David B. Kolesky

3D Bioprinting of Vascularized, Heterogeneous Cell‐Laden Tissue Constructs

A new bioprinting method is reported for fabricating 3D tissue constructs replete with vasculature, multiple types of cells, and extracellular matrix. These intricate, heterogeneous structures are created by precisely co-printing multiple materials, known as bioinks, in three dimensions. These 3D micro-engineered environments open new -avenues for drug screening and fundamental studies of wound healing, angiogenesis, and stem-cell niches.

Embedded 3D Printing of Strain Sensors within Highly Stretchable Elastomers

A new method, embedded-3D printing (e-3DP), is reported for fabricating strain sensors within highly conformal and extensible elastomeric matrices. e-3DP allows soft sensors to be created in nearly arbitrary planar and 3D motifs in a highly programmable and seamless manner. Several embodiments are demonstrated and sensor performance is characterized.

Three-dimensional bioprinting of thick vascularized tissues

Significance Current tissue manufacturing methods fail to recapitulate the geometry, complexity, and longevity of human tissues. We report a multimaterial 3D bioprinting method that enables the creation of thick human tissues (>1 cm) replete with an engineered extracellular matrix, embedded vasculature, and multiple cell types. These 3D vascularized tissues can be actively perfused with growth factors for long durations (>6 wk) to promote differentiation of human mesenchymal stem cells toward an osteogenic lineage in situ. The ability to construct and perfuse 3D tissues that integrate parenchyma, stroma, and endothelium is a foundational step toward creating human tissues for ex vivo and in vivo applications. The advancement of tissue and, ultimately, organ engineering requires the ability to pattern human tissues composed of cells, extracellular matrix, and vasculature with controlled microenvironments that can be sustained over prolonged time periods. To date, bioprinting methods have yielded thin tissues that only survive for short durations. To improve their physiological relevance, we report a method for bioprinting 3D cell-laden, vascularized tissues that exceed 1 cm in thickness and can be perfused on chip for long time periods (>6 wk). Specifically, we integrate parenchyma, stroma, and endothelium into a single thick tissue by coprinting multiple inks composed of human mesenchymal stem cells (hMSCs) and human neonatal dermal fibroblasts (hNDFs) within a customized extracellular matrix alongside embedded vasculature, which is subsequently lined with human umbilical vein endothelial cells (HUVECs). These thick vascularized tissues are actively perfused with growth factors to differentiate hMSCs toward an osteogenic lineage in situ. This longitudinal study of emergent biological phenomena in complex microenvironments represents a foundational step in human tissue generation.

Bioprinting of 3D Convoluted Renal Proximal Tubules on Perfusable Chips

Three-dimensional models of kidney tissue that recapitulate human responses are needed for drug screening, disease modeling, and, ultimately, kidney organ engineering. Here, we report a bioprinting method for creating 3D human renal proximal tubules in vitro that are fully embedded within an extracellular matrix and housed in perfusable tissue chips, allowing them to be maintained for greater than two months. Their convoluted tubular architecture is circumscribed by proximal tubule epithelial cells and actively perfused through the open lumen. These engineered 3D proximal tubules on chip exhibit significantly enhanced epithelial morphology and functional properties relative to the same cells grown on 2D controls with or without perfusion. Upon introducing the nephrotoxin, Cyclosporine A, the epithelial barrier is disrupted in a dose-dependent manner. Our bioprinting method provides a new route for programmably fabricating advanced human kidney tissue models on demand.

High‐Throughput Printing via Microvascular Multinozzle Arrays

Microvascular multinozzle arrays are designed and fabricated for high-throughput printing of functional materials. Ink-flow uniformity within these multigeneration, bifurcating microchannel arrays is characterized by computer modeling and microscopic particle image velocimetry (micro-PIV) measurements. Both single and dual multinozzle printheads are produced to enable rapid printing of multilayered periodic structures over large areas (≈1 m(2)).

Controlling Material Reactivity Using Architecture.

3D-printing methods are used to generate reactive material architectures. Several geometric parameters are observed to influence the resultant flame propagation velocity, indicating that the architecture can be utilized to control reactivity. Two different architectures, channels and hurdles, are generated, and thin films of thermite are deposited onto the surface. The architecture offers an additional route to control, at will, the energy release rate in reactive composite materials.

Additive Micro-Manufacturing of Designer Materials

Material properties are governed by the chemical composition and spatial arrangement of constituent elements at multiple length scales. This fundamentally limits material properties with respect to each other creating trade-offs when selecting materials for a specific application. For example, strength and density are inherently linked so that, in general, the more dense the material, the stronger it is in bulk form. Other coupled material properties include thermal expansion and thermal conductivity, hardness and fracture toughness, strength and thermal expansion, etc. We are combining advanced microstructural design, using flexure and screw theory as well as topology optimization, with new additive micro- and nano-manufacturing techniques to create new material systems with previously unachievable property combinations. Our manufacturing techniques include Projection Microstereolithography (PμSL), Direct Ink Writing (DIW), and Electrophoretic Deposition (EPD). These processes are capable of reliably producing designed architectures that are highly three-dimensional, multi-scale, and often composed of multiple constituent materials.