CN115252869A - 一种促进血管再生的纳米敷料及其制备方法 - Google Patents
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Abstract
本发明属于生物医用材料领域,公开了一种促进血管再生的纳米敷料及其制备方法,包括以四氧化三钴纳米粉作为核,以木质素和单宁酸氧化聚合物作为壳,表面键合亲水性单分子层的纳米颗粒为基础,采用静电纺丝得到促进血管再生的纳米敷料。本发明制备的敷料具有强效激活细胞中缺氧诱导因子1的效果。
Description
技术领域
本发明属于生物医用材料领域,具体涉及一种促进血管再生的纳米敷料及其制备方法。
背景技术
众所周知,血管再生是伤口愈合的重要环节,受到诸多细胞因子的调控。血管内皮生长因子VEGF是目前发现的机体内最主要的血管生长因子之一,缺氧通过增加VEGF表达从而促进血管生成。应用生长因子可以促进血管生成和创面愈合,但是各种促血管生成的生长因子仍然存在一些问题,如保存困难、新生血管质量差、治疗费用大等问题。
最近发现缺氧诱导因子(HIF-1)是参与缺氧应答反应的关键性转录因子,能够调节VEGF的表达,对创面愈合至关重要,已成为慢性创面愈合研究中的新热点。HIF由HIF-1α和HIF-1β构成,其活性主要由HIF-1α决定,HIF核转位后调控细胞因子的转录。在正常创面愈合中,缺氧能导致HIF-1α表达升高,从而促进血管生成和伤口愈合。
常规的创面愈合过程中,伤口暴露在空气中,不能形成低氧环境,进而VEGF不能高表达,使得血管生成受阻,伤口愈合缓慢。
发明内容
有鉴于此,本发明的目的在于提供一种促进血管再生的纳米敷料及其制备方法,该敷料具有强效激活细胞中缺氧诱导因子1的效果。
为了解决上述技术问题,本发明提供了一种促进血管再生的纳米敷料的制备方法,包括如下步骤:
S1.将四氧化三钴纳米粉和杯芳烃加入水中,超声波处理并搅拌,得到氧化钴悬液;
S2.将单宁酸加入步骤S1制备的氧化钴悬液中,充分搅拌,再加入木质素磺酸钠,密闭后放入高压釜中反应,之后离心收集沉淀,干燥得到Co2+-单宁酸/木质素复合物;
S3.将步骤S2制备的Co2+-单宁酸/木质素复合物加入含有巯基高级醇的DMF溶液中,加入热塑性聚氨酯TPU进行静电纺丝制得促进血管再生的纳米敷料。
优选的,步骤S1中,所述四氧化三钴纳米粉平均粒径为50-200nm。
优选的,步骤S1中,所述杯芳烃为4-磺酰杯[6]芳烃、4-磺酸杯[8]芳烃中的一种。
优选的,步骤S1中,所述四氧化三钴纳米粉、杯芳烃和水比例为(1-3)g:(0.02-0.05)g:100mL。
优选的,步骤S2中,所述单宁酸、氧化钴悬液和木质素磺酸钠的比例为(7-10)g:100mL:(0.4-0.8)g。
优选的,步骤S2中,所述反应的温度为160-180℃,反应压力为0.6-1.0MPa,反应时间为4-5h。
优选的,步骤S3中,所述巯基高级醇为16-巯基十六烷-1-醇、11-巯基-1-十一醇中的一种,DMF溶液中巯基高级醇的浓度为1.70-5.67g/L。
优选的,步骤S3中,所述Co2+-单宁酸/木质素复合物、含有巯基高级醇的DMF溶液和热塑性聚氨酯TPU的比例为1.0g:(150-500)mL:(7.5-100)g。
优选的,步骤S3中,所述静电纺丝条件为电压13-18kV、距离10-18cm,注射速率0.6-1.2mL/h。
本发明还提供了一种上述制备方法制备得到的促进血管再生的纳米敷料。
与现有技术相比,本发明具有以下有益效果:
本发明以四氧化三钴作为核心,利用杯芳烃的磺酸基和单宁酸对核心刻蚀,使其粒径进一步缩小,同时溶解的Co2+能够进入杯芳烃的环中,并通过杯芳烃的酚羟基进行配位,稳定其化学性质;而一部分Co3+又能氧化单宁酸和木质素磺酸钠实现分子交联并形成壳层,自身则被还原为Co2+。氧化交联的单宁酸和木质素磺酸钠在壳层表面聚集大量的醌官能团,能够与巯基高级醇接枝,实现粒子的亲水性转变,内部的Co2+也能够在中性溶液环境下缓慢地释放出来。释放的Co2+进入细胞能够持续的上调HIF-1α的表达,促使HIF-1的下游基因调控,其中包括上调VEGF,促进新生血管再生。
附图说明
图1为Westernblot实验的统计结果。
具体实施方式
为了进一步理解本发明,下面结合实施例对本发明优选实施方案进行描述,但是应当理解,这些描述只是为了进一步说明本发明的特征和优点,而不是对本发明权利要求的限制。
实施例1
一种促进血管再生的纳米敷料的制备方法,包括如下步骤:
1.将1.7g平均粒径为120nm的四氧化三钴纳米粉和0.04g4-磺酰杯[6]芳烃加入100mL水中,超声波处理并搅拌,得到氧化钴悬液;
2.将7.8g单宁酸加入步骤1制备的氧化钴悬液中,充分搅拌,再加入0.5g木质素磺酸钠,密闭后放入高压釜中,172℃0.7MPa反应4.7h,之后离心收集沉淀,干燥得到Co2+-单宁酸/木质素复合物;
3.将1g步骤2制备的Co2+-单宁酸/木质素复合物加入240mL含有2.46g/L16-巯基十六烷-1-醇的DMF溶液中,加入25gTPU进行静电纺丝,电压17kV、距离12cm,注射速率0.9mL/h制得促进血管再生的纳米敷料。
实施例2
一种促进血管再生的纳米敷料的制备方法,包括如下步骤:
1.将1g平均粒径为50nm的四氧化三钴纳米粉和0.02g4-磺酰杯[6]芳烃加入100mL水中,超声波处理并搅拌,得到氧化钴悬液;
2.将7g单宁酸加入步骤1制备的氧化钴悬液中,充分搅拌,再加入0.8g木质素磺酸钠,密闭后放入高压釜中,160℃1MPa反应5h,之后离心收集沉淀,干燥得到Co2+-单宁酸/木质素复合物;
3.将1g步骤2制备的Co2+-单宁酸/木质素复合物加入500mL含有5.67g/L11-巯基-1-十一醇的DMF溶液中,加入100gTPU进行静电纺丝,电压18kV、距离10cm,注射速率1.2mL/h制得促进血管再生的纳米敷料。
实施例3
一种促进血管再生的纳米敷料的制备方法,包括如下步骤:
1.将3g平均粒径为200nm的四氧化三钴纳米粉和0.05g4-磺酸杯[8]芳烃加入100mL水中,超声波处理并搅拌,得到氧化钴悬液;
2.将10g单宁酸加入步骤1制备的氧化钴悬液中,充分搅拌,再加入0.4g木质素磺酸钠,密闭后放入高压釜中,180℃0.6MPa反应4h,之后离心收集沉淀,干燥得到Co2+-单宁酸/木质素复合物;
3.将1g步骤2制备的Co2+-单宁酸/木质素复合物加入150mL含有1.70g/L11-巯基-1-十一醇的DMF溶液中,加入7.5gTPU进行静电纺丝,电压13kV、距离18cm,注射速率0.6mL/h制得促进血管再生的纳米敷料。
对比例1
一种敷料的制备方法,包括如下步骤:
1.将1.7g平均粒径为120nm的四氧化三钴纳米粉加入100mL水中,超声波处理并搅拌,得到氧化钴悬液;
2.将7.8g单宁酸加入步骤1制备的氧化钴悬液中,充分搅拌,再加入0.5g木质素磺酸钠,密闭后放入高压釜中,172℃0.7MPa反应4.7h,之后离心收集沉淀,干燥得到单宁酸/木质素复合物;
3.将1g步骤2制备的单宁酸/木质素复合物加入240mL含有2.46g/L16-巯基十六烷-1-醇的DMF溶液中,加入25gTPU进行静电纺丝,电压17kV、距离12cm,注射速率0.9mL/h制得纳米敷料。
对比例2
一种敷料的制备方法,包括如下步骤:
1.将1.7g平均粒径为120nm的四氧化三钴纳米粉和0.04g4-磺酰杯[6]芳烃加入100mL水中,超声波处理并搅拌,得到氧化钴悬液;
2.将7.8g单宁酸加入步骤1制备的氧化钴悬液中,充分搅拌,再加入0.5g木质素磺酸钠,密闭后放入高压釜中,172℃0.7MPa反应4.7h,之后离心收集沉淀,干燥得到Co2+-单宁酸/木质素复合物;
3.将1g步骤2制备的Co2+-单宁酸/木质素复合物加入240mLDMF中,加入25gTPU进行静电纺丝,电压17kV、距离12cm,注射速率0.9mL/h制得纳米敷料。
抗氧化性能测试
利用L929细胞以4×103/孔的接种量接种DMEM高糖完全培养基的24孔板后,加入实施例1-3制备的敷料和对比例1-2制备的敷料,继续37℃,5%CO2培养24h,收集细胞,提取蛋白,采用Westernblot实验检测HIF-1α的表达水平,内标采用β-actin蛋白,以未用敷料处理的为空白对照,统计结果见图1。
根据图1显示,本发明制备的促进血管再生的纳米敷料具有良好的促进HIF-1α表达的作用,对比例的敷料对细胞中HIF-1α表达的促进作用较弱,究其原因,实施例的敷料中缓慢释放的Co2+具有促进HIF-1α的转录和表达。对比例1中由于未加入杯芳烃,刻蚀溶解得到的Co2+无法络合于杯芳烃的环中,复合物不存在可释放的Co2+。对比例2中,由于未与巯基高级醇接枝,获得的Co2+-单宁酸/木质素复合物为疏水性粒子,水不能进入其内部,内部的Co2+不能通过溶液环境释放出来。
本发明提供了一种促进血管再生的纳米敷料及其制备方法的思路及方法,具体实现该技术方案的方法和途径很多,以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。本实施例中未明确的各组成部分均可用现有技术加以实现。
Claims (10)
1.一种促进血管再生的纳米敷料的制备方法,其特征在于,包括如下步骤:
S1.将四氧化三钴纳米粉和杯芳烃加入水中,超声波处理并搅拌,得到氧化钴悬液;
S2.将单宁酸加入步骤S1制备的氧化钴悬液中,充分搅拌,再加入木质素磺酸钠,密闭后放入高压釜中反应,之后离心收集沉淀,干燥得到Co2+-单宁酸/木质素复合物;
S3.将步骤S2制备的Co2+-单宁酸/木质素复合物加入含有巯基高级醇的DMF溶液中,加入热塑性聚氨酯,静电纺丝制得促进血管再生的纳米敷料。
2.根据权利要求1所述的制备方法,其特征在于,步骤S1中,所述四氧化三钴纳米粉平均粒径为50-200nm。
3.根据权利要求1所述的制备方法,其特征在于,步骤S1中,所述杯芳烃为4-磺酰杯[6]芳烃或4-磺酸杯[8]芳烃。
4.根据权利要求1所述的制备方法,其特征在于,步骤S1中,所述四氧化三钴纳米粉、杯芳烃和水的比例为(1-3)g:(0.02-0.05)g:100mL。
5.根据权利要求1所述的制备方法,其特征在于,步骤S2中,所述单宁酸、氧化钴悬液和木质素磺酸钠的比例为(7-10)g:100mL:(0.4-0.8)g。
6.根据权利要求1所述的制备方法,其特征在于,步骤S2中,所述反应的温度为160-180℃,反应压力为0.6-1.0MPa,反应时间为4-5h。
7.根据权利要求1所述的制备方法,其特征在于,步骤S3中,所述巯基高级醇为16-巯基十六烷-1-醇或11-巯基-1-十一醇,所述含有巯基高级醇的DMF溶液中巯基高级醇的浓度为1.70-5.67g/L。
8.根据权利要求1所述的制备方法,其特征在于,步骤S3中,所述Co2+-单宁酸/木质素复合物、含有巯基高级醇的DMF溶液和热塑性聚氨酯的比例为1.0g:(150-500)mL:(7.5-100)g。
9.根据权利要求1所述的制备方法,其特征在于,步骤S3中,所述静电纺丝条件为:电压13-18kV、距离10-18cm,注射速率0.6-1.2mL/h。
10.根据权利要求1~9任一项所述制备方法制备得到促进血管再生的纳米敷料。
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