CN117503984B - 一种富血小板血浆-硫化铜复合纳米纤维敷料及制备方法 - Google Patents
一种富血小板血浆-硫化铜复合纳米纤维敷料及制备方法 Download PDFInfo
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Abstract
本发明实施例提供一种富血小板血浆‑硫化铜复合纳米纤维敷料及制备方法,所述制备方法包括以下步骤:S1:制备富血小板血浆;S2:制备CuS纳米颗粒;S3:制备复合纳米纤维壳层溶液;S4:制备复合纳米纤维核层溶液;S5:将所述壳层溶液与所述核层溶液混合,利用同轴静电纺丝装置原位制备得到具有核壳结构的富血小板血浆‑硫化铜复合纳米纤维敷料。该制备方法利用同轴静电纺丝技术将富血小板血浆及CuS复合纳米纤维材料的优势进行结合,实现血小板快速释放和持续释放的功能,同时具备抗菌、止血、促进创面愈合、防粘连等多种功能。
Description
技术领域
本发明涉及医用敷料技术领域,尤其涉及一种富血小板血浆-硫化铜复合纳米纤维敷料及制备方法。
背景技术
目前,创面治疗和伤口修复是医学领域的重要研究方向。血小板是一种重要的血液成分,其α颗粒中储存了一系列有效的生物活性介质,包括溶酶体酶、凝血因子、免疫和粘附分子、趋化因子和生长因子等,在止血、宿主防御、血管生成和组织修复等方面发挥重要作用,目前富血小板血浆(PRP)作为一种含有丰富生长因子和细胞因子的血浆,已被广泛应用于创面治疗。然而,传统的PRP应用存在血小板不稳定、保存困难、释放速度慢、释放量不可控等问题。
现有的纳米纤维辅料大多功能较为单一,单纯的具备抗菌或止血某一项功能;并且纳米纤维敷料在应用过程中可能会遇到稳定性问题,由于纳米纤维的特殊结构和较大比表面积,容易受到环境条件的影响,如湿度、温度等,这可能导致纳米纤维的聚集、团块化或失去原有的功能。此外由于血小板制品在制备过程中存在不稳定、保存困难以及血小板中生长因子的释放不可控等问题,影响血小板在科研及医疗领域的广泛应用。
发明内容
本发明的目的在于克服现有技术的不足,提供一种富血小板血浆-硫化铜复合纳米纤维敷料及制备方法,利用同轴静电纺丝技术将富血小板血浆与CuS复合纳米纤维进行结合。本发明的富血小板血浆-复合纳米纤维敷料原位纺丝于伤口处,既解决了血小板制品及纳米纤维材料长期储存不稳定的问题,同时将两者的优势进行结合,得到快速抑菌、止血、控制血小板释放和持续释放,促进创面愈合、防粘连等多种功能的复合纳米纤维敷料。
本发明第一方面,提供一种富血小板血浆-硫化铜复合纳米纤维敷料的制备方法,所述制备方法包括以下步骤:
对采集的血液样本,利用离心法分离得到富血小板血浆,并在所述富血小板血浆中加入凝血酶进行血小板活化;
将适量的铜盐和硫化物源分别溶解在适量溶剂中,通过溶剂热法制备得到CuS纳米颗粒;
将所述CuS纳米颗粒分散在混有聚合物的有机溶剂中得到壳层溶液;
将经过凝血酶活化后的富血小板血浆溶于聚乙烯醇溶液得到核层溶液;
将所述壳层溶液与所述核层溶液混合,利用同轴静电纺丝装置原位制备具有核壳结构的富血小板血浆-硫化铜复合纳米纤维敷料。
作为本发明一种优选方案:其中,所述铜盐为硝酸铜,所述硫化物源为硫代乙酰胺,所述溶剂为明胶溶液。
作为本发明一种优选方案:其中,所述硝酸铜与所述硫代乙酰胺的浓度配比为1:1-1:3。
作为本发明一种优选方案:其中,所述聚合物为生物可降解聚合物,所述聚合物包括聚乙酸内酯(PCL)、聚乳酸(PLA)、聚乙烯醇(PVA)、聚乙烯吡咯烷酮(PVP)中的一种或多种。
作为本发明一种优选方案:其中,所述聚合物为聚乙烯吡咯烷酮(PVP),所述有机溶剂为无水乙醇。
作为本发明一种优选方案:其中,在配制的所述壳层溶液中,聚合物浓度为15%,CuS纳米颗粒质量分数为0.1%。
作为本发明一种优选方案:其中,所述聚乙烯醇溶液的浓度为10%,与富血小板血浆的质量配比为1:2。
作为本发明一种优选方案:其中,所述同轴静电纺丝的电压为15kV,核壳流速比为2:1, 同轴静电纺丝针头与接受端之间的距离为15cm,喷嘴直径为0.5mm。
本发明第二方面,提供一种富血小板血浆-硫化铜复合纳米纤维敷料,由第一方面所述的制备方法制备得到。
本发明第三方面,提供一种富血小板血浆-硫化铜复合纳米纤维敷料,所述敷料包括:生物可降解聚合物、CuS纳米颗粒、富血小板血浆。
与现有技术相比,本发明的优点和积极效果是:
1、本发明的制备方法制备得到的敷料操作简单、应用范围广泛、同时具备抑菌、止血、维持血小板相关细胞因子释放、促伤口愈合等多种功能,适用于急性及慢性创伤的修复;
2、本发明采用富血小板血浆与生物可降解聚合物组成的复合纳米纤维材料,使制得的敷料具备快速止血,维持血小板的快速和持续释放,促进创面愈合等多种功能,同时制备过程中添加CuS纳米颗粒,为敷料提供良好的抗菌性能,形成具有一定机械强度的纤维网络,能够支撑和保护伤口,防止外界刺激和细菌侵入,促进伤口修复;
3、本发明制备及使用方便,不受伤口形状、面积、部位等条件的限制,可直接原位纺丝于伤口位置,便于突发紧急创伤的救治,同时血小板成分的持续释放也使其适用于慢性伤口的愈合治疗,此外纳米材料的生物相容性及可降解性,可避免更换敷料对伤口造成的二次创伤;
4、本发明将血小板制品包被于复合纳米纤维材料内部,通过同轴静电纺丝方法制备核壳结构的纳米纤维辅料,解决了血小板制品不稳定、储存困难等问题,扩大了血小板制品的应用范围。
附图说明
为了更清楚地说明本发明实施例中的技术方案,下面将对实施例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为本发明实施例提供的一种富血小板血浆-硫化铜复合纳米纤维敷料的制备方法流程示意图;
图2为本发明实施例制备的富血小板血浆-硫化铜复合纳米纤维敷料扫描电镜SEM图片。
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,本领域技术人员在没有作出创造性劳动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。
本发明提供了一种富血小板血浆-硫化铜复合纳米纤维敷料及制备方法,利用同轴静电纺丝技术将富血小板血浆与CuS复合纳米纤维进行结合,CuS复合纳米纤维具有纳米级别的尺寸,较大的比表面积及良好的生物相容性,有利于与创面接触并释放药物,不会引起明显的毒性反应或免疫排斥反应。CuS纳米颗粒可以与血小板相互作用,促进血小板的活化和聚集,从而加速血小板释放生长因子,促进创面愈合,还可以形成具有一定机械强度的纤维网络,能够支撑和保护伤口,防止外界刺激和细菌侵入,促进伤口的修复。富血小板血浆作为核芯层结构包裹在CuS复合纳米纤维形成的壳层结构内部,实现血小板内部生长因子的快速和持续释放,可适用于急性创伤及慢性伤口的愈合治疗。
本发明提供了一种富血小板血浆-硫化铜复合纳米纤维敷料,所述敷料包括:生物可降解聚合物、CuS纳米颗粒、富血小板血浆。
优选的,所述生物可降解聚合物包括聚乙酸内酯(PCL)、聚乳酸(PLA)、聚乙烯醇(PVA)、聚乙烯吡咯烷酮(PVP)中的一种或多种,由于聚合物的低免疫原性、高生物相容性、无毒环保等特性,为本发明中敷料与伤口的直接接触及避免粘连造成的二次创伤提供可能;
所述CuS纳米颗粒具有较大的比表面积和高活性,有利于与生物体接触和反应,同时通过释放Cu2+离子和产生活性氧来抑制细菌的生长和繁殖,此外还可以与血小板相互作用,促进血小板的活化和聚集,从而加速血小板释放生长因子,促进创面愈合,这使得CuS纳米颗粒在医学领域的伤口敷料和抗菌材料方面具有较大的应用潜力,因此作为本发明中敷料的快速止血、抗菌及促伤口愈合组分;
所述富血小板血浆作为自体生物活性成分,可快速以及持续释放生长因子及细胞因子,高浓度的生长因子和细胞因子能够促进表皮和内皮细胞再生、刺激血管生成和胶原蛋白沉积,加速伤口愈合,作为本发明中敷料的止血及促伤口愈合组分。
本发明实施例提供一种富血小板血浆-硫化铜复合纳米纤维敷料的制备方法,所述制备方法包括以下步骤:
S1:采集血液样本,利用离心法分离得到富血小板血浆,并在所述富血小板血浆中加入凝血酶进行血小板活化;
S2:将适量的铜盐和硫化物源分别溶解在适量溶剂中,通过溶剂热法制备得到CuS纳米颗粒;
S3:将所述CuS纳米颗粒分散在混有聚合物的有机溶剂中得到壳层溶液;
S4:将步骤S1中经凝血酶活化后的富血小板血浆溶于聚乙烯醇溶液得到核层溶液;
S5:将所述壳层溶液与所述核层溶液混合,利用同轴静电纺丝装置原位制备具有核壳结构的富血小板血浆-硫化铜复合纳米纤维敷料。
本发明的上述制备方法利用同轴静电纺丝技术将富血小板血浆及CuS复合纳米纤维材料的优势进行结合,实现血小板快速释放和持续释放的功能,同时具备抗菌、止血、促进创面愈合、防粘连等多种功能。通过同轴静电纺丝装置将富血小板血浆-复合纳米纤维敷料原位纺丝于伤口处,既解决了血小板制品及纳米纤维材料长期储存不稳定的问题,同时将两者的优势进行结合,得到快速抑菌、止血、控制血小板释放和持续释放,促进创面愈合、防粘连等多种功能的复合纳米纤维敷料。
优选的,在一些实施例中:所述步骤S2中,所述铜盐为硝酸铜,所述硫化物源为硫代乙酰胺,所述溶剂为明胶溶液。
优选的,在一些实施例中:所述步骤S2中,所述硝酸铜与所述硫代乙酰胺的浓度配比为1:1-1:3。
优选的,在一些实施例中:所述步骤S3中,其中,所述聚合物为生物可降解聚合物,所述聚合物包括聚乙酸内酯(PCL)、聚乳酸(PLA)、聚乙烯醇(PVA)、聚乙烯吡咯烷酮(PVP)中的一种或多种。
优选的,在一些实施例中:所述步骤S3中,所述聚合物为聚乙烯吡咯烷酮(PVP),所述有机溶剂为无水乙醇。
优选的,在一些实施例中:所述步骤S3中,在所述壳层溶液中,配制聚合物浓度为10%-30%的溶液,加入制备好的CuS纳米粒子,质量分数为0.1~5%,室温下在磁力搅拌器中搅拌12h至均匀稳定,制备成壳层溶液;进一步优选地,聚合物浓度为15%,CuS纳米颗粒质量分数为0.1%。
优选的,在一些实施例中:所述步骤S4中,所述聚乙烯醇溶液的浓度为1%-10%,与富血小板血浆的质量配比为1:1-1:3;进一步优选地,所述聚乙烯醇溶液的浓度为10%,与富血小板血浆的质量配比为1:2。
优选的,在一些实施例中:所述步骤S5中,所述同轴静电纺丝的电压为10-20kV,核壳流速比为1:1-3:1,同轴静电纺丝针头与接受极之间的距离为10-20cm,喷嘴直径为0.4-0.7mm,得到所述富血小板血浆-CuS复合纳米纤维敷料;进一步优选地,所述同轴静电纺丝的电压为15kV,核壳流速比为2:1, 同轴静电纺丝针头与接受端之间的距离为15cm,喷嘴直径为0.5mm。
本发明提供一种富血小板血浆-硫化铜复合纳米纤维敷料,由所述制备方法制备得到。
实施例1:图1示出了本发明实施例1提供的一种富血小板血浆-硫化铜复合纳米纤维敷料的制备方法流程示意图,如图1所示,所述制备方法包括以下步骤:
S1:制备富血小板血浆:采集患者的血液样本,利用离心法分离得到富血小板血浆,调整血小板浓度为1*109/ml,加入终浓度为1U/ml的凝血酶进行血小板活化;
S2:制备CuS纳米颗粒:硝酸铜与硫代乙酰胺的浓度配比为1:1,准备50ml质量分数为1%的明胶溶液,加入5ml 0.2M硝酸铜溶液,室温搅拌1h,将5ml 0.2M硫代乙酰胺溶液缓慢滴加到硝酸铜溶液中搅拌均匀,100℃反应2h,反应完成后,将产生的CuS纳米颗粒收集起来用乙醇和去离子水对材料进行交替洗涤3次,以去除残余的反应物和杂质,最后,将洗涤后的CuS纳米颗粒进行冷冻干燥;
S3:制备复合纳米纤维壳层溶液:称取1.5g聚乙烯吡咯烷酮(PVP)溶于8.5g无水乙醇中,室温下磁力搅拌1h至完全溶解,将制备好的CuS纳米颗粒分散在上述聚合物溶液中,最终质量分数为0.1%,室温下在磁力搅拌器中搅拌12h至均匀稳定,制备成壳层纺丝液;
S4:制备复合纳米纤维核层溶液:称取1g聚乙烯醇溶于9g去离子水中制备成浓度为10%的溶液,与富血小板血浆的质量配比为1:2进行混匀,制备成核层纺丝液;
S5:将配置好的核层纺丝液与壳层纺丝液分别装入同轴静电纺丝装置,原位制备具有核壳结构的富血小板血浆-CuS复合纳米纤维多功能敷料,参数设置电压为15kV,核壳流速比为2:1, 同轴静电纺丝针头与接受端之间的距离为15cm,喷嘴直径为0.5mm。
实施例2:一种富血小板血浆-硫化铜复合纳米纤维敷料,由实施例1制备得到。通过扫描电镜对由上述步骤S1-S5制备得到的富血小板血浆-硫化铜复合纳米纤维敷料进行表征,如图2所示是利用本发明实施例1的制备方法制备得到的富血小板血浆-硫化铜复合纳米纤维敷料扫描电镜SEM图片。
本发明利用同轴静电纺丝技术将富血小板血浆与CuS复合纳米纤维进行结合,CuS复合纳米纤维具有纳米级别的尺寸,较大的比表面积及良好的生物相容性,有利于与创面接触并释放药物,不会引起明显的毒性反应或免疫排斥反应。CuS纳米颗粒可以与血小板相互作用,促进血小板的活化和聚集,从而加速血小板释放生长因子,促进创面愈合,还可以形成具有一定机械强度的纤维网络,能够支撑和保护伤口,防止外界刺激和细菌侵入,促进伤口的修复。富血小板血浆作为核芯层结构包裹在CuS复合纳米纤维形成的壳层结构内部,实现血小板内部生长因子的快速和持续释放,可适用于急性创伤及慢性伤口的愈合治疗。该敷料具备快速止血,维持血小板的快速和持续释放,促进创面愈合等多种功能,同时制备过程中添加CuS纳米颗粒,为敷料提供良好的抗菌性能,形成具有一定机械强度的纤维网络,能够支撑和保护伤口,防止外界刺激和细菌侵入,促进伤口修复。
需要说明的是,对于前述的各方法实施例,为了简单描述,故将其都表述为一系列的动作组合,但是本领域技术人员应该知悉,本公开并不受所描述的动作顺序的限制,因为依据本公开,某些步骤可以采用其他顺序或者同时进行。其次,本领域技术人员也应该知悉,说明书中所描述的实施例均属于可选实施例,所涉及的动作并不一定是本公开所必须的。
虽然已经通过示例对本发明的一些特定实施例进行了详细说明,但是本领域的技术人员应该理解,以上示例仅是为了进行说明,而不是为了限制本发明的范围。本领域的技术人员还应理解,可以对实施例进行多种修改而不脱离本发明的范围和精神。本发明的范围由所附权利要求来限定。
Claims (5)
1.一种富血小板血浆-硫化铜复合纳米纤维敷料的制备方法,其特征在于,所述制备方法包括如下步骤:
对采集的血液样本,利用离心法分离得到富血小板血浆,并在所述富血小板血浆中加入凝血酶进行血小板活化;
将适量的硝酸铜和硫代乙酰胺分别溶解在适量明胶溶剂中,其中,所述硝酸铜与所述硫代乙酰胺的摩尔浓度配比为1:1-1:3,通过溶剂热法制备得到CuS纳米颗粒;
将所述CuS纳米颗粒分散在混有生物可降解聚合物的无水乙醇中得到壳层溶液;其中,在制备所述壳层溶液过程中,先配制聚合物质量浓度为10%-30%的溶液,然后加入制备好的CuS纳米粒子,质量分数为0.1~5%,室温下在磁力搅拌器中搅拌12h至均匀稳定,制备成壳层溶液;
将经过凝血酶活化后的富血小板血浆溶于聚乙烯醇溶液得到核层溶液;
将所述壳层溶液与所述核层溶液混合,利用同轴静电纺丝装置原位制备具有核壳结构的富血小板血浆-硫化铜复合纳米纤维敷料;
其中,所述生物可降解聚合物为聚乙烯吡咯烷酮(PVP)。
2.根据权利要求1所述的制备方法,其特征在于,其中,所述聚乙烯醇溶液的质量浓度为10%,与富血小板血浆的质量配比为1:2。
3.根据权利要求1所述的制备方法,其特征在于,其中,所述同轴静电纺丝的电压为15kV,核壳流速比为2:1, 同轴静电纺丝针头与接受端之间的距离为15cm,喷嘴直径为0.5mm。
4.根据权利要求1所述的制备方法,其特征在于,其中,生物可降解聚合物质量浓度为15%,CuS纳米颗粒质量分数为0.1%。
5.一种富血小板血浆-硫化铜复合纳米纤维敷料,其特征在于,由权利要求1-4任一项所述的制备方法制备得到。
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