本文選題：聚醚醚酮(PEEK) + 多巴胺　； 參考：《蘇州大學》2016年博士論文

【摘要】：第一部分表面載銀的PEEK材料的制備與表征檢測目的:在弱堿性條件下自聚合形成的聚多巴胺涂層沉積在聚醚醚酮(Polyetheretherketone,PEEK)表面,利用聚多巴胺自身的弱還原性,可以將銀氨離子還原為銀納米顆粒沉積在PEEK材料表面,制備出表面載銀的PEEK復合材料,并對其進行檢測與表征。方法:將表面打磨光滑的PEEK圓片用等離子體清洗后,在表面負載大量的羥基,將上述材料浸泡在2 mg/ml多巴胺溶液中(10 m M Tris-HCl溶液,p H=8.5)24小時,在PEEK表面制備出聚多巴胺涂層(PEEK-PDA),然后將含有聚多巴胺涂層的PEEK復合材料浸泡在0.02 mol/L的銀氨溶液中1小時,制備出表面載銀的PEEK復合材料(PEEK-PDA-Ag)。然后對該涂層的形貌特征、化學組成進行檢測,材料表面親水性分析,以及銀離子釋放等表征。結果:檢測顯示,經(jīng)過載銀處理后的PEEK材料顏色上發(fā)生了很明顯的變化,電鏡下可以很明顯觀察到材料表面均勻的沉積了銀顆粒,粒徑約為90 nm,EDS能譜分析顯示載銀量為5.83 wt%。接觸角測試顯示經(jīng)過載銀處理后的PEEK材料表面親水性發(fā)生了很大的改變。當將表面載銀的PEEK復合材料浸泡于緩沖溶液中,涂層能向周圍溶液中持續(xù)釋放銀離子。結論:利用多巴胺的自聚合以及聚多巴胺的還原性,能夠在PEEK材料表面制備出尺寸均勻的銀粒子涂層,該涂層能夠長期向周圍溶液中釋放銀離子。第二部分表面載銀的PEEK材料的抗菌性能檢測目的:探討表面載銀的PEEK復合材料(PEEK-PDA-Ag)對金黃色葡萄球菌的抗菌性能。方法:⑴以PEEK-PDA-Ag和PEEK-PDA作為實驗組,PEEK作為對照組,以金黃色葡萄球菌菌株ATCC25923為實驗菌株。將金黃色葡萄球菌菌株ATCC25923接種與各組材料培養(yǎng)24小時,進行活菌計數(shù),觀察抑菌效果,計算其抑菌率。⑵在各組材料表面接種金黃色葡萄球菌菌珠,經(jīng)過24小時培養(yǎng)后,觀察有無抑菌圈生成,并測量抑菌圈直徑大小。⑶在各組材料表面接種實驗菌株培養(yǎng)24小時后,脫水干燥后,電鏡下觀測菌落形貌。結果:表面抑菌實驗結果提示:培養(yǎng)24小時以后,實驗組(PEEK-PDA-Ag)表現(xiàn)出明顯的抑菌效應,其表面的金黃色葡萄球菌活菌計數(shù)低于接種濃度,抑菌率可達到99.84%,PEEK-PDA組也展現(xiàn)出了一定的抑菌效應,抑菌率為78.6%。實驗組與對照組之間抑菌率具有顯著統(tǒng)計學差異(P㩳0.05)。將金黃色葡萄球菌接種在各組材料表面,培養(yǎng)24小時后,在PEEK-PDA-Ag組樣品周圍產(chǎn)生了明顯的抑菌圈,抑菌圈直徑約為8.8 mm,而在PEEK組和PEEK-PDA組周圍未見明顯的抑菌圈產(chǎn)生。將金黃色葡萄球菌接種在各組材料表面,培養(yǎng)24小時后,經(jīng)過脫水干燥處理后,在電鏡下可以觀察,PEEK-PDA-Ag組樣品表面細菌數(shù)量明顯少于PEEK組和PEEK-PDA組(P㩳0.05)。結論:PEEK-PDA-Ag材料具有明顯的抑菌作用,抑菌率可達到99.84%,同時在PEEK-PDA-Ag材料周圍可以明顯的觀測到抑菌圈的產(chǎn)生,抑菌圈直徑約為8.8 mm。第三部分表面載銀的PEEK材料的生物相容性檢測目的:探討表面載銀的PEEK復合材料(PEEK-PDA-Ag)對成骨細胞MC3T3-E1生物活性的影響,并觀察細胞在材料表面黏附和增殖的情況。用小鼠巨噬細胞(RAW264.7)為動物細胞模型,利用q RT-PCR技術從細胞分子水平上探討幾種材料對小鼠巨噬細胞分泌的相關炎性因子表達的影響。方法:⑴以PEEK-PDA-Ag材料作為實驗組,PEEK材料和PEEK-PDA材料作為對照組,分別按一定的密度接種MC3T3-E1細胞并進行培養(yǎng)。在第1,3,5天,分別用CCK-8試劑檢測各組材料表面細胞增殖情況。⑵在每個時間點,每組取出兩個樣品,經(jīng)過梯度脫水和臨界點干燥后,用掃描電鏡觀察成骨細胞在各組材料表面的黏附鋪展情況。⑶以PEEK-PDA-Ag材料和PEEK-PDA材料作為實驗組,空白板作為對照組,用小鼠巨噬細胞(RAW264.7)為動物細胞模型,檢測IL-4,IL-10,TGF-β抗炎因子的表達以及IL-1β,IL-6,TNF-α等促炎因子的表達情況。結果:CCK-8檢測結果表明:PEEK-PDA-Ag組細胞有明顯的增殖趨勢,在第1,3,5天時間點時,PEEK-PDA-Ag組細胞都有明顯的增殖,對照組PEEK和PEEK-PDA的細胞同樣也有明顯的增殖(P〉0.05);掃描電鏡下觀察細胞形貌及黏附:可以看出,實驗組PEEK-PDA-Ag表面的細胞在第一天時呈細長形,在第3,5天時,材料表面細胞有明顯增殖,材料表面細胞密度明顯增多,且細胞立體性較好,對照組PEEK和PEEK-PDA材料在第1天時,表面的細胞黏附很均勻,且向四周鋪展,第3天時,材料表面細胞密度明顯多于第一天時間點,細胞增殖明顯,第5天時,對照組細胞基本長滿,且細胞均比較飽滿。炎性因子表達的結果顯示,RAW264.7細胞在各組材料上培養(yǎng)72h后,在抗炎因子表達方面,與對照組相比,PEEK-PDA-Ag組細胞中IL-4基因表達量顯著降低(P㩳0.05),IL-10和TGF-β表達量有降低趨勢,但沒達到顯著水平;在促炎因子表達方面,與對照組相比,PEEK-PDA-Ag組細胞中IL-1β基因表達量顯著升高(P㩳0.05),但IL-6與TNF-α表達量無顯著差異。結論:通過聚多巴胺修飾還原制備的表面載銀的PEEK材料(PEEK-PDA-Ag)與MC3T3-E1共培養(yǎng)觀察,顯示在PEEK-PDA-Ag復合材料表面,細胞同樣可以很好地黏附,增殖分化,無嚴重的毒性;通過與小鼠巨噬細胞RAW264.7的共培養(yǎng),檢測相關炎性因子表達情況,材料表面載Ag并無抗炎癥作用,相反還略有輕微促炎作用。
[Abstract]:The preparation and characterization of the first part of the PEEK material carrying silver on the surface of Polyetheretherketone (PEEK) on the surface of the polyether ether ketone (PEEK) under the weak alkali condition, the silver nanoparticles can be reduced to silver nanoparticles on the surface of the PEEK material by using the weak reducibility of the polyamine itself, and the preparation of the silver nanoparticles can be prepared on the surface of the material. The PEEK composite with silver on the surface was detected and characterized. Method: after cleaning the smooth PEEK wafer on the surface with plasma, a large amount of hydroxyl was loaded on the surface, and the above materials were soaked in 2 mg/ml dopamine solution (10 m M Tris-HCl solution, P H=8.5) for 24 hours, and the polydopamine coating was prepared on the PEEK surface (PEEK-P). DA), then the PEEK composite containing polydopamine coating was soaked in a silver ammonia solution of 0.02 mol/L for 1 hours. The surface silver loaded PEEK composite (PEEK-PDA-Ag) was prepared. Then the morphology, chemical composition of the coating, the hydrophilic analysis of the material surface, and the release of silver ions were characterized. The color of the PEEK material after the silver loading has been obviously changed. Under the electron microscope, it can be observed that the silver particles are evenly deposited on the surface of the material, the particle size is about 90 nm. The EDS spectrum analysis shows that the silver load is 5.83 wt%. contact angle, and the hydrophilicity of the surface of the PEEK material after the silver loading has been greatly changed. When the material is treated, the surface hydrophilicity of the material has been greatly changed. The silver coated PEEK composite is soaked in the buffer solution, and the coating can release silver ions continuously in the surrounding solution. Conclusion: using the self polymerization of dopamine and the reducibility of the poly dopamine, a silver particle coating with uniform size can be prepared on the surface of the PEEK material. The coating can release Silver ions in the surrounding solution for a long time. Second parts. Detection of antibacterial properties of PEEK material carrying silver on the surface: To investigate the antibacterial properties of PEEK composite (PEEK-PDA-Ag) on Staphylococcus aureus. Methods: (1) PEEK-PDA-Ag and PEEK-PDA as the experimental group, PEEK as the control group and Staphylococcus aureus strain ATCC25923 as the experimental strain. Plant ATCC25923 inoculation and each group material culture for 24 hours, carry out the living bacteria count, observe the bacteriostasis effect and calculate its bacteriostasis rate. 2. After inoculation of Staphylococcus aureus beads on the surface of each material, after 24 hours culture, the formation of bacteriostatic ring is observed and the diameter of bacteriostasis circle is measured. 3. (3) inoculation of experimental strains on the surface of each material is 24 small. After dehydration and drying, the morphology of the colony was observed under the electron microscope. The results of the surface bacteriostasis experiment showed that after 24 hours of culture, the experimental group (PEEK-PDA-Ag) showed obvious bacteriostasis effect, the count of Staphylococcus aureus on the surface was lower than the inoculation concentration, the bacteriostasis rate could reach 99.84%, and the PEEK-PDA group also showed a certain bacteriostasis effect. The bacteriostasis rate was significant difference between the 78.6%. experimental group and the control group (P? 0.05). The Staphylococcus aureus was inoculated on the surface of each group. After 24 hours culture, the bacteriostatic circle was found around the PEEK-PDA-Ag group, and the diameter of the bacteriostasis was about 8.8 mm, but there was no obvious inhibition around the PEEK group and the PEEK-PDA group. The micrococcus aureus was inoculated on the surface of each material. After 24 hours culture, after dehydration and drying, the number of bacteria on the surface of PEEK-PDA-Ag group was obviously less than that of group PEEK and PEEK-PDA (P? 0.05). Conclusion: PEEK-PDA-Ag material has obvious bacteriostasis, and the rate of bacteriostasis can reach 99.84%. Meanwhile, the rate of bacteriostasis can reach 99.84%. The production of bacteriostasis circle can be observed around the PEEK-PDA-Ag material. The biocompatibility test of the PEEK material containing 8.8 mm. and third parts of silver containing the bacteriostasis diameter is to investigate the effect of the PEEK composite (PEEK-PDA-Ag) on the bioactivity of the osteoblast MC3T3-E1, and to observe the adhesion of the cells to the surface of the material and the adhesion of the cells on the surface of the material. Using mouse macrophage (RAW264.7) as an animal cell model, the effect of several materials on the expression of related inflammatory factors secreted by mouse macrophages was investigated by Q RT-PCR technology. Methods: (1) the PEEK-PDA-Ag material was used as the experimental group, and the PEEK material and the PEEK-PDA material were used as the control group, respectively. The density of MC3T3-E1 cells was inoculated and cultured. On day 1,3,5, CCK-8 reagents were used to detect the proliferation of the surface cells of each group. 2. At each time point, two samples were taken out of each group. After the gradient dehydration and critical point drying, the adhesion and spreading of the osteoblasts on the surface of each group was observed by scanning electron microscope. 3. (3) PEEK-PD A-Ag and PEEK-PDA materials were used as the experimental group, and the blank plate was used as the control group. The expression of IL-4, IL-10, TGF- beta anti-inflammatory factors and the expression of IL-1 beta, IL-6, TNF- alpha and other proinflammatory factors were detected with mouse macrophage (RAW264.7) as the animal cell model. Results: the results of CCK-8 test showed that the PEEK-PDA-Ag group had obvious proliferation trend. At 1,3,5 day time, the cells of group PEEK-PDA-Ag had obvious proliferation, and the cells of PEEK and PEEK-PDA in the control group also had obvious proliferation (P > 0.05). The morphology and adhesion of the cells were observed under the scanning electron microscope. It can be seen that the cells on the surface of PEEK-PDA-Ag in the experimental group were elongated on the first day, and the surface cells of the material were obvious at day 3,5. On the first day of first days, the cell density of the control group was very much more than the first day point, and the cell proliferation was obvious. At the time of fifth days, the cells of the control group were basically full, and the cells were all cells. The expression of inflammatory factors showed that after the RAW264.7 cells were cultured for 72h in each group, the expression of IL-4 gene in the PEEK-PDA-Ag group was significantly lower than that in the control group (P? 0.05). The expression of IL-10 and TGF- beta was reduced, but it did not reach a significant level. Compared with the control group, the expression of IL-1 beta gene in the cells of the PEEK-PDA-Ag group increased significantly (P? 0.05), but there was no significant difference in the expression of IL-6 and TNF- alpha. Conclusion: the PEEK material (PEEK-PDA-Ag), which is prepared by polydopamine modification and reduction (PEEK-PDA-Ag), is co cultured with MC3T3-E1, showing that the cells can also be well on the surface of the PEEK-PDA-Ag composite material. Adhesion, proliferation and differentiation, no serious toxicity; through co culture with RAW264.7 of mouse macrophages, the expression of related inflammatory factors was detected, and the surface of the material had no anti-inflammatory effect on the surface of Ag. On the contrary, it was slightly proinflammatory.

【學位授予單位】：蘇州大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：R318.08;R68

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