Development of Nanosome-Encapsulated Honokiol for Intravenous Therapy Against Experimental Autoimmune Encephalomyelitis

Yai-Ping Hsiao,1 Hui-Ting Chen,1 Yu-Chih Liang,2 Tse-En Wang,1 Kai-Hung Huang,3 Cheng-Chih Hsu,3 Hong-Jen Liang,4 Chung-Hsiung Huang,5 Tong-Rong Jan1 1Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei 10617, Taiwan; 2School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 10617, Taiwan; 3Department of Chemistry, National Taiwan University, Taipei 10617, Taiwan; 4Department of Food Science, Yuanpei University, Hsinchu 30015, Taiwan; 5Department of Food Science, National Taiwan Ocean University, Keelung 20224, TaiwanCorrespondence: Tong-Rong JanDepartment and Graduate Institute of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, No.1, Sec. 4, Roosevelt Road, Taipei 10617, Taiwan, ROCTel +886-2-33661287Fax +886-2-23661475Email tonyjan@ntu.edu.twChung-Hsiung HuangDepartment of Food Science, National Taiwan Ocean University, No.2, Beining Road, Keelung 20224, TaiwanTel +886-2-24622192#5115Email huangch@mail.ntou.edu.twBackground: Honokiol has been reported to possess anti-inflammatory and neuroprotective activities. However, the poor aqueous solubility of honokiol limits its clinical application for systemic administration.Purpose: This study aims to develop a novel formulation of nanosome-encapsulated honokiol (NHNK) for intravenous therapy against mouse experimental autoimmune encephalomyelitis (EAE) that mimics human multiple sclerosis.Methods: Nanosomes and NHNK were prepared by using an ultra-high pressure homogenization (UHPH) method. Mice were treated with NHNK or empty nanosomes during the peak phase of EAE symptoms. Symptoms of EAE were monitored and samples of the spinal cord were obtained for histopathological examinations.Results: The stock of NHNK containing honokiol in the nanosome formulation, which showed the structure of single phospholipid bilayer membranes, was well formulated with the particle size of 48.0 ± 0.1 nm and the encapsulation efficiency 58.1 ± 4.2%. Intravenous administration of NHNK ameliorated the severity of EAE accompanied by a significant reduction of demyelination and inflammation in the spinal cord. Furthermore, NHNK decreased the number of IL-6+, Iba-1+TNF +, Iba-1+IL-12 p40+, and CD3+IFN-γ+ cells infiltrating the spinal cord.Conclusion: The UHPH method simplified the preparation of NHNK with uniformly distributed nanosize and high encapsulation efficiency. Intravenous administration of NHNK ameliorated the severity of EAE by suppressing the infiltration of activated microglia and Th1 cells into the spinal cord. Collectively, these results suggest that the formulation of NHNK is a prospective therapeutic approach for inflammatory CNS diseases, such as multiple sclerosis.Keywords: experimental autoimmune encephalomyelitis, honokiol, nanosome, neuroinflammation, ultra-high pressure homogenization