Development of personalized cell-populated vascular graft in vitro

Aim. To create a personalized cell-populated small-diameter vascular prosthesis in a pulsating bioreactor.

Methods. Tubular grafts were made by electrospinning from mixtures of biodegradable polymers, poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and poly(εcaprolactone) (PCL). The inner surface is modified with fibrin. Tubular scaffolds were colonized with cultured colony-forming endothelial cells and grown under static conditions for 2 days. Then, the cell-populated prostheses continued to be cultivated for 5 days in a pulsating bioreactor system with a final shear stress of 2.85 dynes/cm².

Results. The advantages of the cultivation of cell-populated vascular prostheses in a pulsating bioreactor have been revealed. The selected mode of cultivation of cellpopulated vascular prostheses under conditions of a pulsating flow with a shear stress of 2.85 dynes/cm² did not have a damaging effect on the integrity of the endothelial monolayer. Moving unidirectional mechanical stimuli of chaotic orientation fibers of F-actin changed to a predominant orientation in the direction of flow, and also increased the expression of F-actin, Talin focal adhesion protein, and specific endothelial markers CD309, CD31, vWF.

Conclusion. The creation of a personalized cell-populated small-diameter vascular prosthesis with a functional endothelial monolayer is possible due to the use of autologous endothelial cells, autologous fibrin, and cultivation under conditions of a pulsating flow.

1. Yazdani S.K., Tillman B.W., Berry J.L., Soker S., Geary R.L. The fate of an endothelium layer after preconditioning. J Vasc Surg. 2010; 51 (1): 174–83. doi: 10.1016/j.jvs.2009.08.074.

2. Aper T., Kolster M., Hilfiker A., Teebken O.E., Haverich. A Fibrinogen Preparations for Tissue Engineering Approaches. J Bioengineer & Biomedical Sci. 2012; 2 (3): 115. doi:10.4172/2155-9538.1000115.

3. Matveeva VG, Khanova MYu, Velikanova EA, Antonova LV, Sardin ES, Krutitsky SS, Barbarash OL. Isolation and characteristics of colony-forming endothelial cells from peripheral blood in patients with ischemic heart disease. Cell and Tissue Biology. 2018; 60 (8): 598–608. (In Russian). doi: 10.31116/tsitol.2018.08.03).

4. Burridge K., Wittchen E.S. The tension mounts: Stress fibers as force-generating mechanotransducers. J Cell Biol. 2013; 200 (1): 9–19. doi: 10.1083/jcb.201210090.

5. Gong X., Liu H., Ding X., Liu M., Li X., Zheng L., Jia X., Zhou G., Zou Y., Li J., Huang X., Fan Y. Physiological pulsatile flow culture conditions to generate functional endothelium on a sulfated silk fibroin nanofibrous scaffold. Biomaterials. 2014; 35 (17): 4782—4791. doi: 10.1016 / j.biomaterials.2014.02.050.

6. Shu Chien. Mechanotransduction and endothelial cell homeostasis: the wisdom of the cell. Am J Physiol Heart Circ Physiol. 2007; 292 (3): H1209-24. doi: 10.1152/ajpheart.01047.2006.

7. Ando J., Yamamoto K. Effects of shear stress and stretch on endothelial function. Antioxid Redox Signal. 2011; 15 (5): 1389–403. doi: 10.1089/ars.2010.3361.

8. Leiss M., Beckmann K., Girós A., Costell M., Fässler R. The role of integrin binding sites in fibronectin matrix assembly in vivo. Elsevier. 2008; 20 (5): 502–507. doi: 10.1016/j.ceb.2008.06.001.