A two-compartment bone tumor model to investigate interactions between healthy and tumor cells

Abstract

We produced a novel three-dimensional (3D) bone tumor model (BTM) to study the interactions between healthy and tumor cells in a tumor microenvironment, the migration tendency of the tumor cells, and the efficacy of an anticancer drug, Doxorubicin, on the cancer cells. The model consisted of two compartments: (a) a healthy bone tissue mimic, made of poly(lactic acid-co-glycolic acid) (PLGA)/beta-tricalcium phosphate (β-TCP) sponge seeded with human fetal osteoblastic cells (hFOB) and human umbilical vein endothelial cells (HUVECs), and (b) a tumor mimic, made of lyophilized collagen sponge seeded with human osteosarcoma cells (Saos-2). The tumor mimic component was placed into a central cavity created in the healthy bone mimic and together they constituted the complete 3D bone tumor model (3D-BTM). The porosities of both sponges were higher than 85% and the diameters of the pores were 199 ± 52 μm for the PLGA/TCP and 50–150 μm for the collagen scaffolds. The compression Young’s modulus of the PLGA/TCP and the collagen sponges were determined to be 4.76 MPa and 140 kPa, respectively. Cell proliferation, morphology, calcium phosphate forming capacity and alkaline phosphatase production were studied separately on both the healthy and tumor mimics. All cells demonstrated cellular extensions and spread well in porous scaffolds indicating good cell–material interactions. Confocal microscopy analysis showed direct contact between the cells present in different parts of the 3D-BTM. Migration of HUVECs from the healthy bone mimic to the tumor compartment was confirmed by the increase in the levels of angiogenic factors vascular endothelial growth factor, basic fibroblast growth factor, and interleukin 8 in the tumor component. Doxorubicin (2.7 μg.ml−1) administered to the 3D-BTM caused a seven-fold decrease in the cell number after 24 h of interaction with the anticancer drug. Caspase-3 enzyme activity assay results demonstrated apoptosis of the osteosarcoma cells. This novel 3D-BTM has a high potential for use in studying the metastatic capabilities of cancer cells, and in determining the effective drug types and combinations for personalized treatments.

Enhanced Biomimetics of Three-Dimensional Osteosarcoma Models: A Scoping Review

“…The full search is illustrated in the PRISMA flowchart ( Figure 2 ) . Finally, 70 studies fulfilling the inclusion/exclusion criteria were included in this review [ 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 6...…”

“…Compartmentalised culture models were developed in two studies to enhance OS-healthy cell crosstalk to closely mimic the natural tumour microenvironment. Komez et al (2020) developed a 3D model with two compartments: (1) a bone compartment composed of PLGA/TCL sponges seeded with hFOB and HUVECs and (2) a tumour compartment composed of collagen sponge seeded with Saos-2 OS cells [ 37 ]. This model investigated tumour–bone crosstalk by assessing HUVEC migration from normal bone to tumour compartment via showing increased angiogenic vascular endothelial growth factor (VEGF), fibroblast GF, and IL-8 within the tumour component.…”

“…However, since pure-collagen hydrogels have poor mechanical strength, collagen-based composites are widely used to mimic bone ECM [ 83 ]. In this review, collagen/HA scaffolds [ 32 ] demonstrated high stability and porosity and comparable gene expression to native bone, while collagen/polylactic-co-glycolic acid (PLGA)/TCP [ 37 ] and chitosan/collagen mimicked the complex bone matrix [ 41 ].…”

A 3D Osteosarcoma Model with Bone‐Mimicking Cues Reveals a Critical Role of Bone Mineral and Informs Drug Discovery

“…Toward the effort of developing in vitro OS experimental models with improved physiological relevance, a few recent studies have reported 3D OS models using spheroids, [19] nanoporous hydrogels, [20,21] or macroporous scaffolds originally developed for bone tissue engineering. [22][23][24][25][26][27] While these studies suggest the benefits of 3D models over 2D culture, they rely on only a few well-established cell lines that have been passaged in 2D culture for decades and may therefore no longer reflect the biology and heterogeneity of the disease. Furthermore, evaluation was limited mostly to cell proliferation and morphology with only a few markers related to stemness, osteogenesis, or angiogenesis.…”

“…[27,28] A few studies utilized collagen and hydroxyapatite-containing scaffolds to culture OS cells in vitro. [22][23][24][25] However, the specific role of bone mineral on OS biology and drug response has never been evaluated. Furthermore, the potential of integrating scalable and physiologically relevant 3D OS models with high-throughput sequencing tools for comprehensive interrogation of OS pathology or for identifying potential novel druggable targets remains untapped.…”