Engineering Novel 3D Models to Recreate High‐Grade Osteosarcoma and its Immune and Extracellular Matrix Microenvironment

Abstract

Osteosarcoma (OS) is the most common primary bone cancer, where the overall 5-year surviving rate is below 20% in resistant forms. Accelerating cures for those poor outcome patients remains a challenge. Nevertheless, several studies of agents targeting abnormal cancerous pathways have yielded disappointing results when translated into clinic because of the lack of accurate OS preclinical modeling. So, any effort to design preclinical drug testing may consider all inter-, intra-, and extra-tumoral heterogeneities throughout models mimicking extracellular and immune microenvironment. Therefore, the bioengineering of patient-derived models reproducing the OS heterogeneity, the interaction with tumor-associated macrophages (TAMs), and the modulation of oxygen concentrations additionally to recreation of bone scaffold is proposed here. Eight 2D preclinical models mimicking several OS clinical situations and their TAMs in hypoxic conditions are developed first and, subsequently, the paired 3D models faithfully preserving histological and biological characteristics are generated. It is possible to shape reproducibly M2-like macrophages cultured with all OS patient-derived cell lines in both dimensions. The final 3D models pooling all heterogeneity features are providing accurate proliferation and migration data to understand the mechanisms involved in OS and immune cells/biomatrix interactions and sustained such that engineered 3D preclinical systems will improve personalized medicine.

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...…”

“…Among the 45 3D scaffold-based models, osteosarcoma spheroids were present in 11 of them. Among these, nine cases [ 16 , 24 , 34 , 40 , 41 , 46 , 53 , 59 , 60 ] utilised pre-formed spheroids created through liquid overlay techniques ( n = 6) and hanging drop methods ( n = 3). These pre-formed spheroids were later embedded with the 3D scaffold.…”

“…MG-63 was the most used OS cell line (>30% of studies), followed by U-2 OS (18%) and Saos-2 (16%) ( Figure 3 C). Pierrevelcin et al (2022) used OS patient-derived cells (PDCL) [ 41 ].…”

“…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 ].…”

Managing the immune microenvironment of osteosarcoma: the outlook for osteosarcoma treatment

“…Osteosarcoma tissue is surrounded by massive immune cell infiltration, resulting in the creation of a complex immune microenvironment that allows osteosarcoma cells to grow within the bone by creating an immunosuppressive microenvironment to maintain their survival and proliferation. [35][36][37] A robust immunosuppressive microenvironment is positively correlated with overactivation of molecules associated with immune suppression, such as indoleamine 2,3-dioxygenase (IDO), programmed cell death protein 1 (PD-1), interleukin-10 (IL-10), transforming growth factor-β (TGF-β), vascular endothelial growth factor (VEGF), and signal transducer and activator of transcription 3 (STAT3), due to their immunosuppressive effects mediated by myeloid-derived suppressor cells (MDSCs), TAMs, and regulatory T lymphocytes (Tregs) [38][39][40][41][42][43] (Fig. 2).…”