Procurement and Perfusion-Decellularization of Porcine Vascularized Flaps in a Customized Perfusion Bioreactor

Xu, Michael S.; Karoubi, Golnaz; Waddell, Thomas K.; Haykal, Siba

doi:10.3791/64068

Cited by 2 publications

(9 citation statements)

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“…H&E staining revealed removal of cellular components between native and decellularized tissues, which in combination with DAPI staining and significant reduction of DNA content, confirmed successful decellularization of both omental and TFL free flaps. These findings are in alignment with previous reports of this decellularization protocol 30 . Next, Masson Trichrome (collagen) and Verheoff-Van Gieson (elastin) staining, immunohistochemistry for collagen IV, fibronectin, laminin, and GAG quantification assays were performed.…”

Section: Discussionsupporting

confidence: 93%

“…Vascularized porcine flaps were perfusion-decellularized through the flap vasculature using low concentration anionic SDS detergent within a customized bioreactor, based on a previously published protocol 30 . Decellularization was macroscopically evidenced by the characteristic white/opaque appearance of decellularized tissues (not pictured).…”

Section: Resultsmentioning

confidence: 99%

“…Initial work in free flap bioengineering conducted by our lab demonstrated the feasibility and efficacy of using low-concentration SDS perfusion to generate vascularized acellular scaffolds 30 . The present work supports these results.…”

Section: Discussionmentioning

confidence: 99%

“…Pigs were prepped and draped in the usual sterile fashion prior to flap procurement. Surgical procedure for porcine omentum and TFL flaps procurement were as previously described 30 . Briefly, the omental flap was procured by midline laparotomy and the left gastroepiploic artery and vein was used as the dominant vascular conduit.…”

Section: Methodsmentioning

confidence: 99%

“…Porcine flaps were perfusion-decellularized using low-concentration SDS followed by DNase (Sigma Aldrich) reconstituted to a concentration of 10 mg/mL, as previously described 30 . Cannulated flaps were each connected to a perfusion system to allow antegrade perfusion via the arterial inlet at 2 ml/min, in which solutions: 0.05% SDS followed by 0.1 mg/mL deoxyribonuclease (DNase) were perfused through the flap vasculature with 1 × phosphate buffered saline (PBS) perfusion in between to remove residual detergent.…”

Section: Methodsmentioning

confidence: 99%

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Bioengineering of vascularized porcine flaps using perfusion-recellularization

Xu,

D’Elia,

Hadzimustafic

et al. 2024

Sci Rep

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Large volume soft tissue defects greatly impact patient quality of life and function while suitable repair options remain a challenge in reconstructive surgery. Engineered flaps could represent a clinically translatable option that may circumvent issues related to donor site morbidity and tissue availability. Herein, we describe the regeneration of vascularized porcine flaps, specifically of the omentum and tensor fascia lata (TFL) flaps, using a tissue engineering perfusion-decellularization and recellularization approach. Flaps were decellularized using a low concentration sodium dodecyl sulfate (SDS) detergent perfusion to generate an acellular scaffold with retained extracellular matrix (ECM) components while removing underlying cellular and nuclear contents. A perfusion-recellularization strategy allowed for seeding of acellular flaps with a co-culture of human umbilical vein endothelial cell (HUVEC) and mesenchymal stromal cells (MSC) onto the decellularized omentum and TFL flaps. Our recellularization technique demonstrated evidence of intravascular cell attachment, as well as markers of endothelial and mesenchymal phenotype. Altogether, our findings support the potential of using bioengineered porcine flaps as a novel, clinically-translatable strategy for future application in reconstructive surgery.

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Section: Discussionsupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Bioengineering of vascularized porcine flaps using perfusion-recellularization

Xu,

D’Elia,

Hadzimustafic

et al. 2024

Sci Rep

Self Cite

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Optimized Decellularization of a Porcine Fasciocutaneaous Flap

Lupon,

Acun,

Taveau

et al. 2024

Bioengineering

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Reconstructive techniques to repair severe tissue defects include the use of autologous fasciocutaneous flaps, which may be limited due to donor site availability or lead to complications such as donor site morbidity. A number of synthetic or natural dermal substitutes are in use clinically, but none have the architectural complexity needed to reconstruct deep tissue defects. The perfusion decellularization of fasciocutaneous flaps is an emerging technique that yields a scaffold with the necessary composition and vascular microarchitecture and serves as an alternative to autologous flaps. In this study, we show the perfusion decellularization of porcine fasciocutaneous flaps using sodium dodecyl sulfate (SDS) at three different concentrations, and identify that 0.2% SDS results in a decellularized flap that is efficiently cleared of its cellular material at 86%, has maintained its collagen and glycosaminoglycan content, and preserved its microvasculature architecture. We further demonstrate that the decellularized graft has the porous structure and growth factors that would facilitate repopulation with cells. Finally, we show the biocompatibility of the decellularized flap using human dermal fibroblasts, with cells migrating as deep as 150 µm into the tissue over a 7-day culture period. Overall, our results demonstrate the promise of decellularized porcine flaps as an interesting alternative for reconstructing complex soft tissue defects, circumventing the limitations of autologous skin flaps.

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Procurement and Perfusion-Decellularization of Porcine Vascularized Flaps in a Customized Perfusion Bioreactor

Cited by 2 publications

References 0 publications

Bioengineering of vascularized porcine flaps using perfusion-recellularization

Bioengineering of vascularized porcine flaps using perfusion-recellularization

Optimized Decellularization of a Porcine Fasciocutaneaous Flap

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