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CN105641735B - 一种抗菌性多糖止血棉基纱布的制备方法 - Google Patents

一种抗菌性多糖止血棉基纱布的制备方法 Download PDF

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CN105641735B
CN105641735B CN201610214687.8A CN201610214687A CN105641735B CN 105641735 B CN105641735 B CN 105641735B CN 201610214687 A CN201610214687 A CN 201610214687A CN 105641735 B CN105641735 B CN 105641735B
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徐福建
胡浩
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Beijing University of Chemical Technology
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Abstract

本发明公开了一种抗菌性多糖止血棉基纱布的制备方法。该方法为首先制备具有抗菌功能的季铵化聚乙烯亚胺和醛化多糖交联剂；然后将季铵化聚乙烯亚胺，醛化多糖交联剂与壳聚糖盐酸盐混合发泡，将发泡液均匀涂覆在经过预处理的纱布表面，冷冻干燥，即可得到抗菌性多糖止血棉基纱布。多糖止血棉与经过预处理过的纱布复合后，两者结合紧密，不脱落，强度大，适合伤口处的压迫止血和包扎。并且与普通纱布相比，抗菌性多糖止血棉基纱布在止血的同时，具有抑制微生物的生长，起到抑菌的作用。另外，涂覆在纱布表面的海绵我们采用安全无毒的多糖为交联剂，进一步提升止血棉基纱布的强度和止血效果。

Description

一种抗菌性多糖止血棉基纱布的制备方法
技术领域
本发明涉及一种抗菌性多糖止血棉基纱布的制备方法，属于生物医用材料领域。
背景技术
目前，各种新型医用生物止血材料不断出现。临床常用的生物止血材料有氧化纤维素、明胶海绵、壳聚糖及藻酸钙纤维等。这些止血材料均有不同的止血效果，但功能单一且价格昂贵。
在中国专利CN 202604936 U中，制备了一种明胶海绵止血棉；在中国专利CN102908651 A中，制备了一种氧化纤维素止血材料；这些止血棉有良好的止血效果，但功能相对单一，不具备抗菌消炎等功效。在中国专利CN 101816802 A中，制备了一种壳聚糖基医用敷料，这种止血棉具备抗菌的功效，但是其抗菌能力有限且机械强度低。
发明内容
为弥补现有技术的不足，本发明提供一种抗菌性多糖止血棉基纱布及其制备方法。制得的纱布强度高，价格低廉，在使用的过程当中，止血棉不脱落，并且在添加具有抗菌功能的聚乙烯亚胺后，使得在止血的同时，可以抑制和杀死细菌。
本发明首先制备具有抗菌功能的季铵化聚乙烯亚胺和醛化多糖交联剂；然后将季铵化聚乙烯亚胺，醛化多糖交联剂与壳聚糖盐酸盐混合发泡，将发泡液均匀涂覆在经过预处理的纱布表面，冷冻干燥，即可得到抗菌性多糖止血棉基纱布。
本发明所述的抗菌性多糖止血棉基纱布的制备方法为：
1)将聚乙烯亚胺溶解于四氢呋喃中，聚乙烯亚胺的浓度为100-300g/L；将溴己烷与四氢呋喃按体积比0.5-1:1混合，然后加入聚乙烯亚胺的四氢呋喃溶液中，聚乙烯亚胺与溴己烷的质量比为4-1:1，30-60℃反应6-20h；反应结束后通过旋转蒸发除去四氢呋喃，减压干燥，得到季铵化聚乙烯亚胺；
2)配制浓度为5-50g/L的多糖溶液，然后加入浓度为40-100g/L的高碘酸钠溶液，多糖溶液与高碘酸钠溶液体积比为2:1，反应2-12h后透析，最后冷冻干燥得醛化多糖交联剂；
3)配制质量浓度为1-5％的壳聚糖盐酸盐溶液，加入甘油、浓度为50-80g/L的季铵化聚乙烯亚胺溶液和浓度为20-50g/L的醛化多糖交联剂溶液；将发泡剂溶解到pH值为7-10的磷酸盐缓冲溶液中，发泡剂浓度为5-30g/L，并加入上述壳聚糖盐酸盐溶液中，搅拌发泡30-90min；
4)将硅烷偶联剂溶解在体积比为5-9:1的乙醇和水混合溶剂中，硅烷偶联剂的浓度为0.1-0.5g/L；将纱布浸泡于上述溶液中6-24小时，然后取出，并用去离子水冲洗干净，得到表面预处理的纱布；
5)将步骤3)最终得到的发泡液均匀涂覆在步骤4)得到的表面预处理的纱布表面，低温冷冻6-24h，随后冷冻干燥，辐照灭菌，得到抗菌性多糖止血棉基纱布。
所述的聚乙烯亚胺为支化聚乙烯亚胺，分子量为8000-25000。
所述的多糖选自分子量为100万-200万的透明质酸、分子量为10万-20万的葡聚糖。
所述的发泡剂为泊洛沙姆407。
所述的壳聚糖盐酸盐与醛化多糖交联剂的质量比为10-30：1，优选5-15：1。
所述的壳聚糖盐酸盐与季铵化聚乙烯亚胺的质量比为10-30：1。
所述的壳聚糖盐酸盐与发泡剂的质量比为1-5：1。
所述的壳聚糖盐酸盐与甘油的质量比为0.5-2。
所述的硅烷偶联剂选自KH540，KH550，KH560，KH580。
本发明中所述的抗菌效果来自季铵化的聚乙烯亚胺。聚乙烯亚胺中含有大量的二级胺，通过亲核取代反应使聚乙烯亚胺中的部分二级胺季铵化。大量的季铵化基团会破坏菌体的细胞膜，使细胞内物质泄漏，从而导致细菌死亡。若是在海绵中物理掺杂抗菌分子，在实际使用过程中抗菌分子会渗出，或会引起使用者的不良反应。本发明中使用多糖交联壳聚糖制备海绵的过程中加入季铵化的聚乙烯亚胺，聚乙烯亚胺上的氨基会与多糖交联剂中的醛基反应，从而被固化在海绵当中，赋予海绵接触式杀菌的功能。然而，单纯的止血棉无法达到类似于纱布的强度用来包扎伤口。因此，本发明中将具有抗菌功能的止血棉与纱布复合，旨在制备出一种抗菌性多糖止血棉基纱布。考虑到单纯将止血棉发泡液涂覆在纱布表面所形成的界面结合力小，因此需要在涂覆之前进行预处理。硅烷偶联剂作为一种具有特殊结构的有机硅化合物，在复合材料领域有着重要的应用。使用硅烷偶联剂对纱布进行预处理，硅烷偶联剂可以与纱布上的羟基发生偶联反应，另一端的官能团可以与抗菌性多糖止血棉中的氨基或醛基反应形成共价键，从而将海绵层牢固的粘附在纱布表面。多糖止血棉与经过预处理过的纱布复合后，两者结合紧密，不脱落，强度大，适合伤口处的压迫止血和包扎。并且与普通纱布相比，抗菌性多糖止血棉基纱布在止血的同时，具有抑制微生物的生长，起到抑菌的作用。另外，涂覆在纱布表面的海绵我们采用安全无毒的多糖为交联剂，进一步提升止血棉基纱布的强度和止血效果。
附图说明
图1实施例1制备的抗菌性多糖止血棉基纱布止血效果图。
具体实施方式
下面通过实施例来进一步说明本发明，但并不局限于以下的实施例。在阅读了本发明讲授的内容之后，本领域技术人员可以对本发明作各种改动或修改，但凡在本发明的精神和原则之内，这些等价形式的改动、修改等均应包含在本发明的保护范围之内。
实施例1：
1)制备具有抗菌功能的季铵化聚乙烯亚胺：将3g聚乙烯亚胺溶解于10mL四氢呋喃中；将5mL溴己烷和5mL四氢呋喃的混合溶液加入聚乙烯亚胺的四氢呋喃溶液中，50℃反应12h；反应结束后通过旋转蒸发除去四氢呋喃，减压干燥，得到季铵化聚乙烯亚胺；
2)将5g分子量为120万的透明质酸溶于100mL去离子水中，将1g高碘酸钠溶解到50mL去离子水中；将高碘酸钠溶液加入到透明质酸溶液中，反应10h后用截留分子量为3500的透析袋透析24h，最后冷冻干燥得醛化透明质酸交联剂；
3)称取2g壳聚糖盐酸盐溶解到50mL水中，搅拌溶解，呈现黄色粘稠液体；称取0.3g季铵化聚乙烯亚胺溶解到5mL去离子水中；称取0.2g醛化透明质酸交联剂溶解到5mL去离子水中；称取1g泊洛沙姆407溶解到20mL pH值为8的磷酸盐缓冲溶液中，搅拌直到完全溶解；向壳聚糖盐酸盐溶液中加入3g甘油、季铵化聚乙烯亚胺溶液和醛化透明质酸交联剂溶液，搅拌均匀后加入泊洛沙姆407溶液，搅拌发泡30min；
4)纱布预处理：将0.01g硅烷偶联剂KH550溶解在100mL体积比为8:1的乙醇和水混合溶剂中，将纱布浸泡于上述溶液中6小时，然后取出，并用去离子水冲洗干净，得到表面预处理的纱布；
5)将步骤3)最终得到的发泡液均匀涂覆在步骤4)得到的表面预处理的纱布表面，-30℃冷冻24h，随后冷冻干燥，钴60辐照灭菌，得到抗菌性多糖止血棉基纱布。
聚乙烯亚胺为支化聚乙烯亚胺，分子量为25000。
将上述制备的抗菌性多糖止血棉基纱布进行效果测试：
1.小鼠股动脉止血实验
选取25只4-6周健康小鼠，雌雄各半，随机分为3组，每组5只小鼠，分组依次是加压纱布包扎组和实施例1抗菌性多糖止血棉基纱布组。将小鼠麻醉后固定于手术台上，去毛后露出一侧股动脉，横向切开一半，令其自由喷血8s，然后根据分组进行相应处理，记录止血时间、失血量和90分钟死亡率。
表1小鼠股动脉止血实验结果
2.抗菌实验
检测用菌为大肠杆菌ATCC 25922和金黄葡萄球菌ATCC6538。纱布和抗菌性多糖止血棉基纱布分别取1×1cm的样品置于不同培养皿中。将菌种用1/500营养肉汤稀释成适当浓度的菌悬液(OD600＝1)备用。取0.1mL的菌悬液加入到每组材料中。在35℃，相对湿度不小于90％的条件下培养24h。用5mL的磷酸盐缓冲生理盐水冲洗样品，然后各取0.1mL涂布于培养基平板上，在35℃培养24h，观察结果见表2。
表2抗菌性多糖止血棉基纱布的抗菌率

Claims (9)

1.一种抗菌性多糖止血棉基纱布的制备方法，其特征在于，其具体操作步骤为：
1)将聚乙烯亚胺溶解于四氢呋喃中，聚乙烯亚胺的浓度为100-300g/L；将溴己烷与四氢呋喃按体积比0.5-1:1混合，然后加入聚乙烯亚胺的四氢呋喃溶液中，聚乙烯亚胺与溴己烷的质量比为4-1:1，30-60℃反应6-20h；反应结束后通过旋转蒸发除去四氢呋喃，减压干燥，得到季铵化聚乙烯亚胺；
2)配制浓度为5-50g/L的多糖溶液，然后加入浓度为40-100g/L的高碘酸钠溶液，多糖溶液与高碘酸钠溶液体积比为2:1，反应2-12h后透析，最后冷冻干燥得醛化多糖交联剂；
3)配制质量浓度为1-5％的壳聚糖盐酸盐溶液，加入甘油、浓度为50-80g/L的季铵化聚乙烯亚胺溶液和浓度为20-50g/L的醛化多糖交联剂溶液；将发泡剂溶解到pH值为7-10的磷酸盐缓冲溶液中，发泡剂浓度为5-30g/L，并加入上述壳聚糖盐酸盐溶液中，搅拌发泡30-90min；
4)将硅烷偶联剂溶解在体积比为5-9:1的乙醇和水混合溶剂中，硅烷偶联剂的浓度为0.1-0.5g/L；将纱布浸泡于上述溶液中6-24小时，然后取出，并用去离子水冲洗干净，得到表面预处理的纱布；
5)将步骤3)最终得到的发泡液均匀涂覆在步骤4)得到的表面预处理的纱布表面，低温冷冻6-24h，随后冷冻干燥，辐照灭菌，得到抗菌性多糖止血棉基纱布。
2.根据权利要求1所述的抗菌性多糖止血棉基纱布的制备方法，其特征在于，所述的聚乙烯亚胺为支化聚乙烯亚胺，分子量为8000-25000。
3.根据权利要求1所述的抗菌性多糖止血棉基纱布的制备方法，其特征在于，所述的多糖选自分子量为100万-200万的透明质酸、分子量为10万-20万的葡聚糖。
4.根据权利要求1所述的抗菌性多糖止血棉基纱布的制备方法，其特征在于，所述的发泡剂为泊洛沙姆407。
5.根据权利要求1所述的抗菌性多糖止血棉基纱布的制备方法，其特征在于，所述的壳聚糖盐酸盐与醛化多糖交联剂的质量比为10-30：1。
6.根据权利要求1所述的抗菌性多糖止血棉基纱布的制备方法，其特征在于，所述的壳聚糖盐酸盐与季铵化聚乙烯亚胺的质量比为10-30：1。
7.根据权利要求1所述的抗菌性多糖止血棉基纱布的制备方法，其特征在于，所述的壳聚糖盐酸盐与发泡剂的质量比为1-5：1。
8.根据权利要求1所述的抗菌性多糖止血棉基纱布的制备方法，其特征在于，所述的壳聚糖盐酸盐与甘油的质量比为0.5-2。
9.根据权利要求1所述的抗菌性多糖止血棉基纱布的制备方法，其特征在于，所述的硅烷偶联剂选自KH540，KH550，KH560，KH580。
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