Study On Effect Of Substrate Stiffness On The Primary Human Umbilical Vein Endothelial Cells Infected With Dengue Virus Serotype 2

Objective: To observe the effects of substrate stiffness on dengue virus infected with human umbilical vein endothelial cells(HUVEC). Methods: Prepared polyacrylamide gels for cell culture(0±4Kpa). The cell proliferation on different substrate stiffness was determined by using MTS assay. The cell cycle and cell apoptosis was determined by FCM. DENV-2 virus were propagated and identified by conventional methods.Then by comparing the cell proliferation seeded on the traditional plastic basement with hydrogel after dengue virus infection by MTS assay at an MOI of 10. The method of nitric acid reductase was used to measure the concentration of nitric oxide and expression of ET-1 by double antibody sandwich method ELISA. Results:Young’s modulus E value of the hydrogels was 3030±0.44 Pa.The cell proliferation rate, the level of NO and ET-1,but not cell cycle and cell apoptosis along with substrate stiffness increase.. Conclusion: Substrate stiffness not only influence on the proliferation of HUVEC,but also release of cytokines after infected with DENV-2. Dengue fever has been occured due to the shrinking of vascular active substances. The hydrogel could be a new models in vitro for research of dengue virus infection.