Plastic Surgery Research Council
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Presenter: Karina A Hernandez, Resident
Co-Authors: Reiffel AJ; Campbell R; Pino A; Harper A; Derrick K; Spector JA
Weill Cornell Medical College

BACKGROUND: Although bioartificial tissue scaffolds created using decellularization techniques may obviate patient donor site morbidity associated with reconstructive surgery, these acellular, avascular tissues are dependent on neovascularization for their survival, making them vulnerable to degradation, infection and failure. We sought to develop a bioengineered matrix with a dominant vascular pedicle that would serve as a platform for decellularization, re-seeding and ultimately direct donor-recipient anastomosis.

METHODS: Fasciocutaneous flaps supplied by the superficial inferior epigastric artery (SIEA) were harvested from Sprague Dawley rats. The femoral vessels were cannulated and a decellularization protocol was initiated via sequential perfusion with 4% sodium deoxycholate for 12 hours and DNAse I for 12 hours. Following decellularization, 6mm discs were obtained and equilibrated in standard culture medium for 48 hours. These acellular scaffolds were topically seeded with 2x10^5 RFP-expressing Human Dermal Fibroblasts (HDFn) with media changes performed daily. At 1, 3 and 7 days the scaffolds were fixed and processed for histology. Unseeded scaffolds served as the control.

RESULTS: Histological analysis with H&E staining of unseeded scaffolds confirmed successful decellularization and removal of all cellular material. Cells were observed along the surface of seeded scaffolds. These were confirmed as HDFn via fluorescent microscopy as evidenced by RFP expression. These cells were also seen lining the space where hair follicles were seen prior to decellularization. There were no differences in the degree of cellular invasion into the scaffold between the different time points.

CONCLUSION: We have established a successful method for perfusion-based decellularization of a fasciocutaneous flap with preservation of the inherent micro and macrovasculature. These data demonstrate that biological scaffolds derived from fasciocutaneous tissues can support the adhesion and proliferation of vital dermal components, taking us one step closer in the development of an off-the-shelf replacement tissue construct.

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