Combinatory Multifactor Treatment Effects on Primary Nanofiber Oligodendrocyte Cultures

Abstract

Multiple sclerosis (MS) is a chronic inflammatory demyelinating and neurodegenerative disease of the central nervous system. Neurological deficits are attributed to inflammatory demyelination, which compromises axonal function and survival. These are mitigated in experimental models by rapid and often complete remyelination of affected axons, but in MS this endogenous repair mechanism frequently fails, leaving axons increasingly vulnerable to the detrimental effects of inflammatory and metabolic stress. Understanding the molecular basis of remyelination and remyelination failure is essential to develop improved therapies for this devastating disease. However, recent studies suggest that this is not due to a single dominant mechanism, but rather represents the biological outcome of multiple changes in the lesion microenvironment that combine to disrupt oligodendrocyte differentiation. This identifies a pressing need to develop technical platforms to investigate combinatory and/or synergistic effects of factors differentially expressed in MS lesions on oligodendrocyte proliferation and differentiation. Here we describe protocols using primary oligodendrocyte cultures from Bl6 mice on 384-well nanofiber plates to model changes affecting oligodendrogenesis and differentiation in the complex signaling environment associated with multiple sclerosis lesions. Using platelet-derived growth factor (PDGF–AA), fibroblast growth factor 2 (FGF2), bone morphogenetic protein 2 (BMP2) and bone morphogenetic protein 4 (BMP4) as representative targets, we demonstrate that we can assess their combinatory effects across a wide range of concentrations in a single experiment. This in vitro model is ideal for assessing the combinatory effects of changes in availability of multiple factors, thus more closely modelling the situation in vivo and furthering high-throughput screening possibilities.

Nanomaterial payload delivery to central nervous system glia for neural protection and repair

“…Olig2, O1, MOG, and MBP ( Rasti Boroojeni et al, 2020 ). Primary oligodendrocytes isolated from B16 mice were able to myelinate aligned PCL nanofibers that released PDGF-AA, FGF2, BMP2, and BMP4 ( Enz et al, 2019 ). PCL nanofibers loaded with miR-219, miR-338-3p, and miR-338-5p enhanced the differentiation of primary rat oligodendrocyte progenitor cells and their maturation into RIP+ oligodendrocytes ( Diao et al, 2015 ).…”

An Update of Palmitoylethanolamide and Luteolin Effects in Preclinical and Clinical Studies of Neuroinflammatory Events

“…Moreover, oligodendrocytes cell surfaces have receptors for a wide-ranging of mediators, such as IL-4, IL-6, IL-7, IL-10, IL-11, IL-12, and IL-18 [123]. Recent evidence also demonstrates that oligodendrocyte also express antigen-presenting molecules and co-stimulatory molecules, complement and complement receptor molecules, complement regulatory molecules, tetraspanins, neuroimmune regulatory proteins, extracellular matrix proteins, and many others [124]. Necrosis and apoptosis are events that can activate oligodendrocytes in various ways in the central nervous system.…”

Oligodendrocyte Physiology and Pathology Function

“…To investigate mechanisms operant during de- and regeneration of the axon-oligodendrocyte-myelin compartment, and to develop effective MS treatment options, the following are required: (i) novel, dynamic technical platforms to investigate complex cell–cell interactions in a CNS-like microenvironment such as the oligodendrocyte-nanofiber platform described by Enz and colleagues [ 11 ]; (ii) unbiased evaluation systems to monitor disease progression and successful therapeutic interventions in pre-clinical models such as those described by Zhan an colleagues [ 12 ], Joost and colleagues [ 13 ], and Hochstrasser and colleagues [ 14 ]; and (iii) novel imaging modalities which would allow longitudinal studies as described by Khodanovich and colleagues [ 15 ]. Of note, a better understanding of the axon-oligodendrocyte-myelin compartment might lead to restorative therapies not just in MS but also other neuronal disorders such as Down Syndrome (reviewed by Reiche and colleagues [ 16 ]) or schizophrenia (reviewed by Raabe and colleagues [ 17 ]).…”