CN115252894A - 复合水凝胶支架及在制备修复神经损伤的产品中的应用 - Google Patents
复合水凝胶支架及在制备修复神经损伤的产品中的应用 Download PDFInfo
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Abstract
本发明公开了一种复合水凝胶支架及在制备修复神经损伤的产品中的应用。所述复合水凝胶支架包括结合的羟丁基壳聚糖与聚乙二醇修饰的氧化石墨烯,所述复合水凝胶支架上还负载有药物小分子。本发明还提供了该复合水凝胶支架在制备修复神经损伤的产品中的应用。本发明利用氧化石墨烯较强的力学性能增强羟丁基壳聚糖的力学强度,增强水凝胶支架稳定性,为NSCs的生长提供良好的基质底物,同时利用氧化石墨烯大的比表面积,实现对蛋白类药物小分子的高效负载和局部控缓释放,该复合水凝胶支架克服了羟丁基壳聚糖水凝胶机械性能不足的缺点,具有稳定的支架力学性能,同时具有生物活性,能够促进神经细胞黏附和分化,从而达到修复神经损伤的目的。
Description
技术领域
本发明涉及一种新型复合水凝胶支架,尤其涉及一种新型的用于修复神经损伤的复合水凝胶支架及其制备方法,以及其在制备修复神经损伤的产品中的应用,属于生物医学材料技术领域。
背景技术
神经系统是人体内起主导作用的功能调节系统,对于生命活动具有十分重要的作用。但由于各种外伤或其它条件所造成的脊髓损伤往往会导致损伤部位以下运动和感觉功能的永久性障碍,是一种极具挑战性的疾病。脊髓损伤包括原发性损伤和继发性损伤。目前脊髓损伤的治疗手段主要集中在减缓和阻止继发性损伤的级联式反应,如早期利用减压手术改善脊髓灌注、降低局部温度来减少神经细胞凋亡等,但传统治疗手段对于神经系统功能的恢复并未有明显效果。而且脊髓损伤往往会造成神经细胞损伤,而成年人脊髓中所含的神经干细胞较少;加上损伤部位微环境的改变,一些抑制因子的产生、营养因子的缺乏、胶质瘢痕的形成等,所以当脊髓损伤发生时,患者自身的修复和再生能力有限,脊髓损伤一旦发生,则难以自发修复。
组织工程为脊髓损伤的修复带来了巨大前景。组织工程是将具有生物相容性和组织环境仿生性的生物材料与干细胞相结合,构建出具有生物功能的复合物,从而应用于组织修复或器官重建。用于组织工程的生物支架材料具有多种功能:首先,支架材料可以模拟细胞外微环境,为干细胞的存活提供适宜的条件,协助干细胞的活体移植,或体外药物筛选;其次,生物支架材料具有良好的生物相容性和负载能力,可以作为药物递送的载体;再之,支架材料所具有的可调控的理化性质和三维结构有助于研究不同条件下干细胞的增殖、迁移、分化情况,从而探索干细胞生命活动的内在机理;最后,通过对支架材料功能化,可实现药物的局部富集以及可控释放,减少对目标外组织和器官的毒副作用。在组织工程领域,可注射水凝胶类材料适应微创伤技术发展的要求,具有微创伤修复组织缺损或畸形、组织损伤小、不破坏修复区血供、操作简便易行等优点,具有良好的应用前景。Yin等将NGF加载于PLGA缓释微球中,然后包封于锂藻土与透明质酸原位聚合形成的可注射水凝胶中,从而实现NGF的缓释;但PLGA微球制备过程需要加入有机试剂,且所得水凝胶的成胶条件和降解速率均受pH值很大影响,水凝胶在降解过程中会造成损伤部位pH值的波动,同时体系载药率较低。
目前,现有方法利用高分子水凝胶进行组织损伤修复时通常需要先在体外预成胶,然后再移植入体内,这种移植方法会对组织再次造成损伤;而且无论基于人工合成或天然聚合物的水凝胶,在制备过程中,需加入交联剂或引发剂使高分子通过共价键或者非共价键交联形成网络结构,易造成交联剂或引发剂的残留,不利于细胞的生长。此外,利用水凝胶或其它纳米载体负载生物活性因子,不仅负载率低,而且小分子的释放率受材料降解速率的影响,易突释,半衰期短,不利于神经损伤的修复。
羟丁基壳聚糖(HBC)是一种可降解、生物相容性好、水溶液中溶解性好的壳聚糖衍生物。作为一种常用的水凝胶,它具有良好的温敏性和三维结构,可以模拟细胞外微环境,为细胞生命活动提供适宜的物理支持、黏附位点及物质交换环境。HBC可以通过分子间的氢键作用,在一定浓度、一定温度下自发形成可逆性温敏水凝胶,可以进行体内注射,避免了传统水凝胶化学交联剂和光引发剂的残留。HBC分子链中富含氢键,可以与其它材料相互作用来增加其性能,同时它保留了壳聚糖上的氨基,也可以对其进一步接枝改性,如季铵化反应、与醛基反应等。但将HBC水凝胶用于修复神经损伤则存在力学强度较低,在体内易坍塌等缺点。而加入机械性能优良的增强材料是提高聚合物力学强度最有效的方法之一。
因此,利用机械性能优良的增强材料,制备一种力学强度良好的复合温敏性水凝胶,已然成为业界研究人员长期以来一直努力的方向。
发明内容
本发明的主要目的在于提供一种新型的用于修复神经损伤的复合水凝胶支架及其制备方法,以克服现有技术中的不足。
本发明的另一目的在于提供一种所述复合水凝胶支架在制备修复神经损伤的产品中的应用。
为实现前述发明目的,本发明采用的技术方案包括:
本发明实施例提供了一种复合水凝胶支架,其包括相互结合的羟丁基壳聚糖(以下可简称为HBC)与聚乙二醇修饰的氧化石墨烯(以下可简称为GO-PEG),所述复合水凝胶支架上还负载有药物小分子。
进一步地,所述复合水凝胶支架能够促进神经干细胞黏附和分化。
进一步地,所述复合水凝胶支架能够在37℃条件下快速实现溶液-凝胶的相转变过程。
本发明实施例还提供了一种复合水凝胶支架的制备方法,其包括:
分别提供氧化石墨烯、羟丁基壳聚糖;
以聚乙二醇对氧化石墨烯进行修饰,获得聚乙二醇修饰的氧化石墨烯;
向包含聚乙二醇修饰的氧化石墨烯的溶液中加入药物小分子搅拌均匀,再加入羟丁基壳聚糖继续搅拌,得到负载药物小分子的复合水凝胶前驱体溶液,之后升温孵育,获得所述复合水凝胶支架。
本发明实施例还提供了由前述方法制备的复合水凝胶支架。
本发明实施例还提供了前述复合水凝胶支架在制备修复神经损伤的产品中的应用。
相应的,本发明实施例还提供了一种修复神经损伤的功能产品,其包含前述复合水凝胶支架。
与现有技术相比,本发明的有益效果在于:
1)本发明引入GO作为HBC的增强材料,巧妙地将GO和HBC结合,利用GO较强的力学性能增强HBC的力学强度,增强水凝胶支架稳定性,为NSCs的生长提供良好的基质底物,同时利用GO大的比表面积,实现对蛋白类药物小分子的高效负载和局部控缓释放,该HBC-(GO-PEG)复合水凝胶支架克服了HBC水凝胶机械性能不足的缺点,具有稳定的支架力学性能,同时具有生物活性,能够促进神经干细胞黏附和分化,为NSCs提供了一种良好的生长支架;
2)本发明所提供的HBC-(GO-PEG)复合水凝胶支架能够高效的负载药物小分子,并实现控缓释NT3,从而有效调控NSCs功能,诱导NSCs向神经元分化;
3)本发明所提供的HBC-(GO-PEG)复合水凝胶支架在人体温条件(37℃)下快速实现溶液-凝胶的相转变过程,具有良好的可注射性,不需要化学交联剂和光引发剂,所得复合水凝胶支架安全性高,无细胞毒性,生物相容性良好,制备工艺简单,可以在神经损伤部位进行原位注射。将其原位注射到神经损伤部位后,可在损伤部位原位成胶,避免外科手术的创伤性以及化学交联剂、光引发剂在损伤部位的残留。同时GO独特的理化性质使其具有比其它纳米载体更高的载药率以及药物控缓释能力,有效解决其它纳米载体负载率低,药物小分子容易突释、分散过快、半衰期短等问题,保持生物活性因子较好的生物活性,促进神经干细胞的分化,从而达到修复神经损伤的目的。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明中记载的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1是本发明一典型实施例中复合水凝胶支架的结构示意图;
图2是本发明实施例1中不同GO-PEG浓度的HBC-(GO-PEG)复合水凝胶支架的实物图;
图3是本发明实施例1中不同GO-PEG浓度的HBC-(GO-PEG)复合水凝胶支架的力学性能测定结果图;
图4a-图4e分别是本发明实施例2中在不同氧化石墨烯浓度下制备的HBC-(GO-PEG)复合水凝胶支架的成胶温度测定结果图;
图5a-图5e分别是本发明实施例3中在不同氧化石墨烯浓度下制备的HBC-(GO-PEG)复合水凝胶支架的成胶时间测定结果图;
图6是本发明实施例4中负载NT3的复合水凝胶(HBC-(GO-PEG)-NT3)诱导神经元分化结果示意图。
具体实施方式
鉴于现有技术中的不足,本案发明人经长期研究和大量实践,得以提出本发明的技术方案,其主要是巧妙地将GO和HBC结合,利用GO较强的力学性能增强HBC的力学强度,同时利用GO大的比表面积,实现对蛋白类药物小分子的高效负载和局部控缓释放。如下将对该技术方案、其实施过程及原理等作进一步的解释说明。
作为一种新型二维纳米材料,氧化石墨烯(GO)结构稳定,机械性能良好,是一种可以提高水凝胶力学强度的增强材料。在GO片层边缘和基面上含有大量的含氧基团,如羟基(-OH)、羰基(-C=O)、羧基(-COOH)和环氧基(C-O-C),可以作为界面连接剂,促进应力从聚合物基体向GO的转移。这些含氧基团还大大增加了GO的亲水性,其片层之间的静电排斥使GO可以均匀地分散在水溶液中。同时这些含氧基团不仅为GO的功能化改性提供了丰富的反应位点,还能大大提高GO与聚合物分子间的作用。不仅如此,GO还具有较大的比表面积,对药物分子具有其它纳米载体无法比拟的高负载率;以及优良的生物相容性,能够促进细胞黏附和增殖。因此本发明引入GO作为HBC的增强材料,以及药物小分子的载体。
本发明实施例的一个方面提供的一种复合水凝胶支架,其包括相互结合的羟丁基壳聚糖与聚乙二醇修饰的氧化石墨烯,所述复合水凝胶支架上还负载有药物小分子。
作为优选方案之一,所述羟丁基壳聚糖与聚乙二醇修饰的氧化石墨烯的质量比为60:1-3000:1。
作为优选方案之一,所述复合水凝胶支架上药物小分子的负载量为0.2-1wt%。
作为优选方案之一,所述药物小分子包括蛋白类药物,优选为NT3(神经营养因子3),也可以是其它蛋白类药物,如BDNF(脑源性神经营养因子)、NGF(神经生长因子)等,但不限于此。
作为优选方案之一,本发明提供的是一种可控缓释药物小分子的HBC-(GO-PEG)复合水凝胶支架,用于神经干细胞功能的诱导,从而有效调控NSCs功能,诱导NSCs向神经元分化。通过将聚乙二醇(PEG)修饰的GO与HBC物理共混,来改善HBC水凝胶的力学强度,增强水凝胶支架稳定性,为NSCs的生长提供良好的基质底物。
进一步地,所述复合水凝胶支架的力学强度为40-120Pa。本发明中HBC-(GO-PEG)复合水凝胶支架克服了HBC水凝胶机械性能不足的缺点,具有稳定的支架力学性能,同时具有生物活性,能够促进神经干细胞黏附和分化,为NSCs提供了一种良好的生长支架。
进一步地,所述复合水凝胶支架能够在37℃条件下快速实现溶液-凝胶的相转变过程,具有良好的可注射性,不需要化学交联剂和光引发剂,所得复合水凝胶支架安全性高,无细胞毒性,生物相容性良好,制备工艺简单,可以在神经损伤部位进行原位注射。
将其原位注射到神经损伤部位后,可在损伤部位原位成胶,避免外科手术的创伤性以及化学交联剂、光引发剂在损伤部位的残留。同时GO独特的理化性质使其具有比其它纳米载体更高的载药率以及药物控缓释能力,有效解决其它纳米载体负载率低,药物小分子容易突释、分散过快、半衰期短等问题,保持生物活性因子较好的生物活性,促进神经干细胞的分化,从而达到修复神经损伤的目的。
本发明利用羟丁基壳聚糖(HBC)对温度的高度响应性,实现在损伤部位原位成胶,条件温和,具有微创性。
本发明实施例的另一个方面还提供了一种复合水凝胶支架的制备方法,其包括:
分别提供氧化石墨烯、羟丁基壳聚糖;
以聚乙二醇对氧化石墨烯进行修饰,获得聚乙二醇修饰的氧化石墨烯;
向包含聚乙二醇修饰的氧化石墨烯的溶液中加入药物小分子搅拌均匀,再加入羟丁基壳聚糖继续搅拌,得到负载药物小分子的复合水凝胶前驱体溶液,之后升温孵育,获得所述复合水凝胶支架。
作为优选方案之一,所述的制备方法具体包括:
向包含聚乙二醇修饰的氧化石墨烯的溶液中加入药物小分子搅拌过夜(12h以上),再加入羟丁基壳聚糖继续搅拌,得到混合体系,使所述混合体系置于冰浴中搅拌至羟丁基壳聚糖完全溶解,得到负载药物小分子的复合水凝胶前驱体溶液,之后从0℃升温至37℃孵育5-10min,获得所述复合水凝胶支架。
作为优选方案之一,所述混合体系中聚乙二醇修饰的氧化石墨烯的浓度为0.01-0.5mg/mL。
进一步地,所述混合体系中药物小分子的浓度为0.1-0.5μg/mL。
进一步地,所述混合体系中羟丁基壳聚糖的浓度为3-5wt%。
作为优选方案之一,所述的制备方法具体包括:
向包含氧化石墨烯的溶液中滴加末端氨基化的六臂聚乙二醇混合均匀,之后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)继续搅拌,获得聚乙二醇修饰的氧化石墨烯。
作为优选方案之一,所述药物小分子包括蛋白类药物,优选为NT3,也可以是其它蛋白类药物,如BDNF、NGF等,但不限于此。
作为优选方案之一,所述的制备方法具体包括:
称取精制壳聚糖分散在50%NaOH溶液中,在N2保护下,室温搅拌24h;过滤除去碱液,将碱化好的壳聚糖移至三颈瓶中,加入异丙醇和蒸馏水,室温搅拌24h;充分反应后加入1,2-环氧丁烷,室温搅拌1h后,升温至60℃,冷凝回流,反应24h;反应结束后,用10%HCl溶液调节pH至中性,用截留分子量为8000~14000的透析袋透析三天,过滤除去不溶物,将溶液冷冻干燥即得羟丁基壳聚糖(HBC)。
作为另一种优选方案之一,所述羟丁基壳聚糖的制备方法还可以是其他方法:
如称取10g氢氧化钠加入10mL蒸馏水中,待冷却后加入10g壳聚糖,充分搅拌后放到-20℃冰箱中冷冻过夜;取上述碱化壳聚糖30g,置于干燥的圆底烧瓶中,加入100mL异丙醇和100mL蒸馏水,常温下充分搅拌6h,加入一定量十二烷基硫酸钠,逐滴滴加200mL的环氧丁烷,升温至60℃,搅拌反应12h,待反应结束后调节产物pH至中性,透析3天,过滤除去不溶物,冷冻干燥后即得HBC。
本发明实施例的另一个方面还提供了由前述方法制备的复合水凝胶支架。
本发明实施例的另一个方面还提供了前述复合水凝胶支架在制备修复神经损伤的产品中的应用。
相应的,本发明实施例的另一个方面还提供了一种修复神经损伤的功能产品,其包含前述复合水凝胶支架。
综上所述,本发明所提供的HBC-(GO-PEG)复合水凝胶支架在人体温条件(37℃)下快速实现溶液-凝胶的相转变过程,具有良好的可注射性,不需要化学交联剂和光引发剂,所得复合水凝胶支架安全性高,无细胞毒性,生物相容性良好,制备工艺简单,可以在神经损伤部位进行原位注射。将其原位注射到神经损伤部位后,可在损伤部位原位成胶,避免外科手术的创伤性以及化学交联剂、光引发剂在损伤部位的残留。同时GO独特的理化性质使其具有比其它纳米载体更高的载药率以及药物控缓释能力,有效解决其它纳米载体负载率低,药物小分子容易突释、分散过快、半衰期短等问题,保持生物活性因子较好的生物活性,促进神经干细胞的分化,从而达到修复神经损伤的目的。
以下通过若干实施例进一步详细说明本发明的技术方案。然而,所选的实施例仅用于说明本发明,而不限制本发明的范围。
实施例1
本实施例中制备HBC-(GO-PEG)复合水凝胶支架的具体技术方案如下:
1)精制壳聚糖:称取5g壳聚糖粗品,将其溶于250mL 1%HCl溶液,室温搅拌至壳聚糖完全溶解,过滤去除不溶物,然后用1M NaOH溶液将其pH调至9使沉淀析出,将沉淀物用蒸馏水洗至中性,随后用250mL 70%乙醇洗涤三次,最后用250mL 95%乙醇洗涤三次,收集沉淀,于60℃烘箱中烘干即得精制壳聚糖。
2)制备羟丁基壳聚糖(HBC):称取1g精制壳聚糖分散在20mL 50%NaOH溶液中,在N2保护下,室温搅拌24h;过滤除去碱液,将碱化好的壳聚糖移至100mL三颈瓶中,加入异丙醇和蒸馏水各10mL,室温搅拌24h;充分反应后加入20mL 1,2-环氧丁烷,室温搅拌1h后,升温至60℃,冷凝回流,反应24h;反应结束后,用10%HCl溶液调节pH至中性,用截留分子量为8000~14000的透析袋透析三天,过滤除去不溶物,将溶液冷冻干燥即得HBC。
3)制备氧化石墨烯(GO):首先分别称取1g石墨粉、0.5g K2S208和0.5g P2O5于三颈瓶中搅拌混合均匀,加1.5mL浓H2SO4,80℃反应6h得到预氧化石墨。将其用蒸馏水反复清洗至中性,真空干燥。称取1.2g预氧化的石墨加入到23mL浓H2SO4中混合均匀,冰浴冷却至0℃。将3.6g KMnO4少量多次缓慢加入到混合物中,并保持反应液温度在20℃以下。之后将温度升高至35℃,搅拌2h后,缓慢滴加46mL蒸馏水,继续搅拌15min,最后加入146mL蒸馏水停止反应。在混合物中缓慢滴加2.5mL 30%H2O2。将所得溶液8000rpm离心15min,所得沉淀物用250mL 10%HCl清洗两次,再将产物用蒸馏水反复洗涤至中性。通过细胞粉碎仪在400W条件下超声30min,10000rpm离心25min,收集上清液,即为剥离的GO。将最终获得的GO在蒸馏水中透析,进一步去除杂质,3天后收集。
4)聚乙二醇(PEG)修饰GO:取5mg GO,加蒸馏水稀释到15mL,水浴超声5min充分分散。称取30mg末端氨基化的六臂PEG,溶于0.2mL蒸馏水中,在不断搅拌下滴加至GO溶液中,水浴超声5min混合均匀。在向上述溶液中加入0.2mL EDC(25mg/mL),室温搅拌20min后,再次加入0.2mL EDC(50mg/mL),继续搅拌48h,最后用截留分子量为100000的透析袋透析除去未反应的分子得到聚乙二醇修饰的氧化石墨烯(GO-PEG)。
5)复合水凝胶支架的制备:在步骤4)所获GO-PEG溶液中加入NT3,搅拌过夜,然后将HBC加入到上述溶液中,冰浴搅拌至HBC完全溶解,得到负载NT3的复合水凝胶前溶液。将温度从0℃提升至37℃孵育即得复合水凝胶支架(如图2所示),其力学性能测试结果如图3所示。
其中,步骤5)中NT3浓度为0.5μg/mL,GO-PEG的浓度为0.05mg/mL,HBC的质量分数为3wt%。HBC、HBC-0.01、HBC-0.05、HBC-0.1、HBC-0.5组分别指加入的聚乙二醇修饰的氧化石墨烯的浓度分别为:0、0.01、0.05、0.1、0.5mg/mL。
其中,步骤5)负载的药物小分子可为其它蛋白类药物,如BDNF、NGF等。
实施例2
将HBC加入到GO-PEG溶液中,冰浴搅拌至HBC完全溶解,得到负载NT3的复合水凝胶前溶液。利用旋转流变仪测定复合水凝胶的成胶温度,结果如图4a-图4e所示。成胶温度一般定义为水凝胶前溶液的相转变温度,即G’=G”时。如结果所示,各组水凝胶的成胶温度均在27℃左右,氧化石墨烯的加入并未影响水凝胶的成胶温度。
图4a-图4e中HBC、HBC-0.01、HBC-0.05、HBC-0.1、HBC-0.5组分别指加入的聚乙二醇修饰的氧化石墨烯的浓度分别为:0、0.01、0.05、0.1、0.5mg/mL。
实施例3
将HBC加入到GO-PEG溶液中,冰浴搅拌至HBC完全溶解,得到HBC-(GO-PEG)复合水凝胶前溶液。利用旋转流变仪测定复合水凝胶的成胶时间,结果如图5a-图5e所示。测定温度为37℃,可以看出,各组水凝胶的成胶温度均在一分钟以内,氧化石墨烯的加入并未影响水凝胶的成胶时间。
图5a-图5e中HBC、HBC-0.01、HBC-0.05、HBC-0.1、HBC-0.5组分别指加入的聚乙二醇修饰的氧化石墨烯的浓度分别为:0、0.01、0.05、0.1、0.5mg/mL。
实施例4
向GO-PEG溶液中加入NT3,搅拌过夜,然后将HBC加入到上述溶液中,冰浴搅拌至HBC完全溶解,得到负载NT3的复合水凝胶前溶液。将温度从0℃提升至37℃孵育即得复合水凝胶支架。然后将培养的神经干细胞消化,种于复合水凝胶支架上,分化培养7天。进行免疫荧光染色,统计神经干细胞向神经元的分化比例,结果如图6所示。结果显示,负载了NT3的复合水凝胶支架上的神经干细胞向神经元的分化比例要明显高于其它各组。
综上所述,本发明利用氧化石墨烯较强的力学性能增强羟丁基壳聚糖的力学强度,增强水凝胶支架稳定性,为NSCs的生长提供良好的基质底物,同时利用氧化石墨烯大的比表面积,实现对蛋白类药物小分子的高效负载和局部控缓释放,该复合水凝胶支架克服了羟丁基壳聚糖水凝胶机械性能不足的缺点,具有稳定的支架力学性能,同时具有生物活性,能够促进神经细胞黏附和分化,从而达到修复神经损伤的目的。
应当理解,以上所述的仅是本发明的一些实施方式,应当指出,对于本领域的普通技术人员来说,在不脱离本发明的创造构思的前提下,还可以做出其它变形和改进,这些都属于本发明的保护范围。
Claims (10)
1.一种复合水凝胶支架,其特征在于包括相互结合的羟丁基壳聚糖与聚乙二醇修饰的氧化石墨烯,所述复合水凝胶支架上还负载有药物小分子。
2.根据权利要求1所述的复合水凝胶支架,其特征在于:所述羟丁基壳聚糖与聚乙二醇修饰的氧化石墨烯的质量比为60:1-3000:1。
3.根据权利要求1所述的复合水凝胶支架,其特征在于:所述复合水凝胶支架上药物小分子的负载量为0.2-1wt%。
4.根据权利要求1所述的复合水凝胶支架,其特征在于:所述药物小分子包括蛋白类药物,优选为神经营养因子,更优选为神经营养因子3、脑源性神经营养因子、神经生长因子中的任意一种或两种以上的组合。
5.根据权利要求1所述的复合水凝胶支架,其特征在于:所述复合水凝胶支架的力学强度为40-120Pa;和/或,所述复合水凝胶支架能够促进神经干细胞黏附和分化,和/或,所述复合水凝胶支架能够在37℃条件下快速实现溶液-凝胶的相转变过程。
6.如权利要求1-5中任一项所述复合水凝胶支架的制备方法,其特征在于包括:
分别提供氧化石墨烯、羟丁基壳聚糖;
以聚乙二醇对氧化石墨烯进行修饰,获得聚乙二醇修饰的氧化石墨烯;
向包含聚乙二醇修饰的氧化石墨烯的溶液中加入药物小分子搅拌均匀,再加入羟丁基壳聚糖继续搅拌,得到负载药物小分子的复合水凝胶前驱体溶液,之后升温孵育,获得所述复合水凝胶支架。
7.根据权利要求6所述的制备方法,其特征在于具体包括:
向包含聚乙二醇修饰的氧化石墨烯的溶液中加入药物小分子搅拌过夜,再加入羟丁基壳聚糖继续搅拌,得到混合体系,使所述混合体系置于冰浴中搅拌至羟丁基壳聚糖完全溶解,得到负载药物小分子的复合水凝胶前驱体溶液,之后从0℃升温至37℃孵育5-10min,获得所述复合水凝胶支架。
8.根据权利要求7所述的制备方法,其特征在于:所述混合体系中聚乙二醇修饰的氧化石墨烯的浓度为0.01-0.5mg/mL;和/或,所述混合体系中药物小分子的浓度为0.1-0.5μg/mL;和/或,所述混合体系中羟丁基壳聚糖的浓度为3-5wt%;
和/或,所述制备方法包括:向包含氧化石墨烯的溶液中滴加末端氨基化的六臂聚乙二醇混合均匀,之后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐继续搅拌,获得聚乙二醇修饰的氧化石墨烯。
9.权利要求1-5中任一项所述复合水凝胶支架在制备修复神经损伤的产品中的应用。
10.一种修复神经损伤的功能产品,其特征在于包含权利要求1-5中任一项所述的复合水凝胶支架。
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