CN116785446A - 一种CeO2/Fe3O4杂化多孔碳纳米颗粒及其制备 - Google Patents
一种CeO2/Fe3O4杂化多孔碳纳米颗粒及其制备 Download PDFInfo
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Abstract
本发明公开一种CeO2/Fe3O4杂化多孔碳纳米颗粒及其制备,属于纳米酶技术领域。该纳米颗粒的制备过程包括:先以水热法制备得到的CeO2/Fe3O4纳米微粒作为核结构;再利用表面修饰剂对核材料进行亲水修饰;接着制备出核壳结构的金属有机框架复合物;最后以其作为前驱物进行热解处理得到终产物;所得纳米材料可用于肿瘤的特异性治疗中,其在肿瘤微环境下,可自发产生活性氧物质,可用于肿瘤的化学动力治疗中;同时,该纳米颗粒还能够有效吸收全波段近红外光并将其转化热能,产生热效应,杀死细胞,实现PTT治疗;本申请制备的纳米颗粒可用于肿瘤的联合治疗中,这为开发更多低副作用、高治疗效率的联合治疗系统提供了新的思路。
Description
技术领域
本发明涉及纳米酶技术领域,具体涉及一种CeO2/Fe3O4杂化多孔碳纳米颗粒及其制备。
背景技术
当前针对肿瘤的临床治疗方法主要有手术切除、化学治疗、放射治疗等。但是这些方法普遍存在治疗效率低、副作用大、易复发和易转移等问题。由于单一的肿瘤治疗手段在治疗中表现出较大的局限性,所以将两种或两种以上的治疗方法结合进行联合治疗的这种疗效更优的手段受到越来越多的关注。到目前为止,关于联合治疗的研究已有诸多报道,且多数都显示出良好的抗肿瘤疗效。例如,化疗联合光疗可以提高光疗的疗效;光疗联合免疫治疗不仅可以杀死原发性肿瘤,而且可以抑制转移性肿瘤。但是由于这些治疗方式对正常组织都有一定程度的损伤,联合治疗过程中这一现象往往表现得更为明显,因此开发高治疗效率且低副作用的联合治疗系统才是当前亟需解决的问题。
化学动力疗法(Chemodynamic Therapy,CDT)是以金属基纳米材料为催化剂,利用芬顿化学或类芬顿化学将肿瘤细胞内源性H2O2转化为活性氧(·OH)以治疗癌症的新兴策略。CDT的反应特征与肿瘤微环境中的弱酸性、H2O2过表达特征相契合,可实现肿瘤微环境特异性激活治疗,在一定程度上确保了正常组织的安全,具有逻辑性强,选择性佳和生物安全性高等优点。
肿瘤热疗(Photothermal therapy,PTT)是继手术、放疗、化疗和免疫疗法之后的第五大疗法,是一种绿色治疗手段。其原理是利用物理方法将组织加热到能杀灭肿瘤细胞的温度(42.5℃~43.5℃)、并持续60~120分钟,达到既破坏肿瘤细胞又不损伤正常组织(正常组织细胞的温度安全界限为45℃±1℃)效果的一种方法。热疗不仅对肿瘤细胞有直接的细胞毒效应,还可以增强化疗、放疗的疗效,提高机体的免疫力,抑制肿瘤的转移。大量体外实验和临床资料显示,肿瘤热疗虽然不能取代手术、化疗或放疗作为一种独立的肿瘤治疗方案,但它对于化疗、放疗和手术等肿瘤治疗手段具有明显增效和补充作用。正因为如此,肿瘤热疗近年来发展迅速,成为继手术、放疗、化疗和生物治疗之后又一重要的肿瘤治疗手段。
在现有的热疗技术中,比较常见和主流的是非侵入式的纳米光热热疗,通过特定波长的近红外光照射具有光热转换特性的材料,将光能转化为热能,使该区域的温度升高到42℃以上进而杀死肿瘤细胞的一种疗法,光热疗法作为一种切实有效的低风险肿瘤治疗技术,具有良好的发展前景。
纳米二氧化铈(CeO2)是非常重要的纳米稀土氧化物,随着CeO2的粒度缩小到100nm以下,由于其具有高的表面积与体积比,颗粒表面存在氧空缺,铈原子的电子轨道结构发生改变,+4价和+3价可以互相转化,从而使其具有氧化还原的双重特性。由于其混合价态不稳定而具有抗氧化的作用,具有超氧化物歧化酶、过氧化氢酶模拟活性,且其催化速率常数超过超氧化物歧化酶,在生物医学领域具有较好的应用价值和前途。Fe3O4具有过氧化酶的催化活性,这种无机材料模拟酶与天然酶相比,具有耐温、耐酸碱、易大量制备、成本低、贮存稳定等优异特性。基于这两种氧化物的特性,如能将这两种材料有效结合并成功应用于肿瘤的联合治疗中将有望为肿瘤治疗提供新的思路。
中国专利CN 107445212 B公开一种磁性Fe3O4@CeO2复合纳米微粒的制备方法及其应用,其将CeO2包覆在Fe3O4纳米粒子表面,通过两者过氧化酶的协同机制,得到具有更高的酶催化活性的复合纳米粒子,进一步提高对H2O2的检测灵敏度,该文件公开的复合纳米粒子综合利用了Fe3O4和CeO2的独有活性,但是其制得的复合材料主要用于检测低浓度的过氧化氢和葡萄糖,其并未指出该材料可用于肿瘤的联合治疗中。此类材料在肿瘤联合治疗中的应用还有待进一步探索。
发明内容
本发明的目的在于解决现有技术中存在的问题,提供一种CeO2/Fe3O4杂化多孔碳纳米颗粒及其制备,该纳米颗粒对过氧化氢具有高度的特异性和敏感性。利用这种纳米材料,可以同时实现CDT(化学动力疗法)和PTT(光热疗法),以达到良好的肿瘤治疗效果。
为了实现上述技术目的,本发明是通过以下技术方案来实现的:一种CeO2/Fe3O4杂化多孔碳纳米颗粒的制备方法,包括如下步骤:
1)将铈盐和铁盐溶于水中,加入碱液,室温搅拌,热处理,反应结束后,离心分离沉淀,洗涤沉淀至中性,冻干得CeO2/Fe3O4纳米颗粒;
2)超声辅助下,将冻干粉末分散于表面修饰剂中进行亲水性修饰,得到分散性好的CeO2/Fe3O4纳米微粒;
3)将纳米颗粒的分散液加入含有2-甲基咪唑和六水合硝酸锌的甲醇溶液中,搅拌,静置反应,离心分离沉淀,烘干;
4)在惰性氛围下,高温碳化烘干后的样品,得CeO2/Fe3O4杂化多孔碳纳米颗粒。
进一步地,步骤1)中,所用铈盐为硝酸铈、醋酸铈、氯化铈中的一种或多种;铁盐为氯化亚铁、氯化铁、硝酸铁中的一种或多种,所用碱液为氢氧化钠溶液。
进一步地,步骤1)中,进行热处理时的温度为90~120℃,反应时间2~10h。
进一步地,步骤2)中,表面修饰剂为聚丙烯酰胺·盐酸、聚乙烯亚胺、CTAB(十六烷基三甲基溴化铵)、PVP、壳聚糖中的一种或多种,表面修饰剂与CeO2/Fe3O4纳米微粒的质量比为0.1~1:1。
进一步地,步骤3)中,甲醇溶液中2-甲基咪唑和六水合硝酸锌的质量比为1:1~2;纳米颗粒的分散液与2-甲基咪唑和六水合硝酸锌的甲醇溶液的体积比为1:50~100。
进一步地,步骤4)中高温碳化的温度为600~800℃、加热时间为2~4h。
利用上述方法制备的CeO2/Fe3O4杂化多孔碳纳米颗粒呈现均匀的十二面体结构,粒径在80~100nm;该纳米颗粒是以CeO2/Fe3O4纳米微粒为核的核壳结构的金属有机框架复合物。
本发明的有益效果为:
1、本申请公开的CeO2/Fe3O4杂化多孔碳纳米颗粒是以CeO2/Fe3O4纳米微粒为核、亲水处理并制备出核壳结构的金属有机框架复合物后、以其作为前驱物进行热解处理得到的;所得纳米材料可用于肿瘤的特异性治疗中,其在肿瘤微环境下,可自发产生活性氧物质,因此可作为纳米药物制剂用于肿瘤的化学动力治疗中;同时,该纳米颗粒还能够有效吸收全波段近红外光并将其转化为热能,产生热效应,杀死细胞,可用于肿瘤的光热治疗中;因此本申请制备的纳米颗粒可用于肿瘤的联合治疗中,这为开发更多高治疗效率且低副作用的联合治疗系统提供了新的思路;
2、本申请公开的CeO2/Fe3O4杂化多孔碳纳米颗粒在正常的生理条件下无法产生活性氧物质,因此对正常细胞无毒性,有效减少肿瘤治疗过程中对正常组织的伤害,可实现对肿瘤的特异性治疗;
3、本申请公开的CeO2/Fe3O4杂化多孔碳纳米颗粒的制备方法简单,制备周期短,且所用原料来源广泛,易于获取,成本可控,这为进行大规模的推广使用提供了可能。
附图说明
图1是实施例1制备的CeO2/Fe3O4杂化多孔碳纳米颗粒的透射电子显微镜图谱;
图2是实施例1制备的CeO2/Fe3O4杂化多孔碳纳米颗粒的XRD谱图;
图3是实施例1制备的CeO2/Fe3O4杂化多孔碳纳米颗粒在H2O2条件下产生的化学动力效应图;
图4是实施例1制备的CeO2/Fe3O4杂化多孔碳纳米颗粒的紫外吸收光谱图;
图5是实施例1制备的CeO2/Fe3O4杂化多孔碳纳米颗粒在光照下产生的光热效应图;
图6是实施例1制备的CeO2/Fe3O4杂化多孔碳纳米颗粒在正常细胞3T3中的活死图。
具体实施方式
以下实施例进一步说明本发明的内容,但不应理解为对本发明的限制。在不背离本发明实质的情况下,对本发明方法、步骤或条件所作的修改和替换,均属于本发明的范围。
实施例1、CeO2/Fe3O4杂化多孔碳纳米颗粒的制备
(1)将0.4g CeCl3·7H2O,29.8mg FeCl3和18.2mg FeCl2·H2O溶解在15mL H2O中,加入20mL的氢氧化钠(质量浓度为15mol/L)溶液,并于室温下搅拌两小时,将所得溶液装入高压反应釜中,在115℃的烘箱中反应2小时,待反应结束后,离心分离沉淀,用超纯水洗涤沉淀至中性,冻干处理;
(2)将1mg冻干后的粉末样品分散于1mL聚丙烯酰胺·盐酸(1mg/mL)水溶液中,超声分散30分钟得淡黄色溶液;
(3)将淡黄色溶液按1:100的体积比加入含有2.58g 2-甲基咪唑和2.63g Zn(NO3)2·6H2O的400mL甲醇溶液中,常温搅拌20分钟,静置反应24h,待反应结束后,离心分离沉淀,在真空干燥箱中烘干得白色粉末;
(4)收集烘干的样品,置于高温烘箱中,在氮气保护下、于800℃加热2-4h,收集得到终产物。
针对本实施例所得的CeO2/Fe3O4杂化多孔炭纳米颗粒样品,通过透射电子显微镜对其进行微观上的形态分析。结果如图1所示,从图中可以观察出,所得样品呈现均匀的十二面体结构,粒径在80-100nm。
图2为所得CeO2/Fe3O4杂化多孔炭纳米颗粒的XRD谱图,从图中可以看出CeO2、Fe3O4的特征峰,说明CeO2/Fe3O4纳米颗粒被顺利包覆在金属有机框架里。
对所得CeO2/Fe3O4杂化多孔炭纳米颗粒进行化学动力效应测试:在pH=5的条件下,将1mM的TMB(3,3',5,5'-四甲基联苯胺)、160μM的H2O2和本实施例制备的纳米复合材料混合(0.02mg/mL),观察TMB的变色情况。结果如图3所示,在纳米复合材料的催化下,TMB与H2O2反应生成了在652nm处有吸收的蓝色TMB二聚体,说明纳米颗粒具有化学动力学效应。
对所得CeO2/Fe3O4杂化多孔炭纳米颗粒进行光热效应测试,将纳米复合材料制备成100μg/mL的水溶液,分别以1W/cm2的808nm和1064nm的近红外激光照射6分钟,结果如图5所示,从图中可以看出,在波长为808nm的近红外光照射下,升温达到60℃,在波长为1064nm的近红外光照射下,升温达到70℃,说明该纳米复合材料具有良好的光热特性。
对所得CeO2/Fe3O4杂化多孔炭纳米颗粒进行MTT测试,用DMEM高糖培养基将纳米复合材料制备成不同浓度的溶液(0μg/mL、2μg/mL、4μg/mL、8μg/mL、16μg/mL、32μg/mL、64μg/mL),分别与培养好的3T3细胞共培养24小时,结果如图6所示,从图中可以看出,3T3细胞存活率在85%以上,说明该纳米复合材料对正常细胞无毒性,有效减少肿瘤治疗过程中对正常组织的伤害,可实现对肿瘤的特异性治疗。
实施例2、CeO2/Fe3O4杂化多孔碳纳米颗粒的制备
(1)将0.4g CeCl3·7H2O,29.8mg FeCl3和18.2mg FeCl2·H2O溶解在15mL H2O中,加入20mL的氢氧化钠(质量浓度15mol/L)溶液,并于室温下搅拌两小时,将所得溶液装入高压反应釜,在115℃的烘箱中反应2小时,待反应结束后,离心分离沉淀,用超纯水洗涤沉淀至中性,冻干处理;
(2)将1mg冻干后的粉末样品分散于1mL壳聚糖(1mg/mL)水溶液中,超声分散30分钟得淡黄色溶液;
(3)将淡黄色溶液按1:100的体积比加入含有2.58g 2-甲基咪唑和2.63g Zn(NO3)2·6H2O的400mL甲醇溶液中,常温搅拌20分钟,静置反应24h,待反应结束后,离心分离沉淀,在真空干燥箱中烘干得白色粉末;
(4)收集烘干的样品,置于高温烘箱中,在氮气保护下、于800℃加热2-4h,收集得到终产物。
实施例3、CeO2/Fe3O4杂化多孔碳纳米颗粒的制备
(1)将0.4g CeCl3·7H2O,29.8mg FeCl3和18.2mg FeCl2·H2O溶解在15mL H2O中,加入20mL的氢氧化钠(质量浓度15mol/L)溶液,并于室温下搅拌两小时,将所得溶液装入高压反应釜中,在120℃的烘箱中反应2小时,待反应结束后,离心分离沉淀,用超纯水洗涤沉淀至中性,冻干处理;
(2)将1mg冻干后的粉末样品分散于1mL聚乙烯吡咯烷酮(PVP,1mg/mL)水溶液中,超声分散30分钟得淡黄色溶液;
(3)将淡黄色溶液按1:100的体积比加入含有2.58g 2-甲基咪唑和2.63g Zn(NO3)2·6H2O的400mL甲醇溶液中,常温搅拌20分钟,静置反应24h,待反应结束后,离心分离沉淀,在真空干燥箱中烘干得白色粉末;
(4)收集烘干的样品,置于高温烘箱中,在氮气保护下、于800℃加热2-4h,收集得到终产物。
实施例4、CeO2/Fe3O4杂化多孔碳纳米颗粒的制备
(1)将0.52g Ce(NO3)3,29.8mg FeCl3和18.2mg FeCl2·H2O溶解在15mL H2O中,加入20mL的氢氧化钠(质量浓度为15mol/L)溶液,并于室温下搅拌两小时,将所得溶液装入高压反应釜中,在90℃的烘箱中反应8小时,待反应结束后,离心分离沉淀,用超纯水洗涤沉淀至中性,冻干处理;
(2)将1mg冻干后的粉末样品分散于1mL聚丙烯酰胺·盐酸(1mg/mL)水溶液中,超声分散30分钟得淡黄色溶液;
(3)将淡黄色溶液按1:100的体积比加入含有2.58g 2-甲基咪唑和2.63g Zn(NO3)2·6H2O的400mL甲醇溶液中,常温搅拌20分钟,静置反应24h,待反应结束后,离心分离沉淀,在真空干燥箱中烘干得白色粉末;
(4)收集烘干的样品,置于高温烘箱中,在氮气保护下、于800℃加热2-4h,收集得到终产物。
实施例5、CeO2/Fe3O4杂化多孔碳纳米颗粒的制备
(1)将0.52g Ce(NO3)3,0.1g Fe(NO3)3·9H2O溶解在15mL H2O中,加入20mL的氢氧化钠(质量浓度15mol/L)溶液,并于室温下搅拌两小时,将所得溶液装入高压反应釜中,在115℃的烘箱中反应2小时,待反应结束后,离心分离沉淀,用超纯水洗涤沉淀至中性,冻干处理;
(2)将1mg冻干后的粉末样品分散于1mL聚丙烯酰胺·盐酸(1mg/mL)水溶液中,超声分散30分钟得淡黄色溶液;
(3)将淡黄色溶液按1:100的体积比加入到含有2.58g 2-甲基咪唑和2.63g Zn(NO3)2·6H2O的400mL甲醇溶液中常温搅拌20分钟,静置反应24h,待反应结束后,离心分离沉淀,在真空干燥箱中烘干得白色粉末;
(4)收集烘干的样品,置于高温烘箱中,在氮气保护下、于800℃加热2-4h,收集得到终产物。
以上显示和描述了本发明的基本原理、主要特征及优点。但是以上所述仅为本发明的具体实施例,本发明的技术特征并不局限于此,任何本领域的技术人员在不脱离本发明的技术方案下得出的其他实施方式均应涵盖在本发明的专利范围之中。
Claims (8)
1.一种CeO2/Fe3O4杂化多孔碳纳米颗粒的制备方法,其特征在于,包括如下步骤:
1)将铈盐和铁盐溶于水中,加入碱液,室温搅拌,热处理,反应结束后,离心分离沉淀,洗涤沉淀至中性,冻干得CeO2/Fe3O4纳米颗粒;
2)超声辅助下,将冻干粉末分散于表面修饰剂中进行亲水性修饰,得到分散性好的CeO2/Fe3O4纳米微粒;
3)将纳米颗粒的分散液加入含有2-甲基咪唑和六水合硝酸锌的甲醇溶液中,搅拌,静置反应,离心分离沉淀,烘干;
4)在惰性氛围下,高温碳化烘干后的样品,得CeO2/Fe3O4杂化多孔碳纳米颗粒。
2.如权利要求1所述的一种CeO2/Fe3O4杂化多孔碳纳米颗粒的制备方法,其特征在于,步骤1)中,所用铈盐为硝酸铈、醋酸铈、氯化铈中的一种或多种;铁盐为氯化亚铁、氯化铁、硝酸铁中的一种或多种,所用碱液为氢氧化钠溶液。
3.如权利要求1所述的一种CeO2/Fe3O4杂化多孔碳纳米颗粒的制备方法,其特征在于,步骤1)中,进行热处理时的温度为90~120℃,反应时间2~10h。
4.如权利要求1所述的一种CeO2/Fe3O4杂化多孔碳纳米颗粒的制备方法,其特征在于,步骤2)中,表面修饰剂为聚丙烯酰胺·盐酸、聚乙烯亚胺、CTAB PVP、壳聚糖中的一种或多种,表面修饰剂与CeO2/Fe3O4纳米微粒的质量比为0.1~1:1。
5.如权利要求1所述的一种CeO2/Fe3O4杂化多孔碳纳米颗粒的制备方法,其特征在于,步骤3)中,甲醇溶液中2-甲基咪唑和六水合硝酸锌的质量比为1:1~2;纳米颗粒的分散液与2-甲基咪唑和六水合硝酸锌的甲醇溶液的体积比为1:50~100。
6.如权利要求1所述的一种CeO2/Fe3O4杂化多孔碳纳米颗粒的制备方法,其特征在于,步骤4)中高温碳化的温度为600~800℃、加热时间为2~4h。
7.一种CeO2/Fe3O4杂化多孔碳纳米颗粒,其特征在于,其是根据权利要求1-6中任一项所述的一种CeO2/Fe3O4杂化多孔碳纳米颗粒的制备方法制备得到的,该纳米颗粒呈现均匀的十二面体结构,粒径在80~100nm。
8.如权利要求7所述的一种CeO2/Fe3O4杂化多孔碳纳米颗粒,其特征在于,该纳米颗粒是以CeO2/Fe3O4纳米微粒为核的核壳结构的金属有机框架复合物。
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