鈦氧薄膜表面固定PEG及硒代胱胺構(gòu)建催化活性層的研究
發(fā)布時(shí)間：2018-08-04 09:52
【摘要】：血管支架等血液接觸材料抗凝抗增生性能不足嚴(yán)重制約了其臨床應(yīng)用。NO是心血管系統(tǒng)的信號因子,可抑制血小板的粘附及平滑肌細(xì)胞的增生,其在生物材料改性研究領(lǐng)域的應(yīng)用受到廣泛的關(guān)注。本論文在氧化鈦表面通過PEG偶聯(lián)固定NO的催化分子硒代胱胺,從而構(gòu)建催化活性層。并對催化活性層進(jìn)行了系列表征和抗凝、抗增生等生物學(xué)評價(jià)。 采用非平衡磁控濺射在單晶硅材料表面沉積氧化鈦薄膜,通過在材料表面沉積聚多巴胺的方式引入有機(jī)官能團(tuán),雙端修飾的聚乙二醇可作為偶聯(lián)劑將硒代胱胺固定于材料表面。論文通過FTIR、XPS跟蹤檢測了各改性階段樣品的表面微特征變化,證實(shí)各分子在樣品表面成功固定。稱重實(shí)驗(yàn)比較了兩種工藝制備的聚多巴胺樣品,顯示以搖床震蕩沉積5次得到的聚多巴胺的量最大；氨基定量實(shí)驗(yàn)顯示以1.5mmol/l濃度接枝的PEG樣品表面的氨基量最大。 對改性樣品進(jìn)行體外生物學(xué)評價(jià),GPx酶活力檢測得到最終改性樣品的酶活力為30.43U/cm2。ELISA檢測證實(shí)改性樣品能夠明顯抑制纖維蛋白原的粘附及變性。體外血小板粘附樣品通過熒光染色、SEM等手段進(jìn)行表征。結(jié)果顯示,無供體組中,經(jīng)聚乙二醇修飾的樣品表面的血小板數(shù)量明顯少于對比樣；供體組中,膠原蛋白嚴(yán)重激活了對比樣表面的血小板,而由于NO的作用,Se樣品表面的血小板粘附、聚集、激活狀態(tài)都得到了顯著抑制,cGMP檢測證實(shí)此效應(yīng)是通過NO-cGMP信號通路實(shí)現(xiàn)的。體外SMC實(shí)驗(yàn)中,無供體組顯示經(jīng)修飾的樣品均可在一定程度上抑制SMC的粘附；在供體存在條件下,SMC在Se樣品表面的粘附受到強(qiáng)烈抑制,胞內(nèi)cGMP的含量也明顯升高。 綜上所述,本文通過PEG偶聯(lián)固定硒代胱胺,成功在材料表面構(gòu)建了催化活性層,改性后表面可催化內(nèi)源性NO供體釋放NO,進(jìn)而升高細(xì)胞內(nèi)cGMP含量,發(fā)揮其生物效應(yīng)。在PEG、NO雙重機(jī)制的作用下,材料表面抗凝及抗增生性能得到明顯改善,為血液接觸材料的表面改性提供了一種新途徑。
[Abstract]:The lack of anticoagulant and anti-proliferative properties of blood contact materials such as vascular stents seriously restricts its clinical application. No is a signal factor of cardiovascular system, which can inhibit platelet adhesion and smooth muscle cell proliferation. Its application in the field of biomaterial modification has received extensive attention. In this paper, the catalytic active layer was constructed by coupling the no catalyst molecule selenocysteine on the surface of titanium oxide by PEG. A series of characterization of catalytic active layer and biological evaluation of anticoagulant and anti-proliferation were carried out. Titanium oxide thin films were deposited on the surface of monocrystalline silicon by unbalanced magnetron sputtering. Organic functional groups were introduced by deposition of dopamine on the surface of the materials. The double end modified polyethylene glycol could be used as a coupling agent to immobilize selenocysteamine on the surface of the materials. In this paper, FTIR XPS was used to trace and detect the changes of surface microfeatures of the samples at different modification stages, and it was proved that the molecules were successfully immobilized on the surface of the samples. The results showed that the maximum amount of polydopamine was obtained by shaking deposition 5 times, and the amino content on the surface of PEG grafted with 1.5mmol/l concentration was the largest. The enzyme activity of the modified sample was determined by biological evaluation in vitro. The result showed that the modified sample could inhibit the adhesion and denaturation of fibrinogen by 30.43U/cm2.ELISA. Platelet adhesion in vitro was characterized by fluorescence staining and SEM. The results showed that in the non-donor group, the number of platelet on the surface of the sample modified by polyethylene glycol was significantly less than that on the control group, while in the donor group, the platelet on the surface of the contrast sample was seriously activated by collagen. Because of the effect of no on platelet adhesion, aggregation and activation of se samples, the inhibition of NO-cGMP signaling pathway was demonstrated. In vitro SMC assay, no donor group showed that the modified samples could inhibit the adhesion of SMC to a certain extent, and the adhesion of SMC to the surface of se samples was strongly inhibited in the presence of donor, and the content of intracellular cGMP was also significantly increased. To sum up, the catalytic active layer was successfully constructed on the surface of the material by PEG coupling immobilization of selenocysteine. The modified surface could catalyze the release of no from endogenous no donors, and then increase the content of cGMP in the cells and exert its biological effect. The anticoagulant and anti-proliferative properties of the materials were improved obviously under the dual mechanism of PEGN and no, which provided a new way for the surface modification of blood contact materials.
【學(xué)位授予單位】：西南交通大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R318.08
本文編號：2163489
[Abstract]:The lack of anticoagulant and anti-proliferative properties of blood contact materials such as vascular stents seriously restricts its clinical application. No is a signal factor of cardiovascular system, which can inhibit platelet adhesion and smooth muscle cell proliferation. Its application in the field of biomaterial modification has received extensive attention. In this paper, the catalytic active layer was constructed by coupling the no catalyst molecule selenocysteine on the surface of titanium oxide by PEG. A series of characterization of catalytic active layer and biological evaluation of anticoagulant and anti-proliferation were carried out. Titanium oxide thin films were deposited on the surface of monocrystalline silicon by unbalanced magnetron sputtering. Organic functional groups were introduced by deposition of dopamine on the surface of the materials. The double end modified polyethylene glycol could be used as a coupling agent to immobilize selenocysteamine on the surface of the materials. In this paper, FTIR XPS was used to trace and detect the changes of surface microfeatures of the samples at different modification stages, and it was proved that the molecules were successfully immobilized on the surface of the samples. The results showed that the maximum amount of polydopamine was obtained by shaking deposition 5 times, and the amino content on the surface of PEG grafted with 1.5mmol/l concentration was the largest. The enzyme activity of the modified sample was determined by biological evaluation in vitro. The result showed that the modified sample could inhibit the adhesion and denaturation of fibrinogen by 30.43U/cm2.ELISA. Platelet adhesion in vitro was characterized by fluorescence staining and SEM. The results showed that in the non-donor group, the number of platelet on the surface of the sample modified by polyethylene glycol was significantly less than that on the control group, while in the donor group, the platelet on the surface of the contrast sample was seriously activated by collagen. Because of the effect of no on platelet adhesion, aggregation and activation of se samples, the inhibition of NO-cGMP signaling pathway was demonstrated. In vitro SMC assay, no donor group showed that the modified samples could inhibit the adhesion of SMC to a certain extent, and the adhesion of SMC to the surface of se samples was strongly inhibited in the presence of donor, and the content of intracellular cGMP was also significantly increased. To sum up, the catalytic active layer was successfully constructed on the surface of the material by PEG coupling immobilization of selenocysteine. The modified surface could catalyze the release of no from endogenous no donors, and then increase the content of cGMP in the cells and exert its biological effect. The anticoagulant and anti-proliferative properties of the materials were improved obviously under the dual mechanism of PEGN and no, which provided a new way for the surface modification of blood contact materials.
【學(xué)位授予單位】：西南交通大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R318.08
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