CN115025275B - 多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法 - Google Patents
多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法 Download PDFInfo
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Abstract
本发明提供了一种多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,包括以下步骤:S1.对壳聚糖纤维进行羧基化和邻苯二酚化接枝改性,得到多功能壳聚糖短纤;S2.将多功能壳聚糖短纤与壳聚糖的乙酸水溶液混合后,进行预冻干,然后放入1%‑4%的氢氧化钠水溶液和无水乙醇组成的混合碱洗液中浸泡洗涤,接着水洗后再次冻干,得到多功能壳聚糖短纤/壳聚糖复合止血海绵。本发明将壳聚糖与吸液性能强、止血效果好的邻苯二酚改性羧乙基壳聚糖纤维复合冷冻,然后碱洗促使邻苯二酚被氧化形成醌以及聚多巴胺,并进一步与壳聚糖中的氨基作用,从而显著增强止血海绵的机械性能,最终得到一种强度高、止血性能优异的新型止血海绵。
Description
技术领域
本发明涉及止血材料技术领域,尤其涉及一种多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法。
背景技术
血液是人体非常重要的组成部分之一,它承担着人体全部组织和器官的物质交换,如果人在短时间内出血量超过全部血液的30%以上,就可能会死亡。据统计,在战争中,大约60%的死亡是由于大量出血没有及时止住,导致失血过多而死在阵地上或运回医院的途中。
目前已经成功开发出来一些能运用于局部止血的优秀材料,比如:明胶止血海绵、沸石止血剂、氧化纤维素等,但是它们在实际应用过程中,均存在不同程度的不足,影响了其在临床的广泛应用。因此开发出一种能将战时急救与平时创伤救治相互结合,并且具有止血快、生物相容性好,同时可在体内降解的生物医用新材料,成为很多科研工作者追求的目标。
目前基于壳聚糖开发的各类止血海绵,在降解、止血速度、应用等方面仍存在不足,其中单纯的壳聚糖海绵同样存在止血效果差的问题。专利CN202110772237.1公开了一种羧基化壳聚糖医用海绵及其制备方法,将壳聚糖纤维羧基化后,与壳聚糖溶液制成混合溶液再冻干,得到羧基化壳聚糖止血海绵。但是该方案仅仅依赖壳聚糖纤维上的羧基提高海绵的吸液速率和吸液量,进而提高止血性能,效果仍不够理想;而且这种主要依靠快速吸液中的水分来实现快速止血的海绵,不可避免的会造成止血的同时,出血量也相应有所增大。因此,如何同时降低止血海绵的止血时间和出血量是亟待解决的问题。
有鉴于此,有必要设计一种改进的多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,以解决上述问题。
发明内容
为了克服上述现有技术的不足,本发明的目的在于提供一种多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,将壳聚糖与吸液性能强、止血效果好的邻苯二酚改性羧乙基壳聚糖纤维复合,制备得到一种强度高、止血性能优异的新型止血海绵。
为实现上述发明目的,本发明提供了一种多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,包括以下步骤:
S1.对壳聚糖纤维进行羧基化和邻苯二酚化接枝改性,得到多功能壳聚糖短纤;
S2.将所述多功能壳聚糖短纤与壳聚糖的乙酸水溶液混合后,进行预冻干,然后放入1%-4%的氢氧化钠水溶液和无水乙醇组成的混合碱洗液中浸泡洗涤,接着水洗后再次冻干,得到多功能壳聚糖短纤/壳聚糖复合止血海绵。
作为本发明的进一步改进,步骤S1中,所述多功能壳聚糖短纤上羧基和邻苯二酚的摩尔比为(5-15):1,优选为9:1;羧基和邻苯二酚的总取代度为0.5-0.9,优选为0.6-0.8。
作为本发明的进一步改进,步骤S1中,所述羧基化优选采用丙烯酸对壳聚糖纤维上的氨基进行加成改性;所述邻苯二酚化优选采用3,4-二羟基苯甲醛对壳聚糖纤维上的氨基进行接枝改性。
作为本发明的进一步改进,所述丙烯酸的接枝包括:将丙烯酸加入到无水乙醇中,然后放入壳聚糖纤维,在60-80℃下反应24-50h;反应结束后,将羧基化壳聚糖纤维碱洗至强碱性,再用无水乙醇/去离子水混合溶液洗至中性,最后干燥得到羧基化壳聚糖纤维。
作为本发明的进一步改进,所述3,4-二羟基苯甲醛的接枝包括:将羧基化壳聚糖纤维和3,4-二羟基苯甲醛置于无水乙醇/去离子水混合溶液中,室温下反应8-15h,反应结束后,用无水乙醇/去离子水混合溶液洗2次,再用无水乙醇醇洗3次,最后干燥得到多功能壳聚糖短纤。
作为本发明的进一步改进,步骤S2中,所述多功能壳聚糖短纤的直径为10~20μm,长度为300-600μm。
作为本发明的进一步改进,步骤S2中,所述多功能壳聚糖短纤的质量含量为0.5%-2%,所述壳聚糖的质量含量为2%-3%;所述多功能壳聚糖短纤的质量含量优选为所述壳聚糖的50%-65%。
作为本发明的进一步改进,步骤S2中,在碱洗液中浸泡洗涤至pH为7-8。
作为本发明的进一步改进,所述1%-4%的氢氧化钠水溶液和无水乙醇的体积比为1:(2-4)。
作为本发明的进一步改进,步骤S2中,所述预冻干包括:在4℃的冰箱中冷冻4-8h,之后取出放入-21℃的冰箱中冷冻4-8h,最后放入-41℃的冰柜中冷冻12h以上;所述再次冻干包括:在-41℃的冰柜中冷冻10-15h,然后移入冷冻干燥机中冻干。
本发明的有益效果是:
1.本发明提供的多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,通过选用羧基化和邻苯二酚化接枝改性的多功能壳聚糖短纤(CH)与壳聚糖(CS)溶液复合冻干,能够丰富壳聚糖纤维上的功能基团,使得复合止血海绵在满足快速吸液溶胀而不溶解的同时,还能够利用邻苯二酚的功能止血性,使得血液快速凝固,从而显著减小出血量。进一步的,本发明还通过预冻干、碱洗、再冻干,对多功能壳聚糖短纤上的邻苯二酚结构进行二次调控,从而形成醌和聚多巴胺,形成的醌又可以与壳聚糖中的氨基作用,生成亚胺键,进而增强复合海绵的机械性能。
2.本发明冻干前,将CS溶于乙酸,CS中的氨基在酸的作用下发生质子化,带有正电荷。CH纤维上部分氨基没有发生加成,因此在溶液中只发生溶胀并不溶解,在弱酸性环境中也是溶胀状态;此时,CS分子链和CH纤维的分子链均处于伸展的状态,在搅拌的作用下,分子链之间相互缠结,能够增强海绵的机械性能。接着,经过搅拌以及冷冻处理后的复合海绵,其内部也会发生分子链之间相互缠结,通过碱洗处理,使得分子链进一步缠结定型,得到的海绵结构更加致密。相比CS海绵,CS-CH复合海绵具有柔软、弹性优等特点。
3.本发明制备方法具有工艺成本低以及制备方法简单的特点,相比单纯的CS海绵有着显著增强的机械性能,快速的吸液能力以及优异的止血效果。羧基的引入,使得海绵可以在短时间内吸收液体,浓缩血液,促进凝血;未被氧化的邻苯二酚可以粘附红细胞和血小板,也可以与血液中的铁离子络合,加快血液的凝固,并且能够和伤口组织出的胺基、巯基等基团形成共价键,促进伤口封闭,促进止血。氧化形成的醌也可以与红细胞血小板作用,加上壳聚糖本身的止血性能,两种物质相互协同,加快止血。本发明制备过程安全无毒,具有良好的生物相容性和可降解性。
附图说明
图1为本发明制备的多功能壳聚糖短纤/壳聚糖复合止血海绵的SEM图。
图2中左图为壳聚糖海绵实物图,右图为多功能壳聚糖短纤/壳聚糖复合止血海绵的实物图。
具体实施方式
为了使本发明的目的、技术方案和优点更加清楚,下面结合具体实施例对本发明进行详细描述。
在此,还需要说明的是,为了避免因不必要的细节而模糊了本发明,在具体实施例中仅仅示出了与本发明的方案密切相关的结构和/或处理步骤,而省略了与本发明关系不大的其他细节。
另外,还需要说明的是,术语“包括”、“包含”或者其任何其他变体意在涵盖非排他性的包含,从而使得包括一系列要素的过程、方法、物品或者设备不仅包括那些要素,而且还包括没有明确列出的其他要素,或者是还包括为这种过程、方法、物品或者设备所固有的要素。
本发明提供的一种多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,包括以下步骤:
S1.对壳聚糖纤维进行羧基化和邻苯二酚化接枝改性,得到多功能壳聚糖短纤;
S2.将所述多功能壳聚糖短纤与壳聚糖的乙酸水溶液混合后,进行预冻干,然后放入1%-4%的氢氧化钠水溶液和无水乙醇组成的混合碱洗液中浸泡洗涤,接着水洗后再次冻干,得到多功能壳聚糖短纤/壳聚糖复合止血海绵。CS溶于乙酸,CS中的氨基在酸的作用下发生质子化,带有正电荷。CH纤维上部分氨基没有发生加成,因此在溶液中只发生溶胀并不溶解,在弱酸性环境中也是溶胀状态,此时,CS分子链和CH纤维的分子链均处于伸展的状态,在搅拌的作用下,分子链之间相互缠结,能够增强海绵的机械性能。
如此操作,通过选用羧基化和邻苯二酚化接枝改性的多功能壳聚糖短纤与壳聚糖溶液复合冻干,能够丰富壳聚糖纤维上的功能基团,使得复合止血海绵在满足快速吸液溶胀而不溶解的同时,还能够利用邻苯二酚的功能止血性,使得血液快速凝固,从而显著减小出血量。进一步的,本发明还通过预冻干、碱洗、再冻干,对多功能壳聚糖短纤上的邻苯二酚结构进行二次调控,从而形成醌和聚多巴胺,形成的醌又可以与壳聚糖中的氨基作用,生成亚胺键,进而增强复合海绵的机械性能。
具体地,步骤S1中,所述多功能壳聚糖短纤上羧基和邻苯二酚的摩尔比为(5-15):1,优选为(7.5-10.5):1;更优选为9:1;羧基和邻苯二酚的总取代度为0.5-0.9,优选为0.6-0.8。基于复合止血海绵的止血机理,通过调控多功能壳聚糖短纤上羧基、邻苯二酚及氨基的摩尔比,能够使得多功能壳聚糖短纤在复合海绵中起到更优的增强作用及促进止血作用。
步骤S1中,所述羧基化优选采用丙烯酸对壳聚糖纤维上的氨基进行加成改性;所述邻苯二酚化优选采用3,4-二羟基苯甲醛对壳聚糖纤维上的氨基进行接枝改性。通过适宜量的丙烯酸接枝,使得多功能壳聚糖短纤能够在吸液时主要只在径向发生膨胀而长度方向基本不变化。如此在止血时,能够使得止血海绵吸液后厚度增加,面积基本不变化,因此能够产生更强的压迫止血作用;而且,不会因为海绵中多功能壳聚糖短纤的长度伸长,而使得海绵交联结构受到影响,从而降低吸液后的强度。
所述丙烯酸的接枝包括:将丙烯酸加入到无水乙醇中,然后放入壳聚糖纤维,在60-80℃下反应24-50h;反应结束后,将羧基化壳聚糖纤维碱洗至强碱性,再用无水乙醇/去离子水混合溶液洗至中性,最后干燥得到羧基化壳聚糖纤维。
所述3,4-二羟基苯甲醛的接枝包括:将羧基化壳聚糖纤维和3,4-二羟基苯甲醛置于无水乙醇/去离子水混合溶液中,室温下反应8-15h,反应结束后,用无水乙醇/去离子水混合溶液洗2次,再用无水乙醇醇洗3次,最后干燥得到多功能壳聚糖短纤。
步骤S2中,所述多功能壳聚糖短纤的直径为10~20μm,长度为300-600μm,优选为500μm。
步骤S2中,所述多功能壳聚糖短纤的质量含量为0.5%-2%,所述壳聚糖的质量含量为2%-3%;所述多功能壳聚糖短纤的质量含量优选为所述壳聚糖的50%-65%。通过调控多功能壳聚糖短纤与壳聚糖的含量比,使得止血性能和机械强度最优化。
步骤S2中,在碱洗液中浸泡洗涤至pH为7-8。所述1%-4%的氢氧化钠水溶液和无水乙醇的体积比为1:(2-4)。步骤S2中,所述预冻干包括:在4℃的冰箱中冷冻4-8h,之后取出放入-21℃的冰箱中冷冻4-8h,最后放入-41℃的冰柜中冷冻12h以上;所述再次冻干包括:在-41℃的冰柜中冷冻10-15h,然后移入冷冻干燥机中冻干。
实施例1-4
一种多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,包括以下步骤:
(1)羧基化壳聚糖(CECS)纤维制备过程
取丙烯酸加入到800mL无水乙醇中,将壳聚糖(CS)纤维放入反应溶液中,在60℃恒温水浴中振荡反应48h;反应结束后,用氢氧化钠制备的碱液,碱洗羧基化壳聚糖至强碱性;再用无水乙醇/去离子水(体积比4:1)的混合溶液将碱洗后的壳聚糖洗至中性,再用无水乙醇醇洗3次;最后在50℃的烘箱中烘干,得到羧基化壳聚糖纤维。在此步骤中,丙烯酸的烯烃基与壳聚糖上的氨基发生加成反应,实现接枝。
(2)制备邻苯二酚改性羧基化壳聚糖(CH)纤维
在450mL无水乙醇/去离子水(体积比为4:1)的混合溶液中,加入羧基化壳聚糖(CECS)纤维和3,4-二羟基苯甲醛(HBA),在室温下反应12小时。反应结束后,用无水乙醇/去离子水混合溶液洗2次,再用无水乙醇醇洗3次,再在50℃烘箱中烘干,得到邻苯二酚改性的羧基化壳聚糖(CH)纤维。其中,CH纤维中羧基与邻苯二酚的摩尔比为9:1,两者总取代度为0.7,即与剩余氨基的摩尔比为0.7:0.3。
(3)制备壳聚糖复合(CS-CH)海绵
将CS及CH纤维(短纤,直径为10~20μm,长度为500μm)放入在体积百分比浓度为2%的乙酸水溶液中,其中CS和CH纤维的含量如表1所示,制备得到其混合溶液。静置一段时间以除去气泡,将溶液取出装入一定尺寸的模具中,密封,静置过夜。将装有混合溶液的模具放入4℃的冰箱中冷冻6h,之后取出放入-21℃的冰箱中冷冻6h,最后放入-41℃的冰柜中冷冻12h以上。冷冻后的壳聚糖溶液移入冷冻干燥机中冻干,取出冻干后的产物,进行后处理(即放入质量百分比浓度(下同)为2%NaOH/无水乙醇(体积比1:2)的溶液中浸泡12h,然后用去离子水将其洗涤至pH值为7-8,之后放入去离子水中浸泡6h)。将处理好的海绵放入-41℃的冰柜中冷冻12h,然后移入冷冻干燥机中冻干,制备得到CS-CH海绵。
如图1所示,可以看出,复合止血海绵的孔隙率高,因此吸液量大。从图2可以看出,加入邻苯二酚改性羧基化壳聚糖(CH)纤维的海绵颜色为棕黑色,间接说明接枝成功。
对比例1
一种壳聚糖(CS)止血海绵的制备方法,与实施例1相比,不同之处在于,不包含邻苯二酚改性羧基化壳聚糖(CH)纤维。其他与实施例1大致相同,在此不再赘述。
表1实施例1-4及对比例1的制备参数及相关性能测试
从表1可以看出,当CS含量为2.5%时,随着CH纤维含量的增加,吸液量和机械强度均先增大后减小。其中,CH纤维含量为1.5%时,吸液量最大,且10s的吸液量和饱和吸液量基本相同,说明其能够快速达到饱和吸液量,有助于提高止血速率。该条件下的拉伸强度和压缩强度也最优,有助于提高压迫止血性能。当CH纤维含量继续增大时,饱和吸液量虽然继续增大,但10s的吸液量显著降低,说明对于本发明的复合止血海绵体系,当CH纤维含量过高时,海绵会变得更加致密,导致吸液速率变慢,进而降低止血速度。
选择上述最优的实施例3及其他止血材料进行不同部位出血的止血性能测试,测试方法如下:
动物模型止血试验:
(1)兔子耳动脉模型:兔子通过肌肉注射盐酸塞拉嗪注射液进行麻醉(0.2mL/kg),然后固定四肢。剃掉兔子耳朵背面的兔毛,暴露耳动脉。用75%的医用酒精对耳朵背部消毒后,在距离耳尖顶端7cm将耳动脉切断,血液立刻不断地在伤口处涌出。在自由出血5s后,用灭菌纱布轻轻的将耳朵伤口上的浮血轻轻擦去,并立刻将止血材料(CS海绵、CS-CH-3复合海绵)按压于流血的伤口,并用100克的重物垂直加压,同时开始计时,每隔30s轻轻揭开止血材料,观察是否继续留血,若伤口继续流血,则继续按压止血,则直到出血停止,记录止血时间;对照组只用标准的纱布进行处理。止血完成后称重,计算失血量。3小时后幸存下来的所有兔子被安乐死,每个样品重复实验6次,取平均值。
(2)兔子肝脏止血试验:兔子被麻醉后,背部固定,剃去腹部兔毛,腹部朝上,用75%的医用酒精对腹部消毒后,切开腹腔后小心的将肝脏的左内侧叶从腹腔内拉出,放在已经消毒纱布上。然后,在肝脏切开一个伤口,长约1厘米,深0.3厘米,让其自由出血10s后,轻轻擦去伤口上的血液,并将止血材料(CS海绵,CS-CH-3复合海绵)按压在出血伤口上,50克的重量垂直加压在止血材料上方,并开始计时,每隔30s轻轻揭开止血材料,观察是否继续留血,若伤口继续流血,则继续按压止血,则直到出血停止,记录止血时间。用标准纱布同样操作进行止血处理,止血完成后,每个材料都称重,计算失血量。3小时后幸存下来的所有兔子被安乐死,每个样品重复实验6次,取其平均值。
(3)兔子股动脉出血模型:兔子被麻醉后,背部固定,腹部朝上,剃去后腿根部兔毛,用75%的医用酒精对后腿根部消毒后,找到大腿动脉,然后在大腿根部动脉处切开一个伤口,长约1厘米,深0.5厘米,将股动脉切断,让其自由出血5s后,轻轻擦去伤口上的浮血,并将止血材料(CS海绵、CS-CH-3复合海绵)按压在出血伤口上,200克的重量垂直加压在止血材料上方,并开始计时,每隔30s轻轻揭开止血材料,观察是否继续止血,若伤口继续流血,则继续按压止血,直到出血停止,记录止血时间。用标准纱布同样操作进行止血处理,止血完成后,每个材料都称重,计算失血量。3小时后幸存下来的所有兔子被安乐死,每个样品重复实验6次,取其平均值。
表2兔子耳动脉止血时间和失血量
样品 | 止血时间(s) | 失血量(g) |
纱布 | 178±2.3 | 3.64±0.21 |
对比例1 | 125±1.7 | 2.53±0.19 |
实施例3 | 28±0.7 | 1.03±0.11 |
表3兔肝脏止血时间和出血量
表4兔股动脉止血时间和出血量
样品 | 止血时间(s) | 失血量(g) |
纱布 | 346±3.6 | 4.97±0.23 |
对比例1 | 163±2.3 | 3.15±0.29 |
实施例3 | 42±0.8 | 1.27±0.18 |
从表2-4可以看出,本发明制备的多功能壳聚糖短纤/壳聚糖复合止血海绵的止血时间和出血量均低于市面的止血纱布或壳聚糖海绵。不仅对肝脏出血具有良好的止血效果,对动脉同样具有显著的止血效果,说明其压迫止血性能优异。
实施例5-8
一种多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,与实施例3相比,不同之处在于,羧基与邻苯二酚的摩尔比及总取代度如表5所示,其他与实施例3大致相同,在此不再赘述。
表5实施例5-8的制备参数及相关性能测试
从表5可以看出,当总取代度为0.7时,随着CH纤维中羧基与邻苯二酚摩尔比的增加,饱和吸液量也随之增大,拉伸强度和压缩强度随之降低,可见一定量的邻苯二酚有助于提高海绵机械强度和吸液性能,邻苯二酚过多时,吸液性能下降,过少时,机械性能下降。随着CH纤维中羧基和邻苯二酚总取代度的增加,饱和吸液量、拉伸强度和压缩强度也随之增大,但总取代度过大时,吸液速率下降,因此在本发明复合海绵体系下,通过给定的取代值范围,实现了综合较优的止血性能。
综上所述,本发明提供的多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,通过选用羧基化和邻苯二酚化接枝改性的多功能壳聚糖短纤与壳聚糖溶液复合冻干,能够丰富壳聚糖纤维上的功能基团,使得复合止血海绵在满足快速吸液溶胀而不溶解的同时,还能够利用邻苯二酚的功能止血性,使得血液快速凝固,从而显著减小出血量。进一步的,本发明还通过预冻干、碱洗、再冻干,对多功能壳聚糖短纤上的邻苯二酚结构进行二次调控,从而形成醌和聚多巴胺,形成的醌又可以与壳聚糖中的氨基作用,生成亚胺键,进而增强复合海绵的机械性能。
以上实施例仅用以说明本发明的技术方案而非限制,尽管参照较佳实施例对本发明进行了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的精神和范围。
Claims (8)
1.一种多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,其特征在于,包括以下步骤:
S1.对壳聚糖纤维进行羧基化和邻苯二酚化接枝改性,得到多功能壳聚糖短纤;
S2.将所述多功能壳聚糖短纤与壳聚糖的乙酸水溶液混合后,进行预冻干,然后放入1%~4%的氢氧化钠水溶液和无水乙醇组成的混合碱洗液中浸泡洗涤,接着水洗后再次冻干,得到多功能壳聚糖短纤/壳聚糖复合止血海绵;
步骤S1中,所述多功能壳聚糖短纤上羧基和邻苯二酚的摩尔比为(5-15):1;
羧基和邻苯二酚的总取代度为0.5-0.9;
所述1%-4%的氢氧化钠水溶液和无水乙醇的体积比为1:(2-4);
步骤S2中,所述多功能壳聚糖短纤的质量含量为0.5%~2%,所述壳聚糖的质量含量为2%-3%;所述多功能壳聚糖短纤的质量含量为所述壳聚糖的50%-65%;
通过预冻干、碱洗、再冻干,对多功能壳聚糖短纤上的邻苯二酚结构进行二次调控,从而形成醌和聚多巴胺,形成的醌又可以与壳聚糖中的氨基作用,生成亚胺键,进而增强复合海绵的机械性能。
2.根据权利要求1所述的多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,其特征在于,所述多功能壳聚糖短纤上羧基和邻苯二酚的摩尔比为9:1;羧基和邻苯二酚的总取代度为0.6-0.8。
3.根据权利要求1所述的多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,其特征在于,步骤S1中,所述羧基化采用丙烯酸对壳聚糖纤维上的氨基进行加成改性;所述邻苯二酚化采用3,4-二羟基苯甲醛对壳聚糖纤维上的氨基进行接枝改性。
4.根据权利要求3所述的多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,其特征在于,所述丙烯酸的接枝包括:将丙烯酸加入到无水乙醇中,然后放入壳聚糖纤维,在60-80℃下反应24-50h;反应结束后,将羧基化壳聚糖纤维洗至强碱性,再用无水乙醇/去离子水混合溶液洗至中性,最后干燥得到羧基化壳聚糖纤维。
5.根据权利要求3所述的多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,其特征在于,所述3,4-二羟基苯甲醛的接枝包括:将羧基化壳聚糖纤维和3,4-二羟基苯甲醛置于无水乙醇/去离子水混合溶液中,室温下反应8-15h,反应结束后,用无水乙醇/去离子水混合溶液洗2次,再用无水乙醇洗3次,最后干燥得到多功能壳聚糖短纤。
6.根据权利要求1所述的多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,其特征在于,步骤S2中,所述多功能壳聚糖短纤的直径为10~20μm,长度为300-600μm。
7.根据权利要求1所述的多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,其特征在于,步骤S2中,在碱洗液中浸泡洗涤至pH为7-8。
8.根据权利要求1所述的多功能壳聚糖短纤/壳聚糖复合止血海绵的制备方法,其特征在于,步骤S2中,所述预冻干包括:在4℃的冰箱中冷冻4-8h,之后取出放入-21℃的冰箱中冷冻4-8h,最后放入-41℃的冰柜中冷冻12h以上;所述再次冻干包括:在-41℃的冰柜中冷冻10-15h,然后移入冷冻干燥机中冻干。
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