• Figueiredo, Patricia
• Liu, Dongfei
• Wang, Shiqi
• Li, Jiachen
• Correia, Alexandra Maria Rebelo
• Santos, Hélder A.
• Hirvonen, Jouni
• Li, Wei
• Wiwattanapatapee, Ruedeekorn
• Xia, Bing
• Wannasarit, Saowanee

Abstract

Introduction: Non-resolving inflammation drives the development of many prevalent chronic diseases, including rheumatoid arthritis, atherosclerosis, diabetes and some types of cancer. (1) Recent studies found that natural melanin nanoparticles (MNs) from Sepia ink had superior antioxidant potency due to the reactive oxygen species (ROS) scavenging capability. (2) Considering the prevalent oxidative stress closely associated with chronic non-resolving inflammation associated diseases. (3) MNs from Cephalopod ink, may show promising potential in anti-inflammatory applications. Although traditional MN preparation methods generate homogenous nanoparticles, the lengthy reaction times and unavoidable batch-to-batch variations limit the possibility to tailor particle production towards real-world applications. Herein we demonstrate that monodispersed MNs can be produced in a highly concentrated substrate solution within seconds for the first time, by microfluidic technique due to the precise control of superfast liquid mixing and mass transfer.Methods: The MNs were produced on a glass-capillary microfluidic chip, with a precise control over the particle formation and reaction termination. The particles were further encapsulated in a pH-responsive, endosomolytic polymer (MAP) by microfluidic technique, with anti-inflammatory drug budesonide loaded. The cellular uptake, endosomal escape behaviour, ROS scavenging capability, inflammatory cytokine release profile of these nanocomposites were evaluated on inflamed macrophages. Results: MNs with tunable sizes and monodispersity could be 1000 times faster than traditional bulk methods, confirmed by dynamic light scattering results and transmission electron microscopy images. Further characterizations by Fourier-transform infrared spectroscopy revealed that the MNs formed have similar chemical composition compared with those prepared by bulk method. The MNs show negligible cytotoxicity towards human and murine macrophages and ROS scavenging capacity. The encapsulation in MAP facilitated the endosomal escape and the intracellular delivery of MN, thus enhancing the uptake and ROS scavenging efficacy. The co-delivery of budesonide with MN by MAP reduced the expression of pro-inflammatory cell makers and induced the secretion of immune-suppressive cytokines interleukin-10, thus modulating the phenotype of macrophages from pro-inflammatory to anti-inflammatory.Conclusion: The simple, but advanced preparation of nanoparticles by microfluidic device, along with the endosomal escape capability, pH-controlled drug release profile, and efficient ROS scavenging property of the nanocomposites makes the nanosystems developed here a promising candidate for anti-inflammation therapy.Acknowledgements: This work was supported by grants from Jenny and Antti Wihuri Foundation, Thailand Research Fund, Finnish Culture Foundation, the HiLIFE Research Funds, the Sigrid Jusélius Foundation, the Norwegian Research Council/Nacamed AS, and the European Research Council.References (up to three): (1) S. I. Grivennikov, F. R. Greten, M. Karin, Cell, 2010: 883-899. (2) B. L. L. Seagle, E. M. Gasyna, W. F. Mieler, J. R. Norris, Proc. Natl. Acad. Sci., 2006: 16644-16648. (3) M. Mittal, M. R. Siddiqui, K. Tran, S. P. Reddy, A. B. Malik, Antioxid. Redox Signal., 2014: 1126-1167.

Topics

• nanoparticle
• nanocomposite
• polymer
• Oxygen
• glass
• reactive
• glass
• chemical composition
• transmission electron microscopy
• dynamic light scattering
• infrared spectroscopy