Intestinal mucus is an anisotropic biological structure that acts as a barrier between the external environment and the epithelium.

The liver is a key organ that can communicate with many other districts of the human body. In the last few decades, much interest has focused on the interaction between the liver and the gut microbiota, with their reciprocal influence on biosynthesis pathways and the integrity the intestinal epithelial barrier. Dysbiosis or liver disorders lead to0 epithelial barrier dysfunction, altering membrane permeability to toxins. Clinical and experimental evidence shows that the permeability hence the delivery of neurotoxins such as LPS, ammonia and salsolinol contribute to neurological disorders. These findings suggested multi-organ communication between the gut microbiota, the liver and the brain. With a view to in vitro modeling this liver-based multi-organ communication, we describe the latest advanced liver-on-a-chip devices and discuss the need for new organ-on-a-chip platforms for in vitro modeling the in vivo multi-organ connection pathways in physiological and pathological situations.

The gut microbiota directly impacts the pathophysiology of different human body districts. Consequently, microbiota investigation is an hot topic of research and its in vitro culture has gained extreme interest in different fields. However, the high sensitivity of microbiota to external stimuli, such as sampling procedure, and the physicochemical complexity of the gut environment make its in vitro culture a challenging task. New engineered microfluidic gut‐on‐a‐chip devices have the potential to model some important features of the intestinal structure, but they are usually unable to sustain culture of microbiota over an extended period of time. The integration of gut‐on‐a‐chip devices with bioreactors for continuous bacterial culture would lead to fast advances in the study of microbiota‐host crosstalk. In this review, we summarize the main technologies for the continuous culture of microbiota as upstream systems to be coupled with microfluidic devices to study bacteria‐host cells communication. The engineering of integrated microfluidic platforms, capable of sustaining both anaerobic and aerobic cultures, would be the starting point to unveil complex biological phenomena proper of the microbiota‐host crosstalks, paving to way to multiple research and technological applications.

Alginate is a common component of bioinks due to its well-described ionic crosslinking mechanism and tunable viscoelastic properties. Extrusion-based 3D-printing of alginate inks requires additives, such as gelatin and Pluronic, pre- or post-printing crosslinking processes and/or coextrusion with crosslinkers. In this work, we aim to develop a different printing approach for alginate-based inks, introducing 3D-reactive printing. Indeed, the control over the crosslinking kinetics and the printing time allowed printing different inks while maintaining their final composition unaltered to identify a suitable formulation in terms of printability. Alginate solutions were crosslinked with insoluble calcium salts (CaCO₃) inducing a dynamic modification of their microstructure and viscoelastic properties over time. The monitoring of fiber printability and internal microstructure, at different time points of ink gelation, was performed by means of a well-defined set of rheological tests to obtain a priori ink properties for the a posteriori 3D-printing process. This new perspective allowed 3D-reactive printing of alginate fibers with predetermined properties, without involving post-extrusion crosslinking steps and additives.

Biological gradients profoundly influence many cellular activities, such as adhesion, migration, and differentiation, which are the key to biological processes, such as inflammation, remodeling, and tissue regeneration. Thus, engineered structures containing bioinspired gradients can not only support a better understanding of these phenomena, but also guide and improve the current limits of regenerative medicine. In this review, we outline the challenges behind the engineering of devices containing chemical-physical and biomolecular gradients, classifying them according to gradient-making methods and the finalities of the systems. Different manufacturing processes can generate gradients in either in-vitro systems or scaffolds, which are suitable tools for the study of cellular behavior and for regenerative medicine; within these, rapid prototyping techniques may have a huge impact on the controlled production of gradients. The parallel need to develop characterization techniques is addressed, underlining advantages and weaknesses in the analysis of both chemical and physical gradients.