【摘要】：大量的研究表明桑蠶絲素蛋白具有良好的生物相容性，而柞蠶絲素蛋白除具備桑蠶絲素蛋白在生物醫(yī)藥領(lǐng)域的優(yōu)點(diǎn)外，它還含有較多的對哺乳動物細(xì)胞有特異性相互作用的RGD三肽序列，有望在作為傷口敷料、組織工程支架材料、藥物緩釋微球材料和細(xì)胞培養(yǎng)基質(zhì)等方面有比家蠶絲素更好的生物相容性和效果。本文在研究再生柞蠶絲素溶液性質(zhì)的基礎(chǔ)上，首次制備了再生的柞蠶絲素蛋白微球材料，，研究其對抗癌藥物阿霉素的緩釋效果；同時(shí)采用比較溫和的方法，即通過加入小分子二元醇交聯(lián)柞蠶絲素分子，制備不溶于水、機(jī)械性能優(yōu)良的柞蠶絲素共混多孔材料和柞蠶絲素共混膜。 表面張力測試表明，再生的柞絲素蛋白具有一定的表面活性，能顯著的降低水的表面張力。柞蠶絲素分子可以組裝形成微球結(jié)構(gòu)，在這個過程中絲素顆粒從3nm逐漸增長到1.5μm，其結(jié)構(gòu)逐漸由α-螺旋和無規(guī)線團(tuán)向β-折疊結(jié)構(gòu)轉(zhuǎn)變。在此基礎(chǔ)上僅通過調(diào)節(jié)柞蠶絲素蛋白溶液pH值制備了具有良好分散性和外形規(guī)整性的柞蠶絲素蛋白微球，同時(shí)通過改變絲蛋白濃度制備出粒徑范圍在1-4μm可控的單分散柞蠶絲素微球。整個制備過程無需使用有機(jī)溶劑、表面活性劑、引發(fā)劑以及交聯(lián)劑等，具有極高的生物安全性，制備工藝單，制備的絲素微球穩(wěn)定性好。絲素溶液的pH為4.3左右（接近pI），制備的微球形態(tài)為規(guī)整的球形結(jié)構(gòu)，絲素濃度為10mg/mL絲素微球的平均粒徑為3.5μm。同時(shí)本文還探討了柞蠶絲素微球形成機(jī)理：成核過程→多核成長過程→陳化過程。在此基礎(chǔ)上制備了載阿霉素絲素微球，阿霉素的載藥率以及包封率與阿霉素的投入量有關(guān)。當(dāng)阿霉素的加入量為14%時(shí)，載藥率可以達(dá)到3.4%。載阿霉素絲素微球的釋放具有pH敏感特征，即在低pH（5.2）環(huán)境下釋放較快，之后釋放較緩慢。在23天后仍然有84%左右藥物未釋放，說明載阿霉素絲素微球具有明顯的緩釋性能。 為了開發(fā)適用于人工皮膚、創(chuàng)面覆蓋等的生物材料，采用冷凍干燥方法制備出平均孔徑為300～1000μm、孔隙率為82%～92%，且具有一定力學(xué)性能的柞蠶絲素/丁二醇多孔材料。1,4-丁二醇的加入促使柞蠶絲素蛋白分子聚集態(tài)結(jié)構(gòu)逐漸向β-折疊結(jié)構(gòu)轉(zhuǎn)變，導(dǎo)致多孔材料的溶失率降低至≤2%。柞蠶絲素/丁二醇多孔材料含有高濃度柞蠶絲素蛋白時(shí)，材料具有較高的壓縮強(qiáng)度。 同時(shí)，本論文還加入對機(jī)體無毒副作用的小分子二元醇與柞蠶絲素共混，采用流延法制備了柞蠶絲素/丙二醇、柞蠶絲素/乙二醇共混膜，兼顧解決了絲素蛋白的水溶性以及脆性問題。當(dāng)柞蠶絲素/丙二醇、柞蠶絲素/乙二醇配比在45/55時(shí)，共混膜蛋白不溶于水，制得的共混膜理化性能最優(yōu)，能夠滿足共混膜作為醫(yī)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 偽 -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

本文編號：2402075