Dynamic culture conditions to generate endothelium on polycaprolactone-based tissue engineered vascular grafts
Sonia Abbasi ravasjani,
1 Ghassem amoabediny,
2,* Mohammad reza safari hajat aghaei,
3 Amir esmaeili,
4 Farideh hajiani,
5 Javad mohammadnejad,
6
1. Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran
2. School of chemical Engineering, College of Engineering, University of Tehran, Tehran, Iran
3. Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran
4. School of chemical Engineering, College of Engineering, University of Tehran, Tehran, Iran
5. Faculty of Applied Arts, Textile and Clothing Design, University of Art, Tehran, iran
6. Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran
Abstract
Introduction
The main reason for the long term failure of small-diameter vascular grafts is thrombus formation. the seeding of vascular endothelial cells onto the lumen of vascular grafts in order to yield a thrombo resistant surface is one approach that has been proposed to address this problem. the aim of this study was to find out the best in vitro culture conditions to generate endothelium onto the lumen of electrospun tubular polycaprolactone (pcl) scaffolds.
Methods
Tubular pcl scaffolds were made by electrospinning method. carboxylic groups were created on its surface by hydrolysis with sodium hydroxide solution (3m, 30min). collagen was immobilized on pcl tubs with carboxylic functional groups. human umbilical vein endothelial cells (huvecs) were seeded onto the lumen of pcl tubs and cultured under static and dynamic conditions .alamar blue assay was performed to evaluate the metabolic activity of the cells. the morphology of endothelial cells attached to pcl surface was visualized after 7 days of culture by scanning electron microscopy (sem).
Results
Metabolic activity of huvecs cultured under static condition increased 2 times during 7 days of culture. in dynamic condition, metabolic activity of cells increased 4.5 times from third day up to seventh day of culture. sem micrographs showed spindle shaped morphology in dynamic culture condition compared to static culture controls.
Conclusion
The results demonstrated that tissue engineered vascular grafts under dynamic flow conditions had better outcome than static culture controls in terms of cell proliferation and alignment.
Keywords
Electrosun pcl, dynamic culture, static culture, metabolic activity
