Gelatin methacryloyl and Laponite bioink for 3D bioprinted organotypic tumor modeling

Abstract

Three-dimensional (3D) in vitro tumor models that can capture the pathophysiology of human tumors are essential for cancer biology and drug development. However, simulating the tumor microenvironment is still challenging because it consists of a heterogeneous mixture of various cellular components and biological factors. In this regard, current extracellular matrix (ECM)-mimicking hydrogels used in tumor tissue engineering lack physical interactions that can keep biological factors released by encapsulated cells within the hydrogel and improve paracrine interactions. Here, we developed a nanoengineered ion-covalent cross-linkable bioink to construct 3D bioprinted organotypic tumor models. The bioink was designed to implement the tumor ECM by creating an interpenetrating network composed of gelatin methacryloyl (GelMA), a light cross-linkable polymer, and synthetic nanosilicate (Laponite) that exhibits a unique ionic charge to improve retention of biological factors released by the encapsulated cells and assist in paracrine signals. The physical properties related to printability were evaluated to analyze the effect of Laponite hydrogel on bioink. Low GelMA (5%) with high Laponite (2.5-3.5%) composite hydrogels and high GelMA (10%) with low Laponite (1.0-2.0%) composite hydrogels showed acceptable mechanical properties for 3D printing. However, a low GelMA composite hydrogel with a high Laponite content could not provide acceptable cell viability. Fluorescent cell labeling studies showed that as the proportion of Laponite increased, the cells became more aggregated to form larger 3D tumor structures. RT-qPCR and western blot experiments showed that an increase in the Laponite ratio induces upregulation of growth factor and tissue remodeling-related genes and proteins in tumor cells. In contrast, cell cycle and proliferation-related genes were downregulated. On the other hand, concerning fibroblasts, the increase in the Laponite ratio indicated an overall upregulation of the mesenchymal phenotype-related genes and proteins. Our study may provide a rationale for using Laponite-based hydrogels in 3D cancer modeling.

Development of a 3Din vitrohuman-sized model of cervical dysplasia to evaluate the delivery of ethyl cellulose-ethanol injection for the treatment of cervical dysplasia ablation

“…GelMA has proven to have good transport of nutrients and oxygen that enables cell response over time. (30,31) A recent model showed the encapsulation of human pancreatic cells and stromal fibroblasts in GelMA (8% w/v) with the photoinitiator LAP could mimic cell-ECM interactions and is proven to have a good diffusion of nutrients and oxygen, which allowed the cells to proliferate in the GelMA beads. (32) Using the GelMA hydrogel, we were able to increase the thickness of our first model and still have a higher number of live cells compared to dead cells.…”

Vascularized Liver Tissue Embedded Bioprinting Utilizing GelMA/Nanoclay-based Composite hydrogels

“…Consequently, during the degradation study, this sample absorbs a greater quantity of water containing collagenase enzyme, leading to the highest degradation rate observed among the samples studied. Conversely, the 5G1.5L and 5G2L samples, featuring smaller pore sizes and lower swelling ratios, demonstrate reduced rates of degradation as shown in Figure S2 [69]. This property renders them well-suited for more prolonged in vitro investigations, such as on-chip applications.…”

3D bioprinting of mouse pre-osteoblasts and human MSCs using bioinks consisting of gelatin and decellularized bone particles

“…Furthermore, to improve the printability and shape fidelity of pure GEL, it has been used with other natural and synthetic polymers and successful results have been reported for instance methacrylated GEL [11,12], oxidized alginate (ADA)-GEL [13], GELsilk [14] and GEL-chitosan [15]. In numerous studies, methacrylated GEL has been used due to the easy cross-linkable property using UV light [16]. In another study, Negrini et al demonstrated a GEL biomaterial ink with a different crosslinking approach, N,N ′ -methylene bisacrylamide (MBA) was used as a crosslinker and included in the GEL solution before 3D printing and the hydrogels were printed on a cooled surface [17].…”