Plastic Surgery Research Council
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PSRC 60th Annual Meeting

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Bioprinted Vascularized Tissue-Engineered Constructs for In Vivo Perfusion
Rachel C. Hooper, MD1, Duan Bin, BS2, Alice Harper, BS1, Adam Jacoby, BA1, Anya Laibangyang, BS Candidate2, Jonathan T. Butcher, PhD2, Jason A. Spector, MD, FACS1.
1Weill Cornell Medical College, New York, NY, USA, 2Cornell University, Ithaca, NY, USA.

PURPOSE:
The greatest challenge to contemporary tissue engineers remains the difficulty associated with creating vascular networks within the engineered tissue. Furthermore, any vascularized construct must be designed to allow for anastomosis to the host vascular system. In previous work we synthesized a tissue-engineered scaffold containing an endothelialized internal loop microchannel for microsurgical anastomosis and in vivo perfusion utilizing a sacrificial microfiber technique. Bioprinting is an emerging technology that allows for the consistent and rapid fabrication of three-dimensional constructs comprised of any combination of extrudable polymers and cells with a precise predetermined microarchitecture. Here we describe the fabrication of bioprinted hydrogel constructs for cell seeding and in vivo microanastomosis.
METHODS:
15mm x 15mm x 5mm “loop” (Figure 1) or “diamond” (Figure 2) internal microchannel containing poly(ethylene glycol) diacrylate/ methacrylated gelatin/ alginate hydrogels were bioprinted using the Fab@Home ™ platform based upon a custom designed stereolithography (STL) file. Briefly, the STL file was divided into layers generating fill-paths for each layer of the scaffold; the hydrogel formulation was loaded and extruded along the X-Y paths for each layer at a rate of 5 mm/s. Constructs for microsurgical anastomosis were modified with polyglactin mesh at the inlet and outlet following printing to allow for suture fixation. To confirm cell compatibility with the chosen “bioink”, 8 mm discs were topically seeded with 2.5 x 10 4 cells/mL porcine aortic smooth muscle cells (SMC) and 2.5 x 10 4 cells/mL aortic endothelial cells (EC) and cultured for 7 days. Fixed constructs were processed for histology and immunohistochemical staining
RESULTS:
Bioprinted loop and diamond shaped constructs were successfully fabricated. Patency was confirmed via perfusion of colored buffer solution. Biocompatibility was established via Live/Dead ™staining and revealed adhesion and proliferation of SMC and EC. Immunohistochemical staining demonstrated smooth muscle actin and myosin heavy chain expressing SMC. Polyglactin mesh-modified constructs were successfully anastomosed to the femoral artery and vein of nude rats.
CONCLUSION:
We have successfully created custom bioprinted biocompatible vascularized constructs that support cell adhesion, growth and microsurgical anastomosis for in vivo perfusion. These proof of concept flaps provide a starting point towards the further development of bioprinted vascularized tissues which has the potential to revolutionize the field of tissue engineering.



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