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Study on the Fabrication of Collagen Gradient via Microcontact Printing and Its Effects on the Adhesion and Migration of Endothelial Cells

Author: ChenChong
Tutor: CaiKaiYong
School: Chongqing University
Course: Cell Biology
Keywords: Microcontact printing Collagen Endothelial cells Adhere to Migrate
CLC: Q819
Type: Master's thesis
Year: 2009
Downloads: 72
Quote: 0
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Abstract


Different concentrations of protein transfer to a substrate surface by a single step , we try to improve the traditional micro-contact printing technology . In this paper, the use of micro- contact printing techniques printing four concentrations of collagen ( 50,100,200,300 μg / ml ) , on the same template and the template material was prepared in the same surface of a substrate containing various protein concentrations . Scanning electron microscopy analysis showed : PDMS stamp surface smoothness , graphics complete . Laser scanning confocal microscope to observe the different concentrations of FITC - collagen (50,100,200,300 μg / ml) in the substrate surface adsorption . Using fluorescent analysis software correlation analysis of the protein domain fluorescence intensity and the protein concentration of the substrate surface . Albumin is used to the closed protein not adhesion domain , after sealing the template material surface inoculated with human umbilical vein endothelial cells , and explore the different concentrations of collagen build biological material impact on endothelial cell adhesion and migration . MTT assay was investigated without the concentration of collagen of human umbilical vein endothelial cells 6,24,48,72 h proliferative capacity . Laser scanning confocal microscope detection initial adhesion Phase cytoskeleton spreading situation ; single cell trajectory endothelial cell cultures 24h after the migration rate and net displacement measuring method . Experimental results show that : the template material surface protein domains complete protein domain uniform fluorescence intensity 50,100,200,300 microg / ml in 4 different concentrations of FITC- collagen adsorbed at the surface of the substrate the fluorescence intensity gradually increase , respectively : 38.51 ± 1.63 , 55.21 ± 3.88,73.17 ± 3.59,80.59 ± 1.12. Different concentrations of protein within the skin cytoskeleton spread than the control group spread out fully . Various concentrations of endothelial cell proliferation and net displacement were stronger than the blank group ( P lt; 0.05 ) with increasing concentrations (50μg/ml to 300μg/ml ) the ability of endothelial cell proliferation and net displacement enhanced (P lt ; 0.05 ). In addition to the migration rate of various concentrations of 300μg / m concentration group were less than the control group ( P lt; 0.05 ) . However, with the concentration of the various concentrations between groups increases, the rate of migration of endothelial cell is increased ( P lt ; 0.05 ) . In summary, the use of micro-contact printing techniques on the same surface of the substrate can get different protein concentration domain for endothelial cell adhesion and migration balance regulation , for the the cytochemistry tendency study provides useful information .

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