蚕蛹壳聚糖复合止血材料的制备及性能研究

发布时间：2018-06-04 14:50

本文选题：蚕蛹壳聚糖 + 止血　；参考：《西南大学》2012年硕士论文

【摘要】：在战伤急救及日常手术治疗中,过度失血是致死的主要原因。目前国内广泛使用的止血材料都有各自的不足之处,而国外进口的高效止血材料因其昂贵的价格限制了其在日常止血中的大范围应用。壳聚糖具有良好的抗菌、止血性能和生物相容性,是制备医用敷料的理想材料。而中国蚕业资源丰富,缫丝副产物蚕蛹中蕴含着大量的壳聚糖资源,且蚕蛹壳聚糖还具有某些优于虾蟹壳聚糖的性能,因此,本研究以蚕蛹中提取出的壳聚糖为主要原料,辅加以明胶作为复合剂,并以氯化钙作为促凝剂、戊二醛作为交联剂、甘油作为抗冻塑性剂,通过冷冻干燥法研制出一种新型的海绵状伤口止血材料。本文采用单因素试验及正交试验对蚕蛹壳聚糖复合止血材料的制备工艺进行了优化,并详细探讨了其体外凝血性能和动物活体止血性能,以及材料的促伤口愈合性能、抑菌性能和毒理学生物安全性能,研究结果如下： 1、从蚕蛹中提取甲壳素进而制备壳聚糖,且通过控制反应时间能够得到不同脱乙酰度的蚕蛹壳聚糖,最高脱乙酰度可达到93.13%,适合作为医药级产品原料。将从蚕蛹中提取出的壳聚糖粉末,与明胶共混溶解并添加适当助剂后,通过冷冻真空干燥法制备蚕蛹壳聚糖复合止血材料,制备出的样品呈海绵状,表面及内部孔隙丰富、分布均匀,且具有一定弹性材料触感。 2、通过单因素试验研究了蚕蛹壳聚糖复合止血材料的制备工艺对凝血指数的影响,实验表明,最佳制备工艺参数为：预冻温度-40℃、预冻时间1h。正交试验分析表明,以质量分数为2.5%的蚕蛹壳聚糖、0.1%的氯化钙、0.005%的戊二醛、3.0%的明胶、0.05%的甘油制备的蚕蛹壳聚糖复合止血材料的凝血效果最佳,该复合止血材料中各组分浓度对体外凝血指数的影响大小顺序是：蚕蛹壳聚糖氯化钙戊二醛明胶甘油。 3、壳聚糖是主导蚕蛹壳聚糖复合止血材料凝血性能的关键性因素,样品凝血性能随着蚕蛹壳聚糖浓度的升高而有效提高；与虾蟹壳商品壳聚糖相比,蚕蛹壳聚糖制备的样品凝血效果更好。优化配方下制备的蚕蛹壳聚糖复合止血.材料在体外及动物活体上均具有优良的止血效果：与医用纱布相比较,其体外凝血指数减小了62.71%,使家兔耳动脉出血模型中的出血量减少了92.08%,止血时间缩短了80.55%：与医用明胶海绵组比较,其体外凝血指数减小了35.61%,使家兔耳动脉出血模型中的出血量减少了88.35%,止血时间缩短了57.01%。 4、以家兔烫伤模型研究了蚕蛹壳聚糖复合止血材料对伤口面积和愈合率的影响。结果表明,敷加了蚕蛹壳聚糖复合止血材料的创面在3d、7d、14d及21d时,伤口面积均小于空白对照组,愈合率均高于空白对照组,促伤口愈合效果明显。对烫伤创面组织进行病理切片检查的结果显示,敷加了蚕蛹壳聚糖复合止血材料的创面组织生长活跃,炎性细胞浸润少,表皮无溃疡,与空白对照相比表现出了优良的促伤口愈合性能。 5、以大肠杆菌为革兰氏阴性菌的代表、以金黄色葡萄球菌为革兰氏阳性菌的代表,采用抑菌率和抑菌直径两种指标对蚕蛹壳聚糖复合止血材料的抑菌性能进行了测试。试验结果表明,目前广泛使用的明胶海绵止血材料对上述两种菌种均无抑菌作用,而蚕蛹壳聚糖复合止血材料对上述两种菌种在72h内的抑菌率均达到了100%,且对大肠杆菌出现了直径为1.8-2.1cm的抑菌圈,对金黄色葡萄球菌出现了直径为1.6-1.8cm的抑菌圈。 6、根据国家标准对医用生物材料的要求,对蚕蛹壳聚糖复合止血材料进行了包括溶血试验、原发性皮肤刺激试验、皮内反应试验、热源试验、急性全身毒性试验等一系列毒理学研究。研究结果表明,蚕蛹壳聚糖复合止血材料具有不溶血、对完整和破损皮肤及皮内组织均无刺激性、不含致热源、无急性全身毒性等特点,作为伤口敷料使用具有安全可靠性,是一种组织相容性良好的止血材料。该研究结果,为蚕蛹壳聚糖复合止血材料的临床应用提供了理论依据。
[Abstract]:Excessive bleeding is the main cause of death in the first aid and routine surgical treatment. At present, the widely used hemostat has its own shortcomings, and the high efficient hemostat imported from abroad restricts its wide application in the daily hemostasis because of its expensive price. Chitosan has good antibacterial, hemostatic performance and birth. The material compatibility is an ideal material for preparation of medical dressings. However, the Chinese sericulture resources are rich, and the silk reeling by-products contain a lot of chitosan resources, and the silkworm pupa chitosan has some properties superior to the shrimp and crab chitosan. Therefore, the chitosan as the main raw material and the gelatin as the compound, and the gelatin as the compound, are used as the compound, and the gelatin is used as the compound. A new type of spongy wound hemostat was developed with calcium chloride as a coagulant and glutaraldehyde as a crosslinker and glycerol as a freeze-resistant plastic agent. A single factor test and orthogonal test were used to optimize the preparation process of the compound hemostat of silkworm chrysalis chitosan, and the blood coagulation in vitro was discussed in detail. The results are as follows: the ability of hemostasis in vivo, and the wound healing, antibacterial and toxicological biological safety of materials.
1, chitin is extracted from silkworm pupae and chitosan is prepared and chitosan is obtained by controlling reaction time. The highest deacetylation degree can be obtained by controlling reaction time. The highest deacetylation degree can reach 93.13%. It is suitable to be used as a pharmaceutical product. The chitosan powder extracted from silkworm pupae is dissolved and added with appropriate additives. The compound hemostat of silkworm chrysalis chitosan was prepared by air drying method. The prepared samples were sponge like, the surface and internal pores were rich, distributed evenly, and had a certain elastic material touch.
2, the effect of the preparation process of the silkworm chrysalis composite Hemostat on the blood coagulation index was studied by a single factor test. The experiment showed that the optimum preparation parameters were prefreezing temperature -40 C, and the prefreezing time 1h. orthogonal test showed that the silkworm pupa shell polysugar, 0.1% calcium chloride, 0.005% glutaraldehyde, 3% gelatin, 0 gelatin, 0 were analyzed by the prefreezing time. The blood coagulation effect of the silkworm chrysalis composite hemostat prepared by.05% glycerin is the best. The effect of the concentration of each component in the compound Hemostat on the external coagulation index is: silkworm chrysalis chitosan gelatin glycerol.
3, chitosan is the key factor to lead the clotting performance of the silkworm chrysalis composite hemostat. The coagulation performance of the sample is improved effectively with the increase of the concentration of the silkworm chrysalis chitosan. Compared with the chitosan chitosan, the sample prepared by the silkworm pupa chitosan has better coagulation effect. Both in vitro and in animal living body have excellent hemostatic effect: compared with medical gauze, the blood coagulation index in vitro decreased by 62.71%, which reduced the amount of bleeding in the model of rabbit ear artery bleeding by 92.08% and the time of hemostasis by 80.55%. Compared with the medical gelatin sponge group, the coagulation index in vitro decreased by 35.61%, and the rabbit ear artery was made. The bleeding volume in the hemorrhage model was reduced by 88.35%, and the hemostasis time was shortened by 57.01%.
4, the effect of the compound hemostat of silkworm chrysalis on the wound area and healing rate was studied with the scald rabbit model. The results showed that the wound area was less than that of the blank control group at 3D, 7d, 14d and 21d, and the healing rate was higher than that of the blank control group, and the wound healing effect was obvious. The pathological sections of the facial tissue showed that the wound tissue was active, the infiltration of inflammatory cells was less and the epidermis was not ulcerated, and the wound healing performance was better than that of the blank control.
5, on behalf of the Gram-negative bacteria of Escherichia coli and Staphylococcus aureus as the representative of Gram-positive bacteria, the bacteriostasis performance of the compound hemostat of silkworm chrysalis chitosan was tested by the two indexes of bacteriostasis rate and bacteriostasis diameter. The results showed that the two kinds of bacteria were widely used in the Gelfoam hemostat. There was no bacteriostasis, and the antibacterial rate of the silkworm chrysalis compound hemostat was 100% in 72h, and the diameter of the Escherichia coli was 1.8-2.1cm, and the Bacteriostasis of Staphylococcus aureus was 1.6-1.8cm in diameter.
6, according to the requirements of national standard for medical biomaterials, a series of toxicological studies, including hemolytic test, primary skin irritation test, intradermal reaction test, heat source test, acute systemic toxicity test, were carried out on the silkworm chrysalis composite hemostatic material. The results showed that the sericulture chitosan composite hemostat had no hemolysis, and the results showed that the silkworm chrysalis composite hemostat had no hemolysis and was not hemolytic. The complete and damaged skin and intradermal tissues have no irritation, no heat source and no acute systemic toxicity. As a wound dressing, it is safe and reliable. It is a kind of hemostatic material with good histocompatibility. The result of this study provides a theoretical basis for the clinical application of the compound hemostat of silkworm chrysalis chitosan.
【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.08

【参考文献】