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Background American scholars proposed "tissue engineering" concept in the1980s, tissue engineering is an interdisciplinary subject, including life science, materials science and other disciplines of knowledge. Tissue engineering research includes: seed cells, scaffolds, extracellular matrix. Among them, the scaffold material is a hot research topic in recent years in this field.Chitosan/alginate composite scaffolds was produced by freeze drying.In this study,we adopt a new natural crosslinking agent genipin crosslinking of the materials, to explore the effect of preparation of scaffold by genipin, UV two crosslinking methods.Objective1. To investigate genipin(GP) and ultraviolet rays(UV) crosslinked effects of chitosan/alginate scaffolds for tissue engineering by detecting the change of the chitosan/alginate scaffolds material degradation rate, porosity, water content, cytotoxicity and biomechanics properties.2. we try to explore cross-linking temperature of genipin crosslinked effects of chitosan/alginate scaffolds for tissue engineering.Methods BMSCs were separated and cultured from one Wistar rat.Chitosan/alginate scaffolds was produced via freeze drying.1.①ccording to different cross-linking methods, biomaterials were divided into (GP)group,(UV)group.②he surface structure of two groups was detected by scanning electron microscope(SEM). The material degradation rate, porosity, water content, cytotoxicity, and biomechanics properties were detected.2.We make the chitosan/alginate scaffolds of tissue engineering by freeze-drying, respectively, in4℃,25℃,37℃conditions, at0.5%solution of genipin crosslinked24h, as the experimental group. The scaffolds of not cross-linked with genipin as a control group to evaluate the degradation rate of scaffolds, cytotoxicity, porosity, water content and biomechanics properties.Rssults1.①UV group and GP group showed more pore structure, and there was no difference.②Degradation rate of UV group was higher than that in GP group.③The porosity for GP group and UV group was no difference. The difference of water content is meaningful in Statistics.④GP group and UV group showed low cytotoxicity, good biocompatibility.⑤CP group of biomechanics properties were significantly inproved compared with UV group.2.①The color of scaffolds material from light purple to purplish black, according with the rise of the cross-linking temperature, however, not cross-linked scaffold material is white;⑤The degradation rate after4weeks at4℃group was21.54%±3.07%,25℃group was9.9%±1.2%,37℃group was8.98%±0.79%. Among them, the37℃group of degradation resistant capacity is better than the4℃group (P<0.05). The experimental groups were better than the control group (P<0.01).③Elastic modulus at4℃group was0.84±0.55,25"C group was1.44±0.06,37℃group was1.53±0.02.37℃group and25℃group of tensile properties better than control group (P<0.05).④Forosity,water content,cytotoxicity between the experimental groups had no significant difference.Conclusion1. Genipin crosslinked chitosan/alginate composite scaffold material has good biological characteristics. It is a very potential material in tissue engineering spinal cord field.2. With the cross-linking temperature increasing, scaffold degradation resistant capacity and tensile properties to strengthen, but on the scaffold material structure, cytotoxicity, porosity and water content were not altered significantly.
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