Innovations in Microfluidics, Biofabrication, Synthetic Biology Agenda

Conference Registration, Materials Pick-Up, Morning Coffee and Breakfast Pastries

Networking Lunch in the Exhibit Hall -- Meet the Exhibitors and View Posters

Session Title: Innovations in Synthetic Biology

Development of in vitro Mucosal Interface in a Multi-well Plate
Jungwoo Lee, Assistant Professor, University of Massachusetts-Amherst, United States of America

The gastrointestinal tract (GIT) accounts for more than 75% of the body surface that directly contact with daily ingested foods along with microbes to absorb nutrients and water while keeping essential barrier function. This makes the GIT the prime site for the host-microbiome interactions where the host deploys about 80% of immune cells to protect and maintain homeostasis. The GIT epithelium consists of a monolayer of polarized cells arranged into microscopic features that increase the GIT surface area tremendously. Recent studies suggest that peristaltic motion would be critical for epithelial cell differentiation, polarization and structural arrange-ment into villi and crypts. Some progress has been made in recent times, there is still a growing need to develop model systems to integrate microbiome, epithelial cells, and immune cells in a single platform along with providing sufficient level of biological and mechanical complexity. To this end, we developed a mechanically actuated 3D tissue culture model system that can achieve the aforementioned integration of different cell types with multiple layer complexity in an easily adaptable well plate platform. In this study we demonstrate how this platform was used to mechanically stimulate colon carcinoma cells (HT-29) leading to polarization, vertical growth and 3D morphogenesis of these cells.

Coffee Break and Networking in the Exhibit Hall

The Age of Applications
Ricky Solorzano, CEO, Allevi

In an era where bioprinting continues to hold promise sometimes its hard to understand why and how are they useful. What key applications will allow me to take my research to the next level and stay on the cutting edge. Come and listen to the key ways bioprinting is being most commonly used by researchers around the world.

Regenerative Medicine Research Performed Using EnvisionTEC’s 3D-Bioplotter
Nicole Witzleben, Biomedical Applications Engineer, EnvisionTEC

Within the field of Regenerative Medicine is the growing research branch into bioprinting. The main application of bioprinting is the creation of bone and tissue scaffolds for implantation. Such scaffolds are not meant to be permanently implanted within the body, they are intended to allow the body’s own tissue to regenerate around the scaffold as it dissolves. Five fields of research where bioprinting is widely used include bone regeneration, cartilage regeneration, soft tissue biofabrication, drug release, and organ printing. The 3D-Bioplotter is a rapid prototyping machine specifically designed to print biocompatible material for these applications. Various biomaterials can be selected by the user for the appropriate area of research or even printed simultaneously to fabricate multi-materials parts. The user is able to optimize the inner structure of the scaffold to meet the requirements of their project with different designs, including lines, zig-zags, waves, and honeycombs. This presentation will provide an insight into the developing field of bioprinting and the 3D-Bioplotter’s role in that growth. We will review the timeline in the development of research utilizing EnvisionTEC’s 3D-Bioplotter and reference noteworthy published studies in bioprinting.

Technology Spotlight:
Building the World of Bioprinting: Science, Education, and Community
Erik Gatenholm, CEO, CELLINK, United States of America
Hector Martínez, Chief Technology Officer, CELLINK, United States of America

From the first bioink company in the world to a collaborative partner to an industry leader and scientific pioneer.

Networking Reception with Beer, Wine and Appetizers in the Exhibit Hall. Engage with Colleagues and Visit the Exhibitors

Close of Day 1 of the Conference

Morning Coffee, Tea, Breakfast Pastries and Networking in the Exhibit Hall

Bio-machines and Bio-manufacturing
Xuanhe Zhao, Associate Professor, Massachusetts Institute of Technology (MIT), United States of America

While human tissues are mostly soft, wet and bioactive; machines are commonly hard, dry and biologically inert.  Bridging human-machine interfaces is of imminent importance in addressing grand challenges in health, security, sustainability and joy of living facing our society in the 21st century. However, designing human-machine interfaces is extremely challenging, due to the fundamentally contradictory properties of human and machine. At MIT SAMs Lab, we propose to use tough bioactive hydrogels to bridge human-machine interfaces. On one side, bioactive hydrogels with similar physiological properties as tissues can naturally integrate with human body, playing functions such as scaffolds, catheters, drug reservoirs, and wearable devices. On the other side, the hydrogels embedded with electronic and mechanical components can control and response to external devices and signals. In the talk, I will first present a bioinspired approach and a general framework to design bioactive and robust hydrogels as the matrices for human-machine interfaces. I will then discuss large-scale manufacturing strategies to fabricate robust and bioactive hydrogels and hydrogel electronics and machines, including 3D printing. Prototypes including smart hydrogel band-aids, hydrogel robots and hydrogel circuits will be further demonstrated.

Contracting 3D Printed Microtissues: Solid and Fluid Instabilities
Thomas Angelini, Professor, Department of Mechanical and Aerospace Engineering, University of Florida, United States of America

Living cells are often dispersed in extracellular matrix (ECM) gels like collagen and Matrigel as minimal tissue models. Generally, large-scale contraction of these constructs is observed, in which the degree of contraction and compaction of the entire system correlates with cell density and ECM concentration. The freedom to perform diverse mechanical experiments on these contracting constructs is limited by the challenges of handling and supporting these delicate samples. Here, we present a method to create simple cell-ECM constructs that can be manipulated with significantly reduced experimental limitations. We 3D print mixtures of cells and ECM (collagen-I) into a 3D growth medium made from jammed microgels. With this approach, we design microtissues with controlled dimensions, composition, and material properties. We also control the elastic modulus and yield stress of the jammed microgel medium that envelops these microtissues. Similar to well-established bulk contraction assays, our 3D printed tissues contract. By contrast, the ability to create high aspect ratio objects with controlled composition and boundary conditions allows us to drive these microtissues into different regimes of physical instability. For example, a contracting tissue can be made to buckle as a whole or break up into droplets, depending on composition, size, and shape. These new instabilities may be employed in tissue engineering applications to anticipate the physical evolution of tissue constructs under the forces generated by the cells within.

Digital Biomanufacturing Enabling Multimode 4D Bioprinting
William G Whitford, Life Science Strategic Solutions Leader, DPS Group, United States of America

3D bioprinting is the deposition of microchannels or droplets of a polymer and/or cell dispersion (bioink) to create 3D tissue-like structures that includes living cells.  4D bioprinting adds the extra dimension of time supporting the activity of smart, environmentally responsive biological structures and tissues.  Many types of printing technologies are now used in bioprinting and each require appropriate manufacturing equipment, procedures and materials. Digital biomanufacturing orchestrates such concepts as increased monitoring, data handling, control algorithms, machine-learning and process modeling to a new level of process understanding, prediction and control.  The IIoT, Big Data and Cloud technologies insure that the (historical and real-time) data being collected can be employed productively in richer data management, analysis and model generation.  This leads to such values as more rapid process development as well as more comprehensive process control, automation and self-learning autonomation. Digital biomanufacturing will assist in the modeling and imaging required to recapitulate the (often personalized) complex and heterogeneous architecture of functional tissues and organs.  It will also provide the required coordination and dynamic control of consequent complex tomographic information and models, multimode 3D printing and biofabrication processes, as well as such ancillary procedures as the environmental control of bioinks and nascent constructs.

Coffee, Tea and Networking in the Exhibit Hall

Recombinant Human Collagen Based BioInk for 3D Bioprinting
Nadav Orr, Vice President, R&D, CollPlant, Ltd. Israel

Recombinant human Type I collagen (rhCollagen) from engineered plants was developed as BioInk for 3D-Bioprinting. The BioInk, rhCollagen-MA, is compatible with all major printing technologies. It has superior rheological properties and demonstrated high biocompatibility with different cells types.

Autocatalytic Immune Reactions
Daniel Irimia, Associate Professor, Surgery Department, Massachusetts General Hospital (MGH), Shriners Burns Hospital, and Harvard Medical School, United States of America
Alex Robert Hopke, Research Scientist, Mass General Hospital (MGH), United States of America

Neutrophil swarms protect healthy tissues by sealing off sites of infection. During swarming, neutrophils accumulate fast and in large numbers, under the control of mediators released by neutrophils already at the site.  These mediators stimulate the arrival of additional neutrophils in an autocatalytic reaction that results in an exponential rate of swarm size increase. However, despite the autocatalytic reaction, not all 25 billion neutrophils from one’s body end up in one giant swarm.  Thus, our recent goal was to identify the physiologic mediators that disrupt the autocatalytic reaction and stop the growth of neutrophil swarms. For this, we developed and validated large microscale arrays of microbe clusters, which can trigger the synchronized growth of thousands of swarms at once.  The new tool enabled us to concentrate large amounts of swarm-released mediators in small volumes, and ultimately identify lipoxin A4 (LXA4) is a key mediator that disrupts the autocatalytic reactions, stops the growth of swarms, and ultimately leads to swarm dispersal.  These and other insights from the study of neutrophil swarming will teach us how to design better strategies to combat infections and to control acute and chronic inflammatory diseases.

Hierarchical Biomaterials for Organ-on-a-Chip Devices and Tissue Engineering
Frederik Claeyssens, Senior Lecturer, Materials Science and Engineering, University of Sheffield, United Kingdom

Natural tissues and organs are typically structured in a hierarchical fashion, in which the Extra-Cellular Matrix (ECM) provides a microporosity to optimally support cell growth while larger scale structures (e.g. vasculature and boundary layers) are incorporated to support the function and structure of the tissue and organ. To mimic this multiscale structuring in synthetic biomaterials we combine additive manufacturing with self-assembly. In this structuring technique the internal porosity is governed by self-assembly and the macroscopic structure is constructed by additive manufacturing. Emulsion templating is used as self-assembly technique to produce materials with a high microscale porosity.  These emulsions can subsequently be used as photocurable resins for stereolithography, producing user-defined macroscale structures with a tissue-like microporosity. The mechanical properties of these materials can be varied via the changing the monomer ratio within the resin. Additionally, biodegradable scaffolds can be fabricated via polycaprolactone-based resins. We produce these hierarchical structured material in 3D structured materials such as woodpile-style scaffolds, microspheres with controllable diameter and as 3D microenvironments that can be integrated in standard poly-dimethylsiloxane (PDMS) based microfluidics. These scaffolds we currently investigate as a platform for organ-on-a-chip based devices and tissue engineering.

Networking Lunch in the Exhibit Hall

Active Materials for Remotely Regulating Complex, Multi-drug Delivery Profiles
Stephen Kennedy, Assistant Professor of Biomedical and Chemical Engineering, University of Rhode Island, United States of America

Biological processes are characterized by a high degree of spatiotemporal complexity and often manifest as a highly choreographed sequence of biological events. Biomaterials such as hydrogels can provide the localized deliveries needed to regulate biological processes. However, they are incapable of regulating multi-drug delivery profiles with the level of control required to regulate complex biological processes (i.e., control over the timing, rate, and sequence of multiple therapeutics). Moreover, the timings, rates, and sequences delivered for particular therapies must likely be customized for individual patients and may need to be changed in real-time according to updated patient prognoses. This necessitates the need for on-demand regulation over the timings, rates, and sequences of deliveries. Here, we will present strategies for on-demand, remote control over the timing, rate, and sequence of therapeutic deliveries from hydrogels using electrically, magnetically, and ultrasonically active materials. We will further describe biological scenarios where the deliveries provided by traditional hydrogels can be greatly improved upon using remotely activated materials (e.g., wound healing, tissue engineering, chemotherapies). Finally, we will demonstrate that these remotely active hydrogel materials have the potential to deliver optimized delivery schedules in a wide range of therapeutic strategies.

3D Printing Functional Materials & Devices
Michael McAlpine, Kuhrmeyer Family Chair Professor of Mechanical Engineering, University of Minnesota, United States of America

The development of methods for interfacing high performance functional devices with biology could impact regenerative medicine, smart prosthetics, and human-machine interfaces. Indeed, the ability to three-dimensionally interweave biological and functional materials could enable the creation of devices possessing unique geometries, properties, and functionalities. Yet, most high quality functional materials are two dimensional, hard and brittle, and require high crystallization temperatures for maximal performance. These properties render the corresponding devices incompatible with biology, which is three-dimensional, soft, stretchable, and temperature sensitive. We overcome these dichotomies by: 1) using 3D printing and scanning for customized, interwoven, anatomically accurate device architectures; 2) employing nanotechnology as an enabling route for overcoming mechanical discrepancies while retaining high performance; and 3) 3D printing a range of soft and nanoscale materials to enable the integration of a diverse palette of high quality functional nanomaterials with biology. 3D printing is a multi-scale platform, allowing for the incorporation of functional nanoscale inks, the printing of microscale features, and ultimately the creation of macroscale devices. This three-dimensional blending of functional materials and ‘living’ platforms may enable next-generation 3D printed devices.

Towards Bioprinting Industrialization
Fabien Guillemot, CEO, Poietis, France

Bioprinting has been defined as “the use of computer-aided transfer processes for patterning and assembling living and non-living materials with a prescribed 2D or 3D organization in order to produce bio-engineered structures serving in regenerative medicine, pharmacokinetic and basic cell biology studies”.

From a technological point of view, different bioprinting technologies have been developed so far. Recently, Laser-Assisted Bioprinting (LAB) technology has emerged as an alternative method to inkjet and bioextrusion methods, thereby overcoming some of their limitations (namely clogging of print heads or capillaries) to pattern living cells and biomaterials with a micron-scale resolution and high cell viability.

During this presentation, we will present current development on the industrialization of bioprinting process, namely in the context of manufacturing 3D skin models.

Bioprinting Multicellular Structure to Advance Vascularized 3D Tissue Engineering
Alisa Morss, Associate Professor, Drexel University, United States of America

3D vascularized tissue engineering would enable mechanistic study of healthy and diseased tissues, deliver a powerful platform for drug screening, and potentially provide a replacement for diseased tissues. The standard 3D tissue biofabrication process is layer-by-layer printing of a bioink composed of cells within a matrix material. While this process allows cells to be printed in a specific pattern to guide them towards the desired 3D structure, it relies primarily on cellular self-assembly into a 3D architecture that hopefully recapitulates the in vivo tissue. Unfortunately, cellular self-assembly may take days or weeks, may require complex spatial and temporal environmental cues, or may not occur when two cell types are co-cultured together. Because of these challenges, critical features of the tissue microenvironment cannot be recapitulated, and therefore cell-cell interactions cannot be studied in a physiologically relevant in vitro model. In this research, we created a new bioprinting paradigm in which multicellular structures that recapitulate in vivo architecture were used to create a 3D vascularized breast cancer tissue model. The multicellular structures were either grown in vitro prior to printing, or they were derived from tissues (e.g., breast organoids). Our method decreases the time between bioprinting and experimental assay from weeks to days; increases physiological relevance by allowing the investigator to more precisely control tissue architecture; and enables the use of primary human tissues together with stromal cells and local extracellular matrix.

Stimuli-Responsive Therapeutic Bioinks for 3D Printing
Rachael Oldinski, Assistant Professor, Department of Mechanical Engineering, University of Vermont, United States of America

The development of stimuli-responsive, or ‘smart’ biomaterials is critical to the manipulative manufacturing of custom tissue engineering scaffolds and implants. This talk will focus on the modification of a seaweed derivative, alginate, for the design of sustainable materials for tissue engineering construct development using 3D printing technology.

Distilling Complexity to Advance Cardiac Tissue Engineering via 3D Bioprinting
Brenda Ogle, Associate Professor of Biomedical Engineering, University of Minnesota, United States of America

The promise of cardiac tissue engineering is in the ability to design replacement muscle for clinical therapy.  Refined techniques for deriving cardiac muscle cells from stem cells coupled with advanced scaffolding techniques and 3D bioprinting are paving the way for a new generation of simulated cardiac tissues.  Recent success, ongoing challenges and the role for industrial innovation in this growing field will frame the content of this talk.

Glioblastoma Growth In Vitro 3D Modelling in an Adjustable Nanofiber-Hydrogel Hybrid Synthetic Scaffold
Nikita Grigoryev, Adjunct Professor, NYU Tandon School of Engineering, United States of America

Glioblastoma multiforme (GBM), the most aggressive form of brain cancer that originates from glial type stem cells. Recent discoveries show that glioma growths show a mysterious property of mosaic tumor heterogeneity. Arising from the same common precursor, later stage tumors and individual migrating GBM cells exist in intermingled clonal subpopulations with mutually exclusive gene amplification contributing to an “epigenetic switch” that gives rise to malignant invasive subclones that escape the original tumor, heavily aggravating the disease progression and patient survival. Current in vitro models are typically far from physiological actuality and do not provide necessary 3D mechanobiological environment native to brain tissue. Our unique approach uses a marriage of patented electrospinning/nanofiber coating technique and dipping/cell trapping gelation method to create a layered scaffold construct for GBM in 3D. A hydrogel layer is built over a glass rod by dipping into either an acellular alginate solution (as a niche for cancerous invasion) or a cells-alginate suspension and is crosslinked by dipping into a cation solution. Subsequently, electrospun nanofibers are deposited directly onto each hydrogel layer for controllable nanotopography exterior similar that of native, while the hydrogel, as soft as brain, itself surrounds cells. We have confirmed successful growth, spread and invasion of originally seeded cells into acellular layers of our construct with an appearance of tumor-like structures resembling those found in-vivo. Our proposed adjustable 3D model is completely artificial and provides cells with mechanobiological nanoenvironment comparable to that of native tissue, while also allowing for easy cancerous growth tracking through transparent layers.

Sessile Drops on a Petri Dish for Biocompatible Cell-based Assays
Aishah Prastowo, DPhil student, University of Oxford, United Kingdom

We developed a technique of performing cell-based assays in drop arrays on a standard tissue-culture polystyrene petri dish without further surface modification, and modelled the adsorption of serum in the cell media at the liquid and solid interfaces.

Close of Day 2 of the Conference.