Plastic Surgery Research Council
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Presenter: Rachel Campbell, MD
Co-Authors: Reiffel AJ; Hernandez KA; Garcia D; Delnero P; Boyko T; Spector JA
Weill Cornell Medical College

Introduction: The fabrication of prevascularized scaffolds continues to be a major focus in tissue engineering, as a lack of blood supply predisposes engineered and/or transplanted tissue to necrosis. Previous work in our laboratory demonstrated the successful de novo synthesis and ex vivo microvascular anastomosis of a biodegradable, biocompatible, alginate scaffold containing an internal microchannel. As type I collagen more closely mimics in vivo extracellular matrix components, supporting cell adhesion, growth, and migration, we aimed to fabricate and anastomose in vivo a prevascularized collagen construct.

Methods: Pluronic F127 fibers were embedded in neutralized collagen, creating a central microchannel, 1.5 mm in diameter. Scaffolds contained an inlet and outlet and were seeded with green fluorescent protein labeled human umbilical vein endothelial cells. Cells were allowed to adhere for 1 hour and constructs underwent static culture or dynamic culture via a gravity-driven perfusion bioreactor. After 4 and 7 days, scaffolds were fixed and imaged.

Results: Pluronic F127 proved to be a suitable sacrificial polymer, reliably forming a microchannel within the collagen matrix. Cells adhered to the microchannel under both static and dynamic culture conditions. There was no obvious difference in cell density among the scaffolds cultured in static and dynamic conditions at 4 and 7 days. Additionally, scaffolds with polyglactone reinforced inlet and outlet channels were anastomosed in vivo to rodent femoral artery and vein using standard microsurgical techniques and successfully perfused.

Conclusions: We have successfully created prevascularized biodegradable, biocompatible scaffolds from type I collagen that support endothelialization and microsurgical anastomosis in vivo. With a preformed vascular network these constructs can be directly anastomosed to the host vasculature, providing immediate perfusion to the matrix thus increasing the chances of survival and incorporation. This will open the door to the creation and application of larger, more complex surgically relevant constructs.

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