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2022, Vol. 26 ›› Issue (16): 2516-2521Effects of co-stimulation of material surface chemistry and fluid shear stress on human umbilical vein endothelial cellsQin Zhongjie1, Chen Siqi2, Wu Yimin1, Yang Jiaojiao1, Xia Delin1, 2
Abstract: BACKGROUND: There is a lack of perfect vascular system in tissue-engineered bone, among which vascularization is the key to limit its wide application. Appropriate matrix chemistry and fluid shear stress stimulation can promote the proliferation, differentiation and function of vascular endothelial cells, and guide the design and fabrication of scaffold materials. OBJECTIVE: To explore the effects of different chemical functional groups and fluid shear stress (FSS) on human umbilical vein endothelial cells, in order to find an efficient combination of material chemistry and fluid shear stress on human umbilical vein endothelial cells. METHODS: Self assembled monolayers with OH, CH3 and NH2 as end functional groups were prepared as matrix materials and blank glass as control. Human umbilical vein endothelial cells were seeded on the surface of four groups of glasses. The release of ATP within 15 minutes after culture and NO within 1 hour after culture was detected. The expression of endothelial nitric oxide synthase protein was detected by western blotting. The formation of focal adhesion and F-actin was observed by laser confocal microscopy. Human umbilical vein endothelial cells were seeded on the surface of four groups of glasses. When the cells fused to 80%, they were loaded with 1.5 N/m2 FSS for 1 hour. ATP release was detected within 15 minutes after loading, and NO release was detected within 1 hour after loading. The expression of endothelial nitric oxide synthase was detected by western blotting after loading for 1 hour. RESULTS AND CONCLUSION: (1) Chemical stimulation had no effect on the release of ATP or NO, but FSS could increase the release of ATP and NO, and the expression of endothelial nitric oxide synthase. When FSS and material chemistry acted simultaneously, the expression of ATP and NO was closely related to material chemistry. The release of ATP and NO and the expression of endothelial nitric oxide synthase were highest in NH2-FSS group, followed by FSS group, and those in the CH3-FSS and OH-FSS groups were lowest. (2) Laser confocal microscope showed that a large number of focal adhesions and cytoskeleton protein F-actin could be detected in the NH2 group, followed by control group; those were least in the CH3 and OH groups. (3) The results showed that the combination of NH2 functional group and FSS produced high-efficiency response to shear stress, and the mechanism may be the formation of optimal focal adhesion and F-actin on NH2 surface. Key words: material, scaffold, self-assembled monolayers, surface chemistry, fluid shear stress, human umbilical vein endothelial cells, protein |