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In vitro spinal cord trauma
J D Balentine1, W B Greene, M Bornstein
1Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston.
Abstract:
Fetal mouse spinal cord explants were harvested and allowed to grow in Maximow chambers. Normal appearing matured cultures were subjected to a brief episode of impact trauma by dropping the flat surface of 25 to 105 mg dressmaker's pins from a height of 10 cm directly onto the exposed surface of the culture. Light and electron microscopic studies at selected posttrauma intervals revealed discrete foci of necrosis preceded or accompanied by nerve fiber changes (granular axoplasm, vesicular myelin, pleomorphic spheroids) identical with those documented in spinal cord trauma in vivo. Although no inherent calcification was observed, calcium was added to some of the traumatized cultures and it was subsequently localized by pyroantimonate in the axoplasm; within mitochondria, adjacent to neurofilaments, and in the cytosol. The study indicates that the morphologic sequence of events of spinal cord trauma in vitro are similar to those observed in vivo, and that the latter may occur in the absence of vascular injury.
Insights
This study demonstrates that spinal cord trauma in vitro mirrors in vivo effects, showing similar nerve fiber damage and necrosis. Calcium
Area of Science:
- Neuroscience
- Cell Biology
- Pathology
Background:
- Spinal cord injury (SCI) research often relies on in vivo models.
- Understanding the cellular mechanisms of SCI requires controlled experimental conditions.
- In vitro models offer a way to study SCI without confounding systemic factors.
Purpose of the Study:
- To investigate the in vitro effects of mechanical trauma on fetal mouse spinal cord explants.
- To compare the resulting cellular and morphological changes with those observed in vivo.
- To explore the role of calcium in the cellular response to spinal cord trauma.
Main Methods:
- Fetal mouse spinal cord explants were cultured in Maximow chambers.
- Impact trauma was induced by dropping calibrated weights onto the explant surface.
- Light and electron microscopy were used for morphological analysis at post-trauma intervals.
- Calcium localization was performed using pyroantimonate staining in traumatized cultures.
Main Results:
- Traumatized explants exhibited necrosis and nerve fiber changes (granular axoplasm, vesicular myelin, spheroids) mirroring in vivo SCI.
- These morphological alterations preceded or accompanied necrotic foci.
- Calcium, when added, localized within the axoplasm, mitochondria, and cytosol of traumatized cells.
- No inherent calcification was observed in the absence of added calcium.
Conclusions:
- In vitro mechanical trauma to spinal cord explants recapitulates key pathological features of in vivo spinal cord injury.
- The observed cellular changes suggest that vascular injury is not essential for the initial morphological sequence of SCI.
- This in vitro model provides a valuable tool for studying the cellular pathogenesis of spinal cord trauma and the role of calcium.