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We developed a method to 3D-print milk-based inks at room temperature by changing the rheological properties. The method is based on direct ink writing (DIW) and permits multi-material printing of 3D edible structures.

Gelatin and transglutaminase (TG) ink is increasingly popular in direct ink writing three-dimensional (3D) printing of cellular scaffolds and edible materials. The use of enzymes to crosslink gelatin chains removes the needs for toxic crosslinkers and bypasses undesired side reactions due to the specificity of the enzymes. However, their application in 3D printing remains challenging primarily due to the rapid crosslinking that leads to the short duration of printable time. In this work, we propose the use of gelatin preheated for 7 days to extend the duration of the printing time of the gelatin ink. We first determined the stiffness of freshly prepared gelatin (FG) and preheated gelatin (PG) (5 – 20% w/w) containing 5% w/w TG. We selected gelatin hydrogels made from 7.5% w/w FG and 10% w/w PG that yielded similar stiffness for subsequent studies to determine the duration of the printable time. PG inks exhibited longer time required for gelation and a smaller increase in viscosity with time than FG inks of similar stiffness. Our study suggested the advantage to preheat gelatin to enhance the printability of the ink, which is essential for extrusion-based bioprinting and food printing.

Orange peels are often discarded as food waste despite being a nutritious source of vitamins and antioxidants. These orange peel wastes (OPW) are produced in millions of tons globally every year; discarding them results in detrimental environmental and economical impacts. This paper discusses the application of 3D printing technology to effectively upcycle the OPW into edible, healthy snacks for consumption. We aimed to develop a method to enable OPW to formulate 3D-printable inks for direct ink writing (DIW). Using DIW 3D printing, we successfully created edible constructs of rheologically modified inks containing OPW. The formulated ink possessed an initial viscosity of 22.5 kPa.s, a yield stress of 377 Pa, and a storage modulus of 44.24 kPa. To validate the method, we conducted a biochemical analysis of the OPW at each stage of the fabrication process. This study suggested that our ink formulation and 3D printing process did not affect the content of bioflavonoids and antioxidants of the OPW. The cell viability test using human dermal microvascular endothelium (HMEC-1) suggested that the OPW did not exhibit cytotoxicity throughout the entire process of the ink manipulation. Overall, this study has highlighted a potential scenario to revalorize food waste into the food value chain using 3D printing toward more sustainable and circular food manufacturing and consumption.

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