磺化PP无纺布血液相容性与吸附低密度脂蛋白性能研究
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摘要
本文采用60Co伽马射线共辐射接枝法，在对苯乙烯磺酸钠(SSS)单体水溶液中分别添加N-乙烯基-2-吡咯烷酮(NVP)、丙烯酰胺(AAm)和丙烯酸(AAc)三种单体，以实现SSS在聚丙烯(PP)无纺布上的接枝，从而制备磺化PP无纺布，并对磺化PP无纺布进行表征和亲水性测试以及对其血液相容性、抗凝血效果和吸附低密度脂蛋白(LDL)性能进行考察。
一、共辐射接枝制备磺化PP无纺布
本论文分别系统地考察了吸收剂量、剂量率、二元单体总浓度以及二元单体摩尔浓度比对总接枝率(DGT)和SSS接枝率(DGSSS)的影响。研究结果表明可以通过调整上述参数制备不同接枝率的PP无纺布。
二、表征
全反射傅里叶变换红外光谱：经过poly(NVP-co-SSS)、poly(AAm-co-SSS)和poly(AAc-co-SSS)三种接枝链修饰后的PP无纺布的红外谱图均在1125、1039和1009cm-1附近处出现了属于磺酸基(-SO-3)的特征吸收峰；此外，三种磺化PP无纺布的红外谱图分别出现了属于NVP中内酰胺基(1673cm-1)、AAm中酰胺基(1667cm-1)和AAc中羧基(1715cm-1)的特征吸收峰。X射线光电子能谱：与原PP无纺布的谱图相比，磺化PP无纺布的谱图均在168eV附近处出现了属于硫元素(S2p)的特征吸收峰，同时氧元素(O1s,531eV)的吸收峰强度均明显增强；此外，PP-g-P(NVP-co-SSS)和PP-g-P(AAm-co-SSS)两个样品的谱图中出现了属于氮元素(N1s,399eV)的信号峰。无纺布表面形貌：通过扫描电子显微镜观察可以发现，与原PP无纺布相比，磺化PP无纺布的纤维直径明显增大，并且纤维结构显得更为紧密。亲水性：吸液率测试结果表明，磺化PP无纺布具有优良的亲水性。
三、血液相容性
溶血：磺化PP无纺布的溶血率均低于5%，符合医用材料对溶血的要求。牛血清蛋白(BSA)吸附：三组样品的BSA吸附量均随着接枝率的增加而下降，并且PP-g-P(AAm-co-SSS)样品比其他两组样品具有更好的抑制BSA吸附效果。血浆总蛋白吸附：与另两组样品相比，PP-g-P(AAm-co-SSS)样品具有更好的抗血浆总蛋白吸附效果；此外，PP-g-P(AAc-co-SSS)样品对血浆总蛋白的吸附量较高，且接枝率的高低对吸附量的大小没有明显的影响。血小板黏附数量：三组样品单位面积黏附血小板的数量均随着接枝率的增加而呈现下降的趋势，并且PP-g-P(AAm-co-SSS)样品表现出更好的抑制血小板黏附效果。黏附血小板形态：通过扫描电镜所拍摄的图片可以明显地看出有大量的血小板聚集和黏附于未改性的PP无纺布上，并且血小板形貌发生明显的变形(伸出伪足和胞基质扩展)；然而未能够明显地观察到有血小板黏附于磺化PP无纺布表面，表明磺化PP无纺布具有良好的抑制血小板黏附性能。血液相容性评估结果表明：经过接枝改性，PP无纺布的血液相容性得到了有效地改善。
四、抗凝血效果
本文通过检测活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)和凝血酶时间(TT)来评价磺化PP无纺布的抗凝血效果。结果显示，磺酸基团的引入能够有效地提高PP无纺布的抗凝血效果，尤其是延长内源性凝血系统的凝血时间，即APTT。此外，羧基具有良好的协同抗凝血作用，能够进一步地增强磺化PP无纺布的抗凝血效果，而酰胺基并没有显示出协同抗凝抗凝作用。
五、吸附LDL性能
本文通过测定所制备的磺化PP无纺布对血浆中LDL和高密度脂蛋白(HDL)的吸附率来评价其选择性吸附性能。结果显示，磺化PP无纺布虽然具有一定的吸附LDL的性能，但是磺化PP无纺布对HDL也有一定的吸附作用。因此，所制备的磺化PP无纺布并没有显示出良好的选择性吸附特性，还有待于进一步对工艺和材料进行改进和优化，从而提高其选择性吸附LDL效果，并降低HDL的吸附率。
Sodium styrenesulfonate (SSS) was grafted onto polypropylene (PP) non-wovenfabric via60Co γ-ray simultaneous radiation grafting method with the addition ofN-vinyl-2-pyrrolidone (NVP), acryl amide (AAm) and acrylic acid (AAc),respectively. The sulfonated PP non-woven fabrics were characterized and theirproperties, including hydrophilicity, blood compatibility, anticoagulant effect andadsorption properties of low density lipoprotein (LDL), were evaluated.
1. Preparation of sulfonated PP non-woven fabric via simultaneous radiationinduced graft copolymerization
In this paper, we systematically studied the effect of absorbed dose, dose rate,total molar concentration and molar concentration ratio of binary monomers on theDGTand DGSSS, respectively. The results showed that different DGTand DGSSScouldbe attained by adjustment of the above parameters.
2. Characterization
Attenuated total reflection Fourier transform infrared spectroscopy: As comparedwith the spectrum of pristine PP non-woven fabric, the new bands at about1125,1039,and1009cm-1in the spectra of sulfonated PP non-woven fabrics inculdingPP-g-P(NVP-co-SSS), PP-g-P(AAm-co-SSS) and PP-g-P(AAc-co-SSS), belonged tosulfonate group (-SO3-). Besides, the peaks at about1673,1667and1715cm-1couldbe assigned to the stretching vibration of C=O from NVP, AAm and AAc,respectively. X-ray photoelectron spectroscopy: A fresh peak at168eV (S2p) appeared in the spectra of sulfonated PP non-woven fabrics. The intensity of O1s (531eV) peak of sulfonated PP non-woven fabrics was stronger than that of neat PPnon-woven fabric. Moreover, an original peak at399eV (N1s) emerged in the spectraof PP-g-P(NVP-co-SSS) and PP-g-P(AAm-co-SSS) samples. Suface morphology ofPP non-woven fabrics: According to the picture of scan electron microscope, thediameter of modified fibers was apparently larger than that of virgin fiber. Besides,the configuration of modified fibres seemed to be more compacted. Hydrophilicity:The consequence of adsorption percentage of deionized water and phosphate buffersolution (PBS)(pH,7.4) demonstrated that the sulfonated PP non-woven fabrics hadexcellent hydrophilicity.
3. Blood compatibility
Hemolysis: The hemolytic ratio of sulfonated PP non-woven fabrics was below5%, which met what the medical materials required. Adsorption density of BSA: Theadsorption density of BSA decreased with the increase of DG. Furthermore, the groupof PP-g-P(AAm-co-SSS) samples was more effective to resist the BSA adsorptionthan the other two groups. Adsorption density of total protein: The group ofPP-g-P(AAm-co-SSS) samples was more efficient to limit the total protein adsorptionthan the other two groups. However, the PP-g-P(AAc-co-SSS) samples showed moreattraction to total protein and the DG had little effect on the adsorption density.Number of adhered platelets: The amount of adhered platelets reduced with theraising of DG, and PP-g-P(AAm-co-SSS) samples were more proficient in restrictingthe adhesion of platelets than the other two group of samples. Morphology of adheredplatelets: A lot of platelets aggregated and adhered onto virgin PP non-woven fabric.The deposited platelets exhibited an activated morphology, as demonstrated byextended pseudopodia and surface spreading. On the contrary, there’s few plateletsadhered and aggregated on the surface of sulfonated PP non-woven fabrics, whichillustrated that sulfonated PP non-woven fabrics could effectively restrict the adhesionof platelets. The above results demonstrated that the hemocompatibility of PPnon-woven fabric was greatly improved after modfication.
4. Anticoagulant effect
We evaluated the anticoagulant effect of sulfonated PP non-woven fabrics viaexamination of clotting times, including activated partial thromboplastin time (APTT),prothrombin time (PT) and thrombin time (TT). The results proved that sulfonategroup showed good anticoagulative activity, especially lengthening the APTT(intrinsic coagulant pathway). Besides, the carboxyl group had a synergetic effect toextend the clotting time. However, the amide group did not indicate this kind ofeffect.
5. Adsorption properties of LDL
In this paper, we assessed the selectivity of adsorbing LDL of sulfonated PPnon-woven fabrics by means of determination of adsorption percentage of LDL andhigh density lipoprotein (HDL). The prepared sulfonated PP non-woven fabrics, to acentain degree, presented the property of adsorbing LDL. However, the sulfonated PPnon-woven fabrics also showed adsorbing effect on HDL. Hence, the sulfonated PPnon-woven fabrics did not show good selectivity to adsorbing LDL, which should beameliorated in order to reduce the adsorption percentage of HDL and enhance theselective property of adsorbing LDL.
一、共辐射接枝制备磺化PP无纺布
本论文分别系统地考察了吸收剂量、剂量率、二元单体总浓度以及二元单体摩尔浓度比对总接枝率(DGT)和SSS接枝率(DGSSS)的影响。研究结果表明可以通过调整上述参数制备不同接枝率的PP无纺布。
二、表征
全反射傅里叶变换红外光谱：经过poly(NVP-co-SSS)、poly(AAm-co-SSS)和poly(AAc-co-SSS)三种接枝链修饰后的PP无纺布的红外谱图均在1125、1039和1009cm-1附近处出现了属于磺酸基(-SO-3)的特征吸收峰；此外，三种磺化PP无纺布的红外谱图分别出现了属于NVP中内酰胺基(1673cm-1)、AAm中酰胺基(1667cm-1)和AAc中羧基(1715cm-1)的特征吸收峰。X射线光电子能谱：与原PP无纺布的谱图相比，磺化PP无纺布的谱图均在168eV附近处出现了属于硫元素(S2p)的特征吸收峰，同时氧元素(O1s,531eV)的吸收峰强度均明显增强；此外，PP-g-P(NVP-co-SSS)和PP-g-P(AAm-co-SSS)两个样品的谱图中出现了属于氮元素(N1s,399eV)的信号峰。无纺布表面形貌：通过扫描电子显微镜观察可以发现，与原PP无纺布相比，磺化PP无纺布的纤维直径明显增大，并且纤维结构显得更为紧密。亲水性：吸液率测试结果表明，磺化PP无纺布具有优良的亲水性。
三、血液相容性
溶血：磺化PP无纺布的溶血率均低于5%，符合医用材料对溶血的要求。牛血清蛋白(BSA)吸附：三组样品的BSA吸附量均随着接枝率的增加而下降，并且PP-g-P(AAm-co-SSS)样品比其他两组样品具有更好的抑制BSA吸附效果。血浆总蛋白吸附：与另两组样品相比，PP-g-P(AAm-co-SSS)样品具有更好的抗血浆总蛋白吸附效果；此外，PP-g-P(AAc-co-SSS)样品对血浆总蛋白的吸附量较高，且接枝率的高低对吸附量的大小没有明显的影响。血小板黏附数量：三组样品单位面积黏附血小板的数量均随着接枝率的增加而呈现下降的趋势，并且PP-g-P(AAm-co-SSS)样品表现出更好的抑制血小板黏附效果。黏附血小板形态：通过扫描电镜所拍摄的图片可以明显地看出有大量的血小板聚集和黏附于未改性的PP无纺布上，并且血小板形貌发生明显的变形(伸出伪足和胞基质扩展)；然而未能够明显地观察到有血小板黏附于磺化PP无纺布表面，表明磺化PP无纺布具有良好的抑制血小板黏附性能。血液相容性评估结果表明：经过接枝改性，PP无纺布的血液相容性得到了有效地改善。
四、抗凝血效果
本文通过检测活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)和凝血酶时间(TT)来评价磺化PP无纺布的抗凝血效果。结果显示，磺酸基团的引入能够有效地提高PP无纺布的抗凝血效果，尤其是延长内源性凝血系统的凝血时间，即APTT。此外，羧基具有良好的协同抗凝血作用，能够进一步地增强磺化PP无纺布的抗凝血效果，而酰胺基并没有显示出协同抗凝抗凝作用。
五、吸附LDL性能
本文通过测定所制备的磺化PP无纺布对血浆中LDL和高密度脂蛋白(HDL)的吸附率来评价其选择性吸附性能。结果显示，磺化PP无纺布虽然具有一定的吸附LDL的性能，但是磺化PP无纺布对HDL也有一定的吸附作用。因此，所制备的磺化PP无纺布并没有显示出良好的选择性吸附特性，还有待于进一步对工艺和材料进行改进和优化，从而提高其选择性吸附LDL效果，并降低HDL的吸附率。
Sodium styrenesulfonate (SSS) was grafted onto polypropylene (PP) non-wovenfabric via60Co γ-ray simultaneous radiation grafting method with the addition ofN-vinyl-2-pyrrolidone (NVP), acryl amide (AAm) and acrylic acid (AAc),respectively. The sulfonated PP non-woven fabrics were characterized and theirproperties, including hydrophilicity, blood compatibility, anticoagulant effect andadsorption properties of low density lipoprotein (LDL), were evaluated.
1. Preparation of sulfonated PP non-woven fabric via simultaneous radiationinduced graft copolymerization
In this paper, we systematically studied the effect of absorbed dose, dose rate,total molar concentration and molar concentration ratio of binary monomers on theDGTand DGSSS, respectively. The results showed that different DGTand DGSSScouldbe attained by adjustment of the above parameters.
2. Characterization
Attenuated total reflection Fourier transform infrared spectroscopy: As comparedwith the spectrum of pristine PP non-woven fabric, the new bands at about1125,1039,and1009cm-1in the spectra of sulfonated PP non-woven fabrics inculdingPP-g-P(NVP-co-SSS), PP-g-P(AAm-co-SSS) and PP-g-P(AAc-co-SSS), belonged tosulfonate group (-SO3-). Besides, the peaks at about1673,1667and1715cm-1couldbe assigned to the stretching vibration of C=O from NVP, AAm and AAc,respectively. X-ray photoelectron spectroscopy: A fresh peak at168eV (S2p) appeared in the spectra of sulfonated PP non-woven fabrics. The intensity of O1s (531eV) peak of sulfonated PP non-woven fabrics was stronger than that of neat PPnon-woven fabric. Moreover, an original peak at399eV (N1s) emerged in the spectraof PP-g-P(NVP-co-SSS) and PP-g-P(AAm-co-SSS) samples. Suface morphology ofPP non-woven fabrics: According to the picture of scan electron microscope, thediameter of modified fibers was apparently larger than that of virgin fiber. Besides,the configuration of modified fibres seemed to be more compacted. Hydrophilicity:The consequence of adsorption percentage of deionized water and phosphate buffersolution (PBS)(pH,7.4) demonstrated that the sulfonated PP non-woven fabrics hadexcellent hydrophilicity.
3. Blood compatibility
Hemolysis: The hemolytic ratio of sulfonated PP non-woven fabrics was below5%, which met what the medical materials required. Adsorption density of BSA: Theadsorption density of BSA decreased with the increase of DG. Furthermore, the groupof PP-g-P(AAm-co-SSS) samples was more effective to resist the BSA adsorptionthan the other two groups. Adsorption density of total protein: The group ofPP-g-P(AAm-co-SSS) samples was more efficient to limit the total protein adsorptionthan the other two groups. However, the PP-g-P(AAc-co-SSS) samples showed moreattraction to total protein and the DG had little effect on the adsorption density.Number of adhered platelets: The amount of adhered platelets reduced with theraising of DG, and PP-g-P(AAm-co-SSS) samples were more proficient in restrictingthe adhesion of platelets than the other two group of samples. Morphology of adheredplatelets: A lot of platelets aggregated and adhered onto virgin PP non-woven fabric.The deposited platelets exhibited an activated morphology, as demonstrated byextended pseudopodia and surface spreading. On the contrary, there’s few plateletsadhered and aggregated on the surface of sulfonated PP non-woven fabrics, whichillustrated that sulfonated PP non-woven fabrics could effectively restrict the adhesionof platelets. The above results demonstrated that the hemocompatibility of PPnon-woven fabric was greatly improved after modfication.
4. Anticoagulant effect
We evaluated the anticoagulant effect of sulfonated PP non-woven fabrics viaexamination of clotting times, including activated partial thromboplastin time (APTT),prothrombin time (PT) and thrombin time (TT). The results proved that sulfonategroup showed good anticoagulative activity, especially lengthening the APTT(intrinsic coagulant pathway). Besides, the carboxyl group had a synergetic effect toextend the clotting time. However, the amide group did not indicate this kind ofeffect.
5. Adsorption properties of LDL
In this paper, we assessed the selectivity of adsorbing LDL of sulfonated PPnon-woven fabrics by means of determination of adsorption percentage of LDL andhigh density lipoprotein (HDL). The prepared sulfonated PP non-woven fabrics, to acentain degree, presented the property of adsorbing LDL. However, the sulfonated PPnon-woven fabrics also showed adsorbing effect on HDL. Hence, the sulfonated PPnon-woven fabrics did not show good selectivity to adsorbing LDL, which should beameliorated in order to reduce the adsorption percentage of HDL and enhance theselective property of adsorbing LDL.
引文
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