Magnetically tunable composite hydrogels with cellulose nanocrystals

In this work, a series of biocompatible nanocomposite hydrogels was prepared by UV-initiated polymerization based on 2-hydroxyethyl methacrylate (HEMA), using ethylene glycol dimethacrylate (EGDMA) as a crosslinker and 2-hydroxymethyl-2-methylpropiophenone as a photoinitiatior, containing liquid crystals of cellulose nanocrystals (CNCs) doped with magnetic nanoparticles. The formation of liquid crystals was achieved thanks to the intrinsic property of CNCs to self-assemble above a critical aqueous concentration. By varying the preparation conditions, allowing different times for phase-separation between the nanoparticles and CNCs and exposing the polymerization mixture to small magnetic field, films with different size and orientation of CNC liquid crystal domains were synthesized. Subsequently, the hydrogel films were studied by dynamic mechanical analysis (DMA) to evaluate the effect of these parameters on the mechanical properties, specifically the Young’s modulus and the ultimate strength. Also, the microstructure of the films was studied via polarized optical microscopy (POM) and scanning electron microscopy (SEM). The water uptake capacity was also analyzed. The results indicate that the presence of cellulose nanocrystals modulates the architecture of the prepared hydrogels. Cholesteric microdomains were embedded in PHEMA matrix and their uniaxial alignment was achieved upon exposure to small static magnetic field, already after several hours. Moreover, structural gradient in the distribution of the liquid crystalline microdomains, in dependence on their size, was obtained within the material. This originated from the direct proportionality between the size and the density of liquid crystals. Finally, it was shown that cellulose nanocrystals act as reinforcing structures of the hydrogels, when the degree of their self-assembly is sufficient, and therefore the resulting hydrogel exhibits both higher resistance to elastic deformation and also higher ultimate strength. Finally, we showed that mechanical performance of these nanocomposites can be enhanced by systematic orientation of the liquid crystalline domains

Magnetic microrheometry of tumor-relevant stiffness levels and probabilistic quantification of viscoelasticity differences inside 3D cell culture matrices.

The progression of breast cancer involves cancer-cell invasions of extracellular matrices. To investigate the progression, 3D cell cultures are widely used along with different types of matrices. Currently, the matrices are often characterized using parallel-plate rheometry for matrix viscoelasticity, or liquid-like viscous and stiffness-related elastic characteristics. The characterization reveals averaged information and sample-to-sample variation, yet, it neglects internal heterogeneity within matrices, experienced by cancer cells in 3D culture. Techniques using optical tweezers and magnetic microrheometry have measured heterogeneity in viscoelasticity in 3D culture. However, there is a lack of probabilistic heterogeneity quantification and cell-size-relevant, microscale-viscoelasticity measurements at breast-tumor tissue stiffness up to ≃10 kPa in Young's modulus. Here, we have advanced methods, for the purpose, which use a magnetic microrheometer that applies forces on magnetic spheres within matrices, and detects the spheres displacements. We present probabilistic heterogeneity quantification using microscale-viscoelasticity measurements in 3D culture matrices at breast-tumor-relevant stiffness levels. Bayesian multilevel modeling was employed to distinguish heterogeneity in viscoelasticity from the effects of experimental design and measurement errors. We report about the heterogeneity of breast-tumor-relevant agarose, GrowDex, GrowDex-collagen and fibrin matrices. The degree of heterogeneity differs for stiffness, and phase angle (i.e. ratio between viscous and elastic characteristics). Concerning stiffness, agarose and GrowDex show the lowest and highest heterogeneity, respectively. Concerning phase angle, fibrin and GrowDex-collagen present the lowest and the highest heterogeneity, respectively. While this heterogeneity information involves softer matrices, probed by ≃30 μm magnetic spheres, we employ larger ≃100 μm spheres to increase magnetic forces and acquire a sufficient displacement signal-to-noise ratio in stiffer matrices. Thus, we show pointwise microscale viscoelasticity measurements within agarose matrices up to Young's moduli of 10 kPa. These results establish methods that combine magnetic microrheometry and Bayesian multilevel modeling for enhanced heterogeneity analysis within 3D culture matrices