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The authors would like to acknowledge the Academy of Finland funding; No. 327248 (ValueBiomat) and 327865 (Bioeconomy). This work was a part of the Academy of Finland's Flagship Programme under Projects No. 318890 and 318891 (Competence Center for Materials Bioeconomy, FinnCERES). The authors would also like to thank the Biohybrid Materials Research Group (Aalto University) for providing the HepG2 cells. The current study provides a comprehensive rheology study and a survey on direct ink writing of xanthan gum/cellulose nanocrystal (XG/CNC) bio-inks for developing 3D geometries that mimic soft tissue engineering scaffolds' physical and mechanical properties. The presence of CNC was found to be a critical prerequisite for the printability of XG bio-inks; accordingly, the hybrid XG/CNC bio-inks revealed the excellent viscoelastic properties that enabled precise control of hydrogel shaping and printing of lattice structures composed of up to eleven layers with high fidelity and fair resolution without any deformation after printing. The lyophilized 3D scaffolds presented a porous structure with open and interconnected pores and a porosity higher than 70%, vital features for tissue engineering scaffolds. Moreover, they showed a relatively high swelling of approximately 11 g/g, facilitating oxygen and nutrient exchange. Furthermore, the elastic and compressive moduli of the scaffolds that enhanced significantly upon increasing CNC content were in the range of a few kPa, similar to soft tissues. Finally, no significant cell cytotoxicity was observed against human liver cancer cells (HepG2), highlighting the potential of these developed 3D printed scaffolds for soft tissue engineering applications.

The authors would like to acknowledge the Academy of Finland funding; No. 327248 (ValueBiomat) and 327865 (Bioeconomy). This work was a part of the Academy of Finland's Flagship Programme under Projects No. 318890 and 318891 (Competence Center for Materials Bioeconomy, FinnCERES). The authors would also like to thank Ms. Marja Kärkkäinen for providing TEMPO-CNF and the Biohybrid Materials Research Group (Aalto University) for providing the HepG2 cells. Plant-based hydrogels have attracted great attention in biomedical fields since they are biocompatible and based on natural, sustainable, cost-effective, and widely accessible sources. Here, we introduced new viscoelastic bio-inks composed of quince seed mucilage and cellulose nanofibrils (QSM/CNF) easily extruded into 3D lattice structures through direct ink writing in ambient conditions. The QSM/CNF inks enabled precise control on printing fidelity where CNF endowed objects with shape stability after freeze-drying and with suitable porosity, water uptake capacity, and mechanical strength. The compressive and elastic moduli of samples produced at the highest CNF content were both increased by ~100% (from 5.1 ± 0.2 kPa and 32 ± 1 kPa to 10.7 ± 0.5 and 64 ± 2 kPa, respectively). These values ideally matched those reported for soft tissues; accordingly, the cell compatibility of the printed samples was evaluated against HepG2 cells (human liver cancer). The results confirmed the 3D hydrogels as being non-cytotoxic and suitable to support attachment, survival, and proliferation of the cells. All in all, the newly developed inks allowed sustainable 3D bio-hydrogels fitting the requirements as scaffolds for soft tissue engineering.