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

【摘要】：為探究絲素蛋白膜用作引導(dǎo)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(duì)絲素蛋白膜進(jìn)行材料學(xué)和生物學(xué)性能評(píng)價(jià):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(chǎng)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(cè)試對(duì)絲素膜及共混膜的理化性質(zhì)進(jìn)行表征;研究共混膜對(duì)小鼠成骨細(xì)胞MC3T3-E1和小鼠成纖維細(xì)胞STO的活性、粘附和增殖的影響,探究共混膜的細(xì)胞生物學(xué)相容性;通過SD大鼠皮下移植實(shí)驗(yàn)研究膜材料在體內(nèi)的生物學(xué)相容性。研究表明抄紙工藝可獲得質(zhì)地柔軟、表面粗糙、溶脹力強(qiáng)的多孔膜,同時(shí)保持共混膜內(nèi)蛋白二級(jí)結(jié)構(gòu)(β-折疊)的穩(wěn)定性;細(xì)胞在絲素膜和共混膜表面均能貼附良好,且細(xì)胞形態(tài)舒展;細(xì)胞增殖實(shí)驗(yàn)表明共混膜能夠促進(jìn)細(xì)胞增殖,且增殖實(shí)驗(yàn)的7天內(nèi)細(xì)胞保持快速增殖;大鼠皮下移植實(shí)驗(yàn)結(jié)果表明,膜材料植入動(dòng)物體后,前期機(jī)體內(nèi)可出現(xiàn)正常的炎細(xì)胞浸潤現(xiàn)象,隨時(shí)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(yīng),與市售膠原蛋白膜生物相容性相當(dāng);并且SF2組在術(shù)后9周時(shí)膜材料仍保持相對(duì)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(dǎo)骨再生的絲素膜。采用FE-SEM、ATR-FTIR和拉伸斷裂測(cè)試對(duì)所獲得的絲素膜進(jìn)行表征。在有和無蛋白酶XIV的PBS體系中,評(píng)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(dǎo)膜(成分為不可降解的PCL聚合物)作為對(duì)照,研究絲素膜的引導(dǎo)骨再生能力。結(jié)果表明,較高濃度的乙醇處理可賦予絲素膜較高的結(jié)晶度,提高絲素膜更好的機(jī)械性能并降低其生物降解性。在兔顱骨缺損實(shí)驗(yàn)中,絲素蛋白膜在體內(nèi)移植后可有效阻止結(jié)締組織細(xì)胞遷移到缺損區(qū)域,對(duì)兔顱骨缺損形成保護(hù)空間,促進(jìn)新骨生成,與商業(yè)骨引導(dǎo)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(dǎo)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(xué)位授予單位】：浙江理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TB383.2;R318.08

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