【作者】 王欣；

【导师】 陈国广； 韦萍；

【作者基本信息】 南京工业大学 ， 生物化工， 2005， 硕士

【摘要】 目前,纤维蛋白胶（Fibrin glue）已被广泛用作外科用粘附剂,但它的组成成分纤维蛋白原和凝血酶都是从人类血液中提取的,这就可能感染传染性疾病。通过微生物合成的γ-聚谷氨酸（Gamma-Polyglutamic acid, γ-PGA）具有的优良的生物相容性和生物降解性,使得它可作为一种理想的生物医用材料。本文研究水溶性碳化二亚胺（WSC）作为交联剂,γ-PGA 和明胶交联制备生物胶的情况。论文的主要工作及结论有: 1.γ-PGA-明胶生物胶的制备研究用WSC 交联的γ-PGA-明胶生物胶的制备和组织粘附性。明胶与不同分子量的γ-PGA 交联制备的生物胶,在25℃明胶浓度低于自发胶凝浓度的情况下,胶凝时间随γ-PGA 和明胶的浓度及WSC 的浓度增加而增大,低分子量γ-PGA 与明胶的胶凝时间短于高分子量γ-PGA,在γ-PGA 浓度20 mg.mL^-1、明胶10 mg.mL^-1、交联剂WSC 120 mg.mL^-1时,其胶凝时间小于30s,和纤维蛋白胶相当。WSC 交联的聚谷氨酸-明胶生物胶的粘连强度优于蛋白纤维胶。通过红外光谱、X 衍射分析和DSC 分析对γ-PGA-明胶生物胶的性能进行了表征。2.添加剂对γ-PGA-明胶生物胶的影响在考察的所有添加剂中,尿素是理想的添加剂。当它的浓度大于75 mg.mL^-1时,就具有良好的防止明胶自发胶凝作用。适量的尿素添加后,没有妨碍γ-PGA和明胶之间的交联和生物胶对组织的粘附能力。当明胶是50 mg.mL^-1,PGA 是20 mg.mL^-1,尿素是100 mg.mL^-1,WSC 是80 mg.mL^-1 时,制得的生物胶具有与纤维蛋白胶相当的胶凝时间。添加尿素的生物胶粘附能力仍然优于纤维蛋白胶。3.γ-PGA-明胶生物胶的生物相容性A 胶(PGA 质量浓度20 mg.mL^-1、明胶质量浓度10 mg.mL^-1、交联剂WSC 质量浓度80 mg.mL^-1制备的生物胶)和B 胶(PGA 质量浓度20 mg.mL^-1,明胶质量浓度50 mg.mL^-1,尿素质量浓度100 mg.mL^-1,WSC 质量浓度80 mg.mL^-1制备的生物胶)的细胞毒性为0 级或1 级,急性毒性试验均为阴性,两种生物胶更多还原

【Abstract】 At present, although fibrin glue（FG） has been widely used as a surgical adhesive, its components, fibrinogen and thrombin, obtained from human blood are not completely free from the risk of virus infection due to acquired immune deficiency and hepatitis. Gamma-polyglutamic acid （γ-PGA） synthesized by microorganism is of good biocompatibility and biodegradability,so it can be used as a kind of ideal biomedical material. In the paper, the preparation and biological performance of γ-PGA -gelatin biological glue crosslinked with the aid of water-soluble carbodiimide（WSC） is studied as fo11owing: 1. The preparation of γ-PGA -gelatin biological glue The preparation and the tissue adhesion property of the γ-PGA -gelatin biological glue is studied. The biological glue is prepared from the gelatin whose aqueous solution doesn’t spontaneously set to a gel at 25℃and the different-molecular-weight γ-PGA by the cross-linking reaction of WSC. When gelatin concentration is lower than the incipient gelation concentration, the gelation time by the WSC-cross-linked γ-PGA -gelatin biological glue lengthen with an increase of the concentration of γ-PGA and gelatin and WSC, and the gelation time by the WSC-cross-linked low-molecular-weight γ-PGA -gelatin biological glue is shorter than the gelation time by the WSC-cross-linked high-molecular-weight γ-PGA-gelatin biological glue .when the concentration of γ-PGA is 40 mg.mL^-1、the concentration of gelatin is 10 mg.mL^-1 and the concentration of WSC is 120 mg.mL^-1, the gelation time by the WSC-cross-linked γ-PGA -gelatin biological glue is close to the gelation time by the fibrin glue. The adhesion property of γ-PGA -gelatin biological glue is superior to the adhesion property of fibrin glue. The biological glue is characterized by IR, XRD and DSC. 2. Effect of additives on γ-PGA -gelatin biological glue Urea is a promising additive in all investigated additives. When Urea’s concentration is more than 75 mg.mL-1, it can prevent spontaneous physical gelation of gelatin well. The adding of Urea doesn’t change the characteristics of WSC-induced crosslinking and tissue adhesion of the formed biological glue. When the concentration of γ-PGA is 40 mg.mL-1、the concentration of gelatin is 10 mg.mL-1and the concentration of WSC is 80 mg.mL-1, the gelation time by the WSC-cross-linked γ-PGA -gelatin biological glue is close to the gelation time by the FG. The adhesion property of γ-PGA -gelatin is still superior to the adhesion property of FG. 3.The biocompatibility of γ-PGA -gelatin biological glue The growth and proliferation of the cultured cells is not significantly inhibited by the biological glue A and the biological glue B, whose results are grade zero or grade one in cell toxicity test. The biological glue A and the biological glue B have no hemolytic reaction and their hemolysis rates are no more than 5%. They have no acute systematic toxicity, no skin irritation reaction, no pyrogenic reaction and no derma sensitization reaction too. All results indicate that the γ-PGA -gelatin biological glue has good biocompatibility. 4.The hemostatic property of γ-PGA -gelatin biological glue Ear artery wound and liver resection wound models of the rabbit are made. Compared with the natural hemostasis, the hemostatical time of the FG group and the γ-PGA -gelatin biological glue group is shorter in ear artery wound model and liver resection wound model. So the γ-PGA -gelatin biological glue provides effective hemostasis and can be used as a potential clinical glue.更多还原