Optimizing blood vessel decellularization process to create biological scaffold for vascular tissue engineering
Optimizing blood vessel decellularization process to create biological scaffold for vascular tissue engineering
Hamed Omid,1Mohammad Ali Shokrgozar,2Javad Mohammadi,3,*Shahin Bonakdar,4Nooshin Haghighipour,5
1. Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran. 2. Department of National Cell Bank of Iran, Pasteur Institute of Iran. 3. Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran. 4. Department of National Cell Bank of Iran, Pasteur Institute of Iran. 5. Department of National Cell Bank of Iran, Pasteur Institute of Iran.
Introduction: For decades researchers have employed different types of biomaterials for tissue engineering. However, finding a biomaterial with the exact properties of natural tissues remains a formidable challenge. In tissues like blood vessel, these challenges become more apparent due to low thickness and delicate structure. Any minor defect in this structure causes serious dump in mechanical properties and pathological consequences like aneurysm. Moreover, direct contact with blood has forced researchers to prepare perfect hemocompatible surfaces for this tissue. Besides these problems, geometrically we need a cylindric shape for vascular tissue engineering which makes it worse. Controlling cell growth inside the cylinder is not easy, and overgrowth of cells like smooth muscle cell (SMC), and fibroblast causes intimal hyperplasia followed by hypertrophy, fibromuscular dysplasia and etc.
In this research, we decellularized the ovine blood vessel to reach a biological scaffold (BS). This BS has the most similar properties to the natural blood vessel. Evaluating the BS’s mechanical strength, viability and cellular adhesion revealed a promising future for blood vessel tissue engineering.
Methods: After cleansing the vessels tissue, they were immersed in 70% ethanol for 5 min to sterilize. Then osmotic pressures were exerted on the tissue for physical disruption of the cells without hurting the extracellular matrix (ECM). Finally, 0.025% trypsin and 1% Triton X-100 were employed to remove the cell debris. All the steps have been done at 4˚C. After washing the obtained BS, smooth muscle cell was grown on it for up to 4-day.
Results: The BS showed no cytotoxicity, and SEM images revealed appropriate cell attachment. Histological studies proved cell migration to the BS’s pores has done after 4-day culture. In addition, the optimized decellularized process didn’t have any harmful effects on the mechanical properties of the BS.
Conclusion: Reducing the concentration of the reagents and the temperature of the reactions gentled the kinetics of the decellularization process. It enhances the process control and the quality of the ECM.