Nanoformulated Recombinant Human Myelin Basic Protein and Rituximab Modulate Neuronal Perturbations in Experimental Autoimmune Encephalomyelitis in Mice

Abstract

Muhammed A Saad,1,2 Noha M Eissa,2 Mohammed A Ahmed,3 Aliaa N ElMeshad,3,4 Götz Laible,5– 7 Ahmed S Attia,8 Medhat A Al-Ghobashy,9,10 Rania M Abdelsalam,1,2 Muhammad Y Al-Shorbagy1,11 1Department of Pharmacology and Toxicology, Faculty of Pharmacy, Cairo University, Cairo, Egypt; 2School of Pharmacy, Newgiza University, Giza, Egypt; 3Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Cairo, Egypt; 4Faculty of Nanotechnology for Postgraduate Studies, Cairo University, Giza, Egypt; 5AgResearch, Ruakura Research Centre, Hamilton, New Zealand; 6School of Medical Sciences, University of Auckland, Auckland, New Zealand; 7Maurice Wilkins Centre for Molecular Biodiscovery, Auckland, New Zealand; 8Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt; 9Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt; 10Bioanalysis Research Group, School of Pharmacy, Newgiza University, Giza, Egypt; 11Department of Pharmaceutical Sciences, College of Pharmacy, Gulf Medical University, Ajman, United Arab EmiratesCorrespondence: Ahmed S Attia, Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt, Email [email protected]: Rituximab (RTX) and recombinant human myelin basic protein (rhMBP) were proven to be effective in ameliorating the symptoms of multiple sclerosis (MS). In this study, a nanoformulation containing rhMBP with RTX on its surface (Nano-rhMBP-RTX) was prepared and investigated in comparison with other treatment groups to determine its potential neuro-protective effects on C57BL/6 mice after inducing experimental autoimmune encephalomyelitis (EAE).Methods: EAE was induced in the corresponding mice by injecting 100 μL of an emulsion containing complete Freund’s adjuvant (CFA) and myelin oligodendrocyte glycoprotein (MOG). The subjects were weighed, scored and subjected to behavioural tests. After reaching a clinical score of 3, various treatments were given to corresponding EAE-induced and non-induced groups including rhMBP, RTX, empty nanoparticle prepared by poly (lactide-co-glycolide) (PLGA) or the prepared nanoformulation (Nano-rhMBP-RTX). At the end of the study, biochemical parameters were also determined as interferon-γ (IFN-γ), myeloperoxidase (MPO), interleukin-10 (IL-10), interleukin-4 (IL-4), tumor necrosis factor alpha (TNF-α), nuclear factor kappa B (NF-kB), brain derived neurotrophic factor (BDNF), 2’, 3’ cyclic nucleotide 3’ phosphodiesterase (CNP) and transforming growth factor beta (TGF-β) along with some histopathological analyses.Results: The results of the Nano-rhMBP-RTX group showed promising outcomes in terms of reducing the clinical scores, improving the behavioural responses associated with improved histopathological findings. Elevation in the levels of IL-4, CNP and TGF-β was also noticed along with marked decline in the levels of NF-kB and TNF-α.Conclusion: Nano-rhMBP-RTX treated group ameliorated the adverse effects induced in the EAE model. The effectiveness of this formulation was demonstrated by the normalization of EAE-induced behavioral changes and aberrant levels of specific biochemical markers as well as reduced damage of hippocampal tissues and retaining higher levels of myelination.Keywords: multiple sclerosis, experimental autoimmune encephalomyelitis, nanoparticle, rituximab, recombinant human myelin basic protein

Keywords

• multiple sclerosis
• experimental autoimmune encephalomyelitis
• nanoparticle
• rituximab
• recombinant human myelin basic protein