Biomaterials Research (생체재료학회지)

Korean Society for Biomaterials (한국생체재료학회)
권호 표지

Stimulated Migration and Penetration of Vascular Endothelial Cells Into Poly (L-lactic acid) Scaffolds Under Flow Conditions

  • Koo, Min-Ah (Cellbiocontrol Laboratory, Department of Medical Engineering) ;
  • Kang, Jae Kyeong (Cellbiocontrol Laboratory, Department of Medical Engineering) ;
  • Lee, Mi Hee (Cellbiocontrol Laboratory, Department of Medical Engineering) ;
  • Seo, Hyok Jin (Cellbiocontrol Laboratory, Department of Medical Engineering) ;
  • Kwon, Byeong-Ju (Cellbiocontrol Laboratory, Department of Medical Engineering) ;
  • You, Kyung Eun (Cellbiocontrol Laboratory, Department of Medical Engineering) ;
  • Kim, Min Sung (Cellbiocontrol Laboratory, Department of Medical Engineering) ;
  • Kim, Dohyun (Cellbiocontrol Laboratory, Department of Medical Engineering) ;
  • Park, Jong-Chul (Cellbiocontrol Laboratory, Department of Medical Engineering)
  • Received : 2013.09.04
  • Accepted : 2013.12.05
  • Published : 2014.06.01
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Abstract

Background: The initial procedure of the development of engineered tissues is cell seeding into three-dimensional polymer scaffolds. However, it is hard to make the cells invade into scaffold due to the characteristic of pore and material. Electrospun poly (L-lactic acid) scaffold and flow perfusion system were used to overcome these seeding problems. Materials and Methods: Before starting the experiment, we set up the parallel plate chamber system to observe endothelial cell migration under flow condition. In individual cell migration model, human umbilical endothelial cells started to migrate in the direction of flow at $8dyne/cm^2$ and we observed the cytoskeleton alignment at $8dyne/cm^2$. Results: This study has demonstrated the possibility to evaluate and analyze cell migration using the parallel plate chamber system and we may predict in vivo cell migration under flow condition based on these results. Also the flow perfusion system was established for the effective cell seeding into at three dimensional scaffolds. Moreover, shear stress induced by flow can enhance cell migration into PLLA scaffold that is in the form of cotton. Conclusions: Result indicated that cell penetration was achieved under flow condition better and more than under static condition throughout the matrix.

Keywords

References

  1. Langer R, Vacanti JP: Tissue engineering. Science 1993, 260, 920-926 (1993). https://doi.org/10.1126/science.8493529
  2. Asti A, Visai L, Dorati R, Conti B, Saino E, Sbarra S, Gastaldi G, Benazzo F: Improved cell growth by Bio-Oss/PLA scaffolds for use as a bone substitute. Technol Health Care, 16, 401-413 (2008).
  3. Wendt D, Stroebel S, Jakob M, John GT, Martin I: Uniform tissues engineered by seeding and culturing cells in 3D scaffolds under perfusion at defined oxygen tensions. Biorheology, 43, 481-488 (2006).
  4. Shimizu K, Ito A, Honda H: Enhanced cell-seeding into 3D porous scaffolds by use of magnetite nanoparticles. J Biomed Mater Res B Appl Biomater, 77, 265-272 (2006).
  5. Vunjak-Novakovic G, Obradovic B, Martin I, Bursac PM, Langer R, Freed LE: Dynamic cell seeding of polymer scaffolds for cartilage tissue engineering. Biotechnol Prog, 14, 193-202 (1998). https://doi.org/10.1021/bp970120j
  6. Sprague EA, Luo J, Palmaz JC: Human aortic endothelial cell migration onto stent surfaces under static and flow conditions. J Vasc Interv Radiol, 8(1Pt1), 83-92 (1997). https://doi.org/10.1016/S1051-0443(97)70521-9
  7. Villalona GA, Udelsman B, Duncan DR, McGillicuddy E, Sawh-Martinez RF, Hibino N, Painter C, Mirensky T, Erickson B, Shinoka T, Breuer CK: Cell-seeding techniques in vascular tissue engineering. Tissue Eng Part B Rev, 16, 341-350 (2010). https://doi.org/10.1089/ten.teb.2009.0527
  8. Sailon AM, Allori AC, Davidson EH, Reformat DD, Allen RJ, Warren SM: A novel flow-perfusion bioreactor supports 3D dynamic cell culture. J Biomed Biotechnol, 2009, 873816 (2009).
  9. Li S, Huang NF, Hsu S: Mechanotransduction in endothelial cell migration. J Cell Biochem, 96, 1110-1126 (2005). https://doi.org/10.1002/jcb.20614
  10. Chlen S, Li S, Shyy J. Y-J: Effects of mechanical forces on signal transduction and gene expression in endothelial cells. Hypertension, 31, 162-169 (1998). https://doi.org/10.1161/01.HYP.31.1.162
  11. Davies PF: Flow-mediated endothelial mechanotransduction. Physiol Rev, 75, 519-560 (1995).
  12. Page E, Cura M, Sprague E: Bare-stent technology and its utilization in the treatment of atherosclerotic obstructive disease. Vascular disease management, 7, 95-102 (2010).
  13. Hsu S, Thakar R, Liepmann D, Li S: Effects of shear stress on endothelial cell haptotaxis on micropatterned surfaces. Biochem Biophys Res Commun, 337, 401-409 (2005). https://doi.org/10.1016/j.bbrc.2005.08.272
  14. Li S, Butler P, Wang Y, Hu Y, Han DC, Usami S, Guan JL, Chien S: The role of the dynamics of focal adhesion kinase in the mechanotaxis of endothelial cells. Proc Natl Acad Sci USA, 99, 3546-3551 (2002). https://doi.org/10.1073/pnas.052018099
  15. Hu YL, Li S, Miao H, Tsou TC, Del Pozo MA, Chien S: Roles of microtubule dynamics and small GTPase Rac in endothelial cell migration and lamellipodium formation under flow. J Vasc Res, 39, 465-476 (2002). https://doi.org/10.1159/000067202
  16. Li S, Chen BP, Azuma N, Hu YL, Wu SZ, Sumpio BE, Shyy JY, Chien S: Distinct roles for the small GTPases Cdc42 and Rho in endothelial responses to shear stress. J Clin Invest, 103, 1141-1150 (1999). https://doi.org/10.1172/JCI5367
  17. Ridley AJ: Rho GTPases and cell migration. J Cell Sci, 114, 2713-2722 (2001).
  18. Wojciak-Stothard B, Ridley AJ: Shear stress-induced endothelial cell polarization is mediated by Rho and Rac but not Cdc42 or PI 3-kinases. J Cell Biol, 161(2), 429-439 (2003). https://doi.org/10.1083/jcb.200210135
  19. Malek A, Izumo S: Mechanism of endothelial cell shape change and cytoskeletal remodeling in response to fluid shear stress. J Cell Sci, 109, 713-726 (1996).
  20. Dull RO, Davies PF: Flow modulation of agonist (ATP)-response ($Ca^{2+}$) coupling in vascular endothelial cells. Am J Physiol, 261, 149-154 (1991).
  21. Holy CE, Shoichet MS, Davies JE: Engineering three-dimensional bone tissue in vitro using biodegradable scaffolds: Investigating initial cell-seeding density and culture period. J Biomed Mater Res, 51, 376-382 (2000). https://doi.org/10.1002/1097-4636(20000905)51:3<376::AID-JBM11>3.0.CO;2-G
  22. Li Y, Ma T, Kniss DA, Lasky LC, Yang ST: Effects of filtration seeding on cell density, spatial distribution, and proliferation in nonwoven fibrous matrices. Biotechnol Prog, 17, 935-944 (2001). https://doi.org/10.1021/bp0100878
  23. Vunjak-Novakovic G, Radisic M: Cell seeding of polymer scaffolds. Methods Mol Biol, 238, 131-146 (2004).
  24. Freed LE, Vunjak-Novakovic G, Biron RJ, Eagles DB, Lesnoy DC, Barlow SK, Langer R: Biodegradable polymer scaffolds for tissue engineering. Biotechnology, 12, 689-693 (1994). https://doi.org/10.1038/nbt0794-689
  25. Freed LE, Marquis JC, Nohria A, Emmanual J, Mikos AG, Langer R: Neocartilage formation in vitro and in vivo using cells cultured on synthetic biodegradable polymers. J Biomed Mater Res, 27, 11-23 (1993). https://doi.org/10.1002/jbm.820270104