3D printing, wet spinning and cell culture on carbohydrate low molecular weight supramolecular hydrogels Juliette Fitremann 1 , Faniry Andriamiseza 1 , Mickaël Chabbert 1 , Nadia Kasmi 1 , Christophe Coudret 1 , Laure Gibot 1 , Laurence Vaysse 2 , Isabelle Loubinoux 3 1 CNRS - IMRCP - University of Toulouse, France, 2 RESTORE, INSERM, Toulouse, France, 3 TONIC, INSERM, CHU Purpan, France Low molecular weight supramolecular hydrogels are formed by the self-assembly of small, non-polymer molecules in supramolecular fibers which entraps water. These hydrogels are often mechanically fragile and as such, they are not often considered for cell culture applications and 3D printing. However this fragility can be suitable for growing cells from soft tissues, such as brain (1) . To tackle with applications requiring large quantities of gel, we developed a family of gelators of simple structures, based on long chain amides coupled with sugar polyols. The synthesis is short and gives molecules of high purity suitable for cell culture. On the hydrogels, the cell culture of neural or mesenchymal stem cells, fibroblasts or neural cell lines provides small 3D cell clusters linked together by cell extensions guided by the supramolecular fibers (Fig. b). Specific methods are implemented to observe the 3D cell organization in these fragile hydrogels. We are also developing different gelation methods and notably wet spinning (3,4) or 3D printing (2,4) . Because the hydrogels are not shear-thinning nor thixotropic, usual 3D printing methods by direct injection of the gels are not possible. So we developed a method in which the gelation is triggered by liquid-liquid exchange. The very fast self-assembly of the gelator provides well-resolved 3D printed patterns (Fig. c) and well-shaped gel noodles. Also, by changing slightly the structure of molecular gelator, we got either sacrificial or persistent gels which can be imbricated by 3D printing. The spontaneous dissolution of the sacrificial gel gives supramolecular gel architectures with channels. The gelation mechanism at the liquid-liquid interface has been analyzed by the introduction of fluorescent dyes or colored indicators. In addition, at the microscopic level, in some conditions, the supramolecular fibers are radially organized highlighting diffusion and/or mixing phenomenon at the liquid-liquid interface (Fig. a, ref. 3). References 1. Chalard, A.; et al. Simple Synthetic Molecular Hydrogels from Self-Assembling Alkyl-galactonamides as Scaffold for 3D Neuronal Cell Growth. ACS Appl. Mater. Interfaces 2018 , 17004. 2. Andriamiseza, F. et al. 3D Printing of Biocompatible Low Molecular Weight Gels: Imbricated Structures with Sacrificial and Persistent N-Alkyl-d-Galactonamides. J. Coll. Interf. Sci. 2022 , 617 , 156. 3. Bordignon, D.; et al. Wet Spinning of a Library of Carbohydrate Low Molecular Weight Gels. J. Coll. Interf. Sci. 2021 , 333. 4. Piras, C. C.; Kay, A. G.; Genever, P. G.; Fitremann, J.; Smith, D. K. Self-Assembled Gel Tubes, Filaments and 3D-Printing with in Situ Metal Nanoparticle Formation and Enhanced Stem Cell Growth. Chem. Sci. 2022 , 13 , 1972.
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