Tubular Bioartificial Organs: From Physiological Requirements to Fabrication Processes and Resulting Properties. A Critical Review.

In this featured review manuscript, the aim is to present a critical survey on the processes available for fabricating bioartificial organs (BAOs). The focus will be on hollow tubular organs for the transport of anabolites and catabolites, i.e., vessels, trachea, esophagus, ureter and urethra, and intestine. First, the anatomic hierarchical structures of tubular organs, as well as their principal physiological functions, will be presented, as this constitutes the mandatory requirements for effectively designing and developing physiologically relevant BAOs. Second, 3D bioprinting, solution electrospinning, and melt electrowriting will be introduced, together with their capacity to match the requirements imposed by designing scaffolds compatible with the anatomical and physiologically relevant environment. Finally, the intrinsic correlation between processes, materials, and cells will be critically discussed, and directives defining the strengths, weaknesses, and opportunities offered by each process will be proposed for assisting bioengineers in the selection of the appropriate process for the target BAO and its specific required functions.

Enabling 3D bioprinting of cell-laden pure collagen scaffolds via tannic acid supporting bath.

The fabrication of cell-laden biomimetic scaffolds represents a pillar of tissue engineering and regenerative medicine (TERM) strategies, and collagen is the gold standard matrix for cells to be. In the recent years, extrusion 3D bioprinting introduced new possibilities to increase collagen scaffold performances thanks to the precision, reproducibility, and spatial control. However, the design of pure collagen bioinks represents a challenge, due to the low storage modulus and the long gelation time, which strongly impede the extrusion of a collagen filament and the retention of the desired shape post-printing. In this study, the tannic acid-mediated crosslinking of the outer layer of collagen is proposed as strategy to enable collagen filament extrusion. For this purpose, a tannic acid solution has been used as supporting bath to act exclusively as external crosslinker during the printing process, while allowing the pH- and temperature-driven formation of collagen fibers within the core. Collagen hydrogels (concentration 2-6 mg/mL) were extruded in tannic acid solutions (concentration 5-20 mg/mL). Results proved that external interaction of collagen with tannic acid during 3D printing enables filament extrusion without affecting the bulk properties of the scaffold. The temporary collagen-tannic acid interaction resulted in the formation of a membrane-like external layer that protected the core, where collagen could freely arrange in fibers. The precision of the printed shapes was affected by both tannic acid concentration and needle diameter and can thus be tuned. Altogether, results shown in this study proved that tannic acid bath enables collagen bioprinting, preserves collagen morphology, and allows the manufacture of a cell-laden pure collagen scaffold.

The KERATO Biomechanics Study 1: A Comparative Evaluation Using Brillouin Microscopy and Dynamic Scheimpflug Imaging.

PURPOSE: To assess the corneal biomechanical properties in normal individuals and patients with keratoconus using the Brillouin optical scanning system (Intelon Optics) (BOSS) and compare them with ultra-high-speed Scheimpflug imaging (Corvis ST; Oculus Optikgeräte GmbH). METHODS: Sixty eyes from 60 patients (30 normal and 30 keratoconus) were included in this prospective, single-center, comparative, non-interventional study. Corneal biomechanics were evaluated using the Corvis ST and the BOSS. With the BOSS, each corneal image was acquired three times, measuring 10 locations within an 8-mm diameter. Parameters extracted included mean, maximum, and minimum Brillouin shift. These 10 points were also grouped into superior, central, and inferior regions. BOSS repeatability was assessed using the coefficient of repeatability and coefficient of variation. Furthermore, normal individuals and patients with keratoconus were compared using the Corvis ST and BOSS. RESULTS: The BOSS exhibited good repeatability, with coefficient of repeatability ranging from 0.098 to 0.138 GHz for single points in normal individuals and 0.096 to 0.149 GHz for patients with keratoconus. Statistical analysis revealed significant differences between normal individuals and patients with keratoconus, indicating softer corneas in keratoconus, observed with both the Corvis ST and BOSS. Specifically, the BOSS showed significant differences in mean, inferior, and superior mean, maximum, and minimum Brillouin frequency shift (all P < .05), whereas the Corvis ST displayed highly significant differences in stiffness parameter at first applanation, stress strain index, deformation amplitude ratio, and inverse integrated radius (all P < .001). CONCLUSIONS: Corneal biomechanical measurements proved highly repeatable and effectively demonstrated significant differences between normal individuals and patients with keratoconus using both the BOSS and the Corvis ST. [J Refract Surg. 2024;40(8):e569-e578.].

Development of a hyaluronic acid-collagen bioink for shear-induced fibers and cells alignment.

Human tissues are characterized by complex composition and cellular and extracellular matrix (ECM) organization at microscopic level. In most of human tissues, cells and ECM show an anisotropic arrangement, which confers them specific properties.In vitro, the ability to closely mimic this complexity is limited. However, in the last years, extrusion bioprinting showed a certain potential for aligning cells and biomolecules, due to the application of shear stress during the bio-fabrication process. In this work, we propose a strategy to combine collagen (col) with tyramine-modified hyaluronic acid (THA) to obtain a printable col-THA bioink for extrusion bioprinting, solely-based on natural-derived components. Collagen fibers formation within the hybrid hydrogel, as well as collagen distribution and spatial organization before and after printing, were studied. For the validation of the biological outcome, fibroblasts were selected as cellular model and embedded in the col-THA matrix. Cell metabolic activity and cell viability, as well as cell distribution and alignment, were studied in the bioink before and after bioprinting. Results demonstrated successful collagen fibers formation within the bioink, as well as collagen anisotropic alignment along the printing direction. Furthermore, results revealed suitable biological properties, with a slightly reduced metabolic activity at day 1, fully recovered within the first 3 d post-cell embedding. Finally, results showed fibroblasts elongation and alignment along the bioprinting direction. Altogether, results validated the potential to obtain collagen-based bioprinted constructs, with both cellular and ECM anisotropy, without detrimental effects of the fabrication process on the biological outcome. This bioink can be potentially used for a wide range of applications in tissue engineering and regenerative medicine in which anisotropy is required.

Elastin-like recombinamers in collagen-based tubular gels improve cell-mediated remodeling and viscoelastic properties.

Natural polymers are commonly used as scaffolds for vascular tissue engineering. The recognized biological properties of this class of materials are often counterbalanced by their low mechanical performance. In this work, recombinant elastin-like polypeptides (or elastin-like recombinamers, ELRs) were mixed with collagen gel and cells to produce cellularized tubular constructs in an attempt to recapitulate the mechanical behavior of the vascular extracellular matrix (ECM). The presence of the elastic protein influenced cell-mediated remodeling evaluated in terms of construct compaction, cell proliferation and ECM (collagen, elastin and fibrillin-1) gene expression. The partial substitution of collagen with ELR and the observed differences in cellular behavior synergistically contributed to the superior viscoelastic properties of the constructs containing 30% ELR and 70% of collagen (in mass). This led to the improvement of 40% in the initial elastic modulus, 50% in the equilibrium elastic modulus, and 37% in the tensile strength at break without compromising the strain at break, when compared to a pure collagen scaffold. Suggestions for future research include modifications in the crosslinking technology, ELR composition, polymer concentration, cell seeding density and dynamic stimulation, which have the potential to further improve the mechanical performance of the constructs towards physiological values.