Scaffold-based 3D cellular models mimicking the heterogeneity of osteosarcoma stem cell niche

Abstract

The failure of the osteosarcoma conventional therapies leads to the growing need for novel therapeutic strategies. The lack of specificity for the Cancer Stem Cells (CSCs) population has been recently identified as the main limitation in the current therapies. Moreover, the traditional two-dimensional (2D) in vitro models, employed in the drug testing and screening as well as in the study of cell and molecular biology, are affected by a poor in vitro-in vivo translation ability. To overcome these limitations, this work provides two tumour engineering approaches as new tools to address osteosarcoma and improve therapy outcomes. In detail, two different hydroxyapatite-based bone-mimicking scaffolds were used to recapitulate aspects of the in vivo tumour microenvironment, focusing on CSCs niche. The biological performance of human osteosarcoma cell lines (MG63 and SAOS-2) and enriched-CSCs were deeply analysed in these complex cell culture models. The results highlight the fundamental role of the tumour microenvironment proving the mimicry of osteosarcoma stem cell niche by the use of CSCs together with the biomimetic scaffolds, compared to conventional 2D culture systems. These advanced 3D cell culture in vitro tumour models could improve the predictivity of preclinical studies and strongly enhance the clinical translation.

In vitro three-dimensional cell cultures for bone sarcomas

“…Both hybrids scaffolds resulted in stable CSCs enriched OS spheroid growth without any loss of round morphology compared to 2D. Moreover, an increase of stemness markers including OCT-4, NANOG and SOX-2 was observed that indicated that both types of hybrids scaffolds were able to mimic native environment promoting CSC stimulation [84] . Hydroxyapatite nanoparticle 3D cultures had a strong impact on the survival of OS cells under anti-tumoral oxidative stress therapy [85] .…”

“…The generated spheroids were viable, conserved their pluripotency, and constituted an ideal model for drug screening. CSCs enrichment from MG-63 and SaOS2 spheroids by scaffold-free method was combined with two hybrid scaffolds that mimic ECM and used to analyse the impact of ECM in OS CSCs development [84] . Hybrid scaffold was constituted by Mg-doped hydroxyapatite coupled to collagen fibres and a porous hydroxyapatite substrate.…”

Patient-derived scaffolds as a drug-testing platform for endocrine therapies in breast cancer

“…Moreover, when introducing another ERα-positive cell line, T47D, to the PDS system it was clear that the drug effect was to a large extent the driver of the cellular response, more than the differences between cancer cell lines, which confirms the robustness of the system 14 , 15 . It is well known that cellular 3D growth contributes to more in vivo-like phenotypes 34 . However, the presented data indicated that different 3D models could induce different gene expression profiles of cancer cells in response to the treatments.…”

“…The fact that PDSs and matrigel cultures induced more similar gene expression profiles supports the importance of using cancer tissue-derived materials in drug testing. Since the expression of pluripotency markers were clearly affected differently by the various model systems, the link between the cancer microenvironment and the problematic population of cancer stem cells with pluripotency features indeed needs to be further clarified 34 .…”

3D Cocultures of Osteoblasts and Staphylococcus aureus on Biomimetic Bone Scaffolds as a Tool to Investigate the Host–Pathogen Interface in Osteomyelitis

“…To overcome this limitation, here we propose a composite system in which 3D biomimetic scaffolds made of MgHA/Col mixed matrix are seeded with the murine MC3T3-E1 osteoblast precursor cell line and infected with SA. The MgHA/Col scaffolds have already demonstrated high biocompatibility with different osteogenic cells, such as murine and human mesenchymal stem cells and human osteosarcoma cell lines (MG63 and SAOS-2) [ 37 , 38 , 39 , 53 , 54 ], while to the best of our knowledge the osteoblast cell line MC3T3-E1 derived from primary mouse calvaria has been used here for the first time. In line with previous work on other osteogenic cells, we found that MC3T3-E1 cells adapted to the 3D culture on MgHA/Col scaffolds, proliferated, were metabolically active, and shaped their gene expression according to a more advanced differentiation state by increasing, for instance, the mRNA levels of Bmp2 and Spp1 genes, as compared to 2D conditions ( Figure 1 ).…”