本文選題：絲蛋白 + 慶大霉素��； 參考：《南方醫(yī)科大學(xué)》2017年碩士論文

【摘要】：研究背景現(xiàn)在內(nèi)固定系統(tǒng)已廣泛運(yùn)用于臨床骨折的治療,但是內(nèi)植物感染和材料的不可吸收是骨科內(nèi)固定手術(shù)后需進(jìn)行二次手術(shù)的重要原因。絲蛋白材料已被證明是一種具良好力學(xué)性能和生物相容性的可降解生物材料,然而感染仍然是絲蛋白材料需要面臨的一個(gè)問(wèn)題。因此探究抗菌絲蛋白材料的制備也成為了必然趨勢(shì)。研究目的本研究期望通過(guò)在絲蛋白中加入慶大霉素來(lái)制備具有良好抗菌性能的可吸收絲蛋白骨科內(nèi)固定材料,同時(shí)能保持良好生物相容性及生物力學(xué)性能。我們?cè)敿?xì)探討了新型抗菌材料的抗菌性能,并從生物相容性、生物力學(xué)強(qiáng)度、降解速率及吸水性等角度初步評(píng)價(jià)了抗菌絲蛋白材料相關(guān)性質(zhì)的改變,為后期臨床應(yīng)用提供依據(jù)。研究方法從蠶繭中提取絲蛋白并在六氟異丙醇溶解過(guò)程中加入慶大霉素來(lái)制備不同濃度的抗菌絲蛋白材料。利用抑菌圈、平板計(jì)數(shù)法及掃描電鏡評(píng)估材料在體外對(duì)金黃色葡萄球菌和大腸桿菌的抗菌性能。同時(shí)也對(duì)比了抗菌絲蛋白材料與普通絲蛋白材料相關(guān)性質(zhì)的改變,包括體內(nèi)外的生物相容性和體外的生物力學(xué)性能、膨脹性以及降解性能等。研究結(jié)果普通絲蛋白材料明顯促進(jìn)金黃色葡萄球菌的生長(zhǎng)(抑菌率-306.0±3.6%),加入了慶大霉素(16mg/g)的抗菌絲蛋白材料1小時(shí)后對(duì)金黃色葡萄球菌和大腸桿菌的抑菌率均達(dá)到100%,當(dāng)降低慶大霉素濃度至8mg/g及6mg/g后該時(shí)間均延長(zhǎng)到12小時(shí)。掃描電鏡提示抗菌絲蛋白材料表面兩種細(xì)菌數(shù)量均較少,并且金黃色葡萄球菌黏附形態(tài)受到不同程度影響。此外抗菌絲蛋白材料在酶溶液中降解4周后對(duì)兩種細(xì)菌的抑菌率仍能維持在90%以上。而生物力學(xué)方面,抗菌絲蛋白材料最大彎曲強(qiáng)度為202.8±7.8Mpa、最大剪切強(qiáng)度為35.4±6.1Mpa、彈性模量8.5±0.7Gpa,和普通絲蛋白材料并沒有顯著的差異(p0.05)。同時(shí)在體外抗菌絲蛋白材料未對(duì)MC3T3-E1細(xì)胞的增殖和凋亡產(chǎn)生不良影響,體內(nèi)實(shí)驗(yàn)也證明其并不會(huì)引起明顯的炎癥反應(yīng)和毒性反應(yīng),對(duì)成骨也未產(chǎn)生影響。研究發(fā)現(xiàn)相比普通絲蛋白,抗菌絲蛋白材料的降解速率明顯加快,并且符合三次函數(shù),而其吸水性卻明顯下降。結(jié)論慶大霉素的最適濃度為16mg/g,此時(shí)抗菌絲蛋白材料不僅具有高效、快速、持久的抗菌性,而且能保持其良好的生物力學(xué)性能及生物相容性,更重要的是亦可逐漸被降解。該新型抗菌可吸收絲蛋白內(nèi)固定材料應(yīng)用于臨床有望在一定程度上降低骨科內(nèi)固定手術(shù)后的二次手術(shù)率。
[Abstract]:Background the internal fixation system has been widely used in the treatment of clinical fractures, but the internal plant infection and the non-absorbability of materials are the important reasons for the secondary operation after orthopedic internal fixation. Silk protein material has been proved to be a biodegradable material with good mechanical properties and biocompatibility. However, infection is still a problem for silk protein material. Therefore, it has become an inevitable trend to explore the preparation of antibacterial silk protein materials. Objective to prepare absorbable fibroin orthopedic internal fixation materials with good antibacterial properties by adding gentamicin into silk protein and maintain good biocompatibility and biomechanical properties. The antimicrobial properties of new antibacterial materials were discussed in detail, and the changes of the properties of antibacterial silk protein materials were preliminarily evaluated from the aspects of biocompatibility, biomechanical strength, degradation rate and water absorption, which provided the basis for later clinical application. Methods the silk protein was extracted from silkworm cocoon and gentamicin was added in the dissolution process of hexafluoroisopropanol to prepare different concentrations of antimicrobial silk protein materials. The antibacterial properties of the materials against Staphylococcus aureus and Escherichia coli in vitro were evaluated by bacteriostasis, plate counting and scanning electron microscopy. At the same time, the changes of the properties of antibacterial silk protein material and common silk protein material were compared, including biocompatibility in vivo and in vitro, biomechanical properties in vitro, expansibility and degradability. Results the common silk protein material significantly promoted the growth of Staphylococcus aureus (bacteriostasis rate -306.0 鹵3.6%). After adding gentamicin (16mg / g), the antimicrobial silk protein material reached both Staphylococcus aureus and Escherichia coli inhibition rate 1 hour after adding Gentamicin to Staphylococcus aureus and Escherichia coli. When gentamicin concentration was reduced to 8mg/g and 6mg/g, the time was prolonged to 12 hours. Scanning electron microscope (SEM) showed that the number of two bacteria on the surface of antibacterial silk protein materials was relatively small, and the adhesion morphology of Staphylococcus aureus was affected to some extent. In addition, the bacteriostatic rate of antimicrobial silk protein material to two kinds of bacteria remained above 90% after degradation in enzyme solution for 4 weeks. In biomechanics, the maximum bending strength, shear strength and elastic modulus of antibacterial silk protein material were 202.8 鹵7.8 Mpaand 35.4 鹵6.1 Mpaand 8.5 鹵0.7 Gpa.There was no significant difference between antibacterial silk protein material and common silk protein material. At the same time, the antibacterial silk protein material had no adverse effect on the proliferation and apoptosis of MC3T3-E1 cells in vitro, and the in vivo experiments showed that it did not cause obvious inflammatory and toxic reactions, nor did it affect osteogenesis. It was found that the degradation rate of antibacterial silk protein was faster than that of common silk protein, and the degradation rate of antimicrobial silk protein was in accordance with cubic function, but its water absorption was obviously decreased. Conclusion the optimum concentration of gentamicin is 16 mg / g. The antibacterial silk protein material not only has high efficiency, fast and lasting antibacterial properties, but also can maintain its good biomechanical properties and biocompatibility, and more importantly, it can be degraded gradually. The new antimicrobial absorbable filamentine internal fixation material is expected to reduce the rate of secondary surgery after orthopedic internal fixation to some extent.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R318.08;R68

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