Effect of process conditions on the growth of three-dimensional dermal-equivalent tissue obtained by microtissue precursor assembly.

Bottom-up approach is an appealing strategy to build complex three-dimensional (3D) viable tissues in vitro starting from microtissue precursors (μTP). In this work we biofabricated a thick dermal-like tissue by sequentially combining two steps: a μTPs production and assembly followed by tissue maturation in a purpose-built bioreactor. The μTPs were produced by first seeding bovine primary fibroblasts on gelatine microparticles and then cultivating them in stirring conditions until a thick layer of ~80 μm of de novo synthesized extracellular matrix uniformly covered the microparticle surface. The μTPs were then loaded into a cylindrical chamber (2 mm in depth and 35 mm in diameter) and let to maturate and assemble into a 3D viable biohybrid tissue under specific fluid flow conditions. Several combinations of perfusion and/or tangential fluid flow were applied and their effect on the tissue formation and maturation was assessed. Results show that structural composition and mechanical features of the final 3D bioengineered tissue are strongly affected by the hydrodynamic environment and demonstrate that by optimizing culture conditions a 3D viable tissue with properties similar to that of native derma could be produced.