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Optimizing E x Vivo Model of Traumatic Brain Injury to Test Biomolecules of Therapeutic Interest
Author(s) -
Vig Parminder J.S.,
Ryu Jung,
Lee H. Joon,
Warrington Junie P.,
Janorkar Amol V.,
Pal Pallabi,
Lopez Mariper
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.05654
Subject(s) - traumatic brain injury , in vivo , medicine , excitotoxicity , pathology , pathophysiology , neuroinflammation , lesion , traumatic injury , diffuse axonal injury , neuroprotection , neuroscience , glutamate receptor , biology , surgery , receptor , microbiology and biotechnology , disease , psychiatry
Traumatic brain injury (TBI) is recognized as a major health concern due to its increasing occurrence, broad spectrum of symptoms and disabilities. TBI initiates a cascade of secondary pathophysiological events, such as excitotoxicity, neuroinflammation, and oxidative stress that worsen brain pathology from hours to days following the injury. Apoptotic and necrotic neurons have been observed in regions far from the lesion site in the days and weeks after TBI. To evaluate the complex secondary and severity‐specific injury response, investigators have relied on pre‐clinical rodent models. However, these in vivo TBI models have demonstrated intra‐ and inter‐laboratory variability of clinically relevant functional evaluations that were not consistently correlated with the severity of injury. Here, we describe a novel ex vivo model of traumatic cerebellar injury (TCI) using organotypic cerebellar slice cultures to study trauma‐induced injury, and also to test the effects of hydrogels and Elastin‐Like Polypeptide (ELP) based therapeutic peptides (TPs) on cerebellar injury. The cerebellar slice cultures were prepared from 9–11 day‐old GFP wildtype or parvalbumin‐BrainBow mouse pups. After 7 days of organotypic culture, TCI was induced at day 7 in vitro (7 DIV) using a weight drop device. Live imaging of slices showed a marked loss of GFP‐fluorescence at 24 h post‐injury. At the impact site, we observed a loss of Purkinje cells and interneurons. In some slices where we induced acute TCI, a small cavity‐like defect developed within 72 hr post injury. After 4 DIV post‐injury, the injured area was treated with V120‐collagen hydrogel and the medium replaced with experimental medium (with or without ELP‐TPs). Live slices were observed every 4 DIV. Some TPs were able to progressively reduce the diameter of injured area and the loss of GFP‐fluorescence, emphasizing that a combination of hydrogel and ELP‐TPs is protective. These data suggest that our ex vivo TCI model could be a useful tool to test different pharmacological compounds with potential therapeutic activity on TBI.