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The Study on the Genipin Crosslinked Bombyx Mori Silk Fibroin Materials

Author: WangYiYu
Tutor: LiMingZhong
School: Suzhou University
Course: Textile Material and Textiles Design
Keywords: Silkworm Fibroin Genipin Crosslinked Porous materials Structure Cytocompatibility
CLC: R318.08
Type: Master's thesis
Year: 2010
Downloads: 144
Quote: 2
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Abstract

Silk fibroin has unique physical and chemical properties and good biocompatibility, has been extensively studied using biological materials as tissue repair. But without crosslinked fibroin material soluble in water, which limits the silk fibroin in biomedical applications. At home and abroad, many reported to improve the water resistance of the silk fibroin material, but at the same time improve the material properties of the silk fibroin often its biocompatibility produce negative effects. Genipin of geniposide by β-glucosidase hydrolysis after the product has good biocompatibility. In this paper, genipin as a crosslinking agent preparation of regenerated silk fibroin material, while the silk fibroin structure and physical and chemical properties of the material after the cross-linking system, and explore the crosslinked fibroin film prepared by different methods and L929 cells compatibility. Cast Preparation of genipin crosslinked fibroin film. The failure rate of the Hot Water Test results show that a significant improvement in water resistance of the crosslinked film, while amino acid analysis, infrared spectroscopy results showed that after crosslinking fibroin membrane lysine content decreased, and the emergence of a new covalent bond described genipin can the silk fibroin react to form a new covalent bond to effectively cross-linked silk fibroin. Changes in the fibroin membrane structure before and after the cross-linking, studied by means of infrared spectroscopy, X-ray diffraction, thermal analysis results show that the amorphous structure of the silk fibroin membrane crosslinked reduce aggregation structure tends to be regularized, crystallinity increases, the thermal stability is improved. The degree of crosslinking of the film increased with the increase of the added amount of genipin. Fibroin film after genipin crosslinked, its mechanical properties did not change significantly. Also studied the reaction temperature and the addition of glycerol of crosslinked membrane structure and performance, the test results show that the reaction temperature and the addition of glycerol has little effect on the the crosslinked film's Hot Water loss rate; With the rise of the reaction temperature and glycerol increase in the cross-linked film structure tends to be structured, and improved thermal stability; elevated temperature is possible to improve the degree of crosslinking of the crosslinked film, while no significant effect on the mechanical properties of the crosslinked film; addition of glycerol can significantly increase the cross the elongation at break of the associated membrane, reducing its Young's modulus, and to reduce the brittleness of the crosslinked film. Genipin as a crosslinking agent, freeze-dried prepared porous material. The study showed that after genipin crosslinked fibroin porous material of Hot Water loss rate is significantly lower. IR test results show that the porous material after the crosslinking of a new covalent bond; after crosslinking the porous silk fibroin material is still amorphous structure mainly, but the regular structure of the porous material is increased slightly. The thermal analysis showed a slight increase of the thermal stability of the porous material after the crosslinking. The pore size of the porous material with increasing the concentration of the silk fibroin solution and the freezing temperature reduction decreases, the hole density is increased. By in vitro cell culture L929 cells genipin crosslinked fibroin membrane adhesion and growth of the state, and four fibroin film (ethanol processing fibroin film, 30%, 60% epoxy resin crosslinking fibroin film, 1% glutaraldehyde cross-linked silk fibroin membrane) were compared. Inverted fluorescence microscope to observe the state of cell growth, cell adhesion rate was measured using the MTT assay and Coomassie blue staining method for the determination of cell viability, apoptosis and cell cycle measured by Flow cytometry. The results show that these types of silk fibroin membrane can support the adhesion and proliferation of cells, 20% genipin crosslinked fibroin membrane cell vitality and the ethanol processing fibroin membrane cell viability compared without significant difference, in other cross-linking group. Silk fibroin film 20% genipin crosslinking rate of apoptosis and cell cycle results ethanol processing fibroin film showed no significant difference is superior to other cross-linked group, no significant cytotoxicity.

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