FRESH bioprinting technology for tissue engineering - the influence of printing process and bioink composition on cell behavior and vascularization

The rapid and tailored biofabrication of natural materials is of high interest for the field of tissue engineering and regenerative medicine. Scaffolds require both high biocompatibility and tissue-dependent mechanical strength to function as basis for tissue-engineered implants. Thus, natural hydrogels such as fibrin are promising but their rapid biofabrication remains challenging. Printing of low viscosity and slow polymerizing solutions with good spatial resolution can be achieved by freeform reversible embedding of suspended hydrogels (FRESH) bioprinting of cell-laden natural hydrogels. In this study, fibrin and hyaluronic acid were used as single components as well as blended ink mixtures for the FRESH bioprinting. Rheometry revealed that single materials were less viscous than the blended bioink showing higher values for viscosity over a shear rate of 10–1000 s −1 . While fibrin showed viscosities between 0.1624 and 0.0017 Pa·s, the blended ink containing fibrin and hyaluronic acid were found to be in a range of 0.1–1 Pa·s. In 3D vascularization assays, formation of vascular structures within the printed constructs was investigated indicating that the printing process did not harm cells and allowed formation of vasculature comparable to moulded control samples. Best values for vascularization were achieved in bioinks consisting of 1.0% fibrin-0.5% hyaluronic acid. The vascular structure area and length were three times higher compared to other tested bioinks, and structure volume as well as number of branches revealed almost four times higher values. In this study, we combined the benefits of the FRESH printing technique with in vitro vascularization, showing that it is possible to achieve a mechanically stable small-scale hydrogel construct incorporating vascular network formation

Viability of coaxial atomization for disintegration of cell solutions in cell spray applications

Treating Leukemia with intravenous stem cell transplantation represents a well-established therapy technique. For applications, that require high local cell concentrations, transplantation by conventional intravenous injection is less potent, due to cell distribution with blood circulation. Instead, spraying them directly onto the injured or diseased area shows promising results in various applications, e.g. superficial treatment of topographically challenging wounds, in situ seeding of cells on implants, deposition of cells in tubular organs for stem cell therapy.The present work aims for a basic knowledge about viability boundaries for coaxial cell-spray atomization and the reciprocal influence between cells in solution and primary breakup mechanics. A generic modular nozzle is developed, to ensures reproducible boundary conditions. Investigations are conducted regarding primary breakup and relations between resulting droplet size distribution and cell survival. Measurements are performed, utilizing microscopic high-speed visualization with suitable image post processing. Cell viability is analyzed using phase contrast microscopy prior and after atomization. A relation between Rayleigh-Taylor instability wavelength and droplet size distributions by means of Sauter mean diameter (SMD) and cell survival rate (CSR) is suggested. A power law is presented, exclusively dependent on dimensionless measures (lambda(perpendicular to) similar to Re(-1/2)We(-1/3)) which is found to be proportional to SMD and CSR