The repair of osteochondral (OC) defects requires a tissue engineering approach to fabricate biological tissue that mimics gradient physiological properties such as the transition zone between cartilage and bone. The...

Osteochondral (OC) defects usually involve the damage of both the cartilage and its underneath subchondral bone. In recent years, tissue engineering (TE) has become the most promising method that combines scaffolds, growth factors, and cells for the repair of OC defects. An ideal OC scaffold should have a gradient structure to match the hierarchical mechanical properties of natural OC tissue. To satisfy such requirements, 3D printing, e.g., direct ink writing (DIW), has emerged as a technology for precise and customized scaffold fabrication with optimized structures and mechanical properties.In this study, finite element simulations were applied to investigate the effects of pore geometry on the mechanical properties of 3D printed scaffolds. Scaffold specimens with different lay-down angles, filament diameters, inter-filament spacing, and layer overlaps were simulated in compressive loading conditions. The results showed that Young's moduli of scaffolds decreased linearly with increasing scaffold porosity. The orthotropic characteristics increased as the lay-down angle decreased from 90° to 15°. Moreover, gradient transitions within a wide range of strain magnitudes were achieved in a single construct by assembling layers with different lay-down angles. The results provide quantitative relationships between pore geometry and mechanical properties of lattice scaffolds, and demonstrate that the hierarchical mechanical properties of natural OC tissue can be mimicked by tuning the porosity and local lay-down angles in 3D printed scaffolds.

Backgroundl-Glutamate is an important precursor in the l-arginine (l-Arg) biosynthetic pathway. Various methods, including polyoxyethylene sorbitan monopalmitate (Tween 40) addition and dtsR1 disruption, have been widely used to induce l-glutamate overproduction in Corynebacterium glutamicum. In this study, a novel strategy for l-Arg overproduction through Tween 40 trigger and ΔdtsR1 mutant were proposed in Corynebacterium crenatum.ResultsCorynebacterium crenatum mutant (CCM01) was selected as a host strain, whose argR was lethal via mutagenesis screening, the proB gene was knocked out, and argB was replaced by argB M4 (E19R, H26E, D311R, and D312R) to release l-Arg feedback resistance. After Tween 40 trigger in the logarithmic period, l-Arg production increased from 15.22 to 17.73 g/L in CCM01 strain. When NCgl1221 and dtsR1 disruption (CCM03), l-Arg production drastically increased to 27.45 g/L and then further to 29.97 g/L after Tween 40 trigger. Moreover, the specific activity of α-oxoglutarate dehydrogenase complex (ODHC) decreased, whereas the regeneration of NADP+/NADPH significantly increased after dtsR1 disruption and Tween 40 trigger. Results of real-time PCR showed that the transcriptional levels of odhA, sucB, and lpdA (encoding three subunits of the ODHC complex) were downregulated after Tween 40 trigger or dtsR1 disruption. By contrast, zwf transcription (encoding glucose-6-phosphate dehydrogenase) showed no significant difference among CCM01, CCM02 (ΔNCgl1221), and CCM03 (ΔNCgl1221ΔdtsR1) strains without Tween 40 trigger but evidently increased by 5.50 folds after Tween 40 trigger.ConclusionA novel strategy for l-Arg overproduction by dtsR1 disruption and Tween 40 trigger in C. crenatum was reported. Tween 40 addition exhibited a bifunctional mechanism for l-Arg overproduction, including reduced ODHC activity and enhanced NADPH pools accumulation by downregulated dtsR1 expression and upregulated zwf expression, respectively.

Three-dimensional (3D) printing has been emerging as a new technology for scaffold fabrication to overcome the problems associated with the undesirable microstructure associated with the use of traditional methods. Solvent-based extrusion (SBE) 3D printing is a popular 3D printing method, which enables incorporation of cells during the scaffold printing process. The scaffold can be customized by optimizing the scaffold structure, biomaterial, and cells to mimic the properties of natural tissue. However, several technical challenges prevent SBE 3D printing from translation to clinical use, such as the properties of current biomaterials, the difficulties associated with simultaneous control of multiple biomaterials and cells, and the scaffold-to-scaffold variability of current 3D printed scaffolds. In this review paper, a summary of SBE 3D printing for tissue engineering (TE) is provided. The influences of parameters such as ink biomaterials, ink rheological behavior, cross-linking mechanisms, and printing parameters on scaffold fabrication are considered. The printed scaffold structure, mechanical properties, degradation, and biocompatibility of the scaffolds are summarized. It is believed that a better understanding of the scaffold fabrication process and assessment methods can improve the functionality of SBE-manufactured 3D printed scaffolds.

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