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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

The present protocol describes the porcine fascio-cutaneous flap model and its potential use in vascularized composite tissue research.

Abstract

Vascularized Composite Allografts (VCA) such as hand, face, or penile transplant represents the cutting-edge treatment for devastating skin defects, failed by the first steps of the reconstructive ladder. Despite promising aesthetic and functional outcomes, the main limiting factor remains the need for a drastically applied lifelong immunosuppression and its well-known medical risks, preventing broader indications. Therefore, lifting the immune barrier in VCA is essential to tip the ethical scale and improve patients' quality of life using the most advanced surgical techniques. De novo creation of a patient-specific graft is the upcoming breakthrough in reconstructive transplantation. Using tissue engineering techniques, VCAs can be freed of donor cells and customized for the recipient through perfusion-decellularization-recellularization. To develop these new technologies, a large-scale animal VCA model is necessary. Hence, swine fascio-cutaneous flaps, composed of skin, fat, fascia, and vessels, represent an ideal model for preliminary studies in VCA. Nevertheless, most VCA models described in the literature include muscle and bone. This work reports a reliable and reproducible technique for saphenous fascio-cutaneous flap harvest in swine, a practical tool for various research fields, especially vascularized composite tissue engineering.

Introduction

Vascularized composite allografts (VCA) have revolutionized the treatment of hard-to-repair body part losses, such as hands, face, and penis1,2,3. Unfortunately, the first long-term outcomes4 have shown that lifelong administration of high-dose immunosuppressive agents can lead to severe collateral medical conditions, including diabetes, infections, neoplasia, and reno-vascular dysfunction5. Lately, expert VCA teams have had to manage the risk of chronic rejection leading to graft loss and perform the first face retransplantatio....

Protocol

All animals received human care following the National Institute of Health Guide for the Care and Use of Laboratory Animals. The Institutional Animal Care and Use Committee approved the experimental protocol (IACUC- protocol #2020N000015). Seven female Yorkshire pigs (20-25 kg) were used for all experiments.

1. Preoperative care

  1. Fast the animal for solid food 12 h prior to the surgery.
  2. Sedate the animal with 4.4 mg/kg of Telazol, 2.2 mg/kg of Xylazine, an.......

Representative Results

This work on living animals was preceded by determining the saphenous perforasome on three cadaveric specimens (Figure 2). A colored filling solution was injected into the saphenous artery to opacify the specific vascular network coming from the artery. The solution is composed of 10 mL blue-colored glycerin agent mixed with 10 mL of the diluent agent (see Table of Materials). This generated a colored map of the skin vascularized by the saphenous artery and allowed drawing t.......

Discussion

This article describes a reliable and reproducible fasciocutaneous flap harvested on swine hindlimbs. Following this step-by-step surgical protocol will allow the procurement of two flaps on only one animal in less than 2 h. The most critical step of the surgery is the skeletonization of the vascular pedicle within the gracilis muscle fibers, which requires a thorough dissection by a skilled surgeon. Securing the skin to the fascia using cutaneous sutures is a crucial tip to avoid a shearing effect disrupting the perfora.......

Acknowledgements

This work was funded by Shriners Hospitals for Children grants #85127 (BEU and CLC) and #84702 (AA). The authors would like to thank the "Gueules Cassées" foundation for the salary support to the fellows involved in that project.

....

Materials

NameCompanyCatalog NumberComments
18 G angiocatheterBD Insyte Autoguard381409
20 G angiocatheterBD Insyte Autoguard381411
Adson Tissue Forceps, 11 cm, 1 x 2 Teeth with Tying PlatformASSIASSI.ATK26426
Atropine SulfateAdvaCare212-868
Bipolar cordsASSI228000C
Buprenorphine HClPharmaceutical, Inc42023-179-01
Dilating ForcepsFine science tools (FST)18131-12
Endotrachel tubeJorgensen LabsJO615Xsize from 6 to 15mm depending on the pig weight
Ethilon 3-0 16 mm 3/8EthiconMPVCP683H
EuthasolVirbac AH200-071
Heparin Lock Flush Solution, USP, 100 units/mLBD PosiFlush306424
IsofluranePatterson Veterinary14043-704-06
Jewelers Bipolar Forceps Non Stick 11 cm, straight pointed tip, 0.25 mm tip diameterASSIASSI.BPNS11223
Metzenbaum scissors 180 mmB BraunBC606R
Microfil blueFlow techLMV-120
Microfil dilutionFlow techLMV-112colored filing solution
Monopolar knifeASSI221230C
N°15 scalpel bladeSwann MortonNS11
OmnipaqueGeneral Electric4080358contrast product
Perma-Hand Silk 3-0EthiconA184H
Small LigaclipEthiconMCM20
Stevens scissors 115 mmB BraunBC008R
TelazolZoetis106-111
Xylamed (xylazine)Bimeda200-529

References

  1. Dubernard, J. M., et al. Human hand allograft: Report on first 6 months. The Lancet. 353 (9161), 1315-1320 (1999).
  2. Meningaud, J. P., et al. Procurement of total human face graft for allotransplan....

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PorcineFasciocutaneous FlapVascularized Composite AllograftSaphenous ArteryFemoral VesselsSurgical TechniqueLarge Animal ModelTissue EngineeringImmunology

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