In Situ Generation of Tunable Porosity Gradients in Hydrogel-Based Scaffolds for Microfluidic Cell Culture

Compared with preformed anisotropic matrices, an anisotropic matrix that allows users to alter its properties and structure in situ after synthesis offers the important advantage of being able to mimic dynamic in vivo microenvironments, such as in tissues undergoing morphogenesis or in wounds undergoing tissue repair. In this study, porous gradients are generated in situ in a hydrogel comprising enzymatically crosslinked gelatin hydroxyphenylpropionic acid (GTNHPA) conjugate and carboxylmethyl cellulose tyramine (CMCTYR) conjugate. The GTNHPA component acts as the backbone of the hydrogel, while CMCTYR acts as a biocompatible sacrificial polymer. The hydrogel is then used to immobilize HT1080 human fibrosarcoma cells in a microfluidic chamber. After diffusion of a biocompatible cellulase enzyme through the hydrogel in a spatially controlled manner, selective digestion of the CMC component of the hydrogel by the cellulase gives rise to a porosity gradient in situ instead of requiring its formation during hydrogel synthesis as with other methods. The influence of this in situ tunable porosity gradient on the chemotactic response of cancer cells is subsequently studied both in the absence and presence of chemoattractant. This platform illustrates the potential of hydrogel-based microfluidics to mimic the 3D in vivo microenvironment for tissue engineering and diagnostic applications.

Al-Abboodi, Aswan, Ricky Tjeung, Pauline M Doran, Leslie Y Yeo, James Friend, Yik Chan, Peggy Pui

Advanced Healthcare Materials





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