Influence Of Stress Relaxation Of Cell Culture Substrate On Different Stages Of Differentiation Myofibroblast

Myofibroblasts are heterogeneous cells of diverse origins, which playing essential in wound-healing responses. They are involved in fibrosis course in most tissues in vivo, including: liver fibrosis, pulmonary fibrosis, kidney fibrosis, atherosclerosis, connective tissues and scar formation.Static relaxation was exerted on Myofibroblast from processed New Zealand rabbits (1.5 kg ) using specific approach that Foreign protein have been implanted into rabbits'cavitas abdominalis for a period of time . Simulating the boundary condition and mechanical conditions of cell culture membrane reasonably, a three-dimensional finite element model of the cell culture membrane under tension state was developed by us. According to the analysis of finite element computing results, we have understood some changes of Equivalent (von-Mises) Elastic Strain , Equivalent stress and Total Deformation in the membrane that is made of two kinds of silicon rubber membrane (complete and incomplete) under different stress(such as:3%,6%).The effect of different mechanical courses on the cell morphology, orientation,cytoskeleton rearrangements,and the changes in growth and physiological behavior during was studied by using biomechanics,cell Biology, Immunohistochemistry and computer image processing methods.The abbreviate details are as follow:①Heterologous protein was put into rabbits'cavitas abbsominals after a 3～15 days immunoreactions, Myofibroblast that coated by mesothelial cell can be obtained. Then Myofibroblast can be harvested by digestion and centrifugation.②Myofibroblast of different stages is obtained by controlling the days of heterologous protein implantation: pro-Myofibroblast (less than 3 days), Myofibroblast (5～7 days), high expressionα-SM-actin Myofibroblast (10～15).③Differentiation of Myofibroblast is steping. The cell is regarded as pro-Myofibroblast because noα-SM-actin is detected before 3 days, as Myofibroblast withα-SM-actin appearing during 5～7 days,and then amount ofα-SM-actin appearing and filamentous arrangement during 10～15 days.④Static relaxation of substrate (6%-3%-0 and 6%-0) was exerted on the 5～7-day cell, which can promote the differentiation.⑤Stress plays essential role in the differentiation of Myofibroblast. Because of the prestress of substrate (6% strained before cell planted on it) has not remarkably effects on the differentiation of Myofibroblast .⑥According to the analysis of finite element computing results, when the deformation of substrate sags from 6%-0,the changes of Equivalent stress range from 0.30kPa to 0.34kPa and the changes of Equivalent (von-Mises) Elastic Strain range from 0.030m/m to 0.034m/m; when the deformation of substrate sags from 3%-0,the changes of Equivalent stress range from 0.15kPa to 0.17kPa and the changes of Equivalent (von-Mises) Elastic Strain range from 0.06m/m to 0.068m/m。⑦According to the analysis of finite element computing results, a conclusion that unity degree of stress of Silicone membrane is positively correlated to its completeness while negatively correlated to the size of the deformation is drawn by comparing the complete and un-complete substrate (6%-3%-0 and 6%-0) .