Stability And Cell Compatibility Of Nano-silver/Chitosan Composite Film

Nano-silver has been widely used because of its excellent antibacterial properties and potential selective inhibition of cell proliferation which caused by the interaction between the nano-silver and cells. It also raises concerns about the security of its human use. Preliminary studies have shown that nano-silver fixed in crystal lattice or distributed in solution can both affect the proliferation of cell, but the mechanism is different. To this end, this study intends to have further research in the law of interaction between the cells and nano-silver in the sustained-release state, and looking for the main cause of which affecting the cell proliferation and cell adhesion.In this study, the mechanical mixture of sol and the gel was used to make the composite film. Then study the physical and chemical properties of the nano-silver/chitosan composite film (such as surface energy, composition and distribution of silver nanoparticles and the surface morphology study) and the stability in the different liquid environments (such as SBF, pure water, F12K medium containing FBS, F12K medium). Observed human umbilical vein endothelial cells (HUVEC) and endothelial cells of human amnion cell line (FL) on the surface of composite film by light microscopy, and statistical analysised their adhesion; MTT and CCK-8method were used to evaluate the proliferation of HUVEC and FL on the composite films and their release fluid. The slow releasing mechanism of the nano-silver/chitosan composite films, as well as cell proliferation impact of the composite film were analyzed and discussed.The results showed that:the sol-gel prepared film surface was smooth and dense at micro part, but there were ups and downs on the macroscopic (microns), no cracks、holes or other surface defects. Nano-silver particles evenly dispersed in the composite film with a general way of a physical mixture. There is a linear relationship between film thickness and the mass of film per unit area. The surface eneger of the composite films increased a little when thickness upward.The stability study of the composite films in the four experimental liquid results showed that the films in the early stage of sustained-release (2D) had significant weight loss, and the higher silver content composite film had the higher weight loss, about20%total quality. From4days films entered in a stable period.The overall skeletons of films were still stable after14D soaked in liquid, and there were no holes or cracks on the film surface. The silver releasing was slow, and the releasing rate of silver was only about0.1%after soaking30days, the rest of the nano-silver was retained in the erosion peeling off the chitosan debris.The cytology experimental results showed that the nano-silver/chitosan composite film could affect the adhesion and proliferation of HUVEC and FL cells, and the film thickness and the silver volume were both important factor. When the silver content was the same, the greater the thickness the greater the impact on cell adhesion and proliferation of performance of films; when the film thickness was constant, the higher the silver content the greater the performance impact on cell adhesion and proliferation of film. Two-way ANOVA results showed that the surface properties of the composite films was the main factor about cell proliferation, but the releasing solution of nano-silver, and silver containing in chitosan erosion debris had a certain impact on cell growth. The two cells proliferation responsing on the composite films surface were different, and the FL cells were far more sensitive than HUVEC cells.