3D Disease Modelling of Hard and Soft Cancer Using PHA-Based Scaffolds

SIMPLE SUMMARY: Tumour progression in vivo was able to be well mimicked in 3D culture by utilizing biodegradable 10 mm × 10 mm × 8 mm P(3HO-co-3HD) and P(3HB)-based 3D scaffolds with a pore size of 30 to 300 µm. Both hard (MCF7 and MDA-MB-231) and soft (HCT116) tumour-related cells were successfully grown on the scaffolds, and their growth patterns were studied for 5 days. MDA-MB-231 tend to grow in clusters, and MCF7 cells form an evenly dispersed layer, which covered most of the 3D PHA scaffolds, while HCT116 formed large colonies within the pockets of the 3D PHA scaffold. Epithelial mesenchymal transition (EMT) marker genes, including Wnt-11, E-cadherin, Vim and Snail expression profiles, were like those seen in real tumour samples, which confirmed that the cancer models were exhibiting real tumour-like characteristics with high fidelity. These models are important in mimicking hypoxic tumours and in studying gene expression, cellular signalling, angiogenesis and drug response for translational research. ABSTRACT: Tumour cells are shown to change shape and lose polarity when they are cultured in 3D, a feature typically associated with tumour progression in vivo, thus making it significant to study cancer cells in an environment that mimics the in vivo milieu. In this study we established hard (MCF7 and MDA-MB-231, breast cancer) and soft (HCT116, colon cancer) 3D cancer tumour models utilizing a blend of P(3HO-co-3HD) and P(3HB). P(3HO-co-3HD) and P(3HB) belong to a group of natural biodegradable polyesters, PHAs, that are synthesised by microorganisms. The 3D PHA scaffolds produced, with a pore size of 30 to 300 µm, allow for nutrients to diffuse within the scaffold and provide the cells with the flexibility to distribute evenly within the scaffold and grow within the pores. Interestingly, by Day 5, MDA-MB-231 showed dispersed growth in clusters, and MCF7 cells formed an evenly dispersed dense layer, while HCT116 formed large colonies within the pockets of the 3D PHA scaffolds. Our results show Epithelial Mesenchymal Transition (EMT) marker gene expression profiles in the hard tumour cancer models. In the 3D-based PHA scaffolds, MDA-MB-231 cells expressed higher levels of Wnt-11 and mesenchymal markers, such as Snail and its downstream gene Vim mRNAs, while MCF7 cells exhibited no change in their expression. On the other hand, MCF7 cells exhibited a significantly increased E-Cadherin expression as compared to MDA-MB-231 cells. The expression levels of EMT markers were comparative to their expression reported in the tumour samples, making them good representative of cancer models. In future these models will be helpful in mimicking hypoxic tumours, in studying gene expression, cellular signalling, angiogenesis and drug response more accurately than 2D and perhaps other 3D models.