Design of azobenzene-bearing hydrogel with photoswitchable mechanics driven by photo-induced phase transition for in vitro disease modeling

Abstract

Mechanics of the extracellular matrix (ECM) exhibit changes during many biological events. During disease progression, such as cancer, matrix stiffening or softening occurs due to crosslinking of the collagen matrix or matrix degradation through cell-secreted enzymes. Engineered hydrogels have emerged as a prime in vitro model to mimic such dynamic mechanics during disease progression. Although there have been a variety of engineered hydrogels, few can offer both stiffening and softening properties under the same working principle. In addition, to model individual disease progression, it is desirable to control the kinetics of mechanical changes. To this end, we describe a photoresponsive hydrogel that undergoes stiffness changes by the photo-induced phase transition. The hydrogel was composed of a copolymer of azobenzene acrylate monomer (AzoAA) and N,N-dimethyl acrylamide (DMA). By tuning the amount of azobenzene, the phase transition behavior of this polymer occurs solely by light irradiation, because of the photoisomerization of azobenzene. This phase behavior was confirmed at 37 ^°C by turbidity measurements. In addition, the crosslinked poly(AzoAA-r-DMA) gel undergoes reversible swelling-deswelling upon photoisomerization by ultraviolet or visible light. Furthermore, the poly(AzoAA-r-DMA) sheet gels exhibited modulus changes at different isomerization states of azobenzene. When MCF-7 cells were cultured on the gels, stiffening at different timepoints induced varied responses in the gene expression levels of E-cadherin. Not only did this suggest an adaptive behavior of the cells against changes in mechanics during disease progression, this also demonstrated our material's potential towards in vitro disease modeling. STATEMENT OF SIGNIFICANCE: During disease progression such as cancer, cellular microenvironment called extracellular matrix (ECM) undergoes stiffness changes. Hydrogels, which are swollen network of crosslinked polymers, have been used to model such dynamic mechanical environment of the ECM. However, few could offer both stiffening and softening properties under the same working principle. Herein, we fabricated a novel photoresponsive hydrogel with switchable mechanics, activated by photo-induced structural change of the polymer chains within the hydrogel. When breast cancer cells were cultured on our dynamic hydrogels, gene expression and morphological observation suggested that cells react to changes in stiffness by a transient response, as opposed to a sustained one. The photoresponsive hydrogel offers possibility for use as a patient-specific model of diseases.

Keywords: Azobenzene; Epithelial-mesenchymal transition; Hydrogels; Phase transition; Photoisomerization.