The one-step fabrication of porous hASC-laden GelMa constructs using a handheld printing system

The fabrication of highly porous cell-loaded structures in tissue engineering applications has been a challenging issue because non-porous cell-laden struts can cause severe cell necrosis in the middle region owing to poor transport of nutrients and oxygen. In this study, we propose a versatile handheld 3D printer for the effective fabrication of porous cell-laden methacrylated gelatin (GelMa) with high porosity (approximate to 97%) by air injection and a bubble-making system using mesh filters through which a mixture of air/GelMa bioink is passed. In particular, the pore size and foamability of the cell constructs could be manipulated using various processing parameters (rheological properties of GelMa, filter size and number, and air-bioink volume ratio). To demonstrate the feasibility of the cell construct as a tissue engineering substitute for muscle regeneration, in vitro cellular activities and in vivo regeneration ability of human adipose stem cells were assessed. The in vitro results demonstrated that the human adipose stem cells (hASCs) fabricated using the handheld 3D printer were alive and well-proliferated. Furthermore, the in vivo results showed that the hASCs-constructs directly printed from the handheld 3D printer showed significant restoration of functionality and efficient muscle regeneration in the volumetric muscle loss model of mice. Based on these results, the fabrication method of the porous cell-laden construct could be a promising tool for regenerating muscle tissues.

Automated summary

This study proposes a versatile handheld 3D printer for the fabrication of porous cell-laden methacrylated gelatin (GelMa) with high porosity. The printer utilizes air injection and a bubble-making system using mesh filters to pass a mixture of air/GelMa bioink. The cell constructs' pore size and foamability can be manipulated by adjusting various processing parameters. In vitro and in vivo tests demonstrate the viability and efficacy of the printed cell constructs for muscle regeneration.