CN106381710A - 一种实现pH控释、NIR控释以及光监控药物释放的纳米纤维的制备方法 - Google Patents

一种实现pH控释、NIR控释以及光监控药物释放的纳米纤维的制备方法 Download PDF

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CN106381710A
CN106381710A CN201610858869.9A CN201610858869A CN106381710A CN 106381710 A CN106381710 A CN 106381710A CN 201610858869 A CN201610858869 A CN 201610858869A CN 106381710 A CN106381710 A CN 106381710A
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李翔
刘恒
韩高荣
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Zhejiang University ZJU
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Abstract

本发明公开了一种具有光监控、pH控释和近红外光控释的局部给药系统。此方法主要是将静电纺丝技术和表面改性技术相结合,极大的提高了药物载体的药物加载能力,并且在酸性环境中以及近红外光(808nm)照射下能够促进药物缓释,具有pH控释和近红外光控释的效果。此外,在980nm近红外光照射下,随着药物释放量的改变,其红绿光比会逐渐变化,因此可以通过光学信号的变化检测药物释放过程。本发明制备方法简单,原料成本低廉,整个制备过程在空气气氛中进行,无需特殊装置。

Description

一种实现pH控释、NIR控释以及光监控药物释放的纳米纤维的 制备方法
技术领域
本发明涉及一种通过pH和NIR控制药物释放速率,并能够通过光学信号监控药物释放的纳米纤维的制备方法。
背景技术
化疗目前是目前临床治疗癌症的主要手段之一。可植入型局部纳米给药系统由于能够有效控制药物缓释并在较长时间内保证血药浓度,避免重复给药,因而得到广泛研究。
利用外场响应实现药物控释是近年来研究热点。通过各种外场刺激,控制药物在患者体内释放进而得到更好的治疗效果。常见的外场信号包括pH值、温度、光、磁等等。抗癌药物在人体内的释放和剩余量对于治疗过程是及其重要的,由于个体的不同,其对药物的吸收、分布、代谢也会不同,不同患者用药后血液中紫杉醇浓度可相差4倍左右,这会对患者用药产生一定困扰,因而药物载体的药物释放动力学体内外监控具有重要意义,目前药物检测系统多以纳米载体的发光强度的变化作为追踪信号。为实现个性化治疗,同一体系中实现药物控释及药物释放实时监测显得尤为重要。我们以Yb、Er双掺CaTiO3纳米纤维为基体,通过表面改性,成功实现了pH控释、NIR控释与光监控药物释放,在药物载体领域具有广阔应用前景。
发明内容
本发明基于现有技术的不足,设计了一种可以通过pH和NIR控制药物释放速率,并能够通过光学信号监控药物释放的纳米纤维的制备方法。
本发明的目的是通过以下技术方案实现的,包括以下步骤:一种实现pH控释、NIR控释以及光监控药物释放的纳米纤维的制备方法,包括以下步骤:
(1)将2ml醋酸、6ml乙醇及0.8169g钛酸四丁酯混合搅拌,在搅拌过程中加入0.5ml乙酰丙酮,混合搅拌0.5-1h,得到溶液A;
(2)将0.5668g四水硝酸钙加入溶液A中,搅拌1h得到溶液B;
(3)将0.215g五水硝酸铒和1.94g五水硝酸镱溶于20ml N-N二甲基酰胺中,搅拌1-2h,得到溶液C;
(4)将2ml N-N二甲基酰胺与1ml C溶液加入到步骤2配制的溶液B中并搅拌0.5h,之后加入0.39-0.78g聚乙烯吡咯烷酮,搅拌6-12h后得到静电纺丝液;
(5)用注射器吸取静电纺丝液,以接地的铁丝网作为接收装置进行静电纺丝,其中,流速为0.5ml/h,,电压控制在10-12kV,得到的纤维在80℃下烘干2h后经600-800℃烧结,得到Yb、Er双掺CaTiO3纳米纤维;
(6)取100mg Yb、Er双掺CaTiO3纳米纤维,溶于80ml浓度为5mM的NaOH溶液中搅拌12h,离心干燥后溶于40ml DMF中,加入500μl 3-氨基丙基三乙氧基硅烷,于室温(25℃)下搅拌24h,离心干燥;
(7)取30mg上述样品加入到10ml聚丙烯酸中,搅拌12h,离心并干燥,得到改性的双掺CaTiO3纳米纤维。
进一步地,所述步骤5中,烧结温度为700℃
本发明的有益效果在于:本发明制备方法简单,原料成本低廉,整个制备过程在空气气氛中进行,无需特殊装置,所需设备简单。本发明首次通过溶胶凝胶-静电纺丝方法制备出了可以通过pH和NIR控制药物释放速率,并能够通过光学信号监控药物释放的钛酸钙纳米纤维。在酸性环境中,由于聚丙烯酸分子羧基去质子化,导致聚丙烯酸分子与药物分子之间的静电链接减弱,进而加速药物的释放。在近红外光照射下,局部温度升高,导致聚丙烯酸高分子链震动增快,促进药物释放。同时,在药物释放的过程中,由于药物分子的释放,使得上转换发光的红绿光比逐渐减弱。
附图说明
图1a为CTO纳米纤维扫描电镜表征,b为CTO纳米纤维透射电镜表征。
图2为不同温度下烧结获得的Yb、Er双掺CaTiO3纳米纤维具有不同的物象(a)及上转换过发光强度(b)。
图3为不同pH不同pH值下释放曲线图。
图4为不同NIR照射下的释放曲线图。
图5为不同NIR照射下的细胞活性检测(MTT)。
图6为pH=5.2的条件下的绿红光光比曲线图。
实施例1
(1)将2ml醋酸、6ml乙醇及0.8169g钛酸四丁酯混合搅拌,在搅拌过程中加入0.5ml乙酰丙酮,混合搅拌1h,得到溶液A;
(2)将0.5668g四水硝酸钙加入溶液A中,搅拌1h得到溶液B;
(3)将0.215g五水硝酸铒和1.94g五水硝酸镱溶于20ml N-N二甲基酰胺中,搅拌1-2h,得到溶液C;
(4)将2ml N-N二甲基酰胺与1ml C溶液加入到步骤2配制的溶液B中并搅拌0.5h,之后加入0.78g聚乙烯吡咯烷酮,搅拌12h后得到静电纺丝液;
(5)用注射器吸取静电纺丝液,以接地的铁丝网作为接收装置进行静电纺丝,其中,流速为0.5ml/h,,电压控制在12kV,得到的纤维在80℃下烘干2h后经600-700℃烧结,得到Yb、Er双掺CaTiO3纳米纤维;
(6)取100mg Yb、Er双掺CaTiO3纳米纤维,溶于80ml浓度为 5mM的NaOH溶液中搅拌12h,离心干燥后溶于40ml DMF中,加入500μl 3-氨基丙基三乙氧基硅烷,于室温(25℃)下搅拌24h,离心干燥;
(7)取30mg上述样品加入到10ml聚丙烯酸中,搅拌12h,离心并干燥,得到改性的双掺CaTiO3纳米纤维(CTO纳米纤维)。
图1a为CTO纳米纤维扫描电镜表征,CTO纳米纤维直径在180-250nm之间;b为CTO纳米纤维透射电镜表征,纤维表面呈酥松多孔状,极大的增加了其比表面积,更有利于药物的加载。
图2不同温度下烧结获得的Yb、Er双掺CaTiO3纳米纤维具有不同的物象及上转换过发光强度。(a)由XRD可知,700℃烧结得到的样品对比600℃下烧结得到的样品结晶峰更高,证明样品晶格结构更加完善,结晶性能更好,而800℃烧结样品开始出现Yb2O3杂相;(b)对比600℃与700℃样品,发现700℃下烧结样品具有更好的发光性能,因此我们选用700℃做为最佳烧结温度。
取上述样品30mg于室温下在20μg/mL的DOX溶液中搅拌24h,离心清洗并干燥。置于pH分别为7.4,6.8和5.2的磷酸缓冲盐溶液中,分别检测其药物的释放,如图3所示;由图可知,不同pH值下释放速率不同,并且都能够有效抑制药物的突释,pH为5.2时,在前10h药物释放速率较快,10h的时候释放量达到33%,之后释放速率减缓,100h时释放量可达64%,较之于pH为6.8和5.2的释放曲线可知,低pH环境能够促进药物的释放,进而验证了样品具有pH控释的功能。在酸性环境中,药物释放速率加快,而人体癌症部位整体环境为酸性。样品之所以能够实现药物的pH控释,是因为在酸性环境中,聚丙烯酸分子羧基去质子化,导致聚丙烯酸分子与药物分子之间的静电链接减弱,进而加速药物的释放。
图4为NIR控释药物试验结果图,在不同pH环境下,通过紫外分光光度计来进行药物释放量的检测。从图中可以看出,近红外光照射后,经过PAA改性的样品的药物释放量有了明显提高。在pH=7.4的环境下,经NIR照射的样品在100h后释放量可达到25%,而不经NIR照射的样品释放量只有14%;在pH=5.2的环境下,经NIR照射的样品在100h后释放量可达到64%,而不经NIR照射的样品释放量只有51%。这是因为在近红外光照射下,样品局部温度升高,导致聚丙烯酸高分子链震动增快,进而促进药物释放
进一步通过细胞活性检测(MTT)验证了改性的双掺CaTiO3纳米纤维的NIR控释,如图5所示。取PAA改性之后的样品分为A、B、C三组。A组不加载药物,只加以NIR照射;B组不加以NIR照射,只加载药物;C组既加载药物,又加以NIR照射。利用肝癌细胞对其进行细胞活性实验。通过对比0至48h内不同实验条件下样品的生物活性。只加以NIR照射,在48h内,细胞仍然具有较高的生存率;只加载药物而不加以NIR照射,细胞存活率下降;既加载药物又加以NIR照射,细胞的存活率最低。由此可以验证,NIR照射能够有效促进药物的释放。
图6为药物释放过程中的绿红光光比曲线图,在pH=5.2的条件下进行实验,表现为随药物释放绿红光光比增加。这是因为药物的释放量不断增加,药物DOX的吸收峰为420-550nm,由于荧光共振能量转移效应,在加载药物的时候,绿光(550nm)会被淬灭,随着药物的释放,绿光逐渐恢复。由此可知,通过本发明改性的双掺CaTiO3纳米纤维可以通过绿红光光比来实现对药物释放的实时监控。
实施例2
(1)将2ml醋酸、6ml乙醇及0.8169g钛酸四丁酯混合搅拌,在搅拌过程中加入0.5ml乙酰丙酮,混合搅拌1h,得到溶液A;
(2)将0.5668g四水硝酸钙加入溶液A中,搅拌1h得到溶液B;
(3)将0.215g五水硝酸铒和1.94g五水硝酸镱溶于20ml N-N二甲基酰胺中,搅拌2h,得到溶液C;
(4)将2ml N-N二甲基酰胺与1ml C溶液加入到步骤2配制的溶液B中并搅拌0.5h,之后加入0.78g聚乙烯吡咯烷酮,搅拌12h后得到静电纺丝液;
(5)用注射器吸取静电纺丝液,以接地的铁丝网作为接收装置进行静电纺丝,其中,流速为0.5ml/h,,电压控制在10-12kV,得到的纤维在80℃下烘干2h后经800℃烧结,得到Yb、Er双掺CaTiO3纳米纤维;
(6)取100mg Yb、Er双掺CaTiO3纳米纤维,溶于80ml浓度为 5mM的NaOH溶液中搅拌12h,离心干燥后溶于40ml DMF中,加入500μl 3-氨基丙基三乙氧基硅烷,于室温(25℃)下搅拌24h,离心干燥;
(7)取30mg上述样品加入到10ml聚丙烯酸中,搅拌12h,离心并干燥,得到改性的双掺CaTiO3纳米纤维。
经测试,本实施例制备得到的改性的双掺CaTiO3纳米纤维同样具有较好的pH控释、NIR控释功能,且可以通过绿红光光比来实现对药物释放的实时监控。

Claims (2)

1.一种实现pH控释、NIR控释以及光监控药物释放的纳米纤维的制备方法,其特征在于,包括以下步骤:
(1)将2ml醋酸、6ml乙醇及0.8169g钛酸四丁酯混合搅拌,在搅拌过程中加入0.5ml乙酰丙酮,混合搅拌0.5-1h,得到溶液A;
(2)将0.5668g四水硝酸钙加入溶液A中,搅拌1h得到溶液B;
(3)将0.215g五水硝酸铒和1.94g五水硝酸镱溶于20ml N-N二甲基酰胺中,搅拌1-2h,得到溶液C;
(4)将2ml N-N二甲基酰胺与1ml C溶液加入到步骤2配制的溶液B中并搅拌0.5h,之后加入0.39-0.78g聚乙烯吡咯烷酮,搅拌6-12h后得到静电纺丝液;
(5)用注射器吸取静电纺丝液,以接地的铁丝网作为接收装置进行静电纺丝,其中,流速为0.5ml/h,,电压控制在10-12kV,得到的纤维在80℃下烘干2h后经600-800℃烧结,得到Yb、Er双掺CaTiO3纳米纤维;
(6)取100mg Yb、Er双掺CaTiO3纳米纤维,溶于80ml浓度为5mM的NaOH溶液中搅拌12h,离心干燥后溶于40ml DMF中,加入500μl 3-氨基丙基三乙氧基硅烷,于室温(25℃)下搅拌24h,离心干燥;
(7)取30mg上述样品加入到10ml聚丙烯酸中,搅拌12h,离心并干燥,得到改性的双掺CaTiO3纳米纤维。
2.根据权利要求1所述的方法,其特征在于,所述步骤5中,烧结温度为700℃。
CN201610858869.9A 2016-09-28 2016-09-28 一种实现pH控释、NIR控释以及光监控药物释放的纳米纤维的制备方法 Pending CN106381710A (zh)

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