Strategies for Constructing Osteosarcoma 3D Culture Models and Progress in Their Applications

Changshuai Qin; Na Huang; Chengyi Qin; Cheng Huang; Zonghui Qiu; Jinheng Wang; Wei Fu; Wusong Liu; Wei Peng; Changgong Lan

doi:10.54097/h5e7p918

Authors

Changshuai Qin
Na Huang
Chengyi Qin
Cheng Huang
Zonghui Qiu
Jinheng Wang
Wei Fu
Wusong Liu
Wei Peng
Changgong Lan

DOI:

https://doi.org/10.54097/h5e7p918

Keywords:

Osteosarcoma, Three-dimensional Culture, Spheroid Culture, Scaffold Matrix, Bioprinting, Microfluidic Chips, Drug Screening, Personalized Therapy

Abstract

Osteosarcoma is a highly malignant primary bone tumor, and traditional two-dimensional (2D) culture methods cannot accurately simulate the tumor microenvironment. Three-dimensional (3D) culture better simulates the in vivo tumor microenvironment, providing more realistic experimental data regarding tumor progression, drug resistance, and drug screening. This paper introduces various 3D culture technologies, including scaffold-free culture, scaffold-based culture, microfluidic chips, and bioprinting. Moreover, 3D models show significant advantages in tumor microenvironment simulation, drug screening, and research on osteoblasts and osteoclasts, thus promoting the development of personalized therapy. Overall, 3D culture offers an important tool and platform for a deeper understanding of osteosarcoma's biological mechanisms and improving clinical treatment outcomes.

Downloads

Download data is not yet available.

References

[1] Strauss, S. J., Blay, J. Y., Brodowicz, T., et al. Bone sarcomas: ESMO-EURACAN-GENTURIS-ERNPaedCan Clinical Practice Guideline for diagnosis, treatment and follow-up. Annals of Oncology, 2021. DOI: https://doi.org/ 10.1016/ j. annonc. 2021.08.1995.

[2] Beird, H. C., Stefan, S. B., Adrienne, M. F., et al. Osteosarcoma. Nature Reviews Disease Primers, 2022. DOI: 10.1038/s41572-022-00409-y.

[3] Meltzer, P. S., Helman, L. J., Longo, D. L., et al. New Horizons in the Treatment of Osteosarcoma. The New England Journal of Medicine, 2021, 385(22): 2066-2076. DOI: 10.1056/ NEJMra2103423.

[4] Li, B., Li, Z. P., Lian, Z. G., et al. Evaluation of Tumor Resistance-Related Genes in Assessing Osteosarcoma Neoadjuvant Chemotherapy Resistance. Chinese Medicine, 2023, 18(10): 1522-1526. (in Chinese)

[5] Chen, Y. Study on the Correlation between Changes in TP53 Gene Expression before and after Neoadjuvant Chemotherapy in Osteosarcoma Patients. Xinjiang Medical University, 2023. (in Chinese)

[6] Chen, Y. C., Shu, W. Z., Tang, P. Z. Clinical Efficacy of Neoadjuvant Chemotherapy Combined with Surgery in the Treatment of Osteosarcoma and Its Impact on Prognosis. Cancer Progression, 2023, 21(06): 628-631. (in Chinese)

[7] Chinese Anti-Cancer Association Clinical Chemotherapy Committee, Chinese Society of Clinical Oncology Sarcoma Expert Committee. Expert Consensus on Anthracycline Treatment of Osteosarcoma in China. Cancer, 2022, 41(08): 368-384. (in Chinese)

[8] Edmondson, R., Broglie, J. J., Adcock, A. F., et al. Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors. Assay and Drug Development Technologies, 2014, 12(4): 207-218. DOI: 10. 1089/ adt.2014.573.

[9] Mittal, R., Woo, F. W., Castro, C. S., et al. Organ-on-chip Models: Implications in Drug Discovery and Clinical Applications. Journal of Cellular Physiology, 2019, 234(6): 8352-8380. DOI: 10.1002/jcp.27729.

[10] Fong, E. L. S., Harrington, D. A., Farach-Carson, M. C., et al. Heralding a New Paradigm in 3D Tumor Modeling. Biomaterials, 2016, 108: 197-213. DOI: 10.1016/j. biomaterials. 2016.08.052.

[11] Fong, E. L. S., Lamhamedi-Cherradi, S., Burdett, E., et al. Modeling Ewing Sarcoma Tumors In Vitro with 3D Scaffolds. Proceedings of the National Academy of Sciences, 2013, 110(16): 6500-6505. DOI: 10.1073/pnas.1221403110.

[12] González Díaz, E. C., Lee, A. G., Sayles, L. C., et al. A 3D Osteosarcoma Model with Bone-Mimicking Cues Reveals a Critical Role of Bone Mineral and Informs Drug Discovery. Advanced Healthcare Materials, 2022, 11(17). DOI: 10.1002/ adhm.202200768.

[13] De Luca, A., Raimondi, L., Salamanna, F., et al. Relevance of 3D Culture Systems to Study Osteosarcoma Environment. Journal of Experimental & Clinical Cancer Research, 2018, 37(1). DOI: 10.1186/s13046-017-0663-5.

[14] Rimann, M., Laternser, S., Gvozdenovic, A., et al. An In Vitro Osteosarcoma 3D Microtissue Model for Drug Development. Journal of Biotechnology, 2014, 189: 129-135. DOI: 10. 1016/ j. jbiotec. 2014. 09.005.

[15] Gibbs, C. P., Kukekov, V. G., Reith, J. D., et al. Stem-Like Cells in Bone Sarcomas: Implications for Tumorigenesis. Neoplasia, 2005, 7(11): 967-976. DOI: 10.1593/neo.05394.

[16] Ozturk, S., Gorgun, C., Gokalp, S., et al. Development and Characterization of Cancer Stem Cell-Based Tumoroids as an Osteosarcoma Model. Biotechnology and Bioengineering, 2020, 117(8): 2527-2539. DOI: 10.1002/bit.27381.

[17] González Díaz, E. C., Sinha, S., Avedian, R. S., et al. Tissue-Engineered 3D Models for Elucidating Primary and Metastatic Bone Cancer Progression. Acta Biomaterialia, 2019, 99: 18-32. DOI: 10.1016/j.actbio.2019.08.020.

[18] Jubelin, C., Muñoz-Garcia, J., Cochonneau, D., et al. Liquid Overlay Technique Allows the Generation of Homogeneous Osteosarcoma, Glioblastoma, Lung and Prostate Adenocarcinoma Spheroids for Drug Cytotoxicity Measurements. Frontiers in Bioengineering and Biotechnology, 2023, 11. DOI: 10.3389/fbioe.2023.1260049.

[19] Ohya, S., Kajikuri, J., Endo, K., et al. Calcium-Activated Potassium Channel KCa 1.1 as a Therapeutic Target to Overcome Chemoresistance in Three-Dimensional Sarcoma Spheroid Models. Cancer Science, 2021, 112(9): 3769-3783. DOI: 10.1111/cas.15046.

[20] Mei Li, M. Y. W., Zheng, Y., et al. Nitrogen-Containing, Bisphosphonate-Loaded Micro-Arc Oxidation-Coated Biodegradable Magnesium Alloy Pellets Inhibit Osteosarcoma by Targeting the Mevalonate Pathway. Journal Pre-proof, 2020. DOI: https://doi.org/10.1016/j.actbio.2020.11.019.

[21] Gebhard, C., Miller, I., Hummel, K., et al. Comparative Proteome Analysis of Monolayer and Spheroid Culture of Canine Osteosarcoma Cells. Journal of Proteomics, 2018, 177: 124-136. DOI: 10.1016/j.jprot.2018.01.006.

[22] Massimini, M., De Maria, R., Malatesta, D., et al. Establishment of Three-Dimensional Canine Osteosarcoma Cell Lines Showing Vasculogenic Mimicry and Evaluation of Biological Properties after Treatment with 17-AAG. Veterinary and Comparative Oncology, 2019, 17(3): 376-384. DOI: 10.1111/vco.12482.

[23] Tome, Y., Uehara, F., Kanaya, F., et al. Comparison of "Dimensionality" of Cancer Cell Culture in Gelfoam® Histoculture and Matrigel. In New York, NY: Springer New York, 2018: 205-214.

[24] Lü, Z., Zhang, L., Sun, X., Chen, C. Effect of Matrigel on the Formation of Transplanted Tumors in Human Osteosarcoma Animal Models. Tumor Research and Clinics, 2017. DOI: 10.3760/cma.j.issn.1006-9801.2017.11.001. (in Chinese)

[25] Nie, J. H., Yang, T., Li, H., et al. Frequently Expressed Glypican-3 as a Promising Novel Therapeutic Target for Osteosarcomas. Cancer Science, 2022, 113(10): 3618-3632. DOI: 10.1111/cas.15521.

[26] Chim, L. K., Williams, I. L., Bashor, C. J., et al. Tumor-Associated Macrophages Induce Inflammation and Drug Resistance in a Mechanically Tunable Engineered Model of Osteosarcoma. Biomaterials, 2023, 296: 122076. DOI: 10.1016/j.biomaterials.2023.122076.

[27] He, J., Chen, C., Chen, L., et al. Honeycomb-Like Hydrogel Microspheres for 3D Bulk Construction of Tumor Models. Research, 2022, 2022. DOI: 10.34133/2022/9809763.

[28] Molina, E. R., Chim, L. K., Salazar, M. C., et al. Mechanically Tunable Coaxial Electrospun Models of YAP/TAZ Mechanoresponse and IGF-1R Activation in Osteosarcoma. Acta Biomaterialia, 2019, 100: 38-51. DOI: 10.1016/j. actbio. 2019. 09.029.

[29] Sarkar, S., Peng, C., Tung, Y. Comparison of VEGF-A Secretion from Tumor Cells under Cellular Stresses in Conventional Monolayer Culture and Microfluidic Three-Dimensional Spheroid Models. PLOS ONE, 2020, 15(11): e24 0833. DOI: 10.1371/journal.pone.0240833.

[30] Sarkar, S., Peng, C., Kuo, C. W., et al. Study of Oxygen Tension Variation within Live Tumor Spheroids Using Microfluidic Devices and Multi-Photon Laser Scanning Microscopy. RSC Advances, 2018, 8(53): 30320-30329. DOI: 10. 1039/C8RA05505J.

[31] Fois, M. G., Tahmasebi Birgani, Z. N., López-Iglesias, C., et al. In Vitro Vascularization of 3D Cell Aggregates in Microwells with Integrated Vascular Beds. Materials Today Bio, 2024, 29: 101260. DOI: 10.1016/j.mtbio.2024.101260.

[32] Loi, G., Stucchi, G., Scocozza, F., et al. Characterization of a Bioink Combining Extracellular Matrix-like Hydrogel with Osteosarcoma Cells: Preliminary Results. Gels, 2023, 9(2): 129. DOI: 10.3390/gels9020129.

[33] Lin, Y., Yang, Y., Yuan, K., et al. Multi-Omics Analysis Based on 3D-Bioprinted Models Innovates Therapeutic Target Discovery of Osteosarcoma. Bioactive Materials, 2022, 18: 459-470. DOI: 10.1016/j.bioactmat.2022.03.029.

[34] Lin, Y., Yuan, K., Yang, Y., et al. Osteosarcoma Progression in Biomimetic Matrix with Different Stiffness: Insights from a Three-Dimensional Printed Gelatin Methacrylamide Hydrogel. International Journal of Biological Macromolecules, 2023, 252: 126391. DOI: 10.1016/j. ijbiomac. 2023.126391.

[35] Shahin-Shamsabadi, A., Selvaganapathy, P. R. A Rapid Biofabrication Technique for Self-Assembled Collagen-Based Multicellular and Heterogeneous 3D Tissue Constructs. Acta Biomaterialia, 2019, 92: 172-183. DOI: 10.1016/j.actbio. 2019. 05.024.

[36] Gottfried, E., Kunz-Schughart, L. A., Andreesen, R., et al. Brave Little World: Spheroids as an In Vitro Model to Study Tumor-Immune-Cell Interactions. Cell Cycle, 2006.

[37] Bassi, G., Rossi, A., Campodoni, E., et al. 3D Tumor-Engineered Model Replicating the Osteosarcoma Stem Cell Niche and In Vivo Tumor Complexity. ACS Applied Materials & Interfaces, 2024. DOI: 10.1021/acsami.4c02567.

[38] Villasante, A., Marturano-Kruik, A., Vunjak-Novakovic, G. Bioengineered Human Tumor within a Bone Niche. Biomaterials, 2014, 35(22): 5785-5794. DOI: 10.1016/j. biomaterials. 2014.03.081.

[39] Belka, J., Nickel, J., Kurth, D. G. Growth on Metallo-Supramolecular Coordination Polyelectrolyte (MEPE) Stimulates Osteogenic Differentiation of Human Osteosarcoma Cells (MG63) and Human Bone Marrow Derived Mesenchymal Stem Cells. Polymers, 2019, 11(7): 1090. DOI: 10.3390/polym11071090.

[40] Kihara, T., Umezu, C., Sawada, K., et al. Osteogenic Cells Form Mineralized Particles, a Few μm in Size, in a 3D Collagen Gel Culture. PeerJ, 2019, 7: e7889. DOI: 10.7717/peerj.7889.

[41] Colella, G., Fazioli, F., Gallo, M., et al. Sarcoma Spheroids and Organoids--Promising Tools in the Era of Personalized Medicine. International Journal of Molecular Sciences, 2018, 19(2): 615. DOI: 10.3390/ijms19020615.