The key innovation in Freeform Reversible Embedding of Suspended Hydrogels (FRESH) 3D bioprinting is the extrusion and embedding of an ECM hydrogel within a support bath that holds the extruded gel in place during printing.

In a recent publication in Science we showcased FRESH by 3D printing collagen-I into functional components of the human heart. Multiple bioinks can be printed including alginate, fibrin, collagen-I, methacrylated hyaluronic acid, decellularized ECM, and Matrigel demonstrating our ability to utilize a range of bioinks to tune scaffold composition.

Recently, we created a collagen-based 3D bioprinted vascular microfluidic platform using FRESH bioprinting (CHIPS) as well as the development of a custom perfusion bioreactor for long-term perfusion culture (VAPOR). Together, these systems establish the foundation for design and fabrication to accurately and reproducibly bioprint any 3D fluidic channel design within a collagen-scaffold. At the University of Pittsburgh in the Departments of Bioengineering and Vascular Medicine Institute we collaborate with many investigators from the School of Medicine and Carnegie Mellon University to incorporate physiological readouts, bioelectronics, and real-time fluorescence imaging into our platforms for organ-on-chip systems to study human vascular, kidney, and pancreatic disease.

One of our ongoing research interests lies in utilization of quantitative analysis of live cell fluorescence imaging to investigate basic cell biology and protein trafficking. Dr. Shiwarski is a huge proponent of the phrase “seeing is believing”, and is confident that if we can help aspiring students to see biological processes in action and quantify their results, it will ignite a passion in them scientific discovery.

Recent work in collaboration with the Subramanya and Boyd-Shiwarski Labs at U. Pitt. has focused on leveraging live cell fluorescent imaging and quantitative analysis to study protein phase transitions of a kinase called WNK1. Using fluorescence recovery after photobleaching (FRAP), fluorescence lifetime microscopy (FLIM), and live cell particle tracking, we can measure the kinetics and mobility of WNK1 phase transitions and show that this phenomenon is a conserved and required mechanism the cell uses to regulate the activity of NKCC to achieve osmotic homeostasis.

Additionally, our group continues to utilize image quantification and analysis of fluorescent biosensors to ask novel biological questions about 3D signal propagation and developmental morphogenesis.

Overall, open-source hardware fosters a more inclusive and innovative approach to technology development, benefiting both individuals and the broader community. We have built bi-axial stretchers, 3D bioprinters, alignment systems, heated stages, myography devices, syringe pumps, perfusion systems, bioreactors, with many more to come.