COX-2 inhibition by diclofenac is associated with decreased apoptosis and lesion area after experimental focal penetrating traumatic brain injury in rats

Traumatic brain injury (TBI) is followed by a secondary inflammation in the brain. The inflammatory response includes prostanoid synthesis by the inducible enzyme cyclooxygenase-2 (COX-2). Inhibition of COX-2 is associated with improved functional outcome in experimental TBI models, although central nervous system-specific effects are not fully understood. Animal studies report better outcomes in females than males. The exact mechanisms for this gender dichotomy remain unknown. In an initial study we reported increased COX-2 expression in male rats, compared to female, following experimental TBI. It is possible that COX-2 induction is directly associated with increased cell death after TBI. Therefore, we designed a sequential study to investigate the blocking of COX-2 specifically, using the established COX-2 inhibitor diclofenac. Male Sprague-Dawley rats weighing between 250 and 350 g were exposed to focal penetrating TBI and randomly selected for diclofenac treatment (5 ?g intralesionally, immediately following TBI) (n = 8), controls (n = 8), sham operation (n = 8), and normal (no manipulation) (n = 4). After 24 h, brains were removed, fresh frozen, cut into 14?m coronal sections and subjected to COX-2 immunofluorescence, Fluoro Jade, TUNEL, and lesion area analyses. Diclofenac treatment decreased TUNEL staining indicative of apoptosis with a mean change of 54% (p 0.05) and lesion area with a mean change of 55% (p 0.005). Neuronal degeneration measured by Fluoro Jade and COX-2 protein expression levels were not affected. In conclusion, COX-2 inhibition by diclofenac was associated with decreased apoptosis and lesion area after focal penetrating TBI and may be of interest for further studies of clinical applications

Shear Forces during Blast, Not Abrupt Changes in Pressure Alone, Generate Calcium Activity in Human Brain Cells

Blast-Induced Traumatic Brain Injury (bTBI) describes a spectrum of injuries caused by an explosive force that results in changes in brain function. The mechanism responsible for primary bTBI following a blast shockwave remains unknown. We have developed a pneumatic device that delivers shockwaves, similar to those known to induce bTBI, within a chamber optimal for fluorescence microscopy. Abrupt changes in pressure can be created with and without the presence of shear forces at the surface of cells. In primary cultures of human central nervous system cells, the cellular calcium response to shockwaves alone was negligible. Even when the applied pressure reached 15 atm, there was no damage or excitation, unless concomitant shear forces, peaking between 0.3 to 0.7 Pa, were present at the cell surface. The probability of cellular injury in response to a shockwave was low and cell survival was unaffected 20 hours after shockwave exposure

Neuropeptide and Small Transmitter Coexistence: Fundamental Studies and Relevance to Mental Illness

Neuropeptides are auxiliary messenger molecules that always co-exist in nerve cells with one or more small molecule (classic) neurotransmitters. Neuropeptides act both as transmitters and trophic factors, and play a role particularly when the nervous system is challenged, as by injury, pain or stress. Here neuropeptides and coexistence in mammals are reviewed, but with special focus on the 29/30 amino acid galanin and its three receptors GalR1, -R2 and -R3. In particular, galanin's role as a co-transmitter in both rodent and human noradrenergic locus coeruleus (LC) neurons is addressed. Extensive experimental animal data strongly suggest a role for the galanin system in depression-like behavior. The translational potential of these results was tested by studying the galanin system in postmortem human brains, first in normal brains, and then in a comparison of five regions of brains obtained from depressed people who committed suicide, and from matched controls. The distribution of galanin and the four galanin system transcripts in the normal human brain was determined, and selective and parallel changes in levels of transcripts and DNA methylation for galanin and its three receptors were assessed in depressed patients who committed suicide: upregulation of transcripts, e.g., for galanin and GalR3 in LC, paralleled by a decrease in DNA methylation, suggesting involvement of epigenetic mechanisms. It is hypothesized that, when exposed to severe stress, the noradrenergic LC neurons fire in bursts and release galanin from their soma/dendrites. Galanin then acts on somato-dendritic, inhibitory galanin autoreceptors, opening potassium channels and inhibiting firing. The purpose of these autoreceptors is to act as a 'brake' to prevent overexcitation, a brake that is also part of resilience to stress that protects against depression. Depression then arises when the inhibition is too strong and long lasting - a maladaption, allostatic load, leading to depletion of NA levels in the forebrain. It is suggested that disinhibition by a galanin antagonist may have antidepressant activity by restoring forebrain NA levels. A role of galanin in depression is also supported by a recent candidate gene study, showing that variants in genes for galanin and its three receptors confer increased risk of depression and anxiety in people who experienced childhood adversity or recent negative life events. In summary, galanin, a neuropeptide coexisting in LC neurons, may participate in the mechanism underlying resilience against a serious and common disorder, MDD. Existing and further results may lead to an increased understanding of how this illness develops, which in turn could provide a basis for its treatment

Assessment of Fluid Cavitation Threshold Using a Polymeric Split Hopkinson Bar-Confinement Chamber Apparatus

The authors would like to acknowledge the Natural Sciences and Engineering Research Council of Canada for financial support, and Compute Canada and Sharcnet for providing the necessary computing resources.Mild Traumatic Brain Injury (mTBI) has been associated with blast exposure resulting from the use of improvised explosive devices (IEDs) in recent and past military conflicts. Experimental and numerical models of head blast exposure have demonstrated the potential for high negative pressures occurring within the head at the contre-coup location relative to the blast exposure, and it has been hypothesized that this negative pressure could result in cavitation of Cerebrospinal Fluid (CSF) surrounding the brain, leading to brain tissue damage. The cavitation threshold of CSF, the effect of temperature, and the effect of impurities or dissolved gases are presently unknown. In this study, a novel Polymeric Split Hopkinson Pressure Bar and confinement chamber apparatus were used to generate loading in distilled water similar to the conditions in the vicinity of the CSF during blast exposure. Cavitation was identified using high-speed imaging of the event, and a validated numerical model of the apparatus was applied to determine the pressure in the fluid during the exposure. Increasing the water temperature resulted in a decrease in the 50% probability of cavitation from 21 °C (−3320 kPa ± 3%) to 37 °C (−3195 kPa ± 5%) in agreement with the theoretical values, but was not statistically significant. Importantly, the effect of water treatment had a significant effect on the cavitation pressure for water with wetting agent (−3320 kPa ± 3%), degassed water (−1369 kPa ± 16%) and untreated distilled water (−528 kPa ± 25%). Thus, reducing dissolved gases through degassing or the use of a wetting agent significantly increases the cavitation pressure and reduces the variability of the cavitation pressure threshold

Prolonged but not short-duration blast waves elicit acute inflammation in a rodent model of primary blast limb trauma

BackgroundBlast injuries from conventional and improvised explosive devices account for 75% of injuries from current conflicts; over 70% of injuries involve the limbs. Variable duration and magnitude of blast wave loading occurs in real-life explosions and is hypothesised to cause different injuries. While a number of in vivo models report the inflammatory response to blast injuries, the extent of this response has not been investigated with respect to the duration of the primary blast wave. The relevance is that explosions in open air are of short duration compared to those in confined spaces.MethodsHindlimbs of adult Sprauge-Dawley rats were subjected to focal isolated primary blast waves of varying overpressure (1.8–3.65 kPa) and duration (3.0–11.5 ms), utilising a shock tube and purpose-built experimental rig. Rats were monitored during and after the blast. At 6 and 24 h after exposure, blood, lungs, liver and muscle tissues were collected and prepared for histology and flow cytometry.ResultsAt 6 h, increases in circulating neutrophils and CD43Lo/His48Hi monocytes were observed in rats subjected to longer-duration blast waves. This was accompanied by increases in circulating pro-inflammatory chemo/cytokines KC and IL-6. No changes were observed with shorter-duration blast waves irrespective of overpressure. In all cases, no histological damage was observed in muscle, lung or liver. By 24 h post-blast, all inflammatory parameters had normalised.ConclusionsWe report the development of a rodent model of primary blast limb trauma that is the first to highlight an important role played by blast wave duration and magnitude in initiating acute inflammatory response following limb injury in the absence of limb fracture or penetrating trauma. The combined biological and mechanical method developed can be used to further understand the complex effects of blast waves in a range of different tissues and organs in vivo

Concluding Remarks

In this chapter we summarize the arguments for the use of animal models for neurotrauma experiments. We also stress the need for well-controlled and well-validated models. The use of guidelines for animal experiments is an important way to increase the value of the animal models and facilitate translation to real-life situations

A Sagittal Plane Rotational Injury Rodent Model for Research on Traumatic Brain Injuries

The model presented here produce brain injuries following sagittal plane rearward rotational acceleration in rats. During trauma, a rotating bar, which is tightly secured to the animal head, is impacted by a striker that causes the rotating bar and the animal head to rotate rearward; the acceleration phase is followed by a rotation at constant speed and gentle deceleration when the rotating bar contacts a padded stop. The total head angle change range from 25\ub0 to 30\ub0. By adjusting the air pressure in the air-driven accelerator used to accelerate the striker, a large range of rotational accelerations can be achieved. This model can, depending on the striker velocity, produce subdural bleedings, graded widespread axonal injuries in the corpus callosum, the border between the corpus callosum, cortex, cerebellum, olfactory bulbs, and in some of the tracts in the brain stem. The model has been shown to produce degenerating axons. For lower rotational accelerations no apparent axonal injuries can be observed. The model produces only limited signs of contusion injury, and macrophage invasions, glial fibrillary acidic protein redistribution or hypertrophy, and blood–brain barrier changes are unusual. The model produces distinct S100 and Neurofilament Light serum concentration changes, thus indicating that blood vessel and glia cell injuries may occur. The rotational acceleration trauma model presented can produce graded axonal injury, is repeatable, and produce limited other types of TBIs and as such is useful in the study of injury biomechanics, diagnostics, and treatment strategies following diffuse axonal injury and most likely also following concussion