CN105582573B - 一种纳微米多尺度壳聚糖三维支架及其制备方法 - Google Patents
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Abstract
本发明涉及一种纳微米多尺度壳聚糖三维支架及其制备方法，它采用在一定条件下分别静电纺丝纳米和微米壳聚糖纤维膜，经碱洗、水洗和风干后按一定配比将纳米和微米壳聚糖纤维膜在水中高速剪切成短纤维，分样后，加入微米尺度的盐颗粒作为致孔剂，快速在液氮中冷冻成型并冻干，冻干样品经水置换去除致孔剂，再经冷冻干燥得到纳微米多尺度壳聚糖三维支架。该制备方法工艺简单，支架在纤维直径和孔径同时具有微纳米尺度，赋予支架良好的生物学性能和力学性能。
Description
技术领域
本发明涉及一种纳微米多尺度壳聚糖三维支架及其制备方法，属于生物医用材料技术领域。
背景技术
壳聚糖作为一种阳离子碱性多糖，结构类似于人体内的硫酸软骨素和透明质酸等多糖类物质，在体内可被降解吸收，并具有止痛、止血、抑菌、促进伤口愈合、减小疤痕等优异特性，目前在生物医用材料领域有重要的应用。Sundararajan V等人采用相分离技术首次制备了壳聚糖支架，其结构为数十到数百微米厚度的片状蜂窝结构，具有较好的生物学性能（Biomaterials,1999;20:1133-1142）。胡巧玲等公开了三维有序多孔壳聚糖骨修复支架材料的制备方法（CN101366972），拓展了其应用。壳聚糖具有良好的生物相容性，研究证明了其可作为支架材料，但壳聚糖支架的微观结构也对其综合性能有较大的影响。研究表明纳米纤维支架有较大的比表面积，可提供细胞粘附和分化的更好条件，但单纯的纳米纤维力学强度较低，无法承受支架植入过程的力学载荷。相比而言，微米纤维支架具有较好的力学性能。另外，支架要求具有三维贯通的孔结构，便于营养物质的传递、代谢物质的交换以及细胞的迁移。研究表明，对于营养物质的传递和代谢物质的交换，纳米尺度的孔具有较高的效率，并可促进支架的降解吸收和组织再生，但该尺度的孔阻碍细胞在支架内部的渗透和迁移，微米尺度的孔则对细胞的渗透、迁移以及随之的分化增殖、新血管和神经的长入是比较有利的。微纳米共存的孔结构可以保证细胞生长过程中营养物质的传递、细胞的迁移以及新血管和神经生长所需的空间。
综上，在纤维直径和孔径同时具有微纳米多尺度的三维支架具有良好的生物相容性和力学性能。目前研究者多采用相分离的方法制备壳聚糖三维支架，但无法控制其纤维直径和孔径，或采用静电纺丝的方法获得二维的壳聚糖薄膜，但如何制备纤维直径和孔径同时具有微纳米多尺度的三维壳聚糖支架制备方法尚未见报导。
发明内容
本发明的主要设想是采用静电纺丝的方法分别制备纳米和微米壳聚糖薄膜，按一定配比将纳米和微米壳聚糖纤维膜混合，并将壳聚糖薄膜高速剪切成短纤维，分样后，加入微米尺度的致孔剂，经冷冻干燥成型，水置换去除致孔剂，再经冷冻干燥得到纳微米多尺度壳聚糖三维支架。
本发明中纳微米多尺度壳聚糖三维支架的制备方法具体如下所述。
(1) 静电纺丝纳米壳聚糖纤维膜，配制壳聚糖5.0%的溶液，溶剂为三氟乙酸和二噁烷的混合物，体积比为70:30，纺丝电压为20kV。
(2) 静电纺丝微米壳聚糖纤维膜，配制壳聚糖7.5%的溶液，溶剂为三氟乙酸和二噁烷的混合物，体积比为90:10，纺丝电压为15kV。
(3) 将静电纺丝得到的纳米和微米壳聚糖纤维膜浸泡在0.5mol/L的氢氧化钠溶液中去除未挥发的溶剂，水洗风干后待用。
(4) 分别称取定量的纳米和微米壳聚糖纤维膜，加入到定量的水中，在水中高速剪切成短纤维。
(5) 分样至称量瓶中，加入筛分后具有一定粒度分布的盐作为致孔剂，快速在液氮中冷冻成型并冻干。
(6)冻干后的样品经水置换去除盐，再经冷冻干燥可获得纳微米多尺度壳聚糖三维支架。
以上所使用的壳聚糖分子量在20～40万之间，纳米壳聚糖和微米壳聚糖的质量比在20:80到80:20之间，壳聚糖纳米微米纤维在水中的总浓度在1～3%之间。
本发明制得的纳微米多尺度壳聚糖三维支架外观为白色圆盘，其形状和高度可在分样中采用不同的容器和分样体积调节。制备方法中主要通过静电纺丝获得纳微米尺度的纤维，制备中也可方便地调整纺丝条件得到不同尺度的纳米和微米壳聚糖纤维，通过纳米纤维之间的孔形成纳米尺度的孔，通过微米尺度的盐致孔剂获得微米尺度的孔。该支架在纤维直径和孔径同时具有微纳米尺度，赋予支架良好的生物相容性和力学强度，便于营养物质的输送和细胞的迁移。该制备方法工艺简单，易于得到不同直径的壳聚糖纤维，并可通过盐致孔获得微米尺度的孔，其孔径容易控制。相比于单纯的静电纺丝法，该法还可容易地制备三维结构的支架。
具体实施方式
下面结合具体实施例，对本发明内容作进一步的说明，但本发明的实现方式并不局限于此。
实施例1：分别取7ml的三氟乙酸和3ml的二噁烷，混合均匀，称取0.5g壳聚糖，搅拌溶解，静置12小时。将壳聚糖溶液加入到注射器中，通以20kV的电压纺丝，收集在接地的铝箔上，可获得壳聚糖纤维直径在50-300nm的薄膜。分别取18ml的三氟乙酸和2ml的二噁烷，混合均匀，称取1.5g壳聚糖，搅拌溶解，静置12小时。将壳聚糖溶液加入到注射器中，通以15kV的电压纺丝，收集在接地的铝箔上，可获得壳聚糖纤维直径在1-10μm的薄膜。将静电纺丝得到的纳米和微米纤维膜浸泡在0.5mol/L的氢氧化钠溶液中去除未挥发的溶剂，水洗风干。分别称取0.2g和0.8g纳米和微米壳聚糖纤维膜，在100ml水中高速剪切成短纤维，分样至称量瓶中，加入筛分的氯化钠颗粒，其粒度分布在50-350μm之间，高度与分样的液面相同，快速在液氮中冷冻成型，在冻干机中冻干。冻干后的样品经水置换去除支架中的盐致孔剂，再经冷冻干燥可获得纳微米多尺度壳聚糖三维支架。
实施例2：分别取35ml的三氟乙酸和15ml的二噁烷，混合均匀，称取2.5g壳聚糖，搅拌溶解，静置12小时。将壳聚糖溶液加入到注射器中，通以20kV的电压纺丝，收集在接地的铝箔上，可获得壳聚糖纤维直径在50-300nm的薄膜。分别取9ml的三氟乙酸和1ml的二噁烷，混合均匀，称取0.75g壳聚糖，搅拌溶解，静置12小时。将壳聚糖溶液加入到注射器中，通以15kV的电压纺丝，收集在接地的铝箔上，可获得壳聚糖纤维直径在1-10μm的薄膜。将静电纺丝得到的纳米和微米纤维膜浸泡在0.5mol/L的氢氧化钠溶液中去除未挥发的溶剂，水洗风干。分别称取2.4g和0.6g纳米和微米壳聚糖纤维膜，在100ml水中高速剪切成短纤维，分样至称量瓶中，加入筛分的氯化钠颗粒，其粒度分布在50-350μm之间，高度与分样的液面相同，快速在液氮中冷冻成型，在冻干机中冻干。冻干后的样品经水置换去除支架中的盐致孔剂，再经冷冻干燥可获得纳微米多尺度壳聚糖三维支架。
Claims (6)
1.一种纳微米多尺度壳聚糖三维支架，其特征在于在一定条件下分别静电纺丝纳米和微米壳聚糖纤维膜，在0.5mol/L的氢氧化钠溶液中去除未挥发的溶剂并水洗风干后，按一定配比将纳米和微米壳聚糖纤维膜在水中高速剪切成短纤维，分样后，加入具有一定粒度分布的微米氯化钠致孔剂，快速在液氮中冷冻成型并冻干，冻干样品经水置换去除致孔剂，水置换去除氯化钠致孔剂，再经冷冻干燥得到纳微米多尺度壳聚糖三维支架。
2.根据权利要求1所述的纳微米多尺度壳聚糖三维支架，其特征在于所使用的壳聚糖分子量在20～40万之间。
3.根据权利要求1所述的纳微米多尺度壳聚糖三维支架，其特征在于静电纺丝纳米纤维膜时，壳聚糖浓度为5.0%，溶剂为三氟乙酸和二噁烷的混合物，体积比为70:30，纺丝电压为20kV。
4.根据权利要求1所述的纳微米多尺度壳聚糖三维支架，其特征在于静电纺丝微米壳聚糖纤维膜时，壳聚糖浓度为7.5%，溶剂为三氟乙酸和二噁烷的混合物，体积比为90:10，纺丝电压为15kV。
5.根据权利要求1所述的纳微米多尺度壳聚糖三维支架，其特征在于纳米壳聚糖和微米壳聚糖的质量比在20:80到80:20之间，壳聚糖纳米微米纤维在水中的总浓度在1～3%之间。
6.根据权利要求1所述的纳微米多尺度壳聚糖三维支架，其特征在于使用的氯化钠致孔剂的粒度范围在50～350μm之间。
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