再生柞蚕丝素蛋白生物材料的制备
发布时间：2019-01-05 17:15
【摘要】：大量的研究表明桑蚕丝素蛋白具有良好的生物相容性，而柞蚕丝素蛋白除具备桑蚕丝素蛋白在生物医药领域的优点外，它还含有较多的对哺乳动物细胞有特异性相互作用的RGD三肽序列，有望在作为伤口敷料、组织工程支架材料、药物缓释微球材料和细胞培养基质等方面有比家蚕丝素更好的生物相容性和效果。本文在研究再生柞蚕丝素溶液性质的基础上，首次制备了再生的柞蚕丝素蛋白微球材料，，研究其对抗癌药物阿霉素的缓释效果；同时采用比较温和的方法，即通过加入小分子二元醇交联柞蚕丝素分子，制备不溶于水、机械性能优良的柞蚕丝素共混多孔材料和柞蚕丝素共混膜。 表面张力测试表明，再生的柞丝素蛋白具有一定的表面活性，能显著的降低水的表面张力。柞蚕丝素分子可以组装形成微球结构，在这个过程中丝素颗粒从3nm逐渐增长到1.5μm，其结构逐渐由α-螺旋和无规线团向β-折叠结构转变。在此基础上仅通过调节柞蚕丝素蛋白溶液pH值制备了具有良好分散性和外形规整性的柞蚕丝素蛋白微球，同时通过改变丝蛋白浓度制备出粒径范围在1-4μm可控的单分散柞蚕丝素微球。整个制备过程无需使用有机溶剂、表面活性剂、引发剂以及交联剂等，具有极高的生物安全性，制备工艺单，制备的丝素微球稳定性好。丝素溶液的pH为4.3左右（接近pI），制备的微球形态为规整的球形结构，丝素浓度为10mg/mL丝素微球的平均粒径为3.5μm。同时本文还探讨了柞蚕丝素微球形成机理：成核过程→多核成长过程→陈化过程。在此基础上制备了载阿霉素丝素微球，阿霉素的载药率以及包封率与阿霉素的投入量有关。当阿霉素的加入量为14%时，载药率可以达到3.4%。载阿霉素丝素微球的释放具有pH敏感特征，即在低pH（5.2）环境下释放较快，之后释放较缓慢。在23天后仍然有84%左右药物未释放，说明载阿霉素丝素微球具有明显的缓释性能。 为了开发适用于人工皮肤、创面覆盖等的生物材料，采用冷冻干燥方法制备出平均孔径为300～1000μm、孔隙率为82%～92%，且具有一定力学性能的柞蚕丝素/丁二醇多孔材料。1,4-丁二醇的加入促使柞蚕丝素蛋白分子聚集态结构逐渐向β-折叠结构转变，导致多孔材料的溶失率降低至≤2%。柞蚕丝素/丁二醇多孔材料含有高浓度柞蚕丝素蛋白时，材料具有较高的压缩强度。 同时，本论文还加入对机体无毒副作用的小分子二元醇与柞蚕丝素共混，采用流延法制备了柞蚕丝素/丙二醇、柞蚕丝素/乙二醇共混膜，兼顾解决了丝素蛋白的水溶性以及脆性问题。当柞蚕丝素/丙二醇、柞蚕丝素/乙二醇配比在45/55时，共混膜蛋白不溶于水，制得的共混膜理化性能最优，能够满足共混膜作为医用材料的要求。
[Abstract]:A large number of studies have shown that silk fibroin protein has good biocompatibility, and tussah silk fibroin protein has the advantages of silk fibroin protein in the field of biomedicine. It also contains a large number of RGD tripeptide sequences that specifically interact with mammalian cells and are expected to be used as wound dressings and tissue engineering scaffolds. Drug release microspheres and cell culture matrix have better biocompatibility and effect than silkworm fibroin. On the basis of studying the properties of regenerated tussah silk fibroin solution, the regenerated tussah silk fibroin protein microspheres were prepared for the first time, and the slow-release effect of Antheraea pernyi on adriamycin was studied. At the same time, by adding small molecule diol to crosslink the tussah fibroin molecule, the porous materials and the tussah silk fibroin blend film which are insoluble in water and excellent in mechanical properties were prepared. The surface tension test showed that the regenerated tussah silk protein had a certain surface activity and could significantly reduce the surface tension of water. Antheraea pernyi silk fibroin molecules can be assembled to form microspheres. During this process, silk fibroin particles gradually grow from 3nm to 1.5 渭 m, and their structures gradually change from 伪 -helix and random coils to 尾 -folded structures. On this basis, only by adjusting the pH value of tussah silk fibroin solution, the tussah silk fibroin microspheres with good dispersity and regular appearance were prepared. At the same time, the monodisperse tussah fibroin microspheres were prepared by changing the concentration of silk protein in the range of 1-4 渭 m. There is no need for organic solvents, surfactants, initiators and crosslinkers in the whole preparation process, which has the advantages of high biosafety, single preparation process and good stability of silk fibroin microspheres. The pH of fibroin solution was about 4.3.The morphology of the microspheres close to pI), was regular spherical structure, and the average diameter of fibroin microspheres was 3.5 渭 m when the concentration of fibroin was 10mg/mL fibroin microspheres. The formation mechanism of tussah silk fibroin microspheres was also discussed in this paper. On this basis, doxorubicin microspheres were prepared. The drug loading rate and encapsulation efficiency of doxorubicin were related to the amount of adriamycin. When adriamycin was added to the solution of 14%, the drug loading rate could reach 3.4%. The release of doxorubicin loaded fibroin microspheres was characterized by pH sensitivity, that is, the release was faster in low pH (5.2) environment, and then released slowly. After 23 days, about 84% of the drug was still not released, indicating that doxorubicin fibroin microspheres had obvious slow-release properties. In order to develop biomaterials suitable for artificial skin and wound covering, an average pore diameter of 300 渭 m and a porosity of 82 ~ 92 渭 m were prepared by freeze-drying method. With the addition of 1-4-butanediol, the molecular aggregation structure of tussah silk fibroin gradually changed to 尾 -fold structure, resulting in the solution loss rate of porous material reduced to 鈮
本文编号：2402075
[Abstract]:A large number of studies have shown that silk fibroin protein has good biocompatibility, and tussah silk fibroin protein has the advantages of silk fibroin protein in the field of biomedicine. It also contains a large number of RGD tripeptide sequences that specifically interact with mammalian cells and are expected to be used as wound dressings and tissue engineering scaffolds. Drug release microspheres and cell culture matrix have better biocompatibility and effect than silkworm fibroin. On the basis of studying the properties of regenerated tussah silk fibroin solution, the regenerated tussah silk fibroin protein microspheres were prepared for the first time, and the slow-release effect of Antheraea pernyi on adriamycin was studied. At the same time, by adding small molecule diol to crosslink the tussah fibroin molecule, the porous materials and the tussah silk fibroin blend film which are insoluble in water and excellent in mechanical properties were prepared. The surface tension test showed that the regenerated tussah silk protein had a certain surface activity and could significantly reduce the surface tension of water. Antheraea pernyi silk fibroin molecules can be assembled to form microspheres. During this process, silk fibroin particles gradually grow from 3nm to 1.5 渭 m, and their structures gradually change from 伪 -helix and random coils to 尾 -folded structures. On this basis, only by adjusting the pH value of tussah silk fibroin solution, the tussah silk fibroin microspheres with good dispersity and regular appearance were prepared. At the same time, the monodisperse tussah fibroin microspheres were prepared by changing the concentration of silk protein in the range of 1-4 渭 m. There is no need for organic solvents, surfactants, initiators and crosslinkers in the whole preparation process, which has the advantages of high biosafety, single preparation process and good stability of silk fibroin microspheres. The pH of fibroin solution was about 4.3.The morphology of the microspheres close to pI), was regular spherical structure, and the average diameter of fibroin microspheres was 3.5 渭 m when the concentration of fibroin was 10mg/mL fibroin microspheres. The formation mechanism of tussah silk fibroin microspheres was also discussed in this paper. On this basis, doxorubicin microspheres were prepared. The drug loading rate and encapsulation efficiency of doxorubicin were related to the amount of adriamycin. When adriamycin was added to the solution of 14%, the drug loading rate could reach 3.4%. The release of doxorubicin loaded fibroin microspheres was characterized by pH sensitivity, that is, the release was faster in low pH (5.2) environment, and then released slowly. After 23 days, about 84% of the drug was still not released, indicating that doxorubicin fibroin microspheres had obvious slow-release properties. In order to develop biomaterials suitable for artificial skin and wound covering, an average pore diameter of 300 渭 m and a porosity of 82 ~ 92 渭 m were prepared by freeze-drying method. With the addition of 1-4-butanediol, the molecular aggregation structure of tussah silk fibroin gradually changed to 尾 -fold structure, resulting in the solution loss rate of porous material reduced to 鈮
本文编号：2402075
本文链接：https://www.wllwen.com/yixuelunwen/swyx/2402075.html
