本文選題：GBR膜 + 絲素蛋白膜　； 參考：《浙江理工大學》2017年碩士論文

【摘要】：為探究絲素蛋白膜用作引導骨再生膜(GBR屏障膜)的可行性,本研究提出了兩種新型絲素蛋白膜的制備工藝,并對絲素蛋白膜進行材料學和生物學性能評價:1)利用抄紙工藝制備絲素蛋白膜及絲素蛋白與膠原蛋白共混膜。采用場發(fā)射掃描電子顯微鏡(FE-SEM)、傅立葉變換衰減全反射紅外光譜(ATR-FTIR)、靜態(tài)接觸角及拉伸斷裂測試對絲素膜及共混膜的理化性質進行表征;研究共混膜對小鼠成骨細胞MC3T3-E1和小鼠成纖維細胞STO的活性、粘附和增殖的影響,探究共混膜的細胞生物學相容性;通過SD大鼠皮下移植實驗研究膜材料在體內的生物學相容性。研究表明抄紙工藝可獲得質地柔軟、表面粗糙、溶脹力強的多孔膜,同時保持共混膜內蛋白二級結構(β-折疊)的穩(wěn)定性;細胞在絲素膜和共混膜表面均能貼附良好,且細胞形態(tài)舒展;細胞增殖實驗表明共混膜能夠促進細胞增殖,且增殖實驗的7天內細胞保持快速增殖;大鼠皮下移植實驗結果表明,膜材料植入動物體后,前期機體內可出現(xiàn)正常的炎細胞浸潤現(xiàn)象,隨時間的延長炎癥現(xiàn)象可明顯減緩,其中SF2組在第9周后幾乎無炎癥反應,與市售膠原蛋白膜生物相容性相當;并且SF2組在術后9周時膜材料仍保持相對完整性。綜合比較發(fā)現(xiàn)75:25組膜材料更理想。2)通過冷凍干燥、致密化和乙醇處理制備用于引導骨再生的絲素膜。采用FE-SEM、ATR-FTIR和拉伸斷裂測試對所獲得的絲素膜進行表征。在有和無蛋白酶XIV的PBS體系中,評估絲素膜的生物降解性。在兔顱骨缺損模型中,用豬膠原膜和一種商業(yè)骨引導膜(成分為不可降解的PCL聚合物)作為對照,研究絲素膜的引導骨再生能力。結果表明,較高濃度的乙醇處理可賦予絲素膜較高的結晶度,提高絲素膜更好的機械性能并降低其生物降解性。在兔顱骨缺損實驗中,絲素蛋白膜在體內移植后可有效阻止結締組織細胞遷移到缺損區(qū)域,對兔顱骨缺損形成保護空間,促進新骨生成,與商業(yè)骨引導膜和豬膠原膜相比,冷凍干燥的致密絲素蛋白膜具有用于引導骨組織再生的可行性。
[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.
【學位授予單位】：浙江理工大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：TB383.2;R318.08

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