| Liver is one of the important organs in humam body. However, there are a lot of people died of liver cancer and acute liver failure every year. Therefore, liver transplantation is the best treatment method. Looking for a suitable scaffold is a basic research in the culture of liver organs from human liver cells. As lignin-carbohydrate complexes(LCCs) have good physical strength, biocompatibility, non-toxic and biodegradblity, they are proming biomedical polymer material. In this thesis, the biocompatibility of LCCs in tissue engineering is investigated.LCCs isolated from ginkgo、aspen and wheat straw were crosslinked with glycerol glycidyl ether to obtained gel materials suitable for biological porous carrier. The content of the main component and surface morphology of the biological carrier was also analyzed.The results show that the content of galactose and mannose of LCC gel materials from ginkgo, aspen and wheat straw, which has high biocompatibility with the human liver cells were 2.30% and 21.94%, 3.32% and 3.12%, 0.27% and 10.13% respectively. The three kind of LCCs as raw materials can be processed to be high performance biological porous carrier, The average pore size is about 80-120μm. Biological porous carriers were applied in the culture of human hepatocytes, It is found that the human hepatocytes can successfully grow on LCC gel materials.In the first two days cells grow slowly.Whereas, in the next two days cells grow faster, and reach the peak. Then the cells began to decline, and age and fall off. The experiment with gel materials from LCCs of ginkgo, aspen and wheat straw was compared with that of control experiment. The cells number was higher than control experiment.Cells growth trend is as follows: aspen > ginkgo > wheat straw > control, The hepatocytes proliferation rate and the ALB, BUN, GLUCOSE content show that the hepatocytes with high metabolic activity. The results show that the LCC based gel materials have good biocompatibility.Bleached kraft pulp was oxidized by the TEMPO system and then react with coniferyl alcohol glucoside under the enzyme condition, in order to generate dehydrogenation polymer(DHP). Lignin and polysaccharides is combined by acetal linkage to generate the LCC. Synthesis of LCC material containing galactose and mannose with high biocompatibility to human hepatocytes. The contents were 3.53% and 3.53% respectively. The TEMPO oxided pulp materials and synthesized LCC materials were applied in culture of human hepatocytes in vitro. The hepatocytes proliferation fast in the fouth day with most cells number. Cells growth trend is as follows: synthesis LCC materials > TEMPO oxidation pulp materials > control. Results show that high content of galactose and mannose is more advantageous to cell proliferation. The result of analysis of the metabolism solution also show that the synthetic LCC materials can improve hepatocytes metabolic activity. This also proves that the synthesis LCC has good biological compatibility. There results may become the basis of preparation of LCC and application in tissue engineering.Last, the water-soluble fraction was isolated and applied in the culture of human hepatocytes with different concentration gradient.The water-soluble fraction of the gingko LCCs was characterized by Fourier transform infrared spectrometer(FT-IR)and High Performance Liquid Chromatograph(HPLC). Then gingko water-soluble LCCs of different concentration gradient(0.1%, 0.5%, 1.0%) were used in culture of human hepatocytes L-02 with contious monitor of cell growth. The proliferation and morphology of hepatocytes were also observed by cell counting and inverted microscope. Moreover, metabolic activity of hepatocytes, was detected. The results show the experimental group(LCC concentration: 0.1%-0.5%) of cultured cells grow well in the first three days. However, in the experimental group(concentration: 1.0%) gingko water-soluble LCC, the higher concentration of galactose inhibits the growth of cells. The BUN and GLUCOSE content in third day there was relatively high,... |
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