Name: severe burn injury in a swine model for clinical dressing assessment
Uploaded: 2018-11-06T12:30:00
Description: Watch this Scientific Journal Video about Severe Burn Injury in a Swine Model for Clinical Dressing Assessment at JoVE.com

This study was supported by a grant from the Tri-Service General Hospital; National Defense Medical Center, Taiwan (TSGH-C107-042); Ministry of Science and Technology, Taiwan (MOST 106-2314-B-016 -014); and the National Defense Medical Center (MAB-106-055; MAB-106-010; MAB-107-064).

Procedures involving animal subjects have been approved by the Animal Care Committee at National Defense Medical Center, Taiwan (R.O.C). This study was conducted in the Laboratory Animal Center at the National Defense Medical Center. Swine weighing between 20 and 25 kg has been successfully instrumented using this protocol.
1. Adaptation of the Animals to Human Handling
<ol>
	<li>After arrival in the facility, house the animals solitarily but let them interact with each other.</li>
	<li>Prov...

<ol>
	<li>Sen, C. K., 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=Human+skin+wounds:+a+major+and+snowballing+threat+to+public+health+and+the+economy.">Human skin wounds: a major and snowballing threat to public health and the economy.</a> Wound Repair Regeneration. 17, 763-771 (2009).</li><li>Sen, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+healing+essentials:+let+there+be+oxygen.">Wound healing essentials: let there be oxygen.</a> Wound Repair Regeneration. 17, 1-18 (2009).</li><li>Verhaegen, P. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+methods+for+assessing+epidermal+wound+healing:+the+effects+of+triamcinolone+acetonide+and+polyethelene+film+occlusion.">New methods for assessing epidermal wound healing: the effects of triamcinolone acetonide and polyethelene film occlusion.</a> Journal of Investigative Dermatology. 71, 332-384 (1978).</li><li>Swindle, M. M., Smith, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+anatomy+and+physiology+of+the+pig.">Comparative anatomy and physiology of the pig.</a> Scandinavian Journal of Laboratory Animal Sciences. 25, 1-10 (1998).</li><li>Swindle, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Swine in the Laboratory: Surgery, Anesthesia, Imaging and Experimental Techniques. , (2007).</li><li>Mertz, P. M., 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=IL-1+as+a+potent+inducer+of+wound+re-epithelization.">IL-1 as a potent inducer of wound re-epithelization.</a> Progress in Clinical and Biological Research. 365, 473-480 (1991).</li><li>Eming, S. A., Martin, P., Tomic-Canic, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+repair+and+regeneration:+mechanisms,+signaling,+and+translation.">Wound repair and regeneration: mechanisms, signaling, and translation.</a> Science Translation Medicine. 6, 265-266 (2014).</li><li>Aryza, M. J., Baryza, G. 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+Vancouver+Scar+Scale:+an+administration+tool+and+its+interrater+reliability.">The Vancouver Scar Scale: an administration tool and its interrater reliability.</a> The Journal of Burn &amp; Rehabilitation. 16, 535-538 (1995).</li><li>Bystrom, A., Claesson, R., Sundqvist, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+antibacterial+effect+of+camphorated+paramonochlorophenol,+camphorated+phenol+and+calcium+hydroxide+in+the+treatment+of+infected+root+canals.">The antibacterial effect of camphorated paramonochlorophenol, camphorated phenol and calcium hydroxide in the treatment of infected root canals.</a> Endodontics &amp; Dental Traumatology. 1, 170-175 (1985).</li><li>Reig, A., Tejerina, C., Codina, J., Hidalgo, J., Mirabet, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+a+new+cicatrization+dressing+in+treating+second-degree+burns+and+donor+sites.">Application of a new cicatrization dressing in treating second-degree burns and donor sites.</a> Annals of Burn and Fire Disasters. 4, 174 (1991).</li><li>Hindy, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+study+between+sodium+carboxymethyl-cellulose+silver,+moist+exposed+burn+ointment,+and+saline-soaked+dressing+for+treatment+of+facial+burns.">Comparative study between sodium carboxymethyl-cellulose silver, moist exposed burn ointment, and saline-soaked dressing for treatment of facial burns.</a> Annals of Burn and Fire Disasters. 22, 131-137 (2009).</li><li>Galiano, R. D., 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=Topical+vascular+endothelial+growth+factor+accelerates+diabetic+wound+healing+through+increased+angiogenesis+and+by+mobilizing+and+recruiting+bone+marrow-derived+cells.">Topical vascular endothelial growth factor accelerates diabetic wound healing through increased angiogenesis and by mobilizing and recruiting bone marrow-derived cells.</a> The American Journal of Pathology. 164, 1935-1947 (2004).</li><li>Thangarajah, 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=The+molecular+basis+for+impaired+hypoxia-induced+VEGF+expression+in+diabetic+tissues.">The molecular basis for impaired hypoxia-induced VEGF expression in diabetic tissues.</a> Proceedings of the National Academy of Sciences of the United States of America. , 13505-13510 (2009).</li></ol>

The present study established a swine model of severe burn injury and examined the model using a CAPS-containing dressing. Our results suggest that this swine model can be used for monitoring the clinical performance of experimental dressings, including antibacterial property. Wound healing rate, wound closure, and antibacterial activity were also analyzed using VSS, H&#38;E staining, and antibacterial test. The use of animal burn models has been developed as a valuable tool to review the pathophysiology of burn injury. ...

A burn injury initiates the inflammatory process and induces complex pathological effects, which influence numerous body functions immediately after an accident, resulting in negative impact on patients&#39; quality of life. Impaired wound healing causes significant morbidity and mortality among patients with diabetes mellitus1,2. Most patients with burn injuries experience pain during burn wound debridement, which is known as an excruciating process despite the use of powerful opioid analgesics3.
In contrast to other mammals, swine share several anatomic and physiologic characteristics with humans regarding the process of epithelialization, cellular proliferation, and angiogenesis. This makes swine a potentially better model for certain procedures and studies, and they are often used in subsequent studies that demonstrated promising results in mice. These features have led to the increasing use of swine as a major species in preclinical testing. Recently, a rapid increase in biomedical research with respect to cardiovascular, urinary, integumentary, and digestive systems has been observed4,5,6. This study aims to demonstrate a swine model of severe burn injury that eliminates wound contraction and provides a closer approximation of the human wound healing processes and the formation of new tissue. Six burn wounds were created symmetrically on the dorsum, three on each side of the spine of the swine. Next,anexperimental dressing was examined in a swine model of severe burn injury, which can be adapted to replicate human wound healing (Figure 1). The wounds were covered with a clinical dressing, which is composed of four layers: an inner contact layer of experimental materials, an inner intermediate layer of waterproof film, an outer intermediate layer of gauze, and an outer layer of adhesive plaster. A waterproof film keeps the wound environment moist while preventing bacterial infection and allowing gases to permeate the dressing. The outer intermediate layer of gauze was applied on the waterproof films and secured by an outer layer of adhesive plaster. At the completion of experiments, wound closure, wound area, and Vancouver Scar Scale (VSS) score were examined. The samples of skin resected from each animal post-sacrifice were histologically prepared and stained using hematoxylin and eosin (HE) staining. Antibacterial activity of each dressing in the context of wound healing was also examined in this model. Our previous study has demonstrated wound-healing performance in rat and swine model using a minimally invasive surgical technique7. Since there were six burn wounds on the dorsum within each swine, each experimental dressing was tested and evaluated in all positions to minimize bias related to wound healing process in different spots on the swine dorsum. Therefore, the swine model of severe burn injury established in this study provides a new approach for the evaluation of clinical dressings and facilitates the development of a novel treatment for burn injury. This study provides crucial tools to uncover the pathophysiology of burn wound healing.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr >
			<td>Sedation:</td>
			<td></td>
			<td></td>
			<td></td>	
	</tr>
		<tr >
			<td >Ketamine</td>
			<td >Merial</td>
			<td>2528 ESP</td>
			<td>10 mL vial</td>
			<td ></td>
		</tr>
		<tr >
			<td >Azaperone</td>
			<td >China chemical &#38; pharmaceutical</td>
			<td>47W406</td>
			<td>100 mL vial</td>
			<td ></td>
		</tr>
		<tr >
			<td >Atropine&#160;</td>
			<td >Oriental chemical works</td>
			<td>IN120802</td>
			<td>1 mL vial</td>
			<td ></td>
		</tr>
		<tr >
			<td>Anesthetic:</td>
			<td></td>
			<td></td>
			<td></td>	
	</tr>
		<tr >
			<td >Tiletamine+Zolazepam</td>
			<td >Virbac</td>
			<td>BC91</td>
			<td >5 mL vial</td>
			<td ></td>
		</tr>
		<tr >
			<td >Isoflurane</td>
			<td >Baxter</td>
			<td>N002A225</td>
			<td >100 mL vial</td>
			<td ></td>
		</tr>
		<tr >
			<td>Surgery:</td>
			<td></td>
			<td></td>
			<td></td>
			</tr>
		<tr >
			<td >Hair clippers</td>
			<td >Moser</td>
			<td>-</td>
			<td >-</td>
			<td ></td>
		</tr>
		<tr >
			<td >Povidone iodine scrub solution</td>
			<td >Ever star</td>
			<td>HA161202</td>
			<td >4 L barrel</td>
			<td ></td>
		</tr>
		<tr >
			<td >Modified iron</td>
			<td >-</td>
			<td>-</td>
			<td >-</td>
			<td ></td>
		</tr>
		<tr >
			<td >Electronic thermometer</td>
			<td >Dogger</td>
			<td>A9SA-ST9215C</td>
			<td >- 50~300&#8451;</td>
			<td ></td>
		</tr>
		<tr >
			<td >0.9% saline solution</td>
			<td >CHI SHENG</td>
			<td>KC130</td>
			<td >500 mL vial</td>
			<td ></td>
		</tr>
		<tr >
			<td >Gauze</td>
			<td >China Surgical Dressings Center</td>
			<td>MO15900080</td>
			<td >10 x10 cm</td>
			<td ></td>
		</tr>
		<tr >
			<td >CAPS</td>
			<td >CoreLeader Biotech Co., Ltd, Taipei, Taiwan</td>
			<td>-</td>
			<td >-</td>
			<td ></td>
		</tr>
		<tr >
			<td >Paper tape</td>
			<td >3M</td>
			<td>NDC-8333-1530-01</td>
			<td >2.5 cm x 9.1m</td>
			<td ></td>
		</tr>
		<tr >
			<td >Waterproof film</td>
			<td >3M</td>
			<td>NDC-8333-1600-40</td>
			<td >10 cm x 10 m</td>
			<td ></td>
		</tr>
		<tr >
			<td >Adhesive plaster</td>
			<td >Young chemical</td>
			<td>BH1426015</td>
			<td >10 cm x 10 m</td>
			<td ></td>
		</tr>
		<tr >
			<td>Dissection:</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Pair of standard sharp/blunt straight scissors (14 cm)</td>
			<td >Shinetec instruments</td>
			<td>ST-S114</td>
			<td >-</td>
			<td ></td>
		</tr>
		<tr >
			<td >Halstead-Mosquito (12.5 cm)</td>
			<td >Shinetec instruments</td>
			<td>ST-H012</td>
			<td >-</td>
			<td ></td>
		</tr>
		<tr >
			<td >Handle(# 4)</td>
			<td >Shinetec instruments</td>
			<td>ST-H004</td>
			<td >-</td>
			<td ></td>
		</tr>
		<tr >
			<td >Surgical Blade(#21)</td>
			<td >Shinetec instruments</td>
			<td>ST-B021</td>
			<td >-</td>
			<td ></td>
		</tr>
		<tr >
			<td>Post-Fixation &#38; Storage:</td>
		</tr>
		<tr >
			<td >50 ml Plastic centrifuge tube&#160;</td>
			<td >Falcon</td>
			<td>352070</td>
			<td >-</td>
			<td ></td>
		</tr>
		<tr >
			<td >10% neutral buffered formalin</td>
			<td >Leica</td>
			<td>3800604EG</td>
			<td >-</td>
			<td ></td>
		</tr>
		<tr >
			<td>Bacterial Growth Experiments&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Blood agar plate (BAP) (TSA with 5% sheep blood)&#160;</td>
			<td >CMP</td>
			<td>-</td>
			<td >90 mm Mono</td>
			<td></td>
		</tr>
	</tbody>
</table>

severe burn injury in a swine model for clinical dressing assessment

Wound healing is a dynamic repair process and is the most complex biological process in human life. In response to burn injury, alterations in biological pathways impair the inflammation response, resulting in delayed wound healing. Impaired wound healing frequently occurs in patients with diabetes leading to unfavorable outcomes such as amputation. Hence, dressings having beneficial effect in promoting burn wound repair are needed. However, studies on burn wound treatment are limited due to lack of proper animal models. Our previous study demonstrated wound-healing performance in rat and swine models using a minimally invasive surgical technique. This study aimed to demonstrate a swine model of severe burn injury that eliminates wound contraction and more closely approximates the human processes of re-epithelialization and new tissue formation. This protocol provides a detailed procedure for creating consistent burn wounds and examining the wound-healing performance under the treatment of an experimental dressing in a swine model. Six burn wounds were created symmetrically on the dorsum, which were covered with a clinical dressing composed of four layers: an inner contact layer of experimental materials, an inner intermediate layer of waterproof film, an outer intermediate layer of gauze, and an outer layer of adhesive plaster. Upon the completion of experiments, wound closure, wound area, and Vancouver Scar Scale score were examined. The samples of skin resected from each animal post-sacrifice were histologically prepared and stained using hematoxylin and eosin staining. Antibacterial activity of each dressing in the context of wound healing was also examined. The application of the clinical dressing to the wounds in swine model mimics the biological processes of human wound healing with respect to the processes of epithelialization, cellular proliferation, and angiogenesis. Therefore, this swine model provides an easy-to-learn, cost-effective, and robust method to assess the effect of clinical dressings in severe burn injury.

Burn duration of 30 seconds by hot iron resulted in wounds that were circular with a well-defined margin and uniformly pale with a rim of erythema (Figure 1D). Within each animal, there were six burn wounds on the dorsum. The arrangement of burn wounds was depicted in Figure 1K. Burn wounds were completely covered with CAPS-containing dressing and used to evaluate the depth of scar formation on post-burn days 0, 7, 21, and 42 and...

Watch this Scientific Journal Video about Severe Burn Injury in a Swine Model for Clinical Dressing Assessment at JoVE.com

Severe Burn Injury in a Swine Model for Clinical Dressing Assessment

To closely mimic the mode of burn injuries requires the interplay between clinical observation and studies in animal models. In this study, a swine model of severe burn injury was established to assess an experimental dressing in physiological and pathophysiological settings.

Wound healing is a dynamic repair process and is the most complex biological process in human life. In response to burn injury, alterations in biological ...

This method can help answer key questions in the burn injury field about warfare wound handling or how to treat burn wounds after fire accident. The main advantage of this swine burn injury model is the degree of similarity between the anatomical and the physiological structure of pig and human skin tissue. Though this method can provide insight into the effects of calcium alginate polysaccharide dressings on burn wound healing, it can also be applied to studies of burn management.Before delivering the burn, shave and moisture an approximately 25-centimeter-wide area of skin from the vertebral column all the way to the axilla on both sides of the spine of an anesthetized adult pig. Then, scrub the exposed skin with povidone-iodine for approximately five minutes. Use wet sterile gauze to remove the povidone-iodine soap, and sterilize the clean skin with povidone-iodine lotion.And use a surgical marking pen to demarcate the center of six burn wound locations on the dorsum of the animal. Then, cover the animal with sterile surgical drapes, taking care that the distance between each area is at least greater than the radius of the area. When all of the areas have been marked, fill a modified soldering iron equipped with an approximately nine square centimeter flat area with 50 milliliters of glycerin, and insert an electronic thermometer into the iron to monitor the temperature.After heating the iron to 137 to 139 degrees Celsius on a hot plate, gently press the iron onto each marked area for 30 seconds per area to create six uniform burn wounds. When the heated iron touches the skin, the heat will immediately dissipate across the surface of burnt area, so be sure to preheat the iron and the glycerin for at least 10 minutes before applying the burns. Then, wash the wounds with 0.9%saline solution, and measure and record the wound dimensions by photomicrography.When all the wounds have been measured, apply the dressings directly onto each wound, followed by the application of a waterproof film over the inner dressing layer to serve as a barrier against bacterial penetration. Then, tape a 5-centimeter-thick piece of gauze over each wound to serve as the mid-layer of the dressing, and secure the gauze with an outer layer of adhesive plaster, extending the plaster to the torso to avoid displacement of the dressing. Change the clinical dressings every two days for the first 10 days, then twice a week until day 42, cleaning and measuring the wounds according to the Vancouver Scar Scale before each dressing reapplication.Swab the wound for antibacterial testing on post-burn days zero, seven, 21, and 42, and place the swabs in 100 milliliters of 0.9%sterile saline solution. Gently vortex the swabs to obtain a homogenous suspension, and serially dilute the resulting cell solution. Then, plate 100 microliters of each dilution in selective or nonselective medium, and incubate the wound culture under aerobic conditions at 37 degrees Celsius for 24 to 72 hours.At the end of the incubation, plate 10-microliter aliquots from each dilution culture in triplicate onto blood agar plates supplemented with 5%sheep blood for overnight culture at 37 degrees Celsius. The next morning, use a marker and a small ruler to divide the bottom of each dish into four equal quadrants, and determine the number of colony-forming units in each quadrant under a dissecting microscope. In this representative experiment, burn wounds re-epithelialization was evaluated by gross inspection on post-burn day 42.A significant reduction in wound area was observed compared to day zero. The healing rate was defined as the greatest average wound margin distance from the wound center divided by the time to complete wound closure, demonstrating over 73%wound closure on post-burn day 42. The Vancouver scar scores peaked on post-burn day 21 and decreased to below the initial wound size by post-burn day 42.Histologic examination of skin samples harvested on day 42 confirmed that full-thickness burns were achieved and that the wounds appeared to be fully healed. Necrosis resulting from burns could be observed in the epidermis, dermis, and dermal components of the wound without significantly affecting the underlying muscle. The thickness beneath the experimental dressing was 5.4 millimeters, and dermal sloughing and lymphocytic infiltration were observed.Bacterial cultures of wound swabs collected on post-burn days zero, seven, 21, and 42 demonstrated a slight increase in colony-forming units from days zero to 21 that were significantly increased on day 42, confirming that this swine model of severe burn injury can be used for monitoring the clinical performance of experimental dressings. While attempting this procedure, it's important to remember to know how to handle the animal and then to properly monitor the animal's vital sign during the evental period. Following this procedure, other methods like bacterial growth experiments can be performed to answer additional questions about how natural polysaccharide materials work in cases of severe burn injury.After its development, this technique paved the way for researchers in the field of burn wound healing to explore the effects of natural calcium alginate polysaccharide dressings in human skin healing. Don't forget that working with heated iron and glycerine can be extremely hazardous and that precautions such as functioning equipment should always be taken while performing the procedure.

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Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

A Chronic Cardiac Ischemia Model in Swine Using an Ameroid Constrictor

Cardiovascular disease remains the number one cause of mortality in the United States. There are numerous approaches to treating these diseases, but ...

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of ...

Establishment of a Severe Dry Eye Model Using Complete Dacryoadenectomy in Rabbits

Dry eye disease (DED) is a complex disease with multiple etiologies and variable symptoms, having ocular surface inflammation as its key pathophysiologic ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...