This research presents the development of an innovative paper-supported tumor tissue model incorporating the human breast cancer cell line, MDA-MB-231. Collagen and Matrigel served as the extracellular matrix, suspending the cancer cells to create a biologically relevant microenvironment. To evaluate the functional performance of the device, a tracer dye (tartrazine) experiment was conducted, and the system was also modeled using COMSOL Multiphysics to compare and validate the experimental results against simulated chemical gradient profiles. The cell-laden matrix was distributed onto ten individual paper discs, which were meticulously stacked to construct a three-dimensional architecture within a custom 3D-printed device. Following a 24–48-hour culture period, the layers were de-stacked to evaluate the establishment of biochemical gradients by assessing no of live and dead cell, HIF-1α expression levels, and the extent of apoptosis across the layers. Notably, a progressive increase in cell death, apoptosis, and HIF-1α expression was observed from the top to the bottom layers, reflecting the diffusion-limited conditions of the tumor milieu. These findings underscore the potential of this paper-based tumor model as a sophisticated platform for replicating tumor microenvironments and advancing cancer research.
