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

wbldb

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

Cited Works (11)

Year	Entry
1995	Ellis EF, McKinney JS, Willoughby KA, Liang S, Povlishock JT. A new model for rapid stretch-induced injury of cells in culture: characterization of the model using astrocytes. J Neurotrauma. June 1995;12(3):325-339.
1997	LaPlaca MC, Thibault LE. Anin vitro traumatic injury model to examine the response of neurons to a hydrodynamically-induced deformation. Ann Biomed Eng. July 1997;25(4):665-677.
1982	Gennarelli TA, Thibault LE, Adams JH, Graham DI, Thompson CJ, Marcincin RP. Diffuse axonal injury and traumatic coma in the primate. Ann Neurol. 1982;12(6):564-574.
1989	McIntosh TK, Vink R, Noble L, Yamakami I, Fernyak S, Soares H, Faden AL. Traumatic brain injury in the rat: characterization of a lateral fluid-percussion model. Neuroscience. 1989;28(1):233-244.
1996	Cargill RS II, Thibault LE. Acute alterations in [Ca²⁺]_i in NG108-15 cells subjected to high strain rate deformation and chemical hypoxia: an in vitro model for neural. J Neurotrauma. July 1996;13(7):395-407.
1990	Margulies SS, Thibault LE, Gennarelli TA. Physical model simulations of brain injury in the primate. J Biomech. 1990;23(8):823-836.
1993	Galbraith JA, Thibault LE, Matteson DR. Mechanical and electrical responses of the squid giant axon to simple elongation. J Biomech Eng. February 1993;115(1):13-22.
1995	Smith DH, Soares HD, Pierce JS, Perlman KG, Saatman KE, Meaney DF, Dixon CE, McIntosh TK. A model of parasagittal controlled cortical impact in the mouse: cognitive and histopathologic effects. J Neurotrauma. April 1995;12(2):169-178.
1972	Gennarelli TA, Thibault LE, Ommaya AK. Pathophysiologic responses to rotational and translational accelerations of the head. In: Proceedings of the 16th Stapp Car Crash Conference. November 8-10, 1972; Detroit, MI. Warrendale, PA: Society of Automotive Engineers:296-308. SAE 720970.
1987	Dixon CE, Lyeth BG, Povlishock JT, Findling RL, Hamm RJ, Marmarou A, Young HF, Hayes RL. A fluid percussion model of experimental brain injury in the rat. J Neurosurg. July 1987;67(1):110-119.
1981	Feeney DM, Boyeson MG, Linn RT, Murray HM, Dail WG. Responses to cortical injury, I: methodology and local effects of contusions in the rat. Brain Res. April 27, 1981;211(1):67-77.

Cited By (34)

Year	Entry
2002	Ommaya AK, Goldsmith W, Thibault L. Biomechanics and neuropathology of adult and paediatric head injury. Br J Neurosurg. June 2002;16(3):220-242.
1999	Barbee KA, Yazdi J, Abait B, Croul S, Fijan R, Thibault LE. An in vivo model for the acute response of neural tissue to traumatic loading. In: Proceedings of the 1999 International IRCOBI Conference on the Biomechanics of Impact. September 23-24, 1999; Sitges, Spain.95-105.
2003	King AI, Yang KH, Zhang L, Hardy W, Viano DC. Is head injury caused by linear or angular acceleration? In: Proceedings of the 2003 International IRCOBI Conference on the Biomechanics of Impact. September 25, 2003; Lisbon, Portugal.1-12.
2003	Morrison B III, Cater HL, Wang CC-B, Thomas FC, Hung CT, Ateshian GA, Sundstrom LE. A tissue level tolerance criterion for living brain developed with an in vitro model of traumatic mechanical loading. Stapp Car Crash J. 2003;47:93-105. SAE 2003-22-0006.
2006	Kimpara H, Nakahira Y, Iwamoto M, Miki K, Ichihara K, Kawano S-i, Taguchi T. Investigation of anteroposterior head-neck responses during severe frontal impacts using a brain-spinal cord complex FE model. Stapp Car Crash J. 2006;50:509-544. SAE 2006-22-0019.
2007	Elkin BS, Morrison B. Region-specific tolerance criteria for the living brain. Stapp Car Crash J. 2007;51:127-138. SAE 2007-22-0005.
2006	Cater HL, Sundstrom LE, Morrison B III. Temporal development of hippocampal cell death is dependent on tissue strain but not strain rate. J Biomech. 2006;39(16):2810-2818.
2000	Morrison B III, Meaney DF, Margulies SS, McIntosh TK. Dynamic mechanical stretch of organotypic brain slice cultures induces differential genomic expression: relationship to mechanical parameters. J Biomech Eng. June 2000;122(3):224-230.
2001	Geddes DM, Cargill RS II. An in vitro model of neural trauma: device characterization and calcium response to mechanical stretch. J Biomech Eng. June 2001;123(3):247-255.
1998	Morrison B III, Saatman KE, Meaney DF, McIntosh TK. In vitro central nervous system models of mechanically induced trauma: a review. J Neurotrauma. November 1998;15(11):911-928.
2003	Geddes DM, Cargill RS II, LaPlaca MC. Mechanical stretch to neurons results in a strain rate and magnitude-dependent increase in plasma membrane permeability. J Neurotrauma. October 2003;20(10):1039-1049.
2006	Geddes-Klein DM, Schiffman KB, Meaney DF. Mechanisms and consequences of neuronal stretch injury in vitro differ with the model of trauma. J Neurotrauma. February 2006;23(2):193-204.
2010	Elkin BS. Biomechanics and Biological Consequences of Traumatic Brain Injury [PhD thesis]. Columbia University; 2010.
2008	Kilinc D. Mechanisms and Prevention of Axonal Damage in Response to Mechanical Trauma to Cultured Neurons [PhD thesis]. Drexel University; March 2008.
2014	Dastgheyb RM. Mechanisms of Axonal Pathology in the Context of Traumatic Brain Injury [PhD thesis]. Drexel University; November 2014.
2019	Wofford KL. Utilizing the Innate Immune System to Mitigate Secondary Injury Following Traumatic Brain Injury [PhD thesis]. Drexel University; May 2019.
2002	Geddes DM. Selective Vulnerability of Hippocampal Vs. Cortical Neurons to Mechanically Induced Increases in Plasma Membrane Permeability [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; February 2002.
2002	Tate MC. The Development of Extracellular Matrix Based Neural Stem Cell Transplants for Treatment of Traumatic Brain Injury [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; August 2002.
2004	Prado GR. Neuronal Plasma Membrane Disruption in Traumatic Brain Injury [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; July 2004.
2005	Cullen DK. Traumatically-Induced Degeneration and Reactive Astrogliosis in Three-Dimensional Neural Co-Cultures: Factors Influencing Neural Stem Cell Survival and Integration [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; December 2005.
2007	Stabenfeldt SE. Bioactive Thermoresponsive Hydrogels for Neural Tissue Engineering [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; December 2007.
2008	Simon CM. Investigation of Plasma Membrane Compromise and Citicoline-Mediated Repair After Spinal Cord Injury [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; April 2008.
2008	Lessing MC. The Acute Cellular and Behavioral Response to Mechanical Neuronal Injury [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; December 2008.
2010	Vernekar VN. Optimization of 3-D Neural Culture and Extracellular Electrophysiology for Studying Injury-Induced Morphological and Functional Changes [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; May 2010.
2011	Wester B. Development and Characterization of Mechanically Actuated Microtweezers for Use in a Single-Cell Neural Injury Model [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; May 2011.
2004	Sparrey CJ. The Effect of Impact Velocity on Acute Spinal Cord Injury [Master's thesis]. Vancouver, BC: University of British Columbia; July 2004.
2013	Carlson SW. Evaluation of Insulin-Like Growth Factor-1 as a Therapeutic Approach for the Treatment of Traumatic Brain Injury [PhD thesis]. University of Kentucky; 2013.
2014	Rousseau P. Analysis of Concussion Metrics of Real-World Concussive and Non-Injurious Elbow and Shoulder to Head Collisions in Ice Hockey [PhD thesis]. University of Ottawa; 2014.
1999	Morrison B III. Differential Genomic Expression After Mechanical Injury of Organotypic Brain Slice Cultures: An in Vitro Model of Traumatic Brain Injury [PhD thesis]. Philadelphia, PA: University of Pennsylvania; 1999.
2002	Lusardi TAG. Initiation and Propagation of Mechanically Induced Neuronal Injury [PhD thesis]. Philadelphia, PA: University of Pennsylvania; 2002.
2010	Quinn KP. Integrating Electrophysiological, Mechanical, and Optical Methods to Define the Mechanisms of Painful Facet Joint Injury [PhD thesis]. Philadelphia, PA: University of Pennsylvania; 2010.
2010	von Reyn CR. Calpain Mediated Proteolysis of Voltage Gated Sodium Channels and Its Role in Pathological Development After Traumatic Brain Injury [PhD thesis]. Philadelphia, PA: University of Pennsylvania; 2010.
2012	Maltese MR. Traumatic Brain Injury Thresholds in the Pre-Adolescent Juvenile [PhD thesis]. Philadelphia, PA: University of Pennsylvania; 2012.
2016	Boruah S. A Viscoelastic Model for High Strain Rate Loading of the Human Calvarium [PhD thesis]. Charlottesville, VA: University of Virginia; May 2016.