment efficiency of 50 ± 4.8%. Cell viability was 81  0.17% for Ber-SLNs (10 µM) and 73.22  0.83% for Ber (10 µM) treated cells in MTT assay. Percentage cytotoxicity calculated from LDH release was 58.91  0.54% after Dox, 40.3  1.3% with Ber (10 µM) and 40.7  1.3% with Ber-SLNs (1 µM) (p<0.001). Inflammation and oxidative stress markers were lower with Ber and Ber-SLNs. Attenuation of ROS generation and apoptosis of cardiomyocytes were noted on fluorescence microscopy. Conclusion: Ber loaded SLNs effectively prevented Doxorubicin-induced inflammation and oxidative stress in rat cardiomyocytes. The results demonstrate that microemulsion is a simple, cost-effective technique to prepare Ber-SLNs and may be considered as a drug delivery vehicle for berberine.