Name: A Translational Surgical Porcine Model for Postoperative Intra-Abdominal Adhesion Formation (Video) | JoVE Journal
Uploaded: 2026-03-13T12:55:00.000+00:00
Duration: 3 min 47 s
Description: Explore a translational porcine model for postoperative intra-abdominal adhesion formation using Yucatan mini pigs. This protocol combines laparotomy, bowel resection, and peritoneal abrasion to study fibrosis biology and evaluate anti-adhesion therapeutics.

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5 minutes

The research reported in this publication was supported by the Stanford SPARK program, American College of Surgeons Surgical Adhesions Improvement Project Grant, generously supported by the Carlino family, and the Department of Surgery at Stanford University. We would like to thank Stanford Veterinary Service Center and the staff at the Comparative Animal Pavillion. All ideas, analyses, references, and figures were created and verified by the authors. AI tools were used only for minor language and grammar edits.

Ethics Statement
All porcine studies were conducted under protocols approved by the Stanford University Institutional Animal Care and Use Committee (IACUC) protocol #34052 as previously described28.
1. Animal Husbandry
<ol>
	<li>Obtain Yucatan mini pigs (recommended 9- to 12-kg, 7 to 9 weeks old) from a verified vendor. We recommend&#160;n = 4 pigs per cohort. House animals in pairs in light- and temperature-controlled rooms.</li>
	<li>Provide standard porcine chow ad libitum. The animals should have access to a continuous supply of water to avoid dehydration. 
	NOTE: Animals were fasted for 12 h prior to surgery and terminal harvest.</li>
	<li>Monitor and record body weight and general health at regular intervals per institutional standards.</li>
</ol>
2. Anesthesia and Preoperative Preparation
<ol>
	<li>Sedate pigs in their home enclosures using intramuscular Xylazine (2.2 mg/kg, intramuscular (IM)) and Ketamine (12 mg/kg IM). Obtain intravenous (IV) access. Carefully transport animals to the surgical suite once sedation is deemed adequate by a veterinarian. Please refer to the Supplementary Table for anesthesia and analgesia information, as protocols may vary by institution.</li>
	<li>Once in the surgical suite, secure the animal&#39;s airway via endotracheal intubation. Maintain anesthesia throughout the procedure with isoflurane administered in oxygen via a precision vaporizer through an anesthesia circuit equipped with waste gas scavenging under the supervision of a licensed veterinarian.</li>
	<li>Administer intravenous (IV) fluids, anti-nausea medications (Cerenia 1-2 mg/kg IV), vasoconstricting medications, and inotropes as needed to treat intraoperative hypotension to avoid bowel hypoperfusion and causing potential risk of anastomotic leak. Please see Supplementary Table for full medication details, regimens will vary per institution.</li>
	<li>Apply ophthalmic ointment to lubricate eyes and prevent corneal abrasions. Administer cefazolin (25 mg/kg, IV) or equivalent antibiotic 1 hour prior to incision to prevent a wound infection. Administer an intraoperative analgesic regimen according to institutional veterinary standards. A reference regimen is provided in the Supplementary Table and includes a combination of Carpofen (25 mg/kg IV), Buprenorphine ER (0.18 mg/kg IV).</li>
</ol>
3. Surgical Procedure
<ol>
	<li>Position the animal supine on a heating pad to maintain normothermia. Measure oxygen saturations (SpO2) via pulse oximeter, pulse with a heart rate monitor, and temperature every 10 mins.</li>
	<li>Remove hair from the abdomen using electric clippers, then prepare the skin with a single round of alcohol swabs followed by gauze soaked in povidone-iodine. Blot the excess antiseptic solution away from the abdomen using a sterile towel.</li>
	<li>Drape the abdomen using sterile towels and surgical drapes to maintain sterility throughout the procedure. Infiltrate lidocaine (2 mg/kg subcutaneously (SQ)) into the abdominal wall at the planned incision site using a 25-gauge needle.</li>
	<li>Perform a midline laparotomy by making a full-thickness incision through the skin, subcutaneous tissue, and abdominal wall musculature along the linea alba to enter the peritoneal cavity using a 10-blade scalpel. Place abdominal wall retractors and eviscerate the intestines.</li>
	<li>Identify a region of the bowel ~10-15 cm proximal to the ileocecal valve and resect a 2.5-cm (1-inch) segment of the small bowel using scissors. Resect the associated mesentery with electrocautery for hemostasis of the mesenteric blood vessels. Close the two open bowel ends using interrupted 3-0 silk sutures to achieve a watertight closure. Perform a two-layer, side-to-side primary anastomosis (functional end-to-end). Reconstruct the inner layer of the lumen with a synthetic 3-0 absorbable suture in a running fashion and the outer layer with a running 3-0 non-absorbable suture. Attempt to minimize bowel spillage throughout the procedure.</li>
	<li>Gently abrade a 3 x 5 cm section of the parietal peritoneum along the right abdominal sidewall using sterile gauze or an electrocautery scratch pad until the peritoneal surface appears frayed or there is sub serosal edema in the abdominal wall (10-15 strokes with medium pressure). The left abdominal sidewall can be left unabraded to serve as a technical control although using this model stochastic adhesions form throughout the abdomen in a manner similar to human disease.</li>
	<li>Reposition the bowel into the abdominal cavity and remove the abdominal wall retractors. Irrigate the abdomen with 1 L of warmed sterile saline, evacuate all fluid and confirm hemostasis (no active bleeding) prior to closure.</li>
	<li>Close the fascia with a running 2-0 absorbable suture. Close the skin with a running 2-0 or 3-0 non-absorbable baseball stitch and cover the incision with a sterile dressing. The average duration of the procedure was 1 hour.</li>
	<li>Administer Atipamezole (0.2 mg/kg IV) at the end of the procedure to reverse sedation prior to extubating. Extubate animals upon return of spontaneous respiration. Monitor vital signs (pulse, oxygen saturation, temperature, blood pressure) continuously until fully recovered from anesthesia.</li>
</ol>
4. Postoperative Care
<ol>
	<li>Administer Carprofen (25 - 50 mg) twice a day by mouth (PO) the first 3 days after the surgical procedure and then as needed (PRN) for pain. Maintain the animals on a liquid&#160;high&#160;protein diet (1.5 bottles twice a day) for the first 3 postoperative days. 
	NOTE: Of note, although routine suture removal at 10&#8211;14 days postoperatively is common practice, sutures were left in place in animals without signs of wound infection or dehiscence to avoid an additional anesthetic exposure prior to the planned 4-week harvest.</li>
	<li>Administer amoxicillin (10 mg/kg) twice a day by mouth for one week then PRN for signs of wound infection (redness, warmth, swelling with or without drainage).</li>
	<li>Check on animals twice a day for appetite, activity, bowel function and wound integrity. If concerned about infection, discuss antibiotic dosage and duration with the veterinarian. The sterile dressing should be left on for the first 3 days after the surgical procedure to minimize risk of wound infection. Record weight in 2 - 3 day intervals to ensure weight maintenance postoperatively.</li>
	<li>Transition animals to solid chow beginning on postoperative day 4 (POD4). Bridge solid chow intake with supplemental liquid high protein diet until tolerating sufficient calories from solid chow alone.</li>
</ol>
5. Euthanasia
<ol>
	<li>Euthanize animals at 4 weeks under veterinary supervision prior to necropsy and tissue harvest using an intravenous euthanasia solution according to institutional and AVMA standards.</li>
	<li>Assess adhesions grossly. Immediately collect tissues and blood samples for downstream analyses. Dispose of all biological and pharmaceutical waste according to your institution&#39;s Environment Health and Safety guidelines.</li>
</ol>

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</ol>

The authors have no relevant disclosures.

The proposed porcine adhesion model provides a clinically relevant and reliable method for modeling intra-abdominal adhesions. In contrast to focal or single-site injury models, the combination of midline laparotomy, small bowel resection with anastomosis, and standardized parietal peritoneal abrasion was intentionally selected to recapitulate the multifactorial peritoneal insults encountered during open abdominal surgical procedures. The controlled abrasion of the parietal peritoneum along the abdominal sidewall mimics ...

Postoperative adhesions are a major source of morbidity following abdominal and pelvic surgery, developing in up to 93% of patients1,2, with approximately 20% requiring hospitalization within five years for adhesion-related complications3,4. Peritoneal injury arises from abdominal surgical procedures as well as intra-abdominal irritation (infection or chemical exposure) or chronic inflammatory conditions such as inflammatory bowel disease. The resulting adhesions can lead to a wide range of clinical consequences, including recurrent bowel obstructions sometimes requiring additional surgeries, or causing infertility and chronic pain5,6,7.&#160;Adhesion formation can be particularly severe following multi-visceral resections for malignancy, such as cytoreductive surgeries, chronic inflammation, or bowel perforation, where extensive peritoneal injury amplifies the fibrotic response. Additionally, adhesive disease can significantly complicate subsequent surgical interventions, often making it impossible to proceed with a minimally invasive approach or causing inadvertent iatrogenic injury to the bowel or other organs.
Despite their clinical significance, the development of effective anti-adhesion strategies is limited - likely hindered in part by limitations in preclinical models that recapitulate the anatomy, physiology, and wound-healing responses seen in humans8. Rodent models are widely used due to their availability, experimental tractability, and reliable adhesion generation9,10,11. However, their small peritoneal cavity, thin mesentery, and distinct inflammatory milieu can limit translational relevance8,12. Such models have contributed to the development of anti-adhesion approaches that demonstrated robust efficacy in animal studies but demonstrated limited or context-dependent clinical benefit in humans8,13,14,15,16,17. In addition, human-comparable surgical procedures such as bowel resection and anastomosis are difficult to model in a translationally relevant manner in rodents, and species-specific differences in drug metabolism further limit&#160;clinical extrapolation18.
Large animal models, particularly swine, are widely used to study abdominal processes due to their anatomical and physiological similarity to humans and their comparable surgical handling, providing a platform to study both adhesion formation and test therapeutic interventions19,20,21,22. Existing porcine adhesion models have provided important insights, yet many focus on procedure-specific adhesion formation and often lack standardized procedures limiting their applicability particularly to severe adhesions arising after open intestinal surgery23,24,25,26,27.
To address these limitations, we developed a porcine model of postoperative intra-abdominal adhesions that integrates midline laparotomy, small bowel resection, and a unilateral, parietal peritoneal abrasion. This multifocal peritoneal strategy recapitulates adhesion-promoting insults commonly encountered during abdominal surgical procedures and produces adhesions of different strengths and locations at the four-week timepoint with guaranteed adhesions on the side of the abdomen&#160;where directed parietal abrasion is performed as well as stochastic adhesions throughout the abdomen. By incorporating a clinically relevant intestinal surgical procedure with a defined and reproducible peritoneal injury, this model overcomes key limitations of prior porcine systems and provides a consistent platform for studying postoperative adhesion formation and evaluating anti-adhesion interventions.

<table><tbody><tr><td>&lt;strong&gt;Animals&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Female Yucatan mini pigs, 9 to 12 kg in body weight (7 to 9 weeks old)</td><td>S&amp;amp;S Farms</td><td></td><td>8</td></tr><tr><td>Surgical Supplies</td><td></td><td></td><td></td></tr><tr><td>DMI4000 B microscope</td><td>Leica Microsystems</td><td></td><td>Histology</td></tr><tr><td>&lt;strong&gt;Materials&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Euthasol&amp;nbsp;</td><td>Virbac</td><td></td><td>Euthanasia</td></tr><tr><td>Eosin-Y Alcoholic Stain</td><td>Azer Scientific&amp;nbsp;&amp;nbsp;</td><td>Cat#ES709</td><td>Histology</td></tr><tr><td>Frosted slides</td><td>Fisher Scientific&amp;nbsp;</td><td>Cat#12-550-400</td><td>Histology</td></tr><tr><td>Mayer&amp;prime;s Hematoxylin Solution&amp;nbsp;</td><td>Sigma-Aldrich&amp;nbsp;</td><td>Cat#MHS80&amp;nbsp;</td><td>Histology</td></tr><tr><td>&lt;strong&gt;Medications&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Amoxicillin</td><td>Covetrus</td><td>59629</td><td>Infection prevention&lt;br/&gt; 10mg/kg PO BID x1</td></tr><tr><td>Ancef/Cefazolin</td><td>Covetrus</td><td>84773</td><td>Pre-op infection control&lt;br/&gt; 25 mg/kg IV</td></tr><tr><td>Atipamezole</td><td>Covetrus</td><td>82124</td><td>Sedation Reversal Agent&lt;br/&gt; 0.2 mg/kg&amp;nbsp;</td></tr><tr><td>Atropine</td><td>Covetrus</td><td>74760</td><td>Bradycardia&lt;br/&gt; 0.04mg/kg&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Bupernorphine Extended Release (ER)</td><td>Wedgewood Pharamcy</td><td></td><td>Analgesia&lt;br/&gt; 0.18 mg/kg IV</td></tr><tr><td>Carprofen - intraoperative</td><td>Dechra</td><td></td><td>Analgesia&lt;br/&gt; 25 mg/kg IV&amp;nbsp;</td></tr><tr><td>Carprofen - post-operative</td><td>Dechra</td><td></td><td>Analgesia&lt;br/&gt; 25-50mg PO PRN</td></tr><tr><td>Cerenia</td><td>Zoetis</td><td></td><td>Anti-nausea&lt;br/&gt; 1-2 mg/kg IV once intraoperatively</td></tr><tr><td>Isoflurane</td><td>Dechra</td><td></td><td>Sedation&amp;nbsp;</td></tr><tr><td>Lidocaine 2%</td><td>VetOne</td><td>V1 510212</td><td>Analgesia&lt;br/&gt; 2 mg/kg SQ prior to incision</td></tr><tr><td>Ketamine</td><td>Dechra</td><td></td><td>Anesthsia &amp;amp; Analgesia&lt;br/&gt; 12 mg/kg IM</td></tr><tr><td>Dobutamine</td><td>Hospira&amp;nbsp;</td><td></td><td>Dosing and titration per anesthesiologist&amp;nbsp;</td></tr><tr><td>Povidone-iodine (Betadine)</td><td>ValleyVet</td><td>12265</td><td>Antiseptic</td></tr><tr><td>Xylazine (Rompun)</td><td>Dechra</td><td>GW l400000</td><td>Sedation/Analgesia&lt;br/&gt; 2.2 mg/kg IM</td></tr><tr><td>&lt;strong&gt;Surgical Supplies&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>3-0 PDS (polydioxanone) Suture</td><td>Ethicon</td><td>Cat#Z316H</td><td>Anastamosis&lt;br/&gt; Quantity - 2&amp;nbsp;</td></tr><tr><td>3-0 Silk Suture (Pop-off)&amp;nbsp;</td><td>Ethicon</td><td>Cat#1679H</td><td>Anastamosis&lt;br/&gt; Quantity - 1 pack of pop off</td></tr><tr><td>0-0 PDS (polydioxanone) Suture</td><td>Ethicon</td><td>Cat#Z990G</td><td>Linea Alba closure&lt;br/&gt; Quantity - 1</td></tr><tr><td>3-0 Prolene Suture</td><td>Ethicon</td><td>Cat#8684G</td><td>Skin Closure&lt;br/&gt; Quantity - 1</td></tr><tr><td>Bovie Electrocautery&amp;nbsp;</td><td>Grayline Medical Supply</td><td>Cat#E2516</td><td>Homeostasis and cutting</td></tr><tr><td>Standard set of sterile abomdinal surgical instruments with abdominal wall retractors like Army/Navy&#039;s or Richardson reactors.&amp;nbsp;</td><td></td><td></td><td></td></tr><tr><td>3M Tegaderm + Pad&amp;nbsp;</td><td>3M</td><td>Cat#3593</td><td>Surgical Dressing</td></tr><tr><td>High protein liquid diet</td><td>Ensure</td><td></td><td>Diet&lt;br/&gt; 1.5 bottles BID</td></tr><tr><td>&lt;strong&gt;Software&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>BioRender</td><td>Agreement number SH28Z3OOKI</td><td></td><td>Figure generation&amp;nbsp;</td></tr><tr><td>Adobe Illustrator</td><td>Adobe</td><td>v29.8.1</td><td>Figure generation&amp;nbsp;</td></tr><tr><td>FIJI</td><td>NIH</td><td></td><td>Figure generation&amp;nbsp;</td></tr><tr><td>*Note: AI tools were used only for minor language and grammar edits</td><td></td><td></td><td></td></tr><tr><td>*BID - two times a day,&amp;nbsp; IV - intravenous, IM - intramuscular, PO - per os/ by mouth, SQ - subcutaneous, PRN - as needed</td><td></td><td></td><td></td></tr></tbody></table>

a translational surgical porcine model for postoperative intra-abdominal adhesion formation

A Translational Surgical Porcine Model for Postoperative Intra-Abdominal Adhesion Formation

A porcine model of postoperative intra-abdominal adhesion formation was established using Yucatan mini pigs. The protocol combines midline laparotomy, small bowel resection with two-layer primary anastomosis, and a unilateral, parietal peritoneal abrasion in the format of an open abdominal surgical procedure. Adhesion formation was assessed four weeks postoperatively using established gross and histologic scoring criteria, with evaluations performed by blinded observers. Adhesions developed in all animals using this model and were multifocal, involving bowel loops, between the bowel and abdominal wall, involving the peritoneum overlying other organs in the abdomen (e.g. the liver), and operative sites, with variable severity. Histological analysis at four weeks demonstrated adhesions composed predominantly of extracellular matrix, fibroblasts, and blood vessels, consistent with a remodeling-phase wound healing tissue phenotype. This model is relevant for the study of abdominal adhesion fibrosis biology and/or the translational evaluation of candidate anti-adhesion therapeutics. By integrating a clinically relevant intestinal surgical procedure with a defined peritoneal injury and a standardized assessment strategy, this protocol provides a reproducible approach for inducing and evaluating postoperative intra-abdominal adhesions in a large animal model.

The proposed porcine postoperative adhesion model (Figure 1) was combined with an adapted scoring system to grade adhesions in real time28. Adhesion scoring was completed independently by 3 team members blinded to experimental conditions at the 4-week timepoint on a representative cohort of n=4 pigs; the scores were averaged for a final adhesion score per animal (Figure 2A). Intra-abdominal adhesions formed in 100% of animals evaluated on...

This protocol describes a porcine model of postoperative intra-abdominal adhesion formation using Yucatan mini pigs undergoing open abdominal surgery with bowel resection. The approach reliably induces intra-abdominal adhesions and provides a platform for evaluating adhesion development and testing anti-adhesion strategies.

A porcine model of postoperative intra-abdominal adhesion formation was established using Yucatan mini pigs. The protocol combines midline laparotomy, ...

This study developed a clinically relevant porcine model of postoperative intraabdominal adhesions to study adhesion biology and test anti adhesion therapies. While rodent models provide valuable insights, this protocol more closely replicates human abdominal surgery and reliably produces clinically relevant adhesions. To begin, drape the abdomen of an anesthetized pig using sterile towels and surgical drapes to maintain sterility throughout the procedure.Perform a midline laparotomy by making a full thickness incision through the skin subcutaneous tissue and abdominal wall musculature along the linear alba to enter the peritoneal cavity using a 10 blade scalpel and electrocautery. Then eviscerate the intestines. Identify a region of the bowel, approximately 10 to 15 centimeters proximal to the ileocecal valve, and resect a 2.5-centimeter segment of the small bowel using scissors.Resect the associated mesentary with electrocautery for hemostasis of the mesenteric blood vessels. Perform a two-layer side to side primary anastomosis. Reconstruct the inner layer of the lumen with a synthetic 3-0 absorbable suture in a running fashion and reconstruct the outer layer with a running 3-0 non-absorbable suture.Gently abrade a three by five centimeter section of the parietal peritoneum along the right abdominal sidewall using sterile gauze or an electrocautery scratch pad. Close the fascia with a running 2-0 absorbable suture. Close the skin with a running 2-0 or 3-0 non-absorbable baseball stitch and cover with a sterile dressing.Euthanize the animal after four weeks and assess adhesions grossly. Immediately collect tissues and blood samples for downstream analyses. Intraabdominal adhesions formed in 100%of animals evaluated on both the abraded and non-abraded sides of the abdomen.A braided side adhesions demonstrated significantly higher adhesion scores than the non-abraded side. In all animals, adhesions involved multiple abdominal sites, including the small intestine, colon, liver, and parietal peritoneum. Adhesion tissue was carefully dissected and harvested in isolation for downstream analyses.Histologic evaluation at four weeks revealed adhesions consisted of extracellular matrix dominated by fibroblasts and new blood vessel formation. This protocol allows researchers to study adhesion biology, fibrosis development, and test potential therapies to prevent postoperative adhesions. The most important consideration is performing standardized peritoneal injury and bowel surgery to ensure consistent and reproducible adhesion formation.Researchers can perform gross and histologic adhesion scoring and harvest tissues for downstream molecular, cellular, and fibrosis analyses.

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Explore a translational porcine model for postoperative intra-abdominal adhesion formation using Yucatan mini pigs. This protocol combines laparotomy, bowel resection, and peritoneal abrasion to study fibrosis biology and evaluate anti-adhesion therapeutics.

A Translational Surgical Porcine Model for Postoperative Intra-Abdominal Adhesion Formation

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