Colorectal Cancer Bioengineered Microtissues as a Model to Replicate Tumor-ECM Crosstalk and Assess Drug Delivery Systems In Vitro

Abstract

Current 3D cancer models (in vitro) fail to reproduce complex cancer cell extracellular matrices (ECMs) and the interrelationships occurring (in vivo) in the tumor microenvironment (TME). Herein, we propose 3D in vitro colorectal cancer microtissues (3D CRC μTs), which reproduce the TME more faithfully in vitro. Normal human fibroblasts were seeded onto porous biodegradable gelatin microbeads (GPMs) and were continuously induced to synthesize and assemble their own ECMs (3D Stroma μTs) in a spinner flask bioreactor. Then, human colon cancer cells were dynamically seeded onto the 3D Stroma μTs to achieve the 3D CRC μTs. Morphological characterization of the 3D CRC μTs was performed to assess the presence of different complex macromolecular components that feature in vivo in the ECM. The results showed the 3D CRC μTs recapitulated the TME in terms of ECM remodeling, cell growth, and the activation of normal fibroblasts toward an activated phenotype. Then, the microtissues were assessed as a drug screening platform by evaluating the effect of 5-Fluorouracil (5-FU), curcumin-loaded nanoemulsions (CT-NE-Curc), and the combination of the two. When taken together, the results showed that our microtissues are promising in that they can help clarify complex cancer–ECM interactions and evaluate the efficacy of therapies. Moreover, they may be combined with tissue-on-chip technologies aimed at addressing further studies in cancer progression and drug discovery.

Synthesis of Photoluminescent Carbon Dots Using Hibiscus Tea Waste and Heteroatom Doping for Multi-Metal Ion Sensing: Applications in Cell imaging and Environmental Samples

“…Faint green fluorescence was observed when HCT-116 cells were incubated with NB-CDs. After incubation with Ag + , Cd 2+ , and Cr 3+ , living cells showed blue and strong green fluorescence [53]. NB-CDs exhibited excellent cell membrane permeability in an overlay of fluorescence and phase-contrast images, indicating effective fluorescence signals [54].…”

Recent Advances in 3D Cultures

“…Many 3D cancer in vitro models present several limitations in reproducing the complexity of the tumor ECM and how it can affect the tumor microenvironment in general [23]. La Rocca A. et al [24] proposed a novel 3D colorectal cancer microtissue made of normal human fibroblasts seeded onto porous biodegradable gelatin microbeads cultured in a spinner flask bioreactor. This encouraged spontaneous ECM synthesis; then, colon cancer cells were dynamically seeded upon these microtissues.…”