1The interactions between glioma cells and neurons are important for glioma progression but are rarely mimicked and recapitulated in in vitro three-dimensional (3D) models, which may affect the success rate of relevant drug research and development. In this study, an in vitro bioprinted 3D glioma model consisting of an outer hemispherical shell with neurons and an inner hemisphere with glioma cells is proposed to simulate the natural glioma. This model was produced by extrusion-based 3D bioprinting technology. The cells survival rate, morphology, and intercellular Ca2+ concentration studies were carried out up to 5 days of culturing. It was found that neurons could promote the proliferation of glioma cells around them, associate the morphological changes of glioma cells to be neuron-like, and increase the expression of intracellular Ca2+ of glioma cells. Conversely, the presence of glioma cells could maintain the neuronal survival rate and promote the neurite outgrowth. The results indicated that glioma cells and neurons facilitated each other implying a symbiotic pattern established between two types of cells during the early stage of glioma development, which were seldom found in the present artificial glioma models. The proposed bioprinted glioma model can mimic the natural microenvironment of glioma tissue, provide an in-depth understanding of cell-cell interactions, and enable pathological and pharmacological studies of glioma.
In vitro neurovascular unit (NVU) models are valuable for investigating brain functions and developing drugs. However, it remains challenging to recapitulate the native architectural features and ultra-soft extracellular matrix (ECM) properties of the natural NVU. Cell-laden bioprinting is promising to prepare complex living tissues, but hard to balance the fidelity and cell growth. This study proposes a novel two-stage methodology for biomanufacturing functional 3D neurovascular constructs in vitro with low modulus of ECM. At the shaping stage, a low-viscosity alginate/collagen is printed through an embedded approach; at the culturing stage, the alginate is removed through targeted lysing. The low-viscosity and rapid crosslinking properties provide a printing resolution of ≈10 µm, and the lysis processing can decrease the hydrogels' modulus to ≈1 kPa and adjust the porosity of the microstructure, providing cells with an environment closing to the brain ECM. A 3D hollow coaxial neurovascular model is fabricated, in which the endothelial cells has expressed tight junction proteins and shown selective permeability, and the astrocytes outside of the endothelial layer are found to spread out with branches and directly interact with endothelial cells. The present study offers a promising modeling method for better understanding the NVU function and screening neuro-drugs.
Bioprinting , Endothelial Cells , Bioprinting/methods , Viscosity , Hydrogels/chemistry , Collagen , Alginates/chemistry , Printing, Three-Dimensional , Tissue Scaffolds , Tissue Engineering/methods
In this paper, the initial values of damage parameters in the Gurson-Tvergaard-Needleman (GTN) model are determined by a microscopic test combined with empirical formulas, and the final accurate values are determined by finite element reverse calibration. The original void volume fraction (f0), the volume fraction of potential nucleated voids (fN), the critical void volume fraction (fc), the void volume fraction at the final failure (fF) of material are assigned as 0.006, 0.001, 0.03, 0.06 according to the simulation results, respectively. The hemispherical punch stretching test of commercially pure titanium (TA1) sheet is simulated by a plastic constitutive formula derived from the GTN model. The stress and strain are obtained at the last loading step before crack. The forming limit diagram (FLD) and the forming limit stress diagram (FLSD) of the TA1 sheet under plastic forming conditions are plotted, which are in good agreement with the FLD obtained by the hemispherical punch stretching test and the FLSD obtained by the conversion between stress and strain during the sheet forming process. The results show that the GTN model determined by the finite element reverse calibration method can be used to predict the forming limit of the TA1 sheet metal.
