Investigation of the Viability of Different Cancer Cells on Decellularized Adipose Tissue

Öz

Cancer is one of the most severe diseases diagnosed in millions of people worldwide each year. Despite many studies, there is insufficient information on how tumor formation prevents cancer treatment development. Although clinical trials are the most effective way to examine tumor formation and test anti-tumor drugs, ethical and safety limitations prevent this method from being widely used. This study aims to test the cellular behavior of different cancer cell lines on the platform obtained from decellularized adipose tissue in vitro. Detergent-based decellularization protocol applied to adipose tissue and cancer cell lines were seeded on obtained extracellular matrixes. Cell viability and apoptosis were observed by MTT assay and Acridine orange/Propidium iodide staining, respectively. Also, cell-cell and cell-matrix interactions were investigated via pan-cadherin immunostaining. All cancer cell lines were also seeded in a cell culture dish to compare three-dimensional culture results with two-dimensional culture. As a result, the decellularization protocol allowed the original structure of the tissue scaffold to be preserved. According to cell viability analysis and immunocytochemical staining results, T98G and Hep3B cell lines were observed to have higher adhesion and viability potential than the WiDr cell line on the tissue matrix obtained. With this study, it can be said that different cancer cells have different behaviors on decellularized matrixes. Finally, developing in vitro models as a more economical, scalable, and reproducible way to test drugs and therapeutics is crucial for successful clinical translation.

Anahtar Kelimeler

• Adipose tissue
• cancer cell lines
• decellularization
• extracellular matrix
• three-dimensional culture model

Kaynakça

1. Kocarnik, J. M., Compton, K., et al., 2022, Cancer incidence, mortality, years of life lost, years lived with disability, and disability- adjusted life years for 29 cancer groups from 2010 to 2019: a systematic analysis for the global burden of disease study 2019. JAMA Oncology, 8: 420-444.
2. Han, S. J., Kwon, S., Kim, K. S., 2022, Contribution of mechanical homeostasis to epithelial-mesenchymal transition. Cellular Oncology, 45:1119-1136.
3. Walker, C., Mojares, E., del Río Hernández, A., 2018, Role of extracellular matrix in development and cancer progression. International Journal of Molecular Sciences,19:3028.
4. Sensi, F., D’Angelo, E., et al., 2020, Recellularized colorectal cancer patient-derived scaffolds as in vitro pre-clinical 3D model for drug screening. Cancers, 12:681. Celal Bayar University Journal of Science Volume 19, Issue 2, 2023, p 113-119 Doi: 10.18466/cbayarfbe.1212604.
5. Brancato, V., Oliveira, J.M., et al., 2020, Could 3D models of cancer enhance drug screening?. Biomaterials, 232:119744.
6. Pospelov, A.D., Timofeeva, L.B., et al., 2020, Comparative analysis of two protocols of mouse tissues decellularization for application in experimental oncology. Opera Medica et Physiologica, 7:13.
7. Ferreira, L.P., Gaspar, V.M., 2020, Decellularized extracellular matrix for bioengineering physiomimetic 3D in vitro tumor models. Trends in Biotechnology, 38:1397-1414.
8. Rijal, G., Li, W., 2018, Native-mimicking in vitro microenvironment: an elusive and seductive future for tumor modeling and tissue engineering. Journal of Biological Engineering, 12:1-22.

9. Mavrogonatou, E., Pratsinis, H., et al., 2019, Extracellular matrix alterations in senescent cells and their significance in tissue homeostasis. Matrix Biology, 75:27-42.
10. Theocharis, A.D., Karamanos, N.K., 2019, Proteoglycans remodeling in cancer: Underlying molecular mechanisms. Matrix Biology, 75:220-259.
11. Ajeti, V., Lara-Santiago, J., et al., 2017, Ovarian and breast cancer migration dynamics on laminin and fibronectin bi-directional gradient fibers fabricated via multiphoton excited photochemistry. Cellular and Molecular Bioengineering, 10:295- 311.
12. Sesli, M., Akbay, E., Onur, M. A., 2018, Decellularization of rat adipose tissue, diaphragm, and heart: a comparison of two decellularization methods. Turkish Journal of Biology, 42:537.
13. Akbay, E., Onur, M. A., 2019, Investigation of survival and migration potential of differentiated cardiomyocytes transplanted with decellularized heart scaffold. Journal of Biomedical Materials Research Part A, 107:561-570.
14. Akbay, E., Onur, M. A., 2018, Myocardial Tissue Engineering: A Comparative Study of Different Solutions for Use as a Natural Scaffold Being of Heart a Comparative Study of Different Solutions for Decellularization Heart Scaffold. Biomedical Journal, 2:4.
15. Thevenot, P., Nair, A., et al., 2008, Method to analyze three- dimensional cell distribution and infiltration in degradable scaffolds. Tissue Engineering Part C Methods, 14:319-31.
16. Lim, P.J., Gan, C.S., et al., 2019, Lipid lowering effect of Eurycoma longifolia Jack aqueous root extract in hepatocytes. Kuwait Journal of Science, 46: 2.
17. Wang, L., Johnson, J.A., et al., 2013, Combining decellularized human adipose tissue extracellular matrix and adipose-derived stem cells for adipose tissue engineering. Acta Biomaterialia, 9:8921-31.
18. Dunne, L.W., Huang, Z., et al., 2014, Human decellularized adipose tissue scaffold as a model for breast cancer cell growth and drug treatments. Biomaterials, 35:4940-49.
19. Tıǧlı, S.R., Ghosh, S., et al., 2009, Comparative chondrogenesis of human cell sources in 3D scaffolds. Journal of Tissue Engineering and Regenerative Medicine, 3:348-60.
20. Abdel Wahab, S.I., Abdul, A.B., et al., 2009, In vitro ultramorphological assessment of apoptosis induced by Zerumbone on (HeLa). Journal fo Biomedicine and Biotechnology, 2009.
21. Mosmann, T., 1983, Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 65:55-63.
22. Gubareva, E.A., Sjöqvist, S., et al., 2016, Orthotopic transplantation of a tissue engineered diaphragm in rats. Biomaterials, 77:320-35.
23. Fisher, M.F., Rao, S.S., 2020, Three‐dimensional culture models to study drug resistance in breast cancer. Biotechnology and Bioengineering, 117:2262-78.
24. Chaitin, H., Lu, M.L., et al., 2021, Development of a decellularized porcine esophageal matrix for potential applications in cancer modeling. Cells, 10:1055.
25. Flynn, L.E., 2010, The use of decellularized adipose tissue to provide an inductive microenvironment for the adipogenic differentiation of human adipose-derived stem cells. Biomaterials, 31:4715-24.
26. Brown, B.N., Freund, J.M., et al., 2011, Comparison of three methods for the derivation of a biologic scaffold composed of adipose tissue extracellular matrix. Tissue Engineering Part C Methods, 17:411-21.
27. Chun, S.Y., Lim, J.O., et al., 2019, Preparation and characterization of human adipose tissue-derived extracellular matrix, growth factors, and stem cells: a concise review. Tissue Engineering and Regenerative Medicine, 16:385-93.
28. Varol, C., Sagi, I., 2018, Phagocyte-extracellular matrix crosstalk empowers tumor development and dissemination. The FEBS Journal, 285:734-51.