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Engineering an elastic bioink for 3D Bioprinting of the Urethra

Abstract

There stands considerable opportunity in the medical field for the rapidly expanding scope of additive manufacturing technology such as 3D bioprinting, as the capacity for fabricating patient-unique tissue engineered scaffolds in a rapid, direct manner may expand the availability and applicability of surgical techniques. In this work, a 3D bioprinting technique was used to fabricate tubularized constructs for the urethral tissue regeneration. In particular, an elastic hydrogel-based biomaterials were optimized to be used as bioinks for 3D bioprinting of a construct for urethral reconstruction. Two radially distinct regions were determined to be necessary for this tissue-engineered construct, to provide cellular proliferation and structural support to the graft while de-facto partitioning two distinctive tissue regions in the urethra, one made of gelatin methacryloyl (GelMA) for the urothelium, and another layer made of methacrylated elastin-like polypeptide (mELP) and GelMA for smooth muscle cells (SMCs). GelMA and mELP were selected for the hydrogel blend to ensure mechanical durability while attaining a greater extensibility desirable for the urethral tissue region. Chemical characterization via H-NMR analysis provided evidence of the photocrosslinking mechanism which was utilized to crosslink the bioink and solidify the construct with the degree of crosslinking as 55% for GelMA and 68% for mELP/GelMA. Mechanical characterization allowed us to select an optimal ratio of GelMA/mELP and achieve the mechanical properties close to native urethra. Following this stage, the optimization of printing parameters for each bioinks yielded for temperature 30 and 8 �C, printing speed of 8 mm/s and 5 mm/s, and extrusion pressure 4 psi and 15 psi for the 10% (w/v) GelMA bioink and 15% (w/v) 1:1 mELP/GelMA bioink, respectively. Future studies will focus on in vitro cellular studies to ascertain the cytocompatibility of this material, as well as quantify the construct’s efficacy in future applications surgically.

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