CN116271246A - 一种还原氧化石墨烯神经导管及其制备方法 - Google Patents
一种还原氧化石墨烯神经导管及其制备方法 Download PDFInfo
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Abstract
本发明涉及神经导管技术领域，具体涉及一种还原氧化石墨烯神经导管及其制备方法，包括神经导管主体，所述神经导管主体为三层结构，所述神经导管主体内层为有序导电的纳米纤维，中间层为冻干羊膜，最外层为无序导电纳米纤维；内层的有序纳米纤维有着优异的沿轴向电荷传输性能，有效提高神经生物电信号的传输速度，同时有序纳米纤维与神经长入方向一致，利于神经纤维衣服生长，促进神经损伤的修复与再生；中层的羊膜释放生物活性物质，同时具有良好的神经相容性促进神经组织的再生；外层的无序纳米纤维具备良好的物理机械性能和柔韧性，为神经残端提供有效的力学支持，保证轴突的再生的稳定环境。
Description
技术领域
本发明涉及神经导管技术领域，具体涉及一种还原氧化石墨烯神经导管及其制备方法。
背景技术
外周神经缺损是临床上常见的外伤，各种创伤所导致的外周神经损伤发生率约为1.3-2.8％，其治疗结果较差,有较高的致残率。尽管周围神经在轻中度损伤后能够在一定程度内实现自我修复，但对于长段神经缺损的情况，我们仍然需要通过移植物来重新桥接神经的两个断端。经典的自体神经移植技术在过去很长一段时间内都被认为是治疗此病的金标准。然而这一方法存在诸多缺陷，如成功率不高、移植物来源受限、神经尺寸不匹配以及对供区组织的潜在伤害。因此，寻找替代自体神经的材料，用于神经修复成为当前的研究热点。利用生物材料制造神经导管为提升疗效提供了可能，是神经再生和修复领域的重要研究方向。
周围神经损伤后，通过逆向运输提供的神经营养因子骤减，导致神经元细胞死亡，再生乏力。但是，一旦受损神经元重新长回它所支配的靶组织，上述情形将得到扭转，表明周围神经再生有赖于靶组织合成的神经营养支持。因此，在神经受损部位的微环境中导入外源性神经营养因子，有助于损伤的修复和神经再生。不同于单纯的生长因子，来源于生物体的天然物质羊膜是一种半透膜，含有丰富的细胞因子、酶类等活性成分，允许营养物质渗透，来源广泛、成本低廉，极低的免疫源性，无致瘤性及伦理学争议，其独特的结构使其成为理想的生物材料。
目前神经导管中应用的纳米纤维是直径从微米到纳米尺度不等且排列无序的纤维；有序排列的纳米纤维由于其自身独特的电学、磁学以及光学等方面的物理性质，有着更为优异的导电性能与力学强度。
为此发明人提出一种还原氧化石墨烯神经导管及其制备方法，石墨烯神经导管内层采用有序纳米纤维，通过使用有序纳米纤维替换无序纳米纤维，提高神经导管的导电性使生物电信号传输速度变快，同时使神经导管强度增大，为神经残端提供有效的力学支持。
发明内容
本发明的目的在于提供一种还原氧化石墨烯神经导管及其制备方法，通过使用有序纳米纤维替换无序纳米纤维，提高神经导管的导电性使生物电信号传输速度变快，同时使神经导管强度增大，为神经残端提供有效的力学支持。
为实现上述技术目的，达到上述技术效果，本发明是通过以下技术方案实现：
一种还原氧化石墨烯神经导管，其特征在于，包括神经导管主体，所述神经导管主体为三层结构，所述神经导管主体内层为有序导电的纳米纤维，中间层为冻干羊膜，最外层为无序导电纳米纤维。
一种还原氧化石墨烯神经导管的制备方法，包括如下步骤：
S1：取新鲜胎盘组织，预处理后备用；
S2：选取还原氧化石墨烯溶解于丙酮中，超声溶解后作为溶液A；
S3：选取聚己内酯微球加入溶液A，溶解后得到前驱体溶液；
S4：将前驱体溶液置入注射器，连接静电纺丝装置，调整溶液的流速、接收距离与电压；
S5：先将有序纳米纤维沉积于接地旋转芯轴的铝箔上，其次将冻干羊膜裹于有序纳米纤维外层，最后将无序纳米纤维沉积于冻干羊膜的外层；
S6：得到三层结构的还原氧化石墨烯神经导管。
作为优选，所述步骤S1中的预处理包括如下子步骤：
S1.1：产妇血清学检查HBV、HCV、HIV、梅毒、淋病均呈阴性；
S1.2：羊膜与绒毛膜之间钝性分离，获得光滑、半透明的新鲜羊膜；
S1.3：选用50.0ug/mL青霉素和50.0ug/mL链霉素的平衡盐溶液将新鲜羊膜浸泡20min；
S1.4：置于DMEM培养基中，将新鲜羊膜置于冷冻干燥机工作室，在-50-0℃下预冻1-10小时，开启冷凝室制冷，冷阱温度<-10℃，开启真空泵，真空冷冻干燥，水份含量范围在1-20％。
作为优选，所述步骤S2中的还原氧化石墨烯用量为0.39g，所述步骤S2中的丙酮用量为80-120ml。
作为优选，所述步骤S2中的超声溶解时间为30min。
作为优选，所述步骤S3中的聚己内酯微球用量为8-12g，所述聚己内酯微球的溶解方式为于40℃加热板上搅拌直至完全溶解。
作为优选，所述步骤S3中的前驱体溶液中聚己内酯的质量体积比为10％w/v，前驱体溶液中还原氧化石墨烯的质量比0.5％wt。
作为优选，所述步骤S4中注射器的针头直径为0.7mm、容量为10mL，溶液的流速6.0mL/h，接收距离10.0cm，电压为15kV。
作为优选，所述步骤S5中的有序纳米纤维以15m/s的线性速率沉积在接地的旋转芯轴的铝箔上。
进一步的，提出了一种还原氧化石墨烯神经导管在神经修复中的应用。
本发明的有益效果：
本发明通过静电纺丝法以还原氧化石墨烯与聚己内酯微球的前驱体溶液进行和有序纳米纤维的生成，同时通过包覆羊膜与最外层的无序纳米纤维制备还原氧化石墨烯神经导管；与现有技术相比，本发明优点及积极效果如下：
1、本发明所制备的还原氧化石墨烯神经导管具有良好的生物相容性、生物活性与可降解性；
2、本发明所制备的神经导管为三层机构，内层的有序纳米纤维有着优异的沿轴向电荷传输性能，有效提高神经生物电信号的传输速度，同时有序纳米纤维与神经长入方向一致，利于神经纤维衣服生长，促进神经损伤的修复与再生；中层的羊膜释放生物活性物质，同时具有良好的神经相容性促进神经组织的再生；外层的无序纳米纤维具备良好的物理机械性能和柔韧性，为神经残端提供有效的力学支持，保证轴突的再生的稳定环境。
当然，实施本发明的任一产品并不一定需要同时达到以上所述的所有优点。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其它实施例，都属于本发明保护的范围。
实施例1
本实施例所述的一种还原氧化石墨烯神经导管，其特征在于，包括神经导管主体，所述神经导管主体为三层结构，所述神经导管主体内层为有序导电的纳米纤维，中间层为冻干羊膜，最外层为无序导电纳米纤维。
实施例2
本实施例所述的一种还原氧化石墨烯神经导管的制备方法，包括如下步骤：
S1：取新鲜胎盘组织，预处理后备用；
S2：选取还原氧化石墨烯溶解于丙酮中，超声溶解后作为溶液A；
S3：选取聚己内酯微球加入溶液A，溶解后得到前驱体溶液；
S4：将前驱体溶液置入注射器，连接静电纺丝装置，调整溶液的流速、接收距离与电压；
S5：先将有序纳米纤维沉积于接地旋转芯轴的铝箔上，其次将冻干羊膜裹于有序纳米纤维外层，最后将无序纳米纤维沉积于冻干羊膜的外层；
S6：得到三层结构的还原氧化石墨烯神经导管。
本实施例中，所述步骤S1中的预处理包括如下子步骤：
S1.1：产妇血清学检查HBV、HCV、HIV、梅毒、淋病均呈阴性；
S1.2：羊膜与绒毛膜之间钝性分离，获得光滑、半透明的新鲜羊膜；
S1.3：选用50.0ug/mL青霉素和50.0ug/mL链霉素的平衡盐溶液将新鲜羊膜浸泡20min；
S1.4：置于DMEM培养基中，将新鲜羊膜置于冷冻干燥机工作室，在-50-0℃下预冻1-10小时，开启冷凝室制冷，冷阱温度<-10℃，开启真空泵，真空冷冻干燥，水份含量范围在1-20％。
本实施例中，所述步骤S2中的还原氧化石墨烯用量为0.39g，所述步骤S2中的丙酮用量为100ml。
本实施例中，所述步骤S2中的超声溶解时间为30min。
本实施例中，所述步骤S3中的聚己内酯微球用量为10g，所述聚己内酯微球的溶解方式为于40℃加热板上搅拌直至完全溶解。
本实施例中，所述步骤S3中的前驱体溶液中聚己内酯的质量体积比为10％w/v，前驱体溶液中还原氧化石墨烯的质量比0.5％wt。
本实施例中，所述步骤S4中注射器的针头直径为0.7mm、容量为10mL，溶液的流速6.0mL/h，接收距离10.0cm，电压为15kV。
本实施例中，所述步骤S5中的有序纳米纤维以15m/s的线性速率沉积在接地的旋转芯轴的铝箔上。
实施例3
还原氧化石墨烯神经导管直径测量的原始数据：
本发明人对由本发明所公开的制备方法制备的神经导管分为30组进行直径测量，测得97％的直径数据为3-5mm之间，长度为20-30mm之间。
神经导管直径测量的原始数据：(30项，单位mm，软件Image J测量，Oingin统计)
神经导管长度测量的原始数据：(30项，单位mm，软件Image J测量，Oingin统计)
由本发明所公开的制备方法制备的神经导管，其直径均符合周围神经缺损手术所需。
实施例4
还原氧化石墨烯神经导管力学性能实验数据：
本发明人对由本发明所公开的制备方法制备的神经导管分为30组进行力学性能实验，测得平均数值：
纤维直径100-300nm，孔隙率90％，极限强度5.00MPa，杨氏模量10.0MPa，断裂伸长率200％。
由本发明所公开的制备方法制备的神经导管，其力学性能均满足周围神经缺损手术所需。
实施例5
还原氧化石墨烯神经导管导电性能实验数据：
本发明人对由本发明所公开的制备方法制备的神经导管分为30组进行导电性能实验，测得平均数值：
电导率为4.0-5.0×10-5S/m。
由本发明所公开的制备方法制备的神经导管，其导电性能均满足周围神经缺损手术所需。
实施例6
还原氧化石墨烯神经导管降解性能实验数据：
本发明人对由本发明所公开的制备方法制备的神经导管分为30组进行降解性能实验，测得4周内降解率为30％。
由本发明所公开的制备方法制备的神经导管，其降解性能均满足周围神经缺损手术所需。
综上所述，本发明一种还原氧化石墨烯神经导管及其制备方法，通过使用有序纳米纤维替换无序纳米纤维，提高神经导管的导电性使生物电信号传输速度变快，同时使神经导管强度增大，为神经残端提供有效的力学支持。
在本说明书的描述的具体特征、结构、材料或者特点可以在任何的一个或多个实施例或示例中以合适的方式结合。
尽管已经示出和描述了本发明的实施例，对于本领域的普通技术人员而言，可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型，本发明的范围由所附权利要求及其等同物限定。
Claims (10)
1.一种还原氧化石墨烯神经导管，其特征在于，包括神经导管主体，所述神经导管主体为三层结构，所述神经导管主体内层为有序导电的纳米纤维，中间层为冻干羊膜，最外层为无序导电纳米纤维。
2.一种如权利要求1所述还原氧化石墨烯神经导管的制备方法，其特征在于，包括如下步骤：
S1：取新鲜胎盘组织，预处理后备用；
S2：选取还原氧化石墨烯溶解于丙酮中，超声溶解后作为溶液A；
S3：选取聚己内酯微球加入溶液A，溶解后得到前驱体溶液；
S4：将前驱体溶液置入注射器，连接静电纺丝装置，调整溶液的流速、接收距离与电压；
S5：先将有序纳米纤维沉积于接地旋转芯轴的铝箔上，其次将冻干羊膜裹于有序纳米纤维外层，最后将无序纳米纤维沉积于冻干羊膜的外层；
S6：得到三层结构的还原氧化石墨烯神经导管。
3.如权利要求2所述的一种还原氧化石墨烯神经导管的制备方法，其特征在于：所述步骤S1中的预处理包括如下子步骤：
S1.1：产妇血清学检查HBV、HCV、HIV、梅毒、淋病均呈阴性；
S1.2：羊膜与绒毛膜之间钝性分离，获得光滑、半透明的新鲜羊膜；
S1.3：选用50.0ug/mL青霉素和50.0ug/mL链霉素的平衡盐溶液将新鲜羊膜浸泡20min；
S1.4：置于DMEM培养基中，将新鲜羊膜置于冷冻干燥机工作室，在-50-0℃下预冻1-10小时，开启冷凝室制冷，冷阱温度<-10℃，开启真空泵，真空冷冻干燥，水份含量范围在1-20％。
4.如权利要求2所述的一种还原氧化石墨烯神经导管的制备方法，其特征在于：所述步骤S2中的还原氧化石墨烯用量为0.39g，所述步骤S2中的丙酮用量为80-120ml。
5.如权利要求4所述的一种还原氧化石墨烯神经导管的制备方法，其特征在于：所述步骤S2中的超声溶解时间为30min。
6.如权利要求2所述的一种还原氧化石墨烯神经导管的制备方法，其特征在于：所述步骤S3中的聚己内酯微球用量为8-12g，所述聚己内酯微球的溶解方式为于40℃加热板上搅拌直至完全溶解。
7.如权利要求2所述的一种还原氧化石墨烯神经导管的制备方法，其特征在于：所述步骤S3中的前驱体溶液中聚己内酯的质量体积比为10％w/v，前驱体溶液中还原氧化石墨烯的质量比0.5％wt。
8.如权利要求2所述的一种还原氧化石墨烯神经导管的制备方法，其特征在于：所述步骤S4中注射器的针头直径为0.7mm、容量为10mL，溶液的流速6.0mL/h，接收距离10.0cm，电压为15kV。
9.如权利要求2所述的一种还原氧化石墨烯神经导管的制备方法，其特征在于：所述步骤S5中的有序纳米纤维以15m/s的线性速率沉积在接地的旋转芯轴的铝箔上。
10.如权利要求1-9任一项所述的一种还原氧化石墨烯神经导管在神经修复中的应用。
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| CN117982722A (zh) * | 2023-12-25 | 2024-05-07 | 北京大学口腔医学院 | 一种还原氧化石墨烯神经导管和促进周围神经再生装置 |
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| CN117982722A (zh) * | 2023-12-25 | 2024-05-07 | 北京大学口腔医学院 | 一种还原氧化石墨烯神经导管和促进周围神经再生装置 |
| CN117797316B (zh) * | 2023-12-25 | 2024-05-31 | 江苏益通生物科技有限公司 | 电诱导医用神经修复材料及其制作方法 |
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