Pre-shear bioprinting of highly oriented porous hydrogel microfibers to construct anisotropic tissues

Anisotropic tissues in vivo have special structural characteristics and biological functions. Nowadays, bioprinting is widely used in tissue engineering and an effective way to process cell-laden hydrogels. However, the direct bioprinting of oriented cell-laden hydrogel structures to engineer anisotropic tissues is still difficult. Meanwhile, the inherent dense micropore network after the gelation of hydrogel-based bioinks usually limits the normal growth of encapsulated cells due to the inadequate supply of nutrient/oxygen. Herein, we proposed a pre-shear bioprinting strategy of highly oriented porous hydrogel microfibers to construct anisotropic tissues. Firstly, based on the phase separation of viscous high-molecular compound mixtures, we utilized a general viscous porous bioink paradigm, e.g., mixing a polymer thickener (PEO) with a hydrogel precursor (GelMA) with excellent biological properties. Secondly, based on the shear-oriented property of the viscous porous bioink, we designed the pre-shear in situ coaxial bioprinting of highly oriented porous hydrogel microfibers. The viscous porous bioink (GelMA/PEO) was shear-oriented through an injection tube and pumped into the inner needle of a coaxial nozzle. When GelMA/PEO passed through a transparent glass tube connected to the coaxial nozzle, GelMA can be in situ photo-crosslinked to form highly oriented porous microfibers. In addition, we showed the manufacturing of heterogeneous oriented microfibers and the manual assembly of microfibers, and within oriented microfibers, different cells or co-cultured cells exhibited highly oriented growth behaviors similar to that in vivo. As far as we know, the direct bioprinting of anisotropic tissues through high orientation induced by pre-shearing is firstly reported in our study. We believe that the pre-shear bioprinting strategy of anisotropic tissues will open more avenues for further biomedical research.