3D bioprinted human iPSC-derived somatosensory constructs with functional and highly purified sensory neuron networks.

Engineering three-dimensional (3D) sensible tissue constructs, along with the complex microarchitecture wiring of the sensory nervous system, has been an ongoing challenge in the tissue engineering field. By combining 3D bioprinting and human pluripotent stem cell (hPSC) technologies, sensible tissue constructs could be engineered in a rapid, precise, and controllable manner to replicate 3D microarchitectures and mechanosensory functionalities of the native sensory tissue (e.g. response to external stimuli). Here, we introduce a biofabrication approach to create complex 3D microarchitecture wirings. We develop an hPSC-sensory neuron (SN) laden bioink using highly purified and functional SN populations to 3D bioprint microarchitecture wirings that demonstrate responsiveness to warm/cold sense-inducing chemicals and mechanical stress. Specifically, we tailor a conventional differentiation strategy to our purification method by utilizing p75 cell surface marker and DAPT treatment along with neuronal growth factors in order to selectively differentiate neural crest cells into SNs. To create spatial resolution in 3D architectures and grow SNs in custom patterns and directions, an induced pluripotent stem cell (iPSC)-SN-laden gelatin bioink was printed on laminin-coated substrates using extrusion-based bioprinting technique. Then the printed constructs were covered with a collagen matrix that guided SNs growing in the printed micropattern. Using a sacrificial bioprinting technique, the iPSC-SNs were seeded into the hollow microchannels created by sacrificial gelatin ink printed in the gelatin methacryloyl supporting bath, thereby demonstrating controllability over axon guidance in curved lines up to several tens of centimeters in length on 2D substrates and in straight microchannels in 3D matrices. Therefore, this biofabrication approach could be amenable to incorporate sensible SN networks into the engineered skin equivalents, regenerative skin implants, and augmented somatosensory neuro-prosthetics that have the potential to regenerate sensible functions by connecting host neuron systems in injured areas.

A 3D-Printed Hybrid Nasal Cartilage with Functional Electronic Olfaction.

Advances in biomanufacturing techniques have opened the doors to recapitulate human sensory organs such as the nose and ear in vitro with adequate levels of functionality. Such advancements have enabled simultaneous targeting of two challenges in engineered sensory organs, especially the nose: i) mechanically robust reconstruction of the nasal cartilage with high precision and ii) replication of the nose functionality: odor perception. Hybrid nasal organs can be equipped with remarkable capabilities such as augmented olfactory perception. Herein, a proof-of-concept for an odor-perceptive nose-like hybrid, which is composed of a mechanically robust cartilage-like construct and a biocompatible biosensing platform, is proposed. Specifically, 3D cartilage-like tissue constructs are created by multi-material 3D bioprinting using mechanically tunable chondrocyte-laden bioinks. In addition, by optimizing the composition of stiff and soft bioinks in macro-scale printed constructs, the competence of this system in providing improved viability and recapitulation of chondrocyte cell behavior in mechanically robust 3D constructs is demonstrated. Furthermore, the engineered cartilage-like tissue construct is integrated with an electrochemical biosensing system to bring functional olfactory sensations toward multiple specific airway disease biomarkers, explosives, and toxins under biocompatible conditions. Proposed hybrid constructs can lay the groundwork for functional bionic interfaces and humanoid cyborgs.