CN115054720B - 一种防粘连医用敷料及其制备方法 - Google Patents
一种防粘连医用敷料及其制备方法 Download PDFInfo
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- CN115054720B CN115054720B CN202210874874.4A CN202210874874A CN115054720B CN 115054720 B CN115054720 B CN 115054720B CN 202210874874 A CN202210874874 A CN 202210874874A CN 115054720 B CN115054720 B CN 115054720B
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- adhesion
- functional layer
- medical dressing
- zinc oxide
- nanofiber membrane
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Abstract
本发明涉及一种防粘连医用敷料及其制备方法,采用离子交换法,利用碳酸氢钠溶液将功能层(混有纳米氧化锌的静电纺纳米纤维膜)与吸液层(海藻酸钙无纺布)进行复合制得防粘连医用敷料;功能层中静电纺纳米纤维膜的材质为疏水生物惰性高分子聚合物,平均孔径为4~8μm;功能层中纳米氧化锌为球形纳米颗粒,含量为1~5wt%;制得的防粘连医用敷料的功能层与吸液层之间通过海藻酸钠凝胶的粘结作用复合。本发明制得的防粘连医用敷料安全可靠、性能稳定、具有良好的机械性能、工艺简易、成本适宜,特别地,其结构设计巧妙,对于不同量渗出液伤口均具有良好的防粘连效果,克服了常规敷料容易粘连伤口,引起二次伤害的弊端。
Description
技术领域
本发明属于医用敷料技术领域,涉及一种防粘连医用敷料及其制备方法。
背景技术
在伤口愈合过程中,毛细管的扩张会导致伤口渗出液的产生,其成分与血浆类似,含有蛋白质、炎性细胞、生长因子、坏死组织,以及细菌等生物质。当传统敷料,如棉纱布、常规无纺布等高表面能材料直接接触伤口时,渗出液中的蛋白质、细菌会和敷料表面之间产生氢键或静电力作用而贴合,随着伤口的愈合,渗出物中的水分蒸发后变干,敷料就会和伤口产生粘连。与此同时,新生肉芽组织也会通过细胞粘附长入敷料。当从伤口处剥离敷料时,就会破坏新生的肉芽组织,导致愈合过程的延迟,给患者增加痛苦。因此,开发防粘连医用敷料具有重要意义。
目前开发防粘连敷料主要有两种策略,策略一是利用材料的超亲水性,采用可吸收、可溶解材料或相变材料,使敷料在使用完毕后无需移除可直接洗去。如使用透明质酸(HA)、羧甲基纤维素钠(CMC)、胶原、聚乳酸(PLA)等。CN109925532A中公开了一种将壳聚糖和羧甲基纤维素钠溶液的混合物涂覆在羧甲基化棉织物表面而制备得到防粘连敷料;CN107261198B中公开了一种以类人胶原蛋白(LHC)和壳聚糖(CS)为原料的凝胶型防粘连敷料。但这种策略的缺陷在于材料吸水后机械性能大幅下降,保型性和抗张性差;有些材料还会存在小分子扩散毒性、单体交联剂残留、易氧化等潜在风险。策略二是利用材料的超疏水性,超疏水材料的表面能低,不易粘附其他物质,但仅凭单一医用超疏水基材很难达到理想的防粘连效果,故经常对基材进行表面改性,进一步增大基材的超疏水性、静电空间排斥、水合作用和拓扑结构等,使复合材料具有良好的防粘连功能。如使用聚乙二醇(PEG)、两性离子聚合物等。CN112007200A中公开了一种利用PEG表面改性的防粘连膜;CN109646704A中公开了一种利用有机硅乳液涂层绷带制备的防粘连医用绷带。但这些产品的防粘连效果仍有提升空间,且存在制作工艺复杂、成本高等问题。
因此需要开发更为理想的防粘连医用敷料,使其在安全可靠、性能稳定、良好机械性能、工艺简易、成本适宜的前提下,仍具有较好的防粘连效果,便于后续临床应用。
发明内容
本发明的目的是解决现有技术中存在的上述问题,提供一种防粘连医用敷料及其制备方法。
为达到上述目的,本发明采用的技术方案如下:
一种防粘连医用敷料的制备方法,采用离子交换法,利用碳酸氢钠溶液将功能层与吸液层进行复合制得防粘连医用敷料;
功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;
功能层中纳米氧化锌为球形纳米颗粒,含量为1~5wt%;球形纳米颗粒由于其比表面积更大,故在ZnO表面形成的水化层的相对面积更大,所以球形ZnO颗粒在较小添加量的情况下即可达到现有技术中棒状ZnO颗粒的抗蛋白吸附效果,从而大大降低了功能层中纳米ZnO颗粒的添加量,保证了医用敷料的安全性;
静电纺纳米纤维膜的材质为疏水生物惰性高分子聚合物;静电纺纳米纤维膜平均孔径为4~8μm,采用平均孔径为4~8μm的静电纺纳米纤维膜能够与无纺布层之间形成最优的润湿性梯度,从而具有优异的定向导液能力。
作为优选的技术方案:
如上所述的一种防粘连医用敷料的制备方法,纳米氧化锌的粒径为10~30nm。
如上所述的一种防粘连医用敷料的制备方法,静电纺纳米纤维膜的纤维平均直径为800~1200nm。
如上所述的一种防粘连医用敷料的制备方法,具体步骤如下:
(1)将疏水生物惰性高分子聚合物溶解在溶剂中得到质量分数为5~30%的纺丝液,向纺丝液中加入纳米氧化锌,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干8h以去除残留溶剂,制得混有纳米氧化锌的静电纺纳米纤维膜;
(2)用空气喷枪将质量分数为0.2~1.0M的碳酸氢钠水溶液喷洒在海藻酸钙无纺布的一面,一段时间后将步骤(1)制得的混有纳米氧化锌的静电纺纳米纤维膜覆盖在海藻酸钙无纺布的该面,于恒温恒湿环境中用50N的重物压置12h,制得防粘连医用敷料。
如上所述的一种防粘连医用敷料的制备方法,步骤(1)中疏水生物惰性高分子聚合物为聚偏氟乙烯(PVDF),溶剂为丙酮和二甲基甲酰胺的混合物。
如上所述的一种防粘连医用敷料的制备方法,步骤(1)中静电纺丝的工艺参数为:纺丝电压15~25kV,推注速度0.5~1.5mL/h,接收距离10~20cm,纺丝时间6~14min。
如上所述的一种防粘连医用敷料的制备方法,步骤(2)中海藻酸钙无纺布上碳酸氢钠水溶液喷洒密度为15~30g/m2。
如上所述的一种防粘连医用敷料的制备方法,步骤(2)中一段时间为3~10min。
本发明还提供采用如上所述的方法制得的防粘连医用敷料,由接触皮肤的功能层和外侧的吸液层复合而成,功能层与吸液层之间通过离子交换法形成的海藻酸钠凝胶的粘结作用而复合。
作为优选的技术方案:
如上所述的防粘连医用敷料,功能层的水接触角为145~150°,BSA蛋白吸附量为7.0~7.5ng/cm2,明胶蛋白凝块剥离强力为0.20~0.25N,6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的2.0~3.0%;
功能层与吸液层剥离强力测试中测得的界面复合强度始终大于2N/m;
防粘连医用敷料的单向导湿指数R为900~1100。
本发明的原理如下:
本发明的防粘连医用敷料,利用功能层的疏水性生物惰性高分子聚合物纳米纤维膜和纳米ZnO颗粒的协同作用,与吸液层可吸收伤口渗出液的功能相结合,联合达到良好的防粘连效果。低表面能的疏水性生物惰性高分子聚合物纳米纤维膜本身具有一定的抗生物质粘附功能;氧化锌分子表面会和周围环境中的水分发生水合作用,形成一层水化层,阻碍蛋白质的吸附,降低细胞粘附,达到防粘连的效果;且由于氧化锌纳米颗粒的添加,部分氧化锌纳米颗粒附着于纳米纤维表面,使纳米纤维表面形成很多微小凸起,微观结构更加粗糙,与荷叶表面类似,进一步提高了纤维膜的超疏水性能,协同增强了功能层的防粘连效果。溶有生物质的伤口渗出液依靠敷料的定向导液性能透过纳米纤维膜,被吸液性能优异的海藻酸钙无纺布吸收,及时清理出伤口表面,也降低了伤口处粘连的风险。
现有防粘附技术中使用的纳米ZnO颗粒为截面直径10~30nm、长100nm的棒状,添加量为20~30wt%。本发明的功能层中混有的纳米ZnO为粒径为10~30nm的球形纳米ZnO颗粒,球形纳米颗粒由于其比表面积更大,故在ZnO表面形成的水化层的相对面积更大,所以球形ZnO颗粒在较小添加量的情况下即可达到现有技术中棒状ZnO颗粒的抗蛋白吸附效果,从而大大降低功能层中纳米ZnO颗粒的添加量(添加量为1~5wt%),更大程度上保证了医用敷料的安全性。
作为医用敷料,需要具备优异的定向导液性能,定向导液性能在于两层之间具有润湿性梯度,实验探究发现,功能层静电纺纳米纤维膜的孔径对润湿性梯度具有很大的影响,孔径太小时,水无法导向无纺布层,从而堆积在静电纺丝膜上,而孔径太大时,静电纺丝膜含水率始终低于无纺布层,也不利于垂直导液,而采用平均孔径为4~8μm的静电纺纳米纤维膜能够与无纺布层之间形成最优的润湿性梯度,从而具有优异的定向导液能力。
本发明的防粘连医用敷料,功能层与吸液层的复合采用离子交换法,将碳酸氢钠水溶液喷洒在海藻酸钙无纺布上时,海藻酸钙在钠离子存在的情况下和钠离子置换变成海藻酸钠,其在有少量水存在的情况下变成凝胶状,当静电纺纳米纤维膜覆盖在其上面时,会产生粘接作用,从而实现复合。本发明的这种复合方法绿色安全,复合强力高且均匀,不会出现点胶复合时的强力不均现象,复合效果好。
有益效果:
(1)本发明的一种防粘连医用敷料的制备方法,操作简易、工艺成熟、材料易得、无有毒试剂残留;制备的敷料性能稳定、机械性能良好、成本适宜,便于推广;
(2)本发明的方法制得的防粘连医用敷料具有良好的防粘连功能,利用功能层的疏水生物惰性高分子聚合物和纳米氧化锌颗粒的协同作用,与吸液层的可吸收伤口渗出液的功能相结合,联合达到良好的防粘连效果,且防粘连功能安全可靠、持久稳定;
(3)对本发明的防粘连医用敷料进行防粘连功能评价,通过明胶蛋白凝块剥离测试、细胞粘附测试和蛋白粘附测试得出结论:本防粘连敷料防粘连效果良好,优于目前被广泛证实防粘连效果优良的PEG表面改性的防粘连膜;
(4)本发明的一种防粘连医用敷料,功能层采用的生物惰性高分子聚合物基材是疏水材料,具有良好的弹性和强度,遇水有良好的保型性;纳米纤维膜的结构特征仿生了皮肤细胞外基质中的纳米纤维结构,利于伤口愈合;加入的纳米ZnO还具有优良的抗菌性,避免了使用抗生素抗菌的潜在耐药性;吸液层保证了伤口微环境的清洁和适宜的湿度,利于伤口修复。
附图说明
图1为自制PVDF电纺膜、实施例1和实施例2的防粘连医用敷料功能层的静电纺纳米纤维膜的SEM图;
图2为明胶蛋白凝胶块分别与自制PVDF电纺膜、实施例1和实施例2的防粘连医用敷料功能层以及凡士林纱布的明胶蛋白凝块剥离强力测试结果;
图3为硅片、自制PVDF电纺膜、PEG涂层硅片、实施例1和实施例2的防粘连医用敷料功能层成纤维细胞的粘附结果;
图4为BSA标准曲线(a)及自制PVDF电纺膜、PEG涂层硅片、实施例1和实施例2的防粘连医用敷料功能层的蛋白吸附量结果(b);
图5为本发明实施例1的防粘连敷料侧截面的扫描电子显微镜图;
图6为实施例1、实施例3和对比例1的复合强力曲线图;
图7为实施例1、实施例3和对比例1的剥离强力柱形图;
其中,1、功能层;2、吸液层。
具体实施方式
下面结合具体实施方式,进一步阐述本发明。应理解,这些实施例仅用于说明本发明而不用于限制本发明的范围。此外应理解,在阅读了本发明的内容之后,本领域技术人员可以对本发明作各种改动或修改,这些等价形式同样落于本申请所附权利要求书所限定的范围。
本发明采用的物质来源:
(1)凡士林纱布:源自于商用的振德医用凡士林纱布;
(2)硅片:购自苏州研材微纳科技有限公司;
(3)PEG涂层的硅片:购自西安齐岳生物科技有限公司。
本发明所涉及到的性能指标的测试方法如下:
(1)水接触角:采用接触角测试仪测试功能层的水接触角;
(2)BSA蛋白吸附量:细胞粘附前提是蛋白质的吸附,因此,可以通过测试材料的蛋白吸附性能来预测纳米纤维膜的细胞粘附;将纳米纤维膜裁剪成直径为1.4cm的圆片作为测试样品,用PBS缓冲液冲洗样品三次,每次3min,随后将样品放置在24孔板中,加入提前配制好的BSA蛋白溶液(1mg/mL),放在37℃环境下静置6h,最后用质量分数为2%的SDS缓冲液对样品进行浸泡,去除未吸附的蛋白质,浸泡2h后加入BCA工作液,37℃水浴30min后,测试溶液的吸光度,通过标准曲线(如图4所示)获得溶液中的BSA含量,从而推测出粘附在样品单位面积上的BSA含量;
(3)明胶蛋白凝块剥离强力:采用PVDF模具(内部尺寸为16cm×60cm×3cm)将纳米纤维膜固定,模拟伤口渗出液的成分和粘度来配制40%w/v明胶溶液,将明胶溶液浇铸到内部尺寸为16cm×60cm×3cm的PVDF模具方框中,使溶液均匀平铺后,将整个装置放入烘箱内(烘箱温度为32℃,湿度为75%)来模拟伤口处的温湿度;24h后取出样品,去除PVDF模具,在30min之内,将纳米纤维膜和明胶蛋白凝块分别安装在拉伸仪的夹具上,剥离角度为180°,以100m/min的速度将纳米纤维膜和明胶蛋白凝块完全剥离,记录拉伸曲线;取拉伸曲线上20%~80%断裂伸长区域段的6个最大强力取平均值,为功能层的明胶蛋白凝块剥离强力;
(4)成纤维细胞粘附量:取尺寸为20×20mm的纳米纤维膜或硅片作为测试样品,对样品进行灭菌,然后将样品和皮肤成纤维细胞接种,培养6h后,先加入PBS对样品清洗三次,每次3min,然后用5%Triton X-100溶液浸泡5min,之后再加入PBS清洗三次,每次3min;之后在室温且避光的条件下,在孔板上有样品的每个孔中添加鬼笔环肽工作液,用PBS清洗三次,每次5min,再将样品的每个孔中添加200μL DIPA染料,复染时间为3min,之后用PBS清洗三次,每次3min,在荧光显微镜下观察,计算样品中成纤维细胞粘附量;
(5)抑菌率:采用GB/T 20944.3测试方法,在环氧乙烷灭菌后的纳米纤维膜上接种一定量的细菌,振荡培养18h后,稀释菌液,并将其接种在固体培养基上,24h后进行单菌落计数,计算菌液中活菌浓度,从而计算抑菌率;
(6)溶血率:采用的血液为成年新西兰大白兔(体重约2.5kg)的耳静脉血,采用的抗凝剂为0.109M的柠檬酸盐,抗凝剂与血液的体积比为1:9,采集血液后立即冷藏,并在4h内结束血液相关测试;取一定量的含有抗凝剂的全血,在1000g离心力作用下离心5min,用PBS缓冲液清洗3次,每次3min,取下面的沉淀液按照1:34的比例进行稀释,获得红细胞悬浮液RBCS(Red Blood Cells Suspension),将纳米纤维膜试样放在37℃的水浴锅中恒温培养2h,培养结束后,离心取上清液,测试540nm波长下吸光度值,计算纳米纤维膜的溶血率;
(7)单向导湿指数R:采用液态水分管理测试仪(MMT)测试防粘连医用敷料的单向导湿指数R;
(8)复合强力和剥离强力:将防粘连医用敷料裁剪成1cm×5cm的长条,在恒温恒湿(20℃、RH65%)环境中放置1h,然后将敷料的一端进行初步剥离,分别将功能层和吸液层固定在上下夹头处,调整两夹头的间距为15mm,然后以恒定的拉伸速度(50mm/min)进行剥离,直至功能层和吸液层完全分开,期间测试的拉伸强力为复合强力;期间最大拉伸强力为剥离强力;
(9)吸液量:将敷料裁剪成5cm×5cm后,在恒温恒湿环境中放置24h,使样品的回潮率达到平衡,记录此时样品的质量w0;再将样品浸入PBS缓冲液中(比样品质量重40倍),37℃条件下,放置30min后,夹住敷料,空中悬挂30s后,记录此时的质量wt;通过(wt-w0)/样品面积,得到敷料在单位面积内的吸液量。
本发明中自制PVDF电纺膜的具体制备方法为:将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为18%的纺丝液,进行静电纺丝后将所得纳米纤维膜真空烘干8h以去除残留溶剂,制得PVDF电纺膜,其SEM图如图1所示;其中,静电纺丝的工艺参数为:纺丝电压20kV,推注速度1mL/h,接收距离15cm,纺丝时间10min。
实施例1
一种防粘连医用敷料的制备方法,具体步骤如下:
(1)将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为18%的纺丝液,向纺丝液中加入粒径为10~30nm的球形纳米氧化锌颗粒,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干8h以去除残留溶剂,制得纤维平均直径为1000nm、平均孔径为6μm的混有纳米氧化锌的静电纺纳米纤维膜,即防粘连医用敷料的功能层,其SEM图如图1所示;
其中,静电纺丝的工艺参数为:纺丝电压20kV,推注速度1mL/h,接收距离15cm,纺丝时间10min;
制得的功能层中,纳米氧化锌的含量为5wt%;功能层的水接触角为148°,BSA蛋白吸附量为7.2ng/cm2(如图4所示),明胶蛋白凝块剥离强力为0.22N(如图2所示),6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的2.5%(如图3所示),对金黄色葡萄糖球菌的抑菌率为99.8%,对大肠杆菌的抑菌率为74%,溶血率为1%;
(2)用空气喷枪,将质量分数为0.6M的碳酸氢钠水溶液按照16g/m2的喷洒密度喷洒在海藻酸钙无纺布的一面,3min后将步骤(1)制得的混有纳米氧化锌的静电纺纳米纤维膜覆盖在海藻酸钙无纺布的该面,于恒温恒湿环境中用50N的重物压置12h,制得单向导湿指数R为1100的防粘连医用敷料;
制得的防粘连医用敷料如图5所示,由接触皮肤的功能层1和外侧的吸液层2复合而成,功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;功能层与吸液层之间通过离子交换法形成的海藻酸钠凝胶的粘结作用而复合,如图6~7所示,功能层与吸液层在剥离的过程中复合强度最低为2N/m、最高(即剥离强力)为5N/m;制得的防粘连医用敷料的吸液量为35g/100cm2。
对比例1
一种敷料的制备方法,具体步骤基本同实施例1,不同在于步骤(2)中利用微量注射器,将生物胶水均匀滴加在海藻酸钙非织造布的一端,点胶的密度为1个/cm2,每个点的胶水滴加量为20μL;随后将静电纺丝膜覆盖在海藻酸钙有胶水的一侧;最后,对整个复合膜重压(50N),恒温恒湿环境中放置12h,制得复合敷料;
制得的敷料由接触皮肤的功能层和外侧的吸液层复合而成,功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;功能层与吸液层之间通过生物胶的粘结力作用而复合,功能层与吸液层的复合强力及剥离强力如图6~7所示。
从图6~7可以看出,由于复合原理的不同,离子交换法和点胶法获得的复合强力曲线不同;在点胶法复合中,只在有生物胶渗透的区域产生了足够的复合强力,而在无生物胶渗透的区域,两层膜之间结合力很弱,复合不均匀;在本发明的离子交换法复合中,钠离子和钙离子置换后,形成小区域的海藻酸钠凝胶,复合相对均匀,复合界面上各处的复合强力均保持在一个稳定范围内。
对比例2
一种敷料的制备方法,具体步骤基本同实施例1,不同仅在于将步骤(1)中的球形纳米氧化锌颗粒替换为截面直径相同、长度为100nm的棒状纳米氧化锌颗粒。
制得的功能层的BSA蛋白吸附量为35ng/cm2,明胶蛋白凝块剥离强力为0.4N,6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的15.0%。
将对比例2与实施例1的数据进行对比,可以发现实施例1的功能层BSA蛋白吸附量、明胶蛋白凝块剥离强力和6h功能层成纤维细胞粘附量均明显小于对比例2,这是因为对比例2中使用的截面直径相同、长度为100nm的棒状纳米氧化锌颗粒的比表面积小于实施例1中使用的球形纳米氧化锌颗粒,在添加量相同的情况下,球形纳米氧化锌颗粒形成的水化层的面积大于棒状纳米氧化锌颗粒,所以能够阻止更多的蛋白吸附,从而阻止更多的细胞粘附,防粘连效果更好。
对比例3
一种敷料的制备方法,具体步骤基本同实施例1,不同之处在于步骤(1)如下:
将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为18%的纺丝液,向纺丝液中加入粒径为10~30nm的球形纳米氧化锌颗粒,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干4h以去除残留溶剂,制得纤维平均直径为1000nm、平均孔径为3μm的混有纳米氧化锌的静电纺纳米纤维膜,即敷料的功能层;
其中,静电纺丝的工艺参数为:纺丝电压20kV,推注速度1.0mL/h,接收距离15cm,纺丝时间15min;
制得的敷料的单向导湿指数R为-957;
将对比例3与实施例1的数据进行对比,可以发现对比例3敷料的单向导湿指数R远小于实施例1,敷料单向导水能力远不及实施例1,这是因为对比例3的功能层孔径太小,水无法导向吸液层,堆积在功能层上。
对比例4
一种敷料的制备方法,具体步骤基本同实施例1,不同之处在于步骤(1)如下:
将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为18%的纺丝液,向纺丝液中加入粒径为10~30nm的球形纳米氧化锌颗粒,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干4h以去除残留溶剂,制得纤维平均直径为1000nm、平均孔径为9μm的混有纳米氧化锌的静电纺纳米纤维膜,即敷料的功能层;
其中,静电纺丝的工艺参数为:纺丝电压20kV,推注速度1.0mL/h,接收距离15cm,纺丝时间5min;
制得的敷料的单向导湿指数R为378;
将对比例4与实施例1的数据进行对比,可以发现对比例4敷料的单向导湿指数R远小于实施例1,敷料单向导水能力远不及实施例1,这是因为对比例3的功能层孔径太大,功能层含水率始终低于吸液层,不利于敷料垂直方向上的单向导水。
实施例2
一种防粘连医用敷料的制备方法,具体步骤如下:
(1)将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为18%的纺丝液,向纺丝液中加入粒径为10~30nm的球形纳米氧化锌颗粒,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干8h以去除残留溶剂,制得纤维平均直径为1000nm、平均孔径为6μm的混有纳米氧化锌的静电纺纳米纤维膜,即防粘连医用敷料的功能层,其SEM图如图1所示;
其中,静电纺丝的工艺参数为:纺丝电压20kV,推注速度1mL/h,接收距离15cm,纺丝时间10min;
制得的功能层中,纳米氧化锌的含量为1wt%;功能层的水接触角为145°,BSA蛋白吸附量为7.5ng/cm2(如图4所示),明胶蛋白凝块剥离强力为0.25N(如图2所示),6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的3%(如图3所示),对金黄色葡萄糖球菌的抑菌率为99.2%,对大肠杆菌的抑菌率为31%,溶血率为0.8%;
(2)用空气喷枪,将质量分数为0.6M的碳酸氢钠水溶液按照16g/m2的喷洒密度喷洒在海藻酸钙无纺布的一面,3min后将步骤(1)制得的混有纳米氧化锌的静电纺纳米纤维膜覆盖在海藻酸钙无纺布的该面,于恒温恒湿环境中用50N的重物压置12h,制得单向导湿指数R为1100的防粘连医用敷料;
制得的防粘连医用敷料,由接触皮肤的功能层和外侧的吸液层复合而成,功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;功能层与吸液层之间通过离子交换法形成的海藻酸钠凝胶的粘结作用而复合,功能层与吸液层在剥离的过程中复合强度最低为2N/m、最高(即剥离强力)为5N/m;制得的防粘连医用敷料的吸液量为35g/100cm2。
实施例3
一种防粘连医用敷料的制备方法,具体步骤如下:
(1)将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为18%的纺丝液,向纺丝液中加入粒径为10~30nm的球形纳米氧化锌颗粒,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干8h以去除残留溶剂,制得纤维平均直径为1000nm、平均孔径为6μm的混有纳米氧化锌的静电纺纳米纤维膜,即防粘连医用敷料的功能层;
其中,静电纺丝的工艺参数为:纺丝电压20kV,推注速度1mL/h,接收距离15cm,纺丝时间10min;
制得的功能层中,纳米氧化锌的含量为5wt%;功能层的水接触角为148°,BSA蛋白吸附量为7.2ng/cm2,明胶蛋白凝块剥离强力为0.22N,6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的2.5%,对金黄色葡萄糖球菌的抑菌率为99.8%,对大肠杆菌的抑菌率为74%,溶血率为1%;
(2)用空气喷枪,将质量分数为0.6M的碳酸氢钠水溶液按照24g/m2的喷洒密度喷洒在海藻酸钙无纺布的一面,3min后将步骤(1)制得的混有纳米氧化锌的静电纺纳米纤维膜覆盖在海藻酸钙无纺布的该面,于恒温恒湿环境中用50N的重物压置12h,制得单向导湿指数R为1050的防粘连医用敷料;
制得的防粘连医用敷料,由接触皮肤的功能层和外侧的吸液层复合而成,功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;功能层与吸液层之间通过离子交换法形成的海藻酸钠凝胶的粘结作用而复合,如图6~7所示,功能层与吸液层在剥离的过程中复合强度最低为2.5N/m、最高(即剥离强力)为7N/m;制得的防粘连医用敷料的吸液量为34g/100cm2。
实施例4
一种防粘连医用敷料的制备方法,具体步骤如下:
(1)将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为18%的纺丝液,向纺丝液中加入粒径为10~30nm的球形纳米氧化锌颗粒,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干8h以去除残留溶剂,制得纤维平均直径为800nm、平均孔径为8μm的混有纳米氧化锌的静电纺纳米纤维膜,即防粘连医用敷料的功能层;
其中,静电纺丝的工艺参数为:纺丝电压25kV,推注速度1mL/h,接收距离15cm,纺丝时间6min;
制得的功能层中,纳米氧化锌的含量为5wt%;功能层的水接触角为150°,BSA蛋白吸附量为7ng/cm2,明胶蛋白凝块剥离强力为0.2N,6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的2%,对金黄色葡萄糖球菌的抑菌率为99.8%,对大肠杆菌的抑菌率为74%,溶血率为1%;
(2)用空气喷枪,将质量分数为1M的碳酸氢钠水溶液按照24g/m2的喷洒密度喷洒在海藻酸钙无纺布的一面,3min后将步骤(1)制得的混有纳米氧化锌的静电纺纳米纤维膜覆盖在海藻酸钙无纺布的该面,于恒温恒湿环境中用50N的重物压置12h,制得单向导湿指数R为900的防粘连医用敷料;
制得的防粘连医用敷料,由接触皮肤的功能层和外侧的吸液层复合而成,功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;功能层与吸液层之间通过离子交换法形成的海藻酸钠凝胶的粘结作用而复合,功能层与吸液层在剥离的过程中复合强度最低为5N/m、最高(即剥离强力)为10N/m;制得的防粘连医用敷料的吸液量为34g/100cm2。
实施例5
一种防粘连医用敷料的制备方法,具体步骤如下:
(1)将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为20%的纺丝液,向纺丝液中加入粒径为10~30nm的球形纳米氧化锌颗粒,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干8h以去除残留溶剂,制得纤维平均直径为1100nm、平均孔径为6μm的混有纳米氧化锌的静电纺纳米纤维膜,即防粘连医用敷料的功能层;
其中,静电纺丝的工艺参数为:纺丝电压20kV,推注速度1mL/h,接收距离20cm,纺丝时间10min;
制得的功能层中,纳米氧化锌的含量为5wt%;功能层的水接触角为147°,BSA蛋白吸附量为7.3ng/cm2,明胶蛋白凝块剥离强力为0.23N,6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的2.6%,对金黄色葡萄糖球菌的抑菌率为99.8%,对大肠杆菌的抑菌率为74%,溶血率为1%;
(2)用空气喷枪,将质量分数为0.2M的碳酸氢钠水溶液按照30g/m2的喷洒密度喷洒在海藻酸钙无纺布的一面,5min后将步骤(1)制得的混有纳米氧化锌的静电纺纳米纤维膜覆盖在海藻酸钙无纺布的该面,于恒温恒湿环境中用50N的重物压置12h,制得单向导湿指数R为970的防粘连医用敷料;
制得的防粘连医用敷料,由接触皮肤的功能层和外侧的吸液层复合而成,功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;功能层与吸液层之间通过离子交换法形成的海藻酸钠凝胶的粘结作用而复合,功能层与吸液层在剥离的过程中复合强度最低为2N/m、最高(即剥离强力)为6N/m;制得的防粘连医用敷料的吸液量为34g/100cm2。
实施例6
一种防粘连医用敷料的制备方法,具体步骤如下:
(1)将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为5%的纺丝液,向纺丝液中加入粒径为10~30nm的球形纳米氧化锌颗粒,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干8h以去除残留溶剂,制得纤维平均直径为900nm、平均孔径为4μm的混有纳米氧化锌的静电纺纳米纤维膜,即防粘连医用敷料的功能层;
其中,静电纺丝的工艺参数为:纺丝电压18kV,推注速度1.1mL/h,接收距离15cm,纺丝时间14min;
制得的功能层中,纳米氧化锌的含量为5wt%;功能层的水接触角为149°,BSA蛋白吸附量为7.3ng/cm2,明胶蛋白凝块剥离强力为0.23N,6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的2.7%,对金黄色葡萄糖球菌的抑菌率为99.8%,对大肠杆菌的抑菌率为74%,溶血率为1%;
(2)用空气喷枪,将质量分数为0.8M的碳酸氢钠水溶液按照20g/m2的喷洒密度喷洒在海藻酸钙无纺布的一面,10min后将步骤(1)制得的混有纳米氧化锌的静电纺纳米纤维膜覆盖在海藻酸钙无纺布的该面,于恒温恒湿环境中用50N的重物压置12h,制得单向导湿指数R为920的防粘连医用敷料;
制得的防粘连医用敷料,由接触皮肤的功能层和外侧的吸液层复合而成,功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;功能层与吸液层之间通过离子交换法形成的海藻酸钠凝胶的粘结作用而复合,功能层与吸液层在剥离的过程中复合强度最低为2.7N/m、最高(即剥离强力)为8N/m;制得的防粘连医用敷料的吸液量为35g/100cm2。
实施例7
一种防粘连医用敷料的制备方法,具体步骤如下:
(1)将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为30%的纺丝液,向纺丝液中加入粒径为10~30nm的球形纳米氧化锌颗粒,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干8h以去除残留溶剂,制得纤维平均直径为1100nm、平均孔径为6μm的混有纳米氧化锌的静电纺纳米纤维膜,即防粘连医用敷料的功能层;
其中,静电纺丝的工艺参数为:纺丝电压18kV,推注速度0.5mL/h,接收距离12cm,纺丝时间10min;
制得的功能层中,纳米氧化锌的含量为5wt%;功能层的水接触角为146°,BSA蛋白吸附量为7.3ng/cm2,明胶蛋白凝块剥离强力为0.23N,6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的2.7%,对金黄色葡萄糖球菌的抑菌率为99.8%,对大肠杆菌的抑菌率为74%,溶血率为1%;
(2)用空气喷枪,将质量分数为0.5M的碳酸氢钠水溶液按照20g/m2的喷洒密度喷洒在海藻酸钙无纺布的一面,8min后将步骤(1)制得的混有纳米氧化锌的静电纺纳米纤维膜覆盖在海藻酸钙无纺布的该面,于恒温恒湿环境中用50N的重物压置12h,制得单向导湿指数R为970的防粘连医用敷料;
制得的防粘连医用敷料,由接触皮肤的功能层和外侧的吸液层复合而成,功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;功能层与吸液层之间通过离子交换法形成的海藻酸钠凝胶的粘结作用而复合,功能层与吸液层在剥离的过程中复合强度最低为2.1N/m、最高(即剥离强力)为6.4N/m;制得的防粘连医用敷料的吸液量为36g/100cm2。
实施例8
一种防粘连医用敷料的制备方法,具体步骤如下:
(1)将聚偏氟乙烯(PVDF)溶解在DMF/丙酮(V/V=1:2)中得到质量分数为10%的纺丝液,向纺丝液中加入粒径为10~30nm的球形纳米氧化锌颗粒,超声处理后进行静电纺丝,后将所得纳米纤维膜真空烘干8h以去除残留溶剂,制得纤维平均直径为1200nm、平均孔径为6μm的混有纳米氧化锌的静电纺纳米纤维膜,即防粘连医用敷料的功能层;
其中,静电纺丝的工艺参数为:纺丝电压15kV,推注速度1.5mL/h,接收距离10cm,纺丝时间10min;
制得的功能层中,纳米氧化锌的含量为5wt%;功能层的水接触角为146°,BSA蛋白吸附量为7.3ng/cm2,明胶蛋白凝块剥离强力为0.22N,6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的2.6%,对金黄色葡萄糖球菌的抑菌率为99.8%,对大肠杆菌的抑菌率为74%,溶血率为1%;
(2)用空气喷枪,将质量分数为0.7M的碳酸氢钠水溶液按照20g/m2的喷洒密度喷洒在海藻酸钙无纺布的一面,6min后将步骤(1)制得的混有纳米氧化锌的静电纺纳米纤维膜覆盖在海藻酸钙无纺布的该面,于恒温恒湿环境中用50N的重物压置12h,制得单向导湿指数R为1000的防粘连医用敷料;
制得的防粘连医用敷料,由接触皮肤的功能层和外侧的吸液层复合而成,功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;功能层与吸液层之间通过离子交换法形成的海藻酸钠凝胶的粘结作用而复合,功能层与吸液层在剥离的过程中复合强度最低为2.6N/m、最高(即剥离强力)为7.5N/m;制得的防粘连医用敷料的吸液量为35g/100cm2。
从图1可以看出,随着纳米氧化锌颗粒添加量的增加,纤维平均直径基本保持稳定,PVDF电纺膜纤维表面光滑,随着纳米氧化锌颗粒添加量的增加,纤维逐渐粗糙;
利用40%w/v的明胶蛋白溶液来模拟伤口渗出液的组成,将其浇铸到纳米纤维膜表面,通过测量纳米纤维膜从明胶蛋白凝块上的剥离强力来表征纳米纤维膜和伤口之间的潜在粘连性;凡士林纱布是现在常用的一种防粘连敷料,作为本实验的对照样,模拟伤口渗出液和敷料的接触时间为24h;从图2可以看出,PVDF纳米纤维膜获得的明胶蛋白凝块剥离强力均显著低于凡士林纱布样品,并且PVDF/ZnO纳米纤维膜的明胶蛋白凝块剥离强力低于PVDF纳米纤维膜,表明PVDF/ZnO纳米纤维膜具有很强的防粘连应用前景;
利用成纤维细胞进行体外细胞粘附实验,评价敷料的防粘连性能;由于硅片具有一定的抗细胞粘附性能,PEG涂层的硅片具有较好的抗细胞粘附性能,故釆用硅片和PEG涂层的硅片作为对照;硅片、PEG涂层硅片、PVDF、不同ZnO含量的PVDF/ZnO纳米纤维膜经过6h细胞培养,其细胞染色图片结果如图3所示;鬼笔环肽的染色结果表明,在硅片上粘附有大量且密集的成纤维细胞,部分细胞已经在硅片上铺展开;PEG涂层后的硅片上,成纤维细胞的数量显著降低,证实了PEG良好的抗细胞粘附性能;PVDF纳米纤维膜上细胞的粘附数量较硅片上的细胞数量明显降低,但是比PEG涂层后的硅片上的细胞含量要多,而不同ZnO含量的PVDF/ZnO纳米纤维膜观察到比PEG涂层后的硅片更少的细胞粘附,反映了PVDF/ZnO纳米纤维膜较好的抗粘连特性,其抗细胞粘附性能优于PEG涂层硅片;
当生物材料应用在人体组织中时,第一步都是进行蛋白的吸附;利用具备良好抗蛋白吸附的材料PEG涂层硅片作为对照,通过评价牛血清蛋白(BSA)在材料表面的粘附量来预测或间接反应材料的防粘连性能;从图4中可以看出,PVDF纳米纤维膜对BSA的吸附量要显著高于PEG涂层硅片,而PVDF/ZnO纳米纤维膜对BSA的吸附量很少,明显低于PEG涂层硅片,并且不同纳米氧化锌颗粒负载量的纳米纤维膜之间,对BSA的吸附量无显著性差异;这说明,PVDF/ZnO纳米纤维膜能够有效抵抗BSA的吸附,预测有较好的抗粘连特性。
Claims (10)
1.一种防粘连医用敷料的制备方法,其特征在于:采用离子交换法,利用碳酸氢钠溶液将功能层与吸液层进行复合制得防粘连医用敷料;
功能层为混有纳米氧化锌的静电纺纳米纤维膜,吸液层为海藻酸钙无纺布;
功能层中纳米氧化锌为球形纳米颗粒,含量为1~5wt%;
静电纺纳米纤维膜的材质为疏水生物惰性高分子聚合物;静电纺纳米纤维膜的平均孔径为4~8μm。
2.根据权利要求1所述的一种防粘连医用敷料的制备方法,其特征在于,纳米氧化锌的粒径为10~30nm。
3.根据权利要求1所述的一种防粘连医用敷料的制备方法,其特征在于,静电纺纳米纤维膜的纤维平均直径为800~1200nm。
4.根据权利要求1所述的一种防粘连医用敷料的制备方法,其特征在于,具体步骤如下:
(1)将疏水生物惰性高分子聚合物溶解在溶剂中得到质量分数为5~30%的纺丝液,向纺丝液中加入纳米氧化锌,超声处理后进行静电纺丝,制得混有纳米氧化锌的静电纺纳米纤维膜;
(2)用空气喷枪将质量分数为0.2~1.0M的碳酸氢钠水溶液喷洒在海藻酸钙无纺布的一面,一段时间后将步骤(1)制得的混有纳米氧化锌的静电纺纳米纤维膜覆盖在海藻酸钙无纺布的该面,制得防粘连医用敷料。
5.根据权利要求4所述的一种防粘连医用敷料的制备方法,其特征在于,步骤(1)中疏水生物惰性高分子聚合物为聚偏氟乙烯,溶剂为丙酮和二甲基甲酰胺的混合物。
6.根据权利要求4所述的一种防粘连医用敷料的制备方法,其特征在于,步骤(1)中静电纺丝的工艺参数为:纺丝电压15~25kV,推注速度0.5~1.5mL/h,接收距离10~20cm,纺丝时间6~14min。
7.根据权利要求4所述的一种防粘连医用敷料的制备方法,其特征在于,步骤(2)中海藻酸钙无纺布上碳酸氢钠水溶液喷洒密度为15~30g/m2。
8.根据权利要求4所述的一种防粘连医用敷料的制备方法,其特征在于,步骤(2)中一段时间为3~10min。
9.采用如权利要求1~8任一项所述的方法制得的防粘连医用敷料,由功能层和吸液层复合而成,其特征在于:功能层与吸液层之间通过海藻酸钠凝胶的粘结作用而复合。
10.根据权利要求9所述的防粘连医用敷料,其特征在于,功能层的水接触角为145~150°,BSA蛋白吸附量为7.0~7.5ng/cm2,明胶蛋白凝块剥离强力为0.20~0.25N,6h功能层成纤维细胞粘附量为硅片材料成纤维细胞粘附量的2.0~3.0%;
功能层与吸液层剥离强力测试中测得的界面复合强度始终大于2N/m;防粘连医用敷料的单向导湿指数R为900~1100。
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