CN114573029A - 纳米硫化钼及其作为尿毒症毒素的高效吸附剂的应用 - Google Patents
纳米硫化钼及其作为尿毒症毒素的高效吸附剂的应用 Download PDFInfo
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Abstract
本发明涉及一种纳米硫化钼及其作为尿毒症毒素的高效吸附剂的应用,解决现有技术中可佩戴人工肾的透析液净化装置中吸附材料难储存、吸附效率低和生物相容性差的技术问题。本发明的纳米硫化钼具有二维超薄纳米片结构,层间距为基面存在大量缺陷,这种具有宽层间距和丰富晶格缺陷的二硫化钼可以作为尿毒症毒素的高效吸附剂。本发明首次报道了利用具有宽层间距和丰富晶格缺陷的二硫化钼吸附尿毒症毒素尿素、肌酐和尿酸。与商用MoS2、活性炭、窄层间距有晶格缺陷的MoS2、窄层间距无晶格缺陷的MoS2相比,通过对不同条件下(吸附剂用量、吸附时间、温度等)材料吸附性能的分析研究,获得一种高吸附性、高选择性和高稳定性的透析液净化材料。
Description
技术领域
本发明涉及透析液净化材料技术领域,具体涉及一种纳米硫化钼及其作为尿毒症毒素的高效吸附剂的应用。
背景技术
可佩戴人工肾的透析液净化装置中尿毒症毒素(尿素、肌酐、尿酸等)的去除是公认的具有挑战性的课题。目前研究的吸附剂替代品材料有Ti3C2Tx片层迈科烯(MXene)、二氧化钛(TiO2)电极以及多孔道活性炭等。但是,大多数用于尿毒症毒素去除的吸附剂材料由于其制备复杂、吸附性质单一、无选择性、吸附性质不高及生物相容性差等缺点,严重限制了其应用。
发明内容
本发明要解决现有技术中可佩戴人工肾的透析液净化装置中吸附材料难储存、吸附效率低和生物相容性差的技术问题,提供一种纳米硫化钼及其作为尿毒症毒素的高效吸附剂的应用。
为了解决上述技术问题,本发明的技术方案具体如下:
本发明提供一种纳米硫化钼,其是由下述方法制备得到的:
将(NH4)6Mo7O24·4H2O和CS(NH2)2添加到蒸馏水中,混合物搅拌均匀后转移至高压反应釜中,在140-220℃反应12-48小时,反应结束后产物经乙醇和蒸馏水洗涤后,真空干燥,得到具有宽层间距和丰富晶格缺陷的纳米硫化钼;
进一步优选的是,所述(NH4)6Mo7O24·4H2O和CS(NH2)2的摩尔比为1:30。
进一步优选的是,混合物搅拌均匀所用时间为30min。
进一步优选的是,真空干燥的温度为60℃,时间为24小时。
本发明还提供一种纳米硫化钼作为尿毒症毒素的高效吸附剂的应用,所述纳米硫化钼是具有宽层间距和丰富晶格缺陷的纳米硫化钼。
进一步的,所述纳米硫化钼是由下述方法制备得到:
按照(NH4)6Mo7O24·4H2O和CS(NH2)2的摩尔比为1:30将(NH4)6Mo7O24·4H2O和CS(NH2)2添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在140-220℃反应12-48小时,产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时,得到具有宽层间距和丰富晶格缺陷的纳米硫化钼。
本发明的有益效果是:
本发明首次报道了利用具有宽层间距和丰富晶格缺陷的二硫化钼(WDR-MoS2)吸附尿毒症毒素尿素、肌酐和尿酸,并进一步研究了其吸附性能和生物相容性。与商用MoS2、活性炭(AC)、窄层间距有晶格缺陷的MoS2(DR-MoS2)、窄层间距无晶格缺陷的MoS2(DF-MoS2)相比,通过对不同条件下(吸附剂用量、吸附时间、温度等)材料吸附性能的分析研究,获得一种高吸附性、高选择性和高稳定性的透析液净化材料。然后,本发明采用了模拟透析液和模拟透析装置对材料的吸附性能进行了探究,证明其在动态条件下具有良好的吸附性能。此外,通过细胞实验和血液实验对这种材料的生物相容性进行了研究。其高效的吸附性能和良好的生物相容性,在实际应用中是一种安全有效的尿毒症毒素吸附剂。
附图说明
下面结合附图和具体实施方式对本发明作进一步详细说明。
图1为不同MoS2样品的结构表征图,其中(a)WDR-MoS2的TEM图像;(b)WDR-MoS2的HRTEM图像,插图为对应的SAED图像;商用MoS2的(c)TEM和(d)HRTEM图像;(e)不同MoS2的XRD谱图;(f)不同MoS2的XPS测量总谱;(g)Mo 3d和(h)S 2p的高分辨率XPS谱图。
图2为水溶液中样品对尿毒症毒素的吸附性能图,其中使用不同剂量的AC和制备的二硫化钼对(a)尿素、(b)肌酐和(c)尿酸的去除效率测试图;采用准一级和准二级模型研究了WDR-MoS2对(d)尿素、(e)肌酐和(f)尿酸的吸附动力学;不同温度下WDR-MoS2对(g)尿素、(h)肌酐和(i)尿酸的吸附等温线及Langmuir和Freundlich模型拟合曲线。
图3为不同剂量WDR-MoS2对尿毒症毒素去除效率图,其中(a)模拟透析液和(b)HSA溶液中对尿毒症毒素去除效率图,(c)动态条件下的简易尿毒症毒素吸附系统示意图,WDR-MoS2从水溶液中去除尿毒症毒素的(d)Thomas和(e)Yoon-Nelson模型拟合曲线,WDR-MoS2从模拟透析液中去除尿毒症毒素的(f)Thomas和(g)Yoon-Nelson模型拟合曲线。
图4为样品的生物相容性测试图,其中(a)溶血率测定图像,插图为血液暴露在(A)蒸馏水,(B)活性炭(AC)和PBS溶液,(C)WDR-MoS2和PBS溶液中的照片;(b)凝血性质测定图像;(c)不同浓度范围材料的体外细胞毒性试验;(d)用活性炭孵育4T1细胞24小时后的活/死染色图像;(e)用WDR-MoS2孵育4T1细胞24小时后的活/死染色图像;(f)用顺铂孵育4T1细胞24小时后的活/死染色图像;(g)用活性炭孵育4T1细胞4小时后的流式细胞测试图像;(h)用WDR-MoS2孵育4T1细胞4小时后的流式细胞测试图像;(i)用顺铂孵育4T1细胞4小时后的流式细胞测试图像。
具体实施方式
实施例1
将(NH4)6Mo7O24·4H2O(1mmol)和CS(NH2)2(30mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在180℃反应24小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例2
将(NH4)6Mo7O24·4H2O(2mmol)和CS(NH2)2(60mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在180℃反应24小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例3
将(NH4)6Mo7O24·4H2O(3mmol)和CS(NH2)2(90mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在180℃反应24小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例4
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在180℃反应24小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
对实施例4制备的纳米硫化钼(标记为WDR-MoS2)的结构及性能表征结果如下:
(1)纳米硫化钼的结构分析:
透射电子显微镜(TEM)和高分辨率透射电子显微镜(HRTEM)图像表明,得到的WDR-MoS2具有二维超薄纳米片结构,层间距为基面存在大量缺陷(图1a和图1b)。相反,商用MoS2的TEM和HRTEM图像显示,其层间距较窄,为基面上没有缺陷(图1c和1d)。不同样品的X射线衍射(XRD)图表明,WDR-MoS2在低角度区域(9.3°和18.6°)出现了两个衍射峰,归因于(002)和(004)晶面,这两个衍射峰与其他制备的MoS2不同,进一步证明了其具有较宽的层间距。X射线光电子能谱(XPS)表明,我们所制备的样品为2H相MoS2(图1f,图1g和图1h)。通过TEM、XRD、XPS等综合分析,确定了WDR-MoS2具有宽层间距结构和丰富晶面缺陷,可以显著增加MoS2对尿毒症毒素的吸附位点。因此,WDR-MoS2有望改善尿毒症毒素的去除性能。
(2)纳米硫化钼对尿毒症毒素的吸附性能分析:
尿素吸附性能测定:分别取5,10,20,50,100mg WDR-MoS2吸附20mL尿素溶液(尿素溶液质量浓度为300mg/L),37℃,2h后测其浓度;取50mg WDR-MoS2吸附20mL尿素溶液(尿素溶液质量浓度为300mg/L),在不同时间t(5,10,20,30,40,50,60min)测其浓度,37℃;取50mg WDR-MoS2吸附20mL尿素溶液(尿素溶液质量浓度分别为90,120,150,180,210,240,270,300mg/L),分别在32℃,37℃和42℃,2h后测其浓度。
肌酐吸附性能测定:分别取5,10,20,50,100mg WDR-MoS2吸附30mL肌酐溶液(肌酐溶液的质量浓度为100mg/L),37℃,2h后测其浓度;取20mg WDR-MoS2吸附30mL肌酐溶液(肌酐溶液的质量浓度为100mg/L),在不同时间t(5,10,20,30,40,50,60min)测其浓度,37℃;取20mg WDR-MoS2吸附30mL肌酐溶液(肌酐溶液的质量浓度分别为50,60,70,80,100,120,140mg/L),分别在32℃,37℃和42℃,2h后测其浓度。
尿酸吸附性能测定:分别取5,10,20,50,100mg WDR-MoS2吸附20mL尿酸溶液(尿酸溶液的质量浓度为60mg/L),37℃,2h后测其浓度;取20mg WDR-MoS2吸附20mL尿酸溶液(尿酸溶液的质量浓度为60mg/L),在不同时间t(5,10,20,30,40,50,60min)测其浓度,37℃;取20mg WDR-MoS2吸附20mL尿酸溶液(尿酸溶液的质量浓度分别为10,20,30,40,50mg/L),分别在32℃,37℃和42℃,2h后测其浓度。
从模拟透析液中去除尿毒症毒素吸附性能测定:根据前述尿毒症从水溶液中的吸附实验,进行了模拟透析液中尿毒症毒素的剂量依赖性实验,除了实验溶剂为模拟透析液外,实验过程同水溶液中相同。
从人血清蛋白(HSA)溶液中去除尿毒症毒素吸附性能测定:根据前述尿毒症从水溶液中的吸附实验,除了实验溶剂为血清蛋白溶液外,实验过程同水溶液中相同。
模拟循环性质测定:通过简易再循环透析装置,分别测定1g WDR-MoS2对800mL尿素、肌酐和尿酸溶液和200mL模拟透析液的吸附性质。
生物相容性测定:对于溶血率测量,50mg WDR-MoS2加入200μL新鲜全血和800μLPBS溶液中(pH=7.4)在37℃下再孵育1小时。同样,在没有样品的情况下,用1mLPBS溶液和1mL去离子水孵育的200μL血液分别用作阴性和阳性对照。在4000rpm离心15分钟后,使用酶标仪在545nm处测量溶液的吸光度。
对于凝血性能测定,通过以4000rpm的速度将新鲜血液离心15分钟来采集贫血小板血浆(PPP)。然后,将1mLPPP与吸附剂(50mg)或不使用吸附剂(阴性对照)在37℃孵育30分钟,并以4000rpm离心10分钟。最后,使用自动血液分析仪测量APTT、PT和TT。
通过CCK-8测定法对小鼠乳腺癌细胞(4T1细胞)来测量材料的细胞毒性。分别将不同浓度(0-200μg mL-1)的顺铂、活性炭和WDR-MoS2浸入新鲜培养基中,细胞在37℃和5%CO2加湿气氛中培育24小时。加入CCK-8溶液并孵育0.5小时后,使用酶标仪在490nm处记录吸光度。实验重复三次。
对于活/死细胞染色,细胞首先在培养基中分别用顺铂、活性炭和WDR-MoS2(200μgmL-1)处理24小时。然后将细胞用碘化丙啶(4μM)和calcein-AM(2μM)染色0.5小时。最后,使用荧光显微镜测试图像。
对于细胞凋亡测定,使用FITC Annexin V细胞凋亡检测试剂盒测量。将4T1细胞与顺铂、活性炭和WDR-MoS2(200μg mL-1)一起孵4小时。使用流式细胞仪获取图像。
如图2a所示,在水溶液中随着吸附剂剂量的增加,AC和制备的MoS2对尿素的去除效率都有所提高。与AC、商用MoS2、DF-MoS2和DR-MoS2相比,相同吸附剂剂量下,WDR-MoS2对尿素的去除效率最高,这主要是由于吸附层间距的增大和缺陷的存在。对于肌酐和尿酸的吸附(图2b和图2c),WDR-MoS2在相同吸附剂剂量下也具有最高的去除效率,说明其在去除尿毒症毒素方面具有很大的潜力。随后,利用准一级和准二级模型研究了WDR-MoS2对尿毒症毒素的吸附动力学。可以看出,尿素吸附体系在10分钟内接近平衡,呈现出高效吸附过程,且模型符合准二级动力学方程,表明吸附过程为化学吸附,其吸附容量为59.2mg g-1,是常用活性炭吸附剂的3.4倍(图2d)。同样,肌酐和尿酸的吸附体系也呈现出高效的化学吸附过程,吸附容量分别为77.7和39.1mg g-1(图2e和图2f)。图2g为采用Langmuir和Freundlich模型在不同温度(32℃,37℃,42℃)下WDR-MoS2对尿素的吸附等温线。随着温度的升高,WDR-MoS2对尿素的吸附量增大,证明其吸附过程为吸热反应,且符合Freundlich模型,属于多分子层吸附。同样,WDR-MoS2对肌酐和尿酸的吸附为多分子层的吸附,吸附过程为吸热反应。
图3a为不同剂量WDR-MoS2在模拟透析液中对尿毒症毒素去除效率图。与水溶液相比,WDR-MoS2即使在电解质阳离子存在的情况下,对尿素、肌酐和尿酸的去除效率也相对较高。图3b为不同剂量WDR-MoS2在人血清蛋白(HSA)溶液中对尿毒症毒素去除效率图。与水溶液相比,HSA溶液对尿毒的去除效率没有明显变化,说明拓宽MoS2层间距可以有效阻断蛋白质分子在吸附尿毒过程中的影响。如图3c所示,我们采用自制简易装置对动态条件下的吸附尿毒症毒素进行了研究。结果表明,在水溶液中WDR-MoS2对尿素、肌酐和尿酸的吸附容量分别为38.4、53.1和23.1mg g-1,突破体积分别为38.3、106.9和77.2mL(图3d和图3e)。在模拟透析溶液中WDR-MoS2对尿素、肌酐和尿酸的吸附容量分别为18.4、20.4和9.3mg g-1,突破体积分别为18.3、61.3和46.3mL(图3f和图3g)。上述结果表明,WDR-MoS2在动态条件下具有良好的吸附性能。
(3)纳米硫化钼的生物相容性测试:
图4a为样品的溶血测定图,结果表明WDR-MoS2的存在不会导致任何溶血作用。如图4b所示,通过活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)来测试样品的凝血效果。结果表明,与对照组和AC相比,WDR-MoS2具有最长的APTT,说明其抗凝活性增强是由于其固有凝血级联的作用。但PT值之间无显著差异,表明外源性凝血级联无变化。此外,WDR-MoS2延长的TT显示纤维蛋白原向纤维蛋白的低转化,这在材料炎症反应中起着关键作用。上述结果表明,WDR-MoS2具有良好的血液相容性。
如图4c所示,AC和WDR-MoS2在不同浓度范围(0-200μg mL-1)均表现出较高的细胞存活率,说明AC和WDR-MoS2对4T1细胞无影响。与AC和WDR-MoS2相比,阳性对照材料(顺铂)随着浓度的增加诱导细胞存活率显著降低。图4d-f显示了用不同的材料(200μg mL-1)培养细胞24小时后的荧光显微镜活/死染色图像,进一步证实AC和WDR-MoS2不会破坏细胞活力。此外,如图4g-i所示,对不同材料进行了早期凋亡诱导研究,可以看出,AC和WDR-MoS2均不诱导早期凋亡,而顺铂则显著增加了早期凋亡细胞。总之,上述结果表明,WDR-MoS2具有应用于涉及血液接触的应用的潜力。
实施例5
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在140℃反应24小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例6
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在160℃反应24小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例7
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在200℃反应24小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例8
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在220℃反应24小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例9
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在180℃反应12小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例10
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在180℃反应18小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例11
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在180℃反应24小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例12
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在180℃反应30小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例13
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在180℃反应36小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
实施例14
将(NH4)6Mo7O24·4H2O(6mmol)和CS(NH2)2(180mmol)添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在180℃反应48小时。产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时。
表一 水溶液中纳米硫化钼对尿毒症毒素的吸附容量
表一说明通过调节反应起始物比例,反应温度及反应时间可以有效地提高WDR-MoS2对尿毒症毒素的吸附性能。
显然,上述实施例仅仅是为清楚地说明所作的举例,而并非对实施方式的限定。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
Claims (7)
2.根据权利要求1所述的纳米硫化钼,其特征在于,所述(NH4)6Mo7O24·4H2O和CS(NH2)2的摩尔比为1:30。
3.根据权利要求1所述的纳米硫化钼,其特征在于,混合物搅拌均匀所用时间为30min。
4.根据权利要求1所述的纳米硫化钼,其特征在于,真空干燥的温度为60℃,时间为24小时。
5.一种纳米硫化钼作为尿毒症毒素的高效吸附剂的应用,所述纳米硫化钼是具有宽层间距和丰富晶格缺陷的纳米硫化钼。
7.根据权利要求5所述的应用,其特征在于,所述纳米硫化钼是由下述方法制备得到:
按照(NH4)6Mo7O24·4H2O和CS(NH2)2的摩尔比为1:30将(NH4)6Mo7O24·4H2O和CS(NH2)2添加到225mL蒸馏水中,混合物搅拌30分钟后转移至300mL聚四氟乙烯高压反应釜中,在140-220℃反应12-48小时,产物经乙醇和蒸馏水洗涤后,在60℃真空干燥24小时,得到具有宽层间距和丰富晶格缺陷的纳米硫化钼。
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