3D-Printed proangiogenic patches of photo-crosslinked gelatin and polyurethane hydrogels laden with vascular cells for treating vascular ischemic diseases.

Engineering vascularized tissues remains a promising approach for treating ischemic cardiovascular diseases. The availability of 3D-bioprinted vascular grafts that induce therapeutic angiogenesis can help avoid necrosis and excision of ischemic tissues. Here, using a combination of living cells and biodegradable hydrogels, we fabricated 3D-printed biocompatible proangiogenic patches from endothelial cell-laden photo-crosslinked gelatin (EC-PCG) bioink and smooth muscle cell-encapsulated polyurethane (SMC-PU) bioink. Implantation of 3D-bioprinted proangiogenic patches in a mouse model showed that EC-PCG served as an angiogenic capillary bed, whereas patterned SMC-PU increased the density of microvessels. Moreover, the assembled patterns between EC-PCG and SMC-PU induced the geometrically guided generation of microvessels with blood perfusion. In a rodent model of hindlimb ischemia, the vascular patches rescued blood flow to distal tissues, prevented toe/foot necrosis, promoted muscle remodeling, and increased the capillary density, thereby improving the heat-escape behavior of ischemic animals. Thus, our 3D-printed vascular cell-laden bioinks constitute efficient and scalable biomaterials that facilitate the engineering of vascular patches capable of directing therapeutic angiogenesis for treating ischemic vascular diseases.

Development of double network polyurethane-chitosan composite bioinks for soft neural tissue engineering.

Three-dimensional (3D) bioprinting is an emerging manufacturing technology to print materials with cells for tissue engineering applications. In this study, we prepared novel ternary soft segment-based biodegradable polyurethane (tPU) using waterborne processes. The ternary soft segment included poly(ε-caprolactone) (PCL), polylactide, and poly(3-hydroxybutyrate) (PHB). tPU2 with a soft segment of PCL, poly(D,L-lactide), and PHB in a molar ratio of 0.7 : 0.2 : 0.1 demonstrated lower stiffness (∼2.3 kPa) and a greater tan δ value (∼0.64) and maintained good vitality (91.3%) of neural stem cells (NSCs) among various tPUs. The bioprinted tPU2 constructs facilitated cell proliferation (∼200% in 7 days) and neural differentiation of NSCs. Meanwhile, tPU2 formed double network composite hydrogels with gelatin or agarose, and the composite hydrogels showed good biocompatibility and achieved high-resolution (∼80 µm nozzle) bioprinting. In addition, a new series of double network polyurethane-chitosan composite (PUC) hydrogels were developed by combining tPU2 with a self-healing chitosan hydrogel. The PUC hydrogel demonstrated self-healing properties and bioprintability without the need for a post-crosslinking process. The bioprinted PUC composite hydrogel promoted cell proliferation (∼300% in 7 days) and neural differentiation of NSCs better than the tPU2 bioink. This study revealed new formulae of a polyurethane bioink and a polyurethane-chitosan composite bioink for 3D bioprinting and tissue engineering applications.