A biocompatible thermoset polymer binder for Direct Ink Writing of porous titanium scaffolds for bone tissue engineering.

There is increasing demand for synthetic bone scaffolds for bone tissue engineering as they can counter issues such as potential harvesting morbidity and restrictions in donor sites which hamper autologous bone grafts and address the potential for disease transmission in the case of allografts. Due to their excellent biocompatibility, titanium scaffolds have great potential as bone graft substitutes as they mimic the structure and properties of human cancellous bone. Here we report on a new thermoset bio-polymer which can act as a binder for Direct Ink Writing (DIW) of titanium artificial bone scaffolds. We demonstrate the use of the binder to manufacture porous titanium scaffolds with evenly distributed and highly interconnected porosity ideal for orthopaedic applications. Due to their porous structure, the scaffolds exhibit an effective Young's modulus similar to human cortical bone, alleviating undesirable stress-shielding effects, and possess superior strength. The biocompatibility of the scaffolds was investigated in vitro by cell viability and proliferation assays using human bone-marrow-derived Mesenchymal stem cells (hMSCs). The hMSCs displayed well-spread morphologies, well-organized F-actin and large vinculin complexes confirming their excellent biocompatibility. The vinculin regions had significantly larger Focal Adhesion (FA) area and equivalent FA numbers compared to that of tissue culture plate controls, showing that the scaffolds support cell viability and promote attachment. In conclusion, we have demonstrated the excellent potential of the thermoset bio-polymer as a Direct Ink Writing ready binder for manufacture of porous titanium scaffolds for hard tissue engineering.

Alternating air-medium exposure in rotating bioreactors optimizes cell metabolism in 3D novel tubular scaffold polyurethane foams.

BACKGROUND: In vitro dynamic culture conditions play a pivotal role in developing engineered tissue grafts, where the supply of oxygen and nutrients, and waste removal must be permitted within construct thickness. For tubular scaffolds, mass transfer is enhanced by introducing a convective flow through rotating bioreactors with positive effects on cell proliferation, scaffold colonization and extracellular matrix deposition. We characterized a novel polyurethane-based tubular scaffold and investigated the impact of 3 different culture configurations over cell behavior: dynamic (i) single-phase (medium) rotation and (ii) double-phase exposure (medium-air) rotation; static (iii) single-phase static culture as control. METHODS: A new mixture of polyol was tested to create polyurethane foams (PUFs) as 3D scaffold for tissue engineering. The structure obtained was morphologically and mechanically analyzed tested. Murine fibroblasts were externally seeded on the novel porous PUF scaffold, and cultured under different dynamic conditions. Viability assay, DNA quantification, SEM and histological analyses were performed at different time points. RESULTS: The PUF scaffold presented interesting mechanical properties and morphology adequate to promote cell adhesion, highlighting its potential for tissue engineering purposes. Results showed that constructs under dynamic conditions contain enhanced viability and cell number, exponentially increased for double-phase rotation; under this last configuration, cells uniformly covered both the external surface and the lumen. CONCLUSIONS: The developed 3D structure combined with the alternated exposure to air and medium provided the optimal in vitro biochemical conditioning with adequate nutrient supply for cells. The results highlight a valuable combination of material and dynamic culture for tissue engineering applications.

Hierarchical "As-Electrospun" Self-Assembled Fibrous Scaffolds Deconvolute Impacts of Chemically Defined Extracellular Matrix- and Cell Adhesion-Type Interactions on Stem Cell Haptokinesis.

Controlled self-assembly of diblock copolymers offers the possibility of fabricating multilength scale, three-dimensional (3D) porous/fibrous structures (or scaffolds) with defined internal nano- or microstructure, with opportunities for application in a variety of fields, ranging from energy storage to bioengineering. Traditional methods by which such 3D constructs are produced are time-consuming and tedious, hindering their broader exploitation within larger-scale industrial processes. We report the development of a one-step process to fabricate "as-electrospun" self-assembled diblock copolymer micro- to nanometer-sized fibers incorporating core-shell or lamellar, closely packed spheres or bicontinuous gyroid nanosized structures. Isotropic and anisotropic (aligned) porous mats presenting spatially controlled chemistries, including bioactive (peptide-based) motifs, were successfully made from these hierarchical fibers. When functionalized with peptide sequences derived from a cell adhesion molecule (E-cadherin) and an extracellular matrix glycoprotein (laminin), these novel materials provided new insight into the impacts of such exquisitely tailored contact-guidance cues on the haptokinesis of human mesenchymal stem cells.

Detergent-enzymatic decellularization of swine blood vessels: insight on mechanical properties for vascular tissue engineering.

Small caliber vessels substitutes still remain an unmet clinical need; few autologous substitutes are available, while synthetic grafts show insufficient patency in the long term. Decellularization is the complete removal of all cellular and nuclear matters from a tissue while leaving a preserved extracellular matrix representing a promising tool for the generation of acellular scaffolds for tissue engineering, already used for various tissues with positive outcomes. The aim of this work is to investigate the effect of a detergent-enzymatic decellularization protocol on swine arteries in terms of cell removal, extracellular matrix preservation, and mechanical properties. Furthermore, the effect of storage at -80°C on the mechanical properties of the tissue is evaluated. Swine arteries were harvested, frozen, and decellularized; histological analysis revealed complete cell removal and preserved extracellular matrix. Furthermore, the residual DNA content in decellularized tissues was far low compared to native one. Mechanical testings were performed on native, defrozen, and decellularized tissues; no statistically significant differences were reported for Young's modulus, ultimate stress, compliance, burst pressure, and suture retention strength, while ultimate strain and stress relaxation of decellularized vessels were significantly different from the native ones. Considering the overall results, the process was confirmed to be suitable for the generation of acellular scaffolds for vascular tissue engineering.