丝素蛋白—壳聚糖肛瘘栓组织工程初步研究
发布时间：2018-08-06 09:01
【摘要】：目的:1、制备丝素蛋白-壳聚糖(SFCS)新型高分子材料,通过表征观察,降解实验,研究符合组织工程要求的两者最优配置比例。2、使用丝素蛋白-壳聚糖复合材料进行生物相容性实验,初步评估其作为高分子医用材料应用到肛瘘栓上的可行性。方法:1、丝素蛋白 壳聚糖三维多孔支架的制做;使用氯化钙 醇 水体系(Ca CL2:C2H5OH:H2O=1:2:8)溶解,经透析、浓缩、离心获得2%的丝素蛋白溶液,壳聚糖溶解于醋酸溶液中配制出3%的壳聚糖溶液,将二者以不同的比例混合,装模、冷冻干燥、交联,获得丝素蛋白 壳聚糖三维支架产品。2、材料表征实验;采用扫描电子显微镜测试法,观察支架材料的特征,计算出材料的溶胀率、吸水率、孔隙率。3、材料静态降解实验;在磷酸盐缓冲溶液(PBS溶液)中放入SFCS复合材料,降解温度为37℃的恒温烤箱中,观察丝素蛋白 壳聚糖的降解速率及稳定性。4、丝素蛋白 壳聚糖复合材料的细胞生物相容性评价。将SFCS进行溶血实验,将SFCS支架和橡胶的样条分别植入SD大鼠大腿肌肉,在第7、14、21、28天,四个时间点,分别处死大鼠,分别在创面组织进行HE染色、Masson三色染色。结果:1、通过丝素蛋白与壳聚糖溶液不同质量比相混合后,可以得到不同成分组成的SFCS支架。2、不同配比的SFCS孔隙率值均高于95%。材料的溶胀率和吸水率是随着壳聚糖比例的增加而变大的。3、体外降解实验表明在同一时间点,随着丝素蛋白含量的增高复合材料的降解率依次递减。3、丝素蛋白 壳聚糖复合材料的溶血率小于5%,符合国家相关标准。4、动物实验表明,4周末,SFCS支架相比对照组孔内部炎症细胞几乎完全消失,被正常细胞填满,形成了完整地纤维囊壁结构,胶原含量明显高于对照组,胶原纤维较粗大、紧密,趋向更加明显。结论:1、将丝素蛋白和壳聚糖相混合,成功地制备出了拥有稳定构造的三维多孔支架,其中40%SF 60%CS组可作为生物支架研究的首选配比。2、动物埋植实验中,显示SFCS复合支架材料具有良好的生物相容性,是进行肛瘘栓前期材料研究中比较好的一个选择参考。
[Abstract]:Objective to prepare a new polymer material of silk fibroin chitosan (SFCS). In order to evaluate the feasibility of using fibroin-chitosan composite as a high molecular material for anal fistula suppository, the biocompatibility experiment was carried out with silk fibroin-chitosan composite. Methods 1: 1, preparation of silk fibroin / chitosan three-dimensional porous scaffold, 2% silk fibroin solution was obtained by dialysis, concentration, centrifugation and dissolution with calcium chloride CL2:C2H5OH:H2O=1:2:8 system (Ca CL2:C2H5OH:H2O=1:2:8). Chitosan was dissolved in acetic acid solution and 3% chitosan solution was prepared. The chitosan solution was mixed in different proportion, mold, freeze-drying, crosslinking, silk fibroin protein chitosan three-dimensional scaffold product. The characteristics of the scaffold material were observed by scanning electron microscopy (SEM). The swelling rate, water absorption, porosity, static degradation of the material were calculated, and the SFCS composite was added to the phosphate buffer solution (PBS solution). The degradation rate and stability of fibroin / chitosan were observed in a constant temperature oven at 37 鈩，
本文编号：2167213
[Abstract]:Objective to prepare a new polymer material of silk fibroin chitosan (SFCS). In order to evaluate the feasibility of using fibroin-chitosan composite as a high molecular material for anal fistula suppository, the biocompatibility experiment was carried out with silk fibroin-chitosan composite. Methods 1: 1, preparation of silk fibroin / chitosan three-dimensional porous scaffold, 2% silk fibroin solution was obtained by dialysis, concentration, centrifugation and dissolution with calcium chloride CL2:C2H5OH:H2O=1:2:8 system (Ca CL2:C2H5OH:H2O=1:2:8). Chitosan was dissolved in acetic acid solution and 3% chitosan solution was prepared. The chitosan solution was mixed in different proportion, mold, freeze-drying, crosslinking, silk fibroin protein chitosan three-dimensional scaffold product. The characteristics of the scaffold material were observed by scanning electron microscopy (SEM). The swelling rate, water absorption, porosity, static degradation of the material were calculated, and the SFCS composite was added to the phosphate buffer solution (PBS solution). The degradation rate and stability of fibroin / chitosan were observed in a constant temperature oven at 37 鈩，
本文编号：2167213
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