Cell-printing methods have been used widely in tissue regeneration because they enable fabricating biomimetic 3D structures laden with various cells. To achieve a cell-matrix block, various natural hydrogels that are nontoxic, biocompatible, and printable have been combined to obtain "bioinks." Unfortunately, most bioinks, including those with alginates, show low cell-activating properties. Here, a strategy for obtaining highly bioactive ink, which consisted of collagen/extracellular matrix (ECM) and alginate, for printing 3D porous cell blocks is developed. An in vitro assessment of the 3D porous structures laden with preosteoblasts and human adipose stem cells (hASCs) demonstrates that the cells in the bioinks are viable. Osteogenic activities with the designed bioinks show much higher levels than with the "conventional" alginate-based bioink. Furthermore, the hepatogenic differentiation ability of hASCs with the bioink is evaluated using the liver-specific genes, albumin, and TDO2, under hepatogenic differentiation conditions. The genes are activated within the 3D cell block fabricated using the new bioink. These results demonstrate that the 3D cell-laden structure fabricated using collagen/ECM-based bioinks can provide a novel platform for various tissue engineering applications.

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Electrohydrodynamic (EHD) direct writing has been used in diverse microelectromechanical systems and various supplemental methods for biotechnology and electronics. In this work, we expanded the use of EHD-induced direct writing to fabricate 3D biomedical scaffolds designed as porous structures for bone tissue engineering. To prepare the scaffolds, we modified a grounded target used in conventional EHD direct printing using a poly(ethylene oxide) solution bath, elastically cushioning the plotted struts to prevent crumbling. The fabricated scaffolds were assessed for not only physical properties including surface roughness and water uptake ability but also biological capabilities by culturing osteoblast-like cells (MG63) for the EHD-plotted polycaprolactone (PCL) scaffold. The EHD-scaffolds showed significantly roughened surface and enhanced water-absorption ability (400% increase) compared with the pure rapid-prototyped PCL. The results of cell viability, alkaline phosphatase activity, and mineralization analyses showed significantly enhanced biological properties of the scaffold (20 times the cell viability and 6 times the mineralization) compared with the scaffolds fabricated using RP technology. Because of the results, the modified EHD direct-writing process can be a promising method for fabricating 3D biomedical scaffolds in tissue engineering.

Axon regeneration is regulated by a neuron-intrinsic transcriptional program that is suppressed during development but that can be reactivated following peripheral nerve injury. Here we identifyProm1, which encodes the stem cell marker prominin-1, as a regulator of the axon regeneration program.Prom1expression is developmentally down-regulated, and the genetic deletion ofProm1in mice inhibits axon regeneration in dorsal root ganglion (DRG) cultures and in the sciatic nerve, revealing the neuronal role ofProm1in injury-induced regeneration. Elevating prominin-1 levels in cultured DRG neurons or in mice via adeno-associated virus-mediated gene delivery enhances axon regeneration in vitro and in vivo, allowing outgrowth on an inhibitory substrate.Prom1overexpression induces the consistent down-regulation of cholesterol metabolism-associated genes and a reduction in cellular cholesterol levels in a Smad pathway-dependent manner, which promotes axonal regrowth. We find that prominin-1 interacts with the type I TGF-β receptor ALK4, and that they synergistically induce phosphorylation of Smad2. These results suggest thatProm1and cholesterol metabolism pathways are possible therapeutic targets for the promotion of neural recovery after injury.

We report a cell-dispensing technique, using a core-shell nozzle and an absorbent dispensing stage to form cell-embedded struts. In the shell of the nozzle, a cross-linking agent flowed continuously onto the surface of the dispensed bioink in the core nozzle, so that the bioink struts were rapidly gelled, and any remnant cross-linking solution during the process was rapidly absorbed into the working stage, resulting in high cell-viability in the bioink strut and stable formation of a threedimensional mesh structure. The cell-printing conditions were optimized by manipulating the process conditions to obtain high mechanical stability and high cell viability. The cell density was 1 × 10 7 mL −1 , which was achieved using a 3-wt% solution of alginate in phosphate-buffered saline, a mass fraction of 1.2 wt% of CaCl 2 flowing in the shell nozzle with a fixed flow rate of 0.08 mL min −1 , and a translation velocity of the printing nozzle of 10 mm s −1 . To demonstrate the applicability of the technique, preosteoblasts and human adipose stem cells (hASCs) were used to obtain cell-laden structures with multi-layer porous mesh structures. The fabricated cell-laden mesh structures exhibited reasonable initial cell viabilities for preosteoblasts (93%) and hASCs (92%), and hepatogenic differentiation of hASC was successfully achieved.Tissue engineering has been widely applied to the regeneration of damaged tissues and organs using a combination of cells, an engineered extracellular matrix (or scaffold), and appropriate bioactive growth and differentiation factors 1,2,3 . The scaffold has been shown to be an important factor in cell attachment, growth, and differentiation; however, the mechanisms for the effects of the chemical and biological compositions and the physical structures that are required to encourage proper tissue regeneration are not completely understood 4 .Biomedical scaffolds for tissue engineering should possess various chemical and physical properties, including biocompatibility, with minimal cytotoxic effects to allow high cell attachment and proliferation; should be biodegradable; should have a highly porous structure (appropriate pore size, tortuosity, pore-interconnectivity) to enable easy vascularization and efficient transportation of nutrients and metabolic waste; and should have appropriate mechanical properties to endure the compressive and shear stresses from the micro-environmental conditions 5,6,7 .