PRS PSRC Podium Proofs 2016
Ross H. Weinreb, BS,* Kerry A. Morrison, BA,* Julia Jin, BS,* Xue Dong, BA,* Yoshiko Toyoda, BA,* Adam Jacoby, MD,* Sushmita Mukherjee, PhD,† Jason A. Spector, MD*
From the *Laboratory of Bioregenerative Medicine and Surgery, Weill Cornell Medical Center, New York, N.Y.; and †Department of Biochemistry, Weill Cornell Medical Center, New York, N.Y.
PURPOSE: Fabrication of tissues with an inherent hierarchical vascular network remains the holy grail of tissue engineering. Herein, we fabricate a prevascularized full-thickness cellularized skin equivalent containing a 3-dimensional vascularized network of interconnected macrochemicals and microchannels lined with vascular cells, within a collagen neodermis containing encapsulated fibroblasts and an epidermis composed of human keratinocytes.
METHODS: Pluronic F127 was used for network preparation: 1.5 mm diameter “U”-shaped macrofibers and 100- to 500-μm interwoven microfibers were heat extruded and then embedded within type I collagen into which CFP-tagged human placental pericytes and human foreskin fibroblasts at a density of 1 × 106 cells/mL, respectively, had been encapsulated. After pluronic sacrifice, channels were intraluminally seeded with 5 × 106 cells/mL RFP-tagged human aortic smooth muscle cells, 5 × 106 cells/mL GFP-tagged human umbilical vein endothelial cells (HUVEC-GFP), and the construct was then topically seeded with 1 × 106 cells/mL human epidermal keratinocytes. Multiphoton microscopy and histology were conducted after 7 and 14 days of culture.
RESULTS: Multiphoton microscopic imaging demonstrates a hierarchical vascular network containing macrovessels and microvessels lined by endothelial and smooth muscle cells and supported by perivascular pericytes, all in appropriate microanatomic arrangement. Neodermal human foreskin fibroblasts proliferated throughout the observation period, and the human epidermal keratinocyte neoepidermis remained stable along the superficial aspect of the construct.
CONCLUSIONS: We have successfully fabricated a tissue-engineered prevascularized full-thickness skin flap construct with stable and anatomically appropriate vascularity. Our platform provides tremendous promise in furthering the development of tissue-engineered skin and other types of prevascularized flaps.