Name: assessment of plasma coagulation on liver tissue in a large animal model in vivo
Uploaded: 2018-08-04T13:00:00
Description: Watch this Scientific Journal Video about Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo at JoVE.com

The authors have no acknowledgements.

Rules governed by German legislation for animal studies as well as Principles of Laboratory Animal Care (National Institutes of Health publication ed. 8, 2011) were followed. Official permission is granted from the governmental animal care office (Landesamt f&#252;r Natur, Umwelt und Verbraucherschutz Nordrhein-Westfalen, Recklinghausen, Germany).
1. Animals
<ol>
	<li>Use female German landrace pigs (weighing 25-30 kg) housed in open cages.</li>
	<li>Use 5 animals per group (argon and helium...

<ol>
	<li>Link, W. J., Incropera, F. P., Glover, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+plasma+scalpel:+comparison+of+tissue+damage+and+wound+healing+with+electrosurgical+and+steel+scalpels.">A plasma scalpel: comparison of tissue damage and wound healing with electrosurgical and steel scalpels.</a> ArchSurg. 111, 392-397 (1976).</li><li>Kwon, A. H., Inui, H., Kamiyama, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Successful+laparoscopic+haemostasis+using+an+argon+beam+coagulator+for+blunt+traumatic+splenic+injury.">Successful laparoscopic haemostasis using an argon beam coagulator for blunt traumatic splenic injury.</a> EurJSurg. 167, 316-318 (2001).</li><li>Frilling, A., 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=Effectiveness+of+a+new+carrier-bound+fibrin+sealant+versus+argon+beamer+as+haemostatic+agent+during+liver+resection:+a+randomised+prospective+trial.">Effectiveness of a new carrier-bound fibrin sealant versus argon beamer as haemostatic agent during liver resection: a randomised prospective trial.</a> Langenbecks ArchSurg. 390, 114-120 (2005).</li><li>Raiser, J., Zenker, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argon+plasma+coagulation+for+open+surgical+and+endoscopic+applications:+state+of+the+art.">Argon plasma coagulation for open surgical and endoscopic applications: state of the art.</a> J Phys Appl Phys. 39 (16), 3520-3523 (2006).</li><li>Farin, G., Grund, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technology+of+argon+plasma+coagulation+with+particular+regard+to+endoscopic+applications.">Technology of argon plasma coagulation with particular regard to endoscopic applications.</a> EndoscSurgAllied Technol. 2, 71-77 (1994).</li><li>Grund, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argon+plasma+coagulation+(APC):+ballyhoo+or+breakthrough?.">Argon plasma coagulation (APC): ballyhoo or breakthrough?.</a> Endoscopy. 29, 196-198 (1997).</li><li>Glowka, T. R., Standop, J., Paschenda, P., Czaplik, M., Kalff, J. C., Tolba, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argon+and+helium+plasma+coagulation+of+porcine+liver+tissue.">Argon and helium plasma coagulation of porcine liver tissue.</a> J Int Med Res. , (2017).</li><li>Dowling, R. D., Ochoa, J., Yousem, S. A., Peitzman, A., Udekwu, A. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argon+beam+coagulation+is+superior+to+conventional+techniques+in+repair+of+experimental+splenic+injury.">Argon beam coagulation is superior to conventional techniques in repair of experimental splenic injury.</a> JTrauma. 31, 717-720 (1991).</li><li>Go, P. M., Goodman, G. R., Bruhn, E. W., Hunter, J. 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+argon+beam+coagulator+provides+rapid+hemostasis+of+experimental+hepatic+and+splenic+hemorrhage+in+anticoagulated+dogs.">The argon beam coagulator provides rapid hemostasis of experimental hepatic and splenic hemorrhage in anticoagulated dogs.</a> JTrauma. 31, 1294-1300 (1991).</li><li>Brand, C. U., Blum, A., Schlegel, A., Farin, G., Garbe, C. <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+argon+plasma+coagulation+in+skin+surgery.">Application of argon plasma coagulation in skin surgery.</a> Dermatology. 197, 152-157 (1998).</li><li>Carus, T., Rackebrandt, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collateral+tissue+damage+by+several+types+of+coagulation+(monopolar,+bipolar,+cold+plasma+and+ultrasonic)+in+a+minimally+invasive,+perfused+liver+model.">Collateral tissue damage by several types of coagulation (monopolar, bipolar, cold plasma and ultrasonic) in a minimally invasive, perfused liver model.</a> ISRNSurg. , 518924 (2011).</li><li>Bludau, M., Vallbohmer, D., Gutschow, C., Holscher, A. H., Schroder, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+measurement+of+gastric+mucosal+microcirculation+using+a+combined+laser+Doppler+flowmeter+and+spectrophotometer.">Quantitative measurement of gastric mucosal microcirculation using a combined laser Doppler flowmeter and spectrophotometer.</a> DisEsophagus. , (2008).</li><li>Beckert, S., Witte, M. B., Konigsrainer, A., Coerper, 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+impact+of+the+Micro-Lightguide+O2C+for+the+quantification+of+tissue+ischemia+in+diabetic+foot+ulcers.">The impact of the Micro-Lightguide O2C for the quantification of tissue ischemia in diabetic foot ulcers.</a> Diabetes Care. 27, 2863-2867 (2004).</li><li>Erdogan, D., de Graaf, W., van Gulik, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adhesive+strength+of+fibrinogen-coated+collagen+patch+or+liquid+fibrin+sealant+in+an+experimental+liver+resection+model+in+pigs.">Adhesive strength of fibrinogen-coated collagen patch or liquid fibrin sealant in an experimental liver resection model in pigs.</a> Eur Surg Res Eur Chir Forsch Rech Chir Eur. 41 (3), 298-302 (2008).</li><li>Knobloch, 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=Microcirculation+of+the+sternum+following+harvesting+of+the+left+internal+mammary+artery.">Microcirculation of the sternum following harvesting of the left internal mammary artery.</a> ThoracCardiovascSurg. 51, 255-259 (2003).</li><li>Kanzler, S., 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=Recommendation+for+severity+assessment+following+liver+resection+and+liver+transplantation+in+rats:+Part+I.">Recommendation for severity assessment following liver resection and liver transplantation in rats: Part I.</a> Lab Anim. 50 (6), 459-467 (2016).</li><li>Pehb&#246;ck, D., Dietrich, H., Klima, G., Paal, P., Lindner, K. H., Wenzel, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anesthesia+in+swine+optimizing+a+laboratory+model+to+optimize+translational+research.">Anesthesia in swine optimizing a laboratory model to optimize translational research.</a> Anaesthesist. 64 (1), 65-70 (2015).</li><li>Nykonenko, A., V&#225;vra, P., Zon&#269;a, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomic+Peculiarities+of+Pig+and+Human+Liver.">Anatomic Peculiarities of Pig and Human Liver.</a> Exp Clin Transplant Off J Middle East Soc Organ Transplant. 15 (1), 21-26 (2017).</li><li>Fechner, G., von Pezold, J., Luzar, O., Hauser, S., Tolba, R. H., M&#252;ller, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modified+spectrometry+(O2C+device)+of+intraoperative+microperfusion+predicts+organ+function+after+kidney+transplantation:+a+pilot+study.">Modified spectrometry (O2C device) of intraoperative microperfusion predicts organ function after kidney transplantation: a pilot study.</a> Transplant Proc. 41 (9), 3575-3579 (2009).</li><li>Patterson, E. J., Scudamore, C. H., Owen, D. A., Nagy, A. G., Buczkowski, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiofrequency+ablation+of+porcine+liver+in+vivo:+effects+of+blood+flow+and+treatment+time+on+lesion+size.">Radiofrequency ablation of porcine liver in vivo: effects of blood flow and treatment time on lesion size.</a> AnnSurg. 227, 559-565 (1998).</li></ol>

Rodent models for liver surgery are established for a long time16. Nevertheless, large animal models offer certain advantages: no microsurgical equipment is needed as standard operative equipment for humans can be applied, surgical techniques are comparable to clinical use and standard clinical evaluation methods can be transferred to the experiments. For example, standard clinical blood tests can be carried out without the need for special laboratory test methods (Figure 10</...

Argon plasma coagulation (APC) is a widely used instrument in abdominal surgery for more than three decades1,2. It is a standard technique for the achievement of secondary hemostasis after major hepatectomy by sealing the liver cut surface to prevent later hemorrhages3. Plasma coagulation is a specialized form of radiofrequency electrocautery, which delivers the electrical energy through an arc of ionized gas. Providing monopolar electrothermal hemostasis, this noncontact technique has the advantage of preventing the electrode to stick to the tissue4. The ionized gas beam is automatically directed to the area of the lowest electrical resistance and is turned away when resistance rises due to desiccation to other areas not yet desiccated. This produces a uniform limited depth of coagulation5,6. Factors influencing the coagulation effect are the activation time, the power setting of the coagulation device and the distance from the probe to the tissue. Helium is another carrier gas, which can be used for plasma coagulation7. Recent clinical studies concentrated on clinical outcomes rather than histological and functional findings3,8,9, while experimental studies focused on in vitro investigations10 or experiments on isolated perfused organs11.
The underlying protocol allows the study of the effects of plasma coagulation in a large animal model close to the clinical application using standard human equipment on pigs: Microcirculation is assessed noninvasively by a laser Doppler flowmeter and spectrophotometer, which is a standard clinical tool for this indication12,13. Temperature changes during coagulation are monitored with an infrared thermometer and a thermographic camera. The depth of coagulation is measured on histological hematoxylin/eosin stained sections after harvesting of tissue samples. For the comparison with other means for secondary hemostasis, burst pressure experiments are performed. In contrast to previously described techniques14, these are conducted on explanted organs to exclude blood pressure related effects. In addition to the described investigations on the local effects of plasma coagulation, standard blood tests can also be undertaken in the porcine model.

<table border="0" cellpadding="0" cellspacing="0" width="824">
	<tbody>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Xylazine 20 mg/mL</td>
			<td style="width:184px;">Vetoquinol GmbH</td>
			<td style="width:192px;">Xylapan</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Ketamine 100 mg/mL</td>
			<td style="width:184px;">Ceva GmbH</td>
			<td style="width:192px;">Ceva Ketamine Injection</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Atropine 100 mg / 10 mL</td>
			<td style="width:184px;">Dr. Franz K&#246;hler Chemie GmbH</td>
			<td style="width:192px;">Atropinsulfat K&#246;hler 100mg Amp.</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;">Propofol</td>
			<td style="width:184px;">Fresenius Kabi GmbH</td>
			<td style="width:192px;">Propofol 1% MCT Fresenius</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;">Fentanyl</td>
			<td style="width:184px;">KG Rotexmedica GmbH</td>
			<td style="width:192px;">Fentanyl 0,5mg Rotexmedica</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;">Isoflurane</td>
			<td style="width:184px;">Abbot GmbH</td>
			<td style="width:192px;">Forene 100% (V/V) 250 mL</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;">Ringer&#39;s lactate solution</td>
			<td style="width:184px;">Baxter Deutschland GmbH</td>
			<td style="width:192px;"></td>
			<td style="width:232px;">sodium 131mmol/l, potassium 5 mmol/l, calcium 2 mmol/l, cloride 111 mmol/l, lactate 29 mmol/l</td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Surgical disinfactant</td>
			<td style="width:184px;">Sch&#252;lke &#38; Mayr GmbH</td>
			<td style="width:192px;">Kodan Tinktur forte gef&#228;rbt 1l 104804</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Motorized microscope</td>
			<td style="width:184px;">Nikon Instruments Europe</td>
			<td style="width:192px;">Eclipse TE2000-E</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Microscope camera</td>
			<td style="width:184px;">Nikon Instruments Europe</td>
			<td style="width:192px;">Digitalsight DS-Qi1Mc</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Imaging software</td>
			<td style="width:184px;">Nikon Instruments Europe</td>
			<td style="width:192px;">NIS elements Vers. 4.40</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Plasma coagulator</td>
			<td style="width:184px;">S&#246;ring GmbH</td>
			<td style="width:192px;">CPC-1000</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Argon gas</td>
			<td style="width:184px;">Linde AG</td>
			<td style="width:192px;">Argon 4.8&#160;</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Helium gas</td>
			<td style="width:184px;">Linde AG</td>
			<td style="width:192px;">Helium 4.8</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">O2C</td>
			<td style="width:184px;">LEA Medizintechnik GmbH</td>
			<td style="width:192px;">O2C Version 1212</td>
			<td style="width:232px;">with LF-2 or LF-3 probe</td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;">Infrared thermometer</td>
			<td style="width:184px;">Voltcraft</td>
			<td>VOLTCRAFT IR 260-8S</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Thermographic camera</td>
			<td>InfraTec GmbH</td>
			<td>VarioCAM HD head 820</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Thermographic analysis sofrtware</td>
			<td>InfraTec GmbH</td>
			<td style="width:192px;">IRBIS 3</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Mayer&#39;s Hematoxylin solution</td>
			<td></td>
			<td style="width:192px;">Merck 1.09249</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Eosin solution</td>
			<td style="width:184px;">VWR International GmbH</td>
			<td style="width:192px;">Merck 1.09844</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Rollerpump Masterflex L/S easy Load</td>
			<td style="width:184px;">Cole-Parmer Instrument Company</td>
			<td style="width:192px;">model 7518-10</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Perfusorpump</td>
			<td style="width:184px;">B. Braun Melsungen AG</td>
			<td style="width:192px;">Perfusor secura FT</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Digital pressure meter</td>
			<td style="width:184px;">Greisinger electronic</td>
			<td style="width:192px;">GMH 3161</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Perfusorsyringe, 50 mL</td>
			<td style="width:184px;">B. Braun Melsungen AG</td>
			<td style="width:192px;">REF 8728810 F</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Perfusor line, Type IV Standard, PVC Luer lock</td>
			<td style="width:184px;">B. Braun Melsungen AG</td>
			<td style="width:192px;">REF 8722960</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">3-Way stopcock, Dicofix C35C</td>
			<td style="width:184px;">B. Braun Melsungen AG</td>
			<td style="width:192px;">REF 16494 C</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Silk 2-0. 3 metric</td>
			<td style="width:184px;">Resorba</td>
			<td style="width:192px;">REF H5F</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Vicryl 4-0 Sutupak</td>
			<td style="width:184px;">Ethicon</td>
			<td>V1224H</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">NaCl 0.9 %</td>
			<td style="width:184px;">B. Braun Melsungen AG</td>
			<td style="width:192px;"></td>
			<td style="width:232px;"></td>
		</tr>
	</tbody>
</table>

assessment of plasma coagulation on liver tissue in a large animal model in vivo

Plasma coagulation as a form of electrocautery is used in liver surgery for decades to seal the large liver cut surface after major hepatectomy to prevent hemorrhages at a later stage. The exact effects of plasma coagulation on liver tissue are only poorly examined. In our porcine model, the coagulation effects can be examined close to the clinical application. A combined laser Doppler flowmeter and spectrophotometer documents microcirculation changes during coagulation at 8 mm tissue depth noninvasively, providing quantifiable information about hemostasis beyond the subjective clinical impression. The temperature at coagulation site is assessed with an infrared thermometer prior and post coagulation and with a thermographic camera during coagulation, a measurement of the gas beam temperature is not possible due to the upper threshold of the devices. The depth of coagulation is measured microscopically on hematoxylin/eosin stained sections after calibration with an object micrometer and gives an exact information about the power setting-coagulation depth-relation. The sealing effect is examined on the bile ducts as it is not possible for a plasma coagulator to seal larger blood vessels. Burst pressure experiments are carried out on explanted organs to rule out blood pressure related effects.

Microcirculation: Utilizing the diagnostic device for hemostasis following plasma coagulation can be demonstrated by microcirculation changes. Capillary blood flow (displayed as arbitrary units (AU)) decreases from a baseline value of 142.7 &#177; 76.08 AU to 57.78 &#177; 49.57 AU at 25 W device output power, to 48.5 &#177; 7.26 AU at 50 W and to 5.04 &#177; 1.31 AU at 100 W (Figure 4).
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Watch this Scientific Journal Video about Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo at JoVE.com

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

Here we present a protocol to experimentally assess plasma coagulation in liver tissue in vivo. In a porcine model, microcirculation is examined by laser Doppler, coagulation depth is measured histologically, temperature at coagulation site by infrared thermometer and thermographic camera, and duct sealing effect is documented by burst pressure experiments.

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

Plasma coagulation as a form of electrocautery is used in liver surgery for decades to seal the large liver cut surface after major hepatectomy to prevent ...

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