Name: a contusion model of severe spinal cord injury in rats
Uploaded: 2013-08-17T14:00:00
Description: Watch this Scientific Journal Video about A Contusion Model of Severe Spinal Cord Injury in Rats at JoVE.com

The authors are grateful to Dr. N. Banik and Dr. D. Mitchell for their guidance in the development of this model.

1. Preparation Before Spinal Cord Injury
The surgical instruments required for this procedure are scalpel, pickups with and without teeth, hemostats, self retaining retractors, fine tip rongeurs, needle driver, absorbable sutures, and skin clip applicators. Other surgical supplies required are surgical drapes, sterile sheets for surgical field, gauze sponges, cotton-tip applicators, and metallic foil. Autoclave the surgical instruments and supplies prior to the surgery. Use the individual set of...

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	<li>Blesch, A., Tuszynski, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+injury:+plasticity,+regeneration+and+the+challenge+of+translational+drug+development.">Spinal cord injury: plasticity, regeneration and the challenge of translational drug development.</a> Trends Neurosci. 32, 41-47 (2009).</li><li>Dobkin, B. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Curiosity+and+cure:+translational+research+strategies+for+neural+repair-mediated+rehabilitation.">Curiosity and cure: translational research strategies for neural repair-mediated rehabilitation.</a> Dev. Neurobiol. 67, 1133-1147 (2007).</li><li>Fehlings, M. G., Cadotte, D. W., Fehlings, L. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+series+of+systematic+reviews+on+the+treatment+of+acute+spinal+cord+injury:+a+foundation+for+best+medical+practice.">A series of systematic reviews on the treatment of acute spinal cord injury: a foundation for best medical practice.</a> J. Neurotrauma. 28, 1329-1333 (2011).</li><li>Iseda, T., Okuda, T., Kane-Goldsmith, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single,+high-dose+intraspinal+injection+of+chondroitinase+reduces+glycosaminoglycans+in+injured+spinal+cord+and+promotes+corticospinal+axonal+regrowth+after+hemisection+but+not+contusion.">Single, high-dose intraspinal injection of chondroitinase reduces glycosaminoglycans in injured spinal cord and promotes corticospinal axonal regrowth after hemisection but not contusion.</a> J. Neurotrauma. 25, 334-349 (2008).</li><li>Khan, T., Havey, R. M., Sayers, S. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+spinal+cord+contusion+injuries.">Animal models of spinal cord contusion injuries.</a> Laboratory Animal Science. 49, 161-172 (1999).</li><li>Kim, B. G., Kang, Y. M., Phi, J. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implantation+of+polymer+scaffolds+seeded+with+neural+stem+cells+in+a+canine+spinal+cord+injury+model.">Implantation of polymer scaffolds seeded with neural stem cells in a canine spinal cord injury model.</a> Cytotherapy. 12, 841-845 (2010).</li><li>Kim, J. H., Tu, T. W., Bayly, P. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+speed+does+not+determine+severity+of+spinal+cord+injury+in+mice+with+fixed+impact+displacement.">Impact speed does not determine severity of spinal cord injury in mice with fixed impact displacement.</a> Journal of Neurotrauma. 26, 1395-1404 (2009).</li><li>Kwon, B. K., Hillyer, J., Tetzlaff, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Translational+research+in+spinal+cord+injury:+a+survey+of+opinion+from+the+SCI+community.">Translational research in spinal cord injury: a survey of opinion from the SCI community.</a> J. Neurotrauma. 27, 21-33 (2010).</li><li>Kwon, B. K., Okon, E. B., Tsai, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+grading+system+to+evaluate+objectively+the+strength+of+pre-clinical+data+of+acute+neuroprotective+therapies+for+clinical+translation+in+spinal+cord+injury.">A grading system to evaluate objectively the strength of pre-clinical data of acute neuroprotective therapies for clinical translation in spinal cord injury.</a> J. Neurotrauma. 28, 1525-1543 (2011).</li><li>Norenberg, M. D., Smith, J., Marcillo, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pathology+of+human+spinal+cord+injury:+defining+the+problems.">The pathology of human spinal cord injury: defining the problems.</a> J. Neurotrauma. 21, 429-440 (2004).</li><li>Siegenthaler, M. M., Tu, M. K., Keirstead, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+extent+of+myelin+pathology+differs+following+contusion+and+transection+spinal+cord+injury.">The extent of myelin pathology differs following contusion and transection spinal cord injury.</a> J. Neurotrauma. 24, 1631-1646 (2007).</li><li>Talac, R., Friedman, J. A., Moore, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+spinal+cord+injury+for+evaluation+of+tissue+engineering+treatment+strategies.">Animal models of spinal cord injury for evaluation of tissue engineering treatment strategies.</a> Biomaterials. 25, 1505-1510 (2004).</li><li>Tator, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review+of+treatment+trials+in+human+spinal+cord+injury:+issues,+difficulties,+and+recommendations.">Review of treatment trials in human spinal cord injury: issues, difficulties, and recommendations.</a> Neurosurgery. 59, 957-982 (2006).</li></ol>

Several novel treatments have recently shown early promise in the field of SCI research.3 Careful evaluation of these treatments is essential in clinically relevant model of SCI to select strategies with maximum translational potential. A scheme of grading was recently developed to evaluate the strength of preclinical studies.9 This scheme emphasized the importance of utilizing contusion model of severe SCI. Here we describe such a contusion model of severe SCI with consistent lesion volumes and fun...

Choice of appropriate injury model is crucial for preclinical evaluation of novel treatments for spinal cord injury (SCI).1,2,13 In a recent survey of physicians and scientists in the field of neurotrauma contusion model, as opposed to hemisection or complete transection models, was universally accepted to be clinically relevant.8 This opinion is based on the observation that majority of spinal cord injury in humans is contusive in nature.10 The biology of contusion also appears to be different from hemisection or transection models.11 Iseda, et al. compared the effect of intraspinal chondroitinase ABC injection on neuroregeneration separately in hemisection and contusion models.4 Axonal regeneration was observed in the neuronal bridge in hemisection but not the contusion SCI group. The hemisection or complete transection models also create conditions known to exist in only a very small subset of clinical circumstances. For example, several investigators have employed scaffold-based interventions for implantation in the lesion cavity after hemisection or complete transection to promote regeneration.6 This approach becomes clinically irrelevant because creation of a cavity within injured spinal cord is impractical and probably unethical. 
Variability in functional recovery remains a major challenge for the contusion models.5,12 This variability can be minimized by the use of computer-controlled impactor and stabilization of spine before impact for uniform force delivery across the spinal cord volume especially the ventrally located motor pathways. It must be noted that plasticity and collateral contribution from surviving axons is the predominant mechanism of recovery after spinal cord injury.1 Therefore even minor variations in contusion technique may yield significantly different results. To this end we have developed a model of severe spinal cord injury which yields consistent contusion volume and functional recovery comparable with transection models. This model may be utilized for investigating both the neuroprotection and neuroregeneration strategies as a proof of concept for the treatment efficacy.

<table border="1">
<tbody>
 <tr>
 <td>Instrument/Drugs</td>
 <td>Company</td>
 <td>Cat #</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Computer controlled impactor</td>
 <td>Leica or the Infinite Horizons (formerly OSU) impactor</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Surgical instruments </td>
 </tr>
 <tr>
 <td>Scissors</td>
 <td>Fine Science Tools Inc
 </td>
 <td>14094-11 or 14060-09</td>
 <td></td>
 </tr>
 <tr>
 <td>Forceps</td>
 <td>Fine Science Tools Inc </td>
 <td>11006-12 and 11027-12 or 			11506-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Hemostats</td>
 <td>Fine Science Tools Inc </td>
 <td>13009-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Retractors</td>
 <td>Fine Science Tools Inc </td>
 <td>17011-10</td>
 <td></td>
 </tr>
 <tr>
 <td>Rongeurs</td>
 <td>Fine Science Tools Inc </td>
 <td>16020-14</td>
 <td></td>
 </tr>
 <tr>
 <td>Needle driver</td>
 <td>Fine Science Tools Inc </td>
 <td>12001-13</td>
 <td></td>
 </tr>
 <tr>
 <td>Stereotactic frame</td>
 <td> Leica or RWD Life Science Co. or
 TSE systems</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Buprinorphine</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Baytril</td>
 <td>Bayer</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Ketamine</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 </tbody>
</table>

a contusion model of severe spinal cord injury in rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

Lesion Volume
We have obtained large and consistent lesion volumes by following the technique described above. Using a Luxol fast blue staining a mean lesion volume of 2.04 mm3 (95% CI 1.9-2.18) (n = 5 animals) was obtained. Figure 2 shows mean lesion volume with a representative staining using Luxol fast blue through the lesion epicenter.
Functional Scores
The behavioral scores as measured by the Basso, Beattie, Bre...

Watch this Scientific Journal Video about A Contusion Model of Severe Spinal Cord Injury in Rats at JoVE.com

A Contusion Model of Severe Spinal Cord Injury in Rats

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

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