An in vitro model of traumatic neuronal injury: loading rate-dependent changes in acute cytosolic calcium and lactate dehydrogenase release

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LaPlaca, Michelle C.¹; Lee, Virginia M.-Y.²; Thibault, Lawrence E.³

An in vitro model of traumatic neuronal injury: loading rate-dependent changes in acute cytosolic calcium and lactate dehydrogenase release

J Neurotrauma. June 1997;14(5):355-368

Affiliations

¹Department of Bioengineering, University of Pennsylvania, Philadelphia, PA

²Department of Laboratory Medicine, University of Pennsylvania, Philadelphia, PA

³Department of Neurosurgery, Division of Bioengineering, Allegheny University of the Health Sciences, Philadelphia, PA
Abstract and keywords

We developed a new in vitro model of neuronal injury using NT2-N cells to examine the effects of hydrodynamic loading rate on intraneuronal calcium dynamics and lactate dehydrogenase (LDH) release. Our apparatus consisted of a parallel disk viscometer which induced fluid shear stress with well-defined magnitudes and loading rates to cultured cells. We found that the deformation response of the cells was dependent on the severity of the insult, with increased cellular strains generated for higher shear stresses at a constant loading rate. Peak intracellular free calcium concentration correlated with strain, suggesting that mechanical deformation may regulate calcium response. Slowly applied fluid shear stress elicited no response, whereas high loading rates resulted in peak calcium increases 2.9 to 3.6 times baseline values as injury severity was increased. LDH release measured within 5 min after the insult correlated with loading rate. In addition, LDH release continued to increase out to 24 h following high loading rate conditions, demonstrating that the application of fluid shear stress led to prolonged cell damage. The acute response in NT2-N cells subjected to an insult with the CSID is dependent on the loading rate, and these results suggest that initial membrane deformation may trigger subsequent events.

Keywords: cellular injury; intracellular free calcium; lactate dehydrogenase; mechanical injury; NT2-N cells; traumatic brain injury models
Links

DOI: 10.1089/neu.1997.14.355

PubMed: 9219851

WoS: A1997XJ83600001

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