Abstract
Abstract MERCURY aimed to develop multidimensional in vitro models (3D-MMo) as advanced cell culture systems capable of closely reproducing the biological complexity of osteosarcoma. The project addressed one of the major challenges in biomedical research: the limited predictive power of conventional preclinical models. While two-dimensional (2D) cell cultures remain valuable for high-throughput studies, they fail to mimic the three-dimensional architecture, extracellular matrix (ECM), and dynamic cell–cell interactions found in native tissues. Similarly, animal models often show limited ability to predict human responses because of interspecies differences and raise ethical concerns. To overcome these limitations, MERCURY focused on engineering biomimetic tumour microenvironments that reproduce key features of osteosarcoma biology. Particular attention was devoted to the interactions between Cancer Stem Cells (CSCs) and Cancer-Associated Fibroblasts (CAFs), which are known to contribute to tumour progression, multidrug resistance, and disease recurrence. Achieved Results The first phase of the project focused on the development of hybrid biomaterials designed to mimic the physicochemical and structural properties of pathological bone tissue. Hybrid scaffolds composed of type I collagen and magnesium-doped hydroxyapatite (MgHA) or low-crystallinity carbonated calcium phosphate (CaP) were successfully produced using freeze-drying techniques. The resulting materials exhibited highly porous and interconnected architectures closely resembling trabecular bone. To further reproduce the complexity of the osteosarcoma extracellular matrix, the scaffolds were enriched with specific bioactive motifs capable of triggering targeted biological responses. This was achieved through the development of amphiphilic PLA-PEG copolymers functionalized with cyclic-RGDyK peptide. These nanosystems also proved suitable for drug delivery applications by incorporating doxorubicin, preserving its antitumour activity while enabling sustained drug release. This work led to a peer-reviewed publication in the International Journal of Pharmaceutics (2024). Following scaffold optimization, three complementary osteosarcoma models were established. The first consisted of a co-culture system of osteosarcoma cells and mesenchymal stem cells (MSCs) cultured within collagen–MgHA scaffolds. This model enabled the study of tumour–stromal interactions in a biomimetic bone environment and demonstrated increased extracellular matrix production together with modulation of inflammatory signalling, ultimately generating a tumour-supportive niche. These findings resulted in a publication in Biomedicine & Pharmacotherapy (2026). The second model focused on the role of cancer stem cells in osteosarcoma progression. CSC-enriched spheroids were incorporated into the biomimetic scaffolds and co-cultured with MSCs. Molecular analyses showed stromal activation and the acquisition of CAF-like phenotypes, indicating the establishment of cellular interactions that resemble those observed in aggressive tumours. The third model aimed to reproduce the biochemical complexity of the pathological extracellular matrix by incorporating glycosaminoglycan-like molecules, including hyaluronic acid and heparin. When seeded with the highly aggressive 143B osteosarcoma cell line, these scaffolds supported robust tumour cell adhesion, proliferation, and colonization. Finally, two of the developed models were evaluated in immunodeficient mice as a proof of concept. The implanted constructs showed significant tumour formation and scaffold colonization, confirming both the tumorigenic potential of the engineered systems and the ability of the biomimetic scaffolds to sustain osteosarcoma growth in vivo. Conclusions Overall, MERCURY successfully established innovative and biologically relevant platforms that reproduce essential aspects of the osteosarcoma microenvironment. These models provide valuable tools for investigating tumour biology, understanding mechanisms of chemoresistance, and improving preclinical drug screening. By bridging the gap between conventional cell cultures and animal studies, MERCURY offers a promising foundation for the development of more effective therapeutic strategies against bone cancers. References RGD-tagging of star-shaped PLA-PEG micellar nanoassemblies enhances doxorubicin efficacy against osteosarcoma. Oliva R, Torcasio SM, Coulembier O, Piperno A, Mazzaglia A, Scalese S, Rossi A, Bassi G, Panseri S, Montesi M, Scala A.Int J Pharm. 2024 May 25;657:124183. doi: 10.1016/j.ijpharm.2024.124183. Epub 2024 Apr 30.PMID: 38692500 Deciphering the interaction between osteosarcoma and mesenchymal stem cells in a 3D bone-mimetic co-culture model. Bassi G, Saqawa M, Apolloni L, Díaz-Prado S, Ollivier E, Levergeois R, Sandri M, Campodoni E, Cochonneau D, Panseri S, Heymann D, Montesi M. Biomed Pharmacother. 2026 Feb;195:118956. doi: 10.1016/j.biopha.2025.118956. Epub 2026 Jan 6.PMID: 41496360
Dettagli del progetto
Responsabile scientifico: Donatella Canistro
Strutture Unibo coinvolte:
Dipartimento di Farmacia e Biotecnologie
Coordinatore:
CNR - Consiglio Nazionale delle Ricerche(Italy)
Contributo totale Unibo: Euro (EUR) 37.482,00
Durata del progetto in mesi: 24
Data di inizio
30/11/2023
Data di fine:
28/02/2026
