新型GBR丝素蛋白屏障膜的制备及其生物相容性评价

发布时间：2018-05-27 14:47

本文选题：GBR膜 + 丝素蛋白膜　；参考：《浙江理工大学》2017年硕士论文

【摘要】：为探究丝素蛋白膜用作引导骨再生膜(GBR屏障膜)的可行性,本研究提出了两种新型丝素蛋白膜的制备工艺,并对丝素蛋白膜进行材料学和生物学性能评价:1)利用抄纸工艺制备丝素蛋白膜及丝素蛋白与胶原蛋白共混膜。采用场发射扫描电子显微镜(FE-SEM)、傅立叶变换衰减全反射红外光谱(ATR-FTIR)、静态接触角及拉伸断裂测试对丝素膜及共混膜的理化性质进行表征;研究共混膜对小鼠成骨细胞MC3T3-E1和小鼠成纤维细胞STO的活性、粘附和增殖的影响,探究共混膜的细胞生物学相容性;通过SD大鼠皮下移植实验研究膜材料在体内的生物学相容性。研究表明抄纸工艺可获得质地柔软、表面粗糙、溶胀力强的多孔膜,同时保持共混膜内蛋白二级结构(β-折叠)的稳定性;细胞在丝素膜和共混膜表面均能贴附良好,且细胞形态舒展;细胞增殖实验表明共混膜能够促进细胞增殖,且增殖实验的7天内细胞保持快速增殖;大鼠皮下移植实验结果表明,膜材料植入动物体后,前期机体内可出现正常的炎细胞浸润现象,随时间的延长炎症现象可明显减缓,其中SF2组在第9周后几乎无炎症反应,与市售胶原蛋白膜生物相容性相当;并且SF2组在术后9周时膜材料仍保持相对完整性。综合比较发现75:25组膜材料更理想。2)通过冷冻干燥、致密化和乙醇处理制备用于引导骨再生的丝素膜。采用FE-SEM、ATR-FTIR和拉伸断裂测试对所获得的丝素膜进行表征。在有和无蛋白酶XIV的PBS体系中,评估丝素膜的生物降解性。在兔颅骨缺损模型中,用猪胶原膜和一种商业骨引导膜(成分为不可降解的PCL聚合物)作为对照,研究丝素膜的引导骨再生能力。结果表明,较高浓度的乙醇处理可赋予丝素膜较高的结晶度,提高丝素膜更好的机械性能并降低其生物降解性。在兔颅骨缺损实验中,丝素蛋白膜在体内移植后可有效阻止结缔组织细胞迁移到缺损区域,对兔颅骨缺损形成保护空间,促进新骨生成,与商业骨引导膜和猪胶原膜相比,冷冻干燥的致密丝素蛋白膜具有用于引导骨组织再生的可行性。
[Abstract]:In order to explore the feasibility of silk fibroin membrane as the barrier membrane for guiding bone regeneration membrane (GBR), two new preparation techniques of silk fibroin membrane were proposed in this study. The silk fibroin membrane and its blend with collagen were prepared by paper-making process. The physical and chemical properties of silk fibroin film and blend film were characterized by field emission scanning electron microscope (SEM), Fourier transform attenuated total reflectance infrared spectroscopy (FTR-FTIR), static contact angle and tensile fracture test. To study the effects of the blend membrane on the activity, adhesion and proliferation of mouse osteoblast MC3T3-E1 and mouse fibroblast STO, to explore the cell biocompatibility of the blend membrane, and to study the biocompatibility of the membrane material in vivo by subcutaneous transplantation of SD rats. The results showed that the paper-making process could obtain the porous membrane with soft texture, rough surface and strong swelling force, while maintaining the stability of the protein secondary structure (尾 -fold) in the blend membrane, and the cells could be attached well on the surface of silk fibroin film and blend membrane. Cell proliferation test showed that the blend membrane could promote cell proliferation and maintain rapid proliferation within 7 days. The results of subcutaneous transplantation in rats showed that the membrane material was implanted into animal body. Normal inflammatory cell infiltration could occur in the early stage of the body, and the inflammatory phenomenon could be slowed down with the prolongation of time. In the SF2 group, there was almost no inflammatory reaction after 9 weeks, which was similar to the biocompatibility of the commercial collagen membrane. In SF2 group, the membrane material remained relatively intact at 9 weeks after operation. It was found that 75:25 membrane material was more ideal. 2) Silk fibroin membrane for bone regeneration was prepared by freeze-drying densification and ethanol treatment. The obtained fibroin films were characterized by FE-SEMN ATR-FTIR and tensile fracture test. The biodegradability of silk fibroin membranes was evaluated in PBS systems with and without protease XIV. In rabbit skull defect model, the ability of leading bone regeneration of silk fibroin membrane was studied by using porcine collagen membrane and a commercial bone guided membrane (composed of non-degradable PCL polymer) as control. The results showed that higher concentration of ethanol could increase the crystallinity of silk fibroin membrane, improve the mechanical properties of silk fibroin membrane and decrease its biodegradability. In the experiment of rabbit skull defect, silk fibroin membrane can effectively prevent connective tissue cells from migrating to the defect area after transplantation in vivo, which can protect the rabbit skull defect and promote the formation of new bone. Compared with commercial bone guiding membrane and porcine collagen membrane, silk fibroin membrane can effectively prevent the migration of connective tissue cells to the defect area. Freeze-dried dense silk fibroin membrane has the feasibility of guiding bone regeneration.
【学位授予单位】：浙江理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TB383.2;R318.08

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