CN115282334A - 一种压电式氨基酸生物支架及其制备方法 - Google Patents
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Abstract
本发明涉及一种压电式氨基酸生物支架的制备方法,步骤包括:将聚合物溶于二元溶剂中,配置成纺丝液;通过静电纺丝技术,由所述纺丝液制得直径为0.5μm‑3.0μm的纳米纤维;通过动态水浴加捻技术,由所述纳米纤维制得纳米纤维支架;将所述纳米纤维支架浸渍于氨基酸溶液中,于低温中将所述纳米纤维支架析出,冷冻干燥后即得所述压电式氨基酸生物支架。本发明的压电式氨基酸生物支架能够满足肌腱作为一种胶原组织所表现出的压电性质,人体在运动时会产生机械刺激,当其作用于压电式生物支架后,支架产生一定强度的电刺激,导致毛细血管在短期内增加,成纤维细胞和肌腱细胞通过增加DNA含量和蛋白表达来进行大规模增殖,从而促进肌腱修复。
Description
技术领域
本发明涉及生物支架技术领域,尤其涉及一种压电式氨基酸生物支架及其制备方法。
背景技术
随着交通运输、运动体育以及老龄化人口的增加,各类急、慢性损伤导致的肌腱损伤、肌腱病的发病率呈逐渐上升趋势;据不完全统计,全球每年大约有3000万的肌腱损伤病例。肌腱的基本生物学功能是将肌肉收缩的力传递到骨骼,从而引导肢体运动,其主要由胶原纤维构成,其呈现纳米级且各向异性结构。由于静电纺丝技术所制备的纤维直径处于亚微米级及纳米级,具有更大的比表面积、更高的孔隙率、更优异的力学性能等优势,从而广泛应用于组织工程领域。通过调控静电纺丝纳米纤维表面形态可以增加细胞粘附位点,调节细胞内控制转录活性和基因表达的信号路径,对纳米纤维排列也可通过改变接收装置进行调控,当其高度取向时可引导肌腱成纤维细胞定向排列,从而从各个角度调控细胞行为。
由于胶原蛋白具有压电性,即胶原蛋白在应变时会产生电荷(直接压电效应),因此肌腱作为一种胶原组织也表现为压电性质。肌腱的直接压电效应与肌腱在受到定向应力时的生长和重塑能力有关,应用于损伤肌腱的微电流可增强愈合过程,在电刺激作用下,毛细血管在短期内增加,成纤维细胞通过增加DNA含量和蛋白表达来进行大规模增殖,从而促进肌腱愈合。组织工程中现有压电式电活性材料主要分为:
(1)压电陶瓷(钛酸钡、铌酸锶钡、PZT、ZNO等):广泛应用于生物传感器、医学成像等方面,压电系数高,各方面性能稳定,机械性能好,但由于生物相容性有待验证而未广泛应用于体内组织工程;
(2)压电聚合物(PVDF、PLLA等):此种材料是构成生物体的生物分子,生物相容性较好,但由于其压电系数较小,体内不易降解等问题限制了其应用;
(3)生物分子压电材料(氨基酸、多肽、蛋白质等):这些材料是人体活组织中存在的生物分子,并且通过调节构象某些生物分子压电材料可以显示出与传统压电材料相当的压电效应。
对于肌腱修复方向的研究大多集中注意力于模仿肌腱的层级结构和取向结构,来达到仿生的效果。然而对比结构调控,采用电活性生物支架通过电刺激①可以更显著的影响细胞取向生长,达到肌腱的各向异性结构;②可以使得毛细血管激增导致所需营养素增加,从而协助再生和修复过程;③可以增加成纤维细胞和肌腱细胞的数量,从而促进胶原合成,加速受损肌腱的修复;④可以加速疤痕组织修复。
目前压电式电活性生物支架主要应用于神经、心肌、肌肉、骨骼组织的修复,用于肌腱组织工程支架的研究鲜有报道,并且目前用于体内组织工程的压电材料种类较少,较好的生物性能和较优异的压电效应难以两全,如何从结构、微环境、压电性能等多角度仿生肌腱组织是肌腱修复组织工程中亟待解决的问题。
发明内容
本发明的目的是针对现有技术中的不足,提供一种压电式氨基酸生物支架及其制备方法,该支架克服现有技术无法仿生肌腱组织具有压电效应的特性,选用具有生物相容性的生物分子氨基酸作为压电材料,同时支架中的纳米纤维具有微孔结构增加细胞粘附位点且高度取向排列,在保证生物相容性的基础上提高了支架的机械拉伸强度,从多角度仿生肌腱结构,促进肌腱修复进程。
为实现上述目的,本发明采取的技术方案是:
本发明的第一方面是提供一种压电式氨基酸生物支架的制备方法,步骤包括:
S1、将聚合物溶于二元溶剂中,配置成纺丝液;
S2、通过静电纺丝技术,由所述纺丝液制得直径为0.5μm-3.0μm的纳米纤维;通过动态水浴加捻技术,由所述纳米纤维制得纳米纤维支架;
S3、将所述纳米纤维支架浸渍于氨基酸溶液中,于低温中将所述纳米纤维支架析出,冷冻干燥后即得所述压电式氨基酸生物支架。
优选地,所述聚合物选自:聚乳酸、聚己内酯或乳酸-己内酯共聚物中的至少一种。
优选地,所述二元溶剂选自:二氯甲烷/二甲基亚砜、二氯甲烷/N,N-二甲基甲酰胺或二氯甲烷/丙酮中的至少一种。
优选地,所述静电纺丝技术包括:将所述纺丝液加入注射器,施加15KV-20KV的正负高压,以1.0mL/h-1.2mL/h的速度推进泵,接收距离为10cm-15cm,接收辊水平方向的移动速度为1000cm/min-1200cm/min,接收辊的转速为100rpm-120rpm。
优选地,氨基酸选自:γ-甘氨酸或γ-酪氨酸中的至少一种。
优选地,所述氨基酸溶液的质量浓度为20%-25%。
优选地,所述浸渍包括:将所述氨基酸溶液于50℃的水浴锅中加热溶解,将所述纳米纤维支架浸入所述氨基酸溶液中30min。
优选地,所述低温不高于-80℃。
本发明的第二方面是提供一种采用如上所述制备方法制得的压电式氨基酸生物支架。
本发明采用以上技术方案,与现有技术相比,具有如下技术效果:
本发明的压电式氨基酸生物支架能够满足肌腱作为一种胶原组织所表现出的压电性质,人体在运动时会产生机械刺激,当其作用于压电式生物支架后,支架产生一定强度的电刺激,导致毛细血管在短期内增加,成纤维细胞和肌腱细胞通过增加DNA含量和蛋白表达来进行大规模增殖,从而促进肌腱修复;
本发明的压电式氨基酸生物支架模仿天然肌腱组织的结构形态,具有一定取向排列的纳米纤维纱,配合电刺激进一步诱导细胞取向迁移和排列;同时取向结构还可以增强纤维轴向方向的拉伸强度;
本发明的压电式氨基酸生物支架采用动态水浴加捻装置将纳米纤维进行缠绕加捻,使纤维之间相互抱和,能够模拟天然肌腱的应力-应变曲线,低应变时为非线性趾区,高应变时为线性区,以满足肌腱在使用时对于力学性能的要求;
本发明的压电式氨基酸生物支架,在纤维表面具有微孔结构,为营养物质的输入和代谢废物的排放提供了通道,保证新陈代谢功能;由于微孔结构增加了纤维的比表面积,使得细胞粘附位点增加,促进细胞粘附增殖;
本发明的压电式氨基酸生物支架根据临床需求,可在纺丝液的制备过程中加入抗炎、防止组织粘连的药物,例如布洛芬、姜黄素等,以用于减少炎症反应、防止组织粘连,更多角度的促进肌腱组织再生。
附图说明
图1为本发明中压电式氨基酸生物支架制备过程中所采用静电纺丝装置以及动态水浴加捻装置的结构示意图;
图2为本发明中压电式氨基酸生物支架在浸渍氨基酸溶液前的扫描电镜图;
图3为本发明中压电式氨基酸生物支架在浸渍氨基酸溶液后的扫描电镜图;
图4为本发明中压电式氨基酸生物支架的应力-应变曲线图;
图5为本发明中压电式氨基酸生物支架的电压-时间曲线图。
具体实施方式
下面将结合本发明实施例中的附图,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的前提下所获得的所有其他实施例,都属于本发明保护的范围。
需要说明的是,在不冲突的情况下,本发明中的实施例及实施例中的特征可以相互组合。
下面结合附图和具体实施例对本发明作进一步说明,但不作为本发明的限定。
实施例
本实施例提供一种压电式氨基酸生物支架,其制备的步骤包括:
S1、将1.2g聚左旋乳酸(分子量约为二十一万,购于济南岱罡生物科技有限公司)溶于10mL二氯甲烷/二甲基亚砜(8:1,购于上海凌峰化学试剂有限公司)中,配置成质量浓度为12%的纺丝液;
S2、如图1所示,将所述纺丝液加入注射器,施加15KV的正负高压,以1.2mL/h的速度推进泵,接收距离为10cm,接收辊水平方向的移动速度为1000cm/min,接收辊的转速为100rpm,制得纳米纤维支架,其表面形态的扫描电镜结果如图2所示;
S3、将25gγ-甘氨酸溶于100mL去离子水中,于50℃的水浴锅中加热溶解,配置成质量浓度为25%的氨基酸溶液;将所述纳米纤维支架浸渍于所述氨基酸溶液中30min,于-80℃的冰箱中将所述纳米纤维支架析出,冷冻干燥后即得所述压电式氨基酸生物支架,其表面形态的扫描电镜结果如图3所示;
所述压电式氨基酸生物支架呈正方体,厚度约为0.8mm-1mm,修复人类肌腱组织时,支架面积可根据实际受损肌腱的尺寸定制。
将所述压电式氨基酸生物支架置于万能测试机上进行力学拉伸测试,试样横截面面积为1.0mm×0.8mm,试样标距为2.0cm,拉伸速率为20.0mm/min;应力-应变曲线如图4所示,结果显示所述压电式氨基酸生物支架最大的拉伸强度为12.95MPa,断裂伸长率为49.66%,杨氏模量为514.85MPa,力学性能较好。
将所述压电式氨基酸生物支架置于原子电荷捕捉器上进行测试,试样面积为10mm×10mm,试样标距为2.0cm,设置施加压力的频率为120次/min,压力为10N;电压-时间曲线如图5所示,结果显示所述压电式氨基酸生物支架100mm2面积可产生6V的电压,符合人体内肌腱压电性质。
以上所述仅为本发明较佳的实施例,并非因此限制本发明的实施方式及保护范围,对于本领域技术人员而言,应当能够意识到凡运用本发明说明书及图示内容所作出的等同替换和显而易见的变化所得到的方案,均应当包含在本发明的保护范围内。
Claims (9)
1.一种压电式氨基酸生物支架的制备方法,其特征在于,步骤包括:
S1、将聚合物溶于二元溶剂中,配置成纺丝液;
S2、通过静电纺丝技术,由所述纺丝液制得直径为0.5μm-3.0μm的纳米纤维;通过动态水浴加捻技术,由所述纳米纤维制得纳米纤维支架;
S3、将所述纳米纤维支架浸渍于氨基酸溶液中,于低温中将所述纳米纤维支架析出,冷冻干燥后即得所述压电式氨基酸生物支架。
2.根据权利要求1所述的制备方法,其特征在于,所述聚合物选自:聚乳酸、聚己内酯或乳酸-己内酯共聚物中的至少一种。
3.根据权利要求1所述的制备方法,其特征在于,所述二元溶剂选自:二氯甲烷/二甲基亚砜、二氯甲烷/N,N-二甲基甲酰胺或二氯甲烷/丙酮中的至少一种。
4.根据权利要求1所述的制备方法,其特征在于,所述静电纺丝技术包括:将所述纺丝液加入注射器,施加15KV-20KV的正负高压,以1.0mL/h-1.2mL/h的速度推进泵,接收距离为10cm-15cm,接收辊水平方向的移动速度为1000cm/min-1200cm/min,接收辊的转速为100rpm-120rpm。
5.根据权利要求1所述的制备方法,其特征在于,氨基酸选自:γ-甘氨酸或γ-酪氨酸中的至少一种。
6.根据权利要求1所述的制备方法,其特征在于,所述氨基酸溶液的质量浓度为20%-25%。
7.根据权利要求1所述的制备方法,其特征在于,所述浸渍包括:将所述氨基酸溶液于50℃的水浴锅中加热溶解,将所述纳米纤维支架浸入所述氨基酸溶液中30min。
8.根据权利要求1所述的制备方法,其特征在于,所述低温不高于-80℃。
9.一种采用如权利要求1-8任一项所述制备方法制得的压电式氨基酸生物支架。
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