透明丝素蛋白膜作为角膜细胞载体的研究
发布时间：2018-12-16 16:22
【摘要】：蚕丝蛋白作为一种优异的天然高分子纤维蛋白已广泛应用于生物医用材料领域。其来源可靠,获取方便,具有良好的力学性能,良好的细胞相容性,组织相容性。本课题探讨了透明丝素蛋白膜作为角膜修复材料的可行性。采用酰胺类小分子与丝素蛋白复合,研究丝素蛋白复合膜的机械性能,透光性能,以及细胞相容性。通过X-射线衍射法研究了丝素蛋白复合膜的结构,结果表明当酰胺添加量较低时,复合膜主要为Silk I结晶结构,当酰胺添加量较多时,复合膜转变为Silk II结晶结构。力学性能测试表明,复合膜的断裂强度虽然比纯丝素膜有所降低,但是酰胺的加入改善了其柔韧性,使得其断裂伸长率增大。溶失率以及溶胀度测定的结果表明,酰胺的加入极大地降低了复合膜的溶失率,并且具有较低的溶胀度。采用酶标仪检测了复合膜的透光性能,并对复合膜的透光性进行了长时间的追踪检测,结果表明,酰胺/丝素蛋白膜具有较长时间的透光稳定性。提取家兔角膜基质细胞,通过在丝素蛋白膜上的体外培养,研究了酰胺/丝素蛋白复合膜与角膜基质细胞的相容性,并与乙醇处理的纯丝素蛋白膜进行对比。采用血球计数板测定细胞在不同膜上的黏附率,使用阿尔玛蓝测定细胞在不同膜上生长不同天后的荧光值,利用荧光倒置显微镜观察角膜基质细胞生长形态,Masson染色观察细胞外胶原蛋白的分泌。结果表明,酰胺/丝素膜与纯丝素膜之间并无显著差异,说明酰胺/丝素膜能够支持角膜基质细胞正常生长,酰胺/丝素复合膜有望用于角膜组织的修复。
[Abstract]:Silk protein as an excellent natural polymer fibrin has been widely used in the field of biomedical materials. Its source is reliable, easy to obtain, with good mechanical properties, good cytocompatibility and histocompatibility. In this paper, the feasibility of hyaline fibroin membrane as corneal repair material was discussed. The mechanical properties, transmittance and cytocompatibility of silk fibroin composite membrane were studied by using amides and silk fibroin. The structure of silk fibroin composite membrane was studied by X-ray diffraction. The results showed that the composite film was mainly Silk I crystalline structure when the amount of amide was low, and the composite film changed into Silk II crystal structure when the amount of amide was more. The mechanical properties test showed that the fracture strength of the composite membrane was lower than that of the pure silk fibroin film, but the addition of amide improved its flexibility and increased its elongation at break. The results of dissolution loss and swelling degree measurement showed that the addition of amide greatly reduced the dissolution rate of the composite membrane and had a lower swelling degree. The transmittance of the composite membrane was measured by enzyme labeling instrument, and the light transmittance of the composite membrane was measured for a long time. The results showed that the amide / silk fibroin film had a long time transmittance stability. Rabbit corneal stromal cells were extracted and cultured on fibroin membrane in vitro. The compatibility of amide-fibroin composite membrane with corneal stromal cells was studied and compared with the pure fibroin membrane treated with ethanol. The adhesion rate of cells on different membranes was measured by blood cell counter, the fluorescence value of cells growing on different membranes was measured by Alma blue, and the morphology of corneal stromal cells was observed by fluorescence inverted microscope. The secretion of extracellular collagen was observed by Masson staining. The results showed that there was no significant difference between the amide / silk fibroin membrane and the pure silk fibroin membrane, which indicated that the amide / silk fibroin membrane could support the normal growth of corneal stromal cells, and the amide / silk fibroin composite membrane could be used in corneal tissue repair.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R318.08
本文编号：2382689
[Abstract]:Silk protein as an excellent natural polymer fibrin has been widely used in the field of biomedical materials. Its source is reliable, easy to obtain, with good mechanical properties, good cytocompatibility and histocompatibility. In this paper, the feasibility of hyaline fibroin membrane as corneal repair material was discussed. The mechanical properties, transmittance and cytocompatibility of silk fibroin composite membrane were studied by using amides and silk fibroin. The structure of silk fibroin composite membrane was studied by X-ray diffraction. The results showed that the composite film was mainly Silk I crystalline structure when the amount of amide was low, and the composite film changed into Silk II crystal structure when the amount of amide was more. The mechanical properties test showed that the fracture strength of the composite membrane was lower than that of the pure silk fibroin film, but the addition of amide improved its flexibility and increased its elongation at break. The results of dissolution loss and swelling degree measurement showed that the addition of amide greatly reduced the dissolution rate of the composite membrane and had a lower swelling degree. The transmittance of the composite membrane was measured by enzyme labeling instrument, and the light transmittance of the composite membrane was measured for a long time. The results showed that the amide / silk fibroin film had a long time transmittance stability. Rabbit corneal stromal cells were extracted and cultured on fibroin membrane in vitro. The compatibility of amide-fibroin composite membrane with corneal stromal cells was studied and compared with the pure fibroin membrane treated with ethanol. The adhesion rate of cells on different membranes was measured by blood cell counter, the fluorescence value of cells growing on different membranes was measured by Alma blue, and the morphology of corneal stromal cells was observed by fluorescence inverted microscope. The secretion of extracellular collagen was observed by Masson staining. The results showed that there was no significant difference between the amide / silk fibroin membrane and the pure silk fibroin membrane, which indicated that the amide / silk fibroin membrane could support the normal growth of corneal stromal cells, and the amide / silk fibroin composite membrane could be used in corneal tissue repair.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R318.08
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