Dolaşımdaki tümör hücreleri araştırmalarında kullanılmak üzere sirkülasyonlu mikroakışkan biyoreaktörün tasarımı ve hemodinamik kayma gerilimi kuvvetlerinin meme kanseri (MDA-MB-231) hücre canlılığı üzerine etkisinin incelenmesi

Kanser metastazı sırasında dolaşımdaki tümör hücreleri (CTC) mikro ortamlarında hidrostatik basınç ve kayma gerilimi gibi biyomekanik kuvvetlere maruz kalmaktadırlar. Bu faktörler kanser hücresinin heterojenitesinde önemli bir rol oynarken, hücre canlılığını da büyük oranda etkilemektedir. Bu çalışmada, CTC hücrelerinin metastaz sürecinde maruz kaldıkları hemodinamik kayma gerilimini taklit etmek için sirkülasyonlu mikroakışkan biyoreaktör geliştirilmiş ve mikroakışkan biyoreaktörün hız, duvar basıncı ve kayma gerilimleri için teorik hesaplamalar gerçekleştirilmiştir. Daha sonra, mikroakışkan kanal boyutları sabit tutularak üç farklı akış hızında (6, 9 ve 12 mL dk-1) artan hemodinamik kayma gerilimlerinde ve sirkülasyon zamanlarında (6, 12 ve 24 saat) MDA-MB-231 meme kanseri hücrelerinin canlılıkları MTT ve Canlı/Ölü testi ile incelenmiştir. Deneysel sonuçlar, statik koşullarda kültür edilen meme kanseri hücrelerine kıyasla, hücreler üzerindeki hemodinamik kayma gerilimi ve sirkülasyon süresi arttırıldıkça hücre canlılığının azaldığını göstermiştir. Bununla birlikte yüksek hemodinamik kayma gerilimi (66 dyn cm-2) ve uzun sirkülasyon süresinde (24 saat) hücre canlılığının %20’ ye kadar düştüğü bildirilmiştir. Bu çalışma ile geliştrilen sirkülasyonlu mikroakışkan biyoreaktör ile kanser hücrelerinin hemodinamik akış koşullarında canlılıkları değerlendirilebileceği gibi fenotipik ve genotipik değişimler de incelenebilecektir.

Anahtar Kelimeler:

Sirkülasyonlu mikroakışkan biyoreaktör, kayma gerilimi, meme kanseri, hücre canlılığı

PDF

___

1. Mitchell, M.J. and M.R. King, Computational and experimental models of cancer cell response to fluid shear stress. Frontiers in oncology, 2013. 3.
2. Ermis, M., et al., A high throughput approach for analysis of cell nuclear deformability at single cell level. Scientific reports, 2016. 6(1): p. 1-13.
3. Shieh, A.C., Biomechanical forces shape the tumor microenvironment. Annals of biomedical engineering, 2011. 39(5): p. 1379-1389.
4. Dixon, J.B., et al., Lymph flow, shear stress, and lymphocyte velocity in rat mesenteric prenodal lymphatics. Microcirculation, 2006. 13(7): p. 597-610.
5. Schmid-Schonbein, G.W., Microlymphatics and lymph flow. Physiological reviews, 1990. 70(4): p. 987-1028. 6. Dafni, H., et al., Overexpression of vascular endothelial growth factor 165 drives peritumor interstitial convection and induces lymphatic drain: magnetic resonance imaging, confocal microscopy, and histological tracking of triple-labeled albumin. Cancer research, 2002. 62(22): p. 6731-6739.
7. Pedersen, J.A., F. Boschetti, and M.A. Swartz, Effects of extracellular fiber architecture on cell membrane shear stress in a 3D fibrous matrix. Journal of biomechanics, 2007. 40(7): p. 1484-1492.
8. Tarbell, J.M. and Z.-D. Shi, Effect of the glycocalyx layer on transmission of interstitial flow shear stress to embedded cells. Biomechanics and modeling in mechanobiology, 2013. 12(1): p. 111-121.
9. Kocal, G.C., et al., Dynamic Microenvironment Induces Phenotypic Plasticity of Esophageal Cancer Cells Under Flow. Scientific Reports, 2016. 6: p. 38221.
10. Ip, C.K., et al., Stemness and chemoresistance in epithelial ovarian carcinoma cells under shear stress. Scientific reports, 2016. 6: p. 26788.
11. Gold, K., A.K. Gaharwar, and A. Jain, Emerging trends in multiscale modeling of vascular pathophysiology: Organ-on-a-chip and 3D printing. Biomaterials, 2019. 196: p. 2-17.
12. Rothbauer, M., et al., Tomorrow today: organ-on-a-chip advances towards clinically relevant pharmaceutical and medical in vitro models. Current opinion in biotechnology, 2019. 55: p. 81-86.
13. Jain, R.K., J.D. Martin, and T. Stylianopoulos, The role of mechanical forces in tumor growth and therapy. Annual review of biomedical engineering, 2014. 16: p. 321-346.
14. Rizvi, I., et al., Flow induces epithelial-mesenchymal transition, cellular heterogeneity and biomarker modulation in 3D ovarian cancer nodules. Proceedings of the National Academy of Sciences, 2013. 110(22): p. E1974-E1983.
15. Malek, A.M., S.L. Alper, and S. Izumo, Hemodynamic shear stress and its role in atherosclerosis. Jama, 1999. 282(21): p. 2035-2042.
16. Rana, K., J.L. Liesveld, and M.R. King, Delivery of apoptotic signal to rolling cancer cells: A novel biomimetic technique using immobilized TRAIL and E‐selectin. Biotechnology and bioengineering, 2009. 102(6): p. 1692-1702.
17. Regmi, S., A. Fu, and K.Q. Luo, High shear stresses under exercise condition destroy circulating tumor cells in a microfluidic system. Scientific reports, 2017. 7: p. 39975.
18. Xin, Y., et al., Mechanics and Actomyosin-Dependent Survival/Chemoresistance of Suspended Tumor Cells in Shear Flow. Biophysical journal, 2019. 116(10): p. 1803-1814.
19. Fan, R., et al., Circulatory shear flow alters the viability and proliferation of circulating colon cancer cells. Scientific reports, 2016. 6: p. 27073.
20. Onstenk, W., et al., Gene expression profiles of circulating tumor cells versus primary tumors in metastatic breast cancer. Cancer letters, 2015. 362(1): p. 36-44.
21. Potdar, P.D. and N.K. Lotey, Role of circulating tumor cells in future diagnosis and therapy of cancer. J Cancer Metastasis Treat, 2015. 1: p. 44-56.
22. Cornelius, V.J., et al. Preparation of SMART wound dressings based on colloidal microgels and textile fibres. in Smart Materials, Nano-and Micro-Smart Systems. 2006. International Society for Optics and Photonics.
23. Selmi, M., H. Belmabrouk, and A. Bajahzar, Numerical Study of the Blood Flow in a Deformable Human Aorta. Applied Sciences, 2019. 9(6): p. 1216.
24. Calamak, S. and K. Ulubayram, Controlled synthesis of multi-branched gold nanodendrites by dynamic microfluidic flow system. Journal of materials science, 2019. 54(10): p. 7541-7552.
25. ÖZSUNAR, A., Ş. BAŞKAYA, and M. SİVRİOĞLU, Dikdörtgen Kesitli Bir Kanalda Laminer Karışık Konveksiyon Şartlarındaki Akışın Sayısal Olarak İncelenmesi. Gazi Üniversitesi Mühendislik-Mimarlık Fakültesi Dergisi, 2000. 15(1).
26. Gürkan, K. and Ş. BAŞKAYA, Havalandırılan bir ofis odasında hava hareketinin sayısal analizi. Gazi Üniversitesi Mühendislik Mimarlık Fakültesi Dergisi, 2002. 17(2): p. 35-52.
27. Akpek, A., Stereolitografi ile üç boyutlu (3B) olarak biyobaskılanmış yapay kalp kapakçıklarının biyouyumluluk özelliklerinin analizi. Gazi Üniversitesi Mühendislik-Mimarlık Fakültesi Dergisi, 2018. 33(3): p. 929-938.
28. Lyczkowski, R.W., et al., Application of multiphase computational fluid dynamics to analyze monocyte adhesion. Annals of biomedical engineering, 2009. 37(8): p. 1516-1533.
29. Karimi, A., S. Yazdi, and A. Ardekani, Hydrodynamic mechanisms of cell and particle trapping in microfluidics. Biomicrofluidics, 2013. 7(2): p. 021501.
30. D’Avino, G. and P.L. Maffettone, Particle dynamics in viscoelastic liquids. Journal of Non-Newtonian Fluid Mechanics, 2015. 215: p. 80-104.
31. Segre, G. and A. Silberberg, Radial particle displacements in Poiseuille flow of suspensions. Nature, 1961. 189(4760): p. 209-210.
32. Seo, K.W., et al., Particle migration and single-line particle focusing in microscale pipe flow of viscoelastic fluids. RSC advances, 2014. 4(7): p. 3512-3520.
33. Leshansky, A.M., et al., Tunable nonlinear viscoelastic “focusing” in a microfluidic device. Physical review letters, 2007. 98(23): p. 234501.
34. D'Avino, G., et al., Single line particle focusing induced by viscoelasticity of the suspending liquid: theory, experiments and simulations to design a micropipe flow-focuser. Lab on a Chip, 2012. 12(9): p. 1638-1645.
35. Shashni, B., et al., Size-based differentiation of cancer and normal cells by a particle size analyzer assisted by a cell-recognition PC software. Biological and Pharmaceutical Bulletin, 2018. 41(4): p. 487-503.
36. Fu, A., et al., High expression of MnSOD promotes survival of circulating breast cancer cells and increases their resistance to doxorubicin. Oncotarget, 2016. 7(31): p. 50239.
37. Mitchell, M.J. and M.R. King, Fluid shear stress sensitizes cancer cells to receptor-mediated apoptosis via trimeric death receptors. New journal of physics, 2013. 15(1): p. 015008.
38. Egan, K., N. Cooke, and D. Kenny, Living in shear: platelets protect cancer cells from shear induced damage. Clinical & experimental metastasis, 2014. 31(6): p. 697-704.
39. Lo, K.-Y., et al., Effects of shear stresses and antioxidant concentrations on the production of reactive oxygen species in lung cancer cells. Biomicrofluidics, 2013. 7(6): p. 064108.
40. Luo, C.-W., C.-C. Wu, and H.-J. Ch'ang, Radiation sensitization of tumor cells induced by shear stress: The roles of integrins and FAK. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research, 2014. 1843(9): p. 2129-2137.
41. Calamak, S., et al., A Circulating Bioreactor Reprograms Cancer Cells Toward a More Mesenchymal Niche. Advanced Biosystems, 2020. 4(2): p. 1900139.
42. Galanzha, E.I., J.W. Kim, and V.P. Zharov, Nanotechnology‐based molecular photoacoustic and photothermal flow cytometry platform for in‐vivo detection and killing of circulating cancer stem cells. Journal of biophotonics, 2009. 2(12): p. 725-735.
43. Galanzha, E.I. and V.P. Zharov, Circulating tumor cell detection and capture by photoacoustic flow cytometry in vivo and ex vivo. Cancers, 2013. 5(4): p. 1691-1738.
44. Mitchell, M.J. and M.R. King, Computational and experimental models of cancer cell response to fluid shear stress. Frontiers in oncology, 2013. 3: p. 44.
45. Hamza, B., et al., Optofluidic real-time cell sorter for longitudinal CTC studies in mouse models of cancer. Proceedings of the National Academy of Sciences, 2019. 116(6): p. 2232-2236.
46. Helzer, K.T., et al., Circulating tumor cells are transcriptionally similar to the primary tumor in a murine prostate model. Cancer research, 2009. 69(19): p. 7860-7866.