CN117258030B - 一种牛角瓜纤维基水凝胶敷料及其制备方法和用途 - Google Patents
一种牛角瓜纤维基水凝胶敷料及其制备方法和用途 Download PDFInfo
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Abstract
本申请涉及一种牛角瓜纤维基水凝胶敷料及其制备方法和用途，组分包括牛角瓜纤维、海藻酸钠以及载药丝素蛋白微球，所述牛角瓜纤维基水凝胶敷料由所述组分经交联得到。本发明的水凝胶敷料通过在水凝胶中添加牛角瓜纤维(CGF)及载药丝素蛋白(SF)微球以实现分散应力，牛角瓜纤维(CGF)及载药丝素蛋白(SF)微球能够显著提高水凝胶的力学性能；此外，搭载黄连素等抗菌药物的SF微球缓释效果好，与水凝胶结合使敷料具备更佳的抗炎抗菌促愈合功能。
Description
技术领域
本发明涉及一种生物医用材料，具体地涉及一种牛角瓜纤维基水凝胶敷料及其制备方法和用途。
背景技术
皮肤创伤是指由物理、化学或生理等因素所导致的皮肤正常组织结构及功能的破坏。目前，皮肤创伤仍是威胁人类健康及经济发展的主要因素之一。皮肤创伤的愈合过程大致可分为三个时期：炎症期、增殖期和重塑期，其中：炎症期的主要过程包括对细胞碎片和创面微生物的清除；增殖期的主要过程包括成纤维细胞和角质形成细胞的增殖及迁移、微血管的再生和胶原的合成等；重塑期的主要过程包括创面多余成纤维细胞和免疫细胞等的凋亡、胶原蛋白的降解及重塑等。良好的创面敷料需满足三个主要条件：（1）具有一定的抗菌作用；（2）促进细胞的增殖；（3）维持湿润的愈合环境。
水凝胶是一类良好的皮肤创伤敷料，因其具有多孔的结构，可搭载多种不同功能的药物以实现抗菌、促愈合及抗瘢痕的效果，并能维持创面湿润愈合环境。然而，现有的用于皮肤创面修复的医用水凝胶大多仅有抗菌功能或促进细胞增殖功能，产品强力较差，药物不具备缓慢释放功能。
发明内容
本发明提供了一种牛角瓜纤维基水凝胶敷料，组分包括牛角瓜纤维（CalotropisGigantea fiber，CGF）、可成胶的高分子材料及载药丝素蛋白微球（silk fibroinmicrosphere particle，SFMP），所述牛角瓜纤维基水凝胶敷料由所述组分经交联得到。
进一步，所述载药丝素蛋白微球（SFMP）为抗菌药物负载的丝素蛋白微球，抗菌药物优选为黄连素、壳聚糖、甲壳质、芦荟、黄芩或金银花中的一种或多种。
进一步，所述可成胶的高分子材料为海藻酸钠、壳聚糖、纤维蛋白、胶原中的一种或多种。
进一步，所述可成胶的高分子材料为海藻酸钠，所述牛角瓜纤维基水凝胶敷料还包括交联剂氯化钙。
本发明还提供了一种所述的牛角瓜纤维基水凝胶敷料的制备方法，将牛角瓜纤维（CGF）、可成胶的高分子材料以及载药丝素蛋白微球（SFMP）经交联制备得到。
进一步，所述牛角瓜纤维基水凝胶敷料的包括如下步骤：先将牛角瓜纤维（CGF）与载药丝素蛋白微球（SFMP）分散在水中形成分散液，再与可成胶的高分子材料溶液混合，通过交联反应制备得到牛角瓜纤维基水凝胶敷料。
进一步，所述载药丝素蛋白微球（SFMP）为搭载抗菌药物的丝素蛋白微球，抗菌药物优选为黄连素、壳聚糖、甲壳质、芦荟、黄芩或金银花中的一种或多种，所述丝素蛋白优选为采用溴化锂法制备。
进一步，所述可成胶的高分子材料为海藻酸钠、壳聚糖、纤维蛋白、胶原中的一种或多种。
进一步，先将牛角瓜纤维（CGF）与载药丝素蛋白微球（SFMP）分散在水中形成分散液，再与海藻酸钠溶液混合，加入交联剂氯化钙，通过交联反应制备得到牛角瓜纤维基水凝胶敷料。
进一步，所述搭载抗菌药物的丝素蛋白微球由抗菌药物与丝素蛋白纳微米颗粒反应得到，优选为将黄连素溶液和丝素蛋白纳微米颗粒溶液混合，避光反应，经洗涤、重悬、冻干，得到搭载抗菌药物的丝素蛋白微球。
进一步，所述丝素蛋白纳微米颗粒的制备方法包括如下步骤：将丝素蛋白溶液与磷酸盐溶液混合，经冷藏、静置、离心，将沉淀加入去离子水后超声，洗涤、最后重悬、冻干，得到丝素蛋白纳微米颗粒。
本发明还提供了一种上述牛角瓜纤维基水凝胶敷料在制备皮肤创面修复材料中的用途。
本发明的有益效果是：本发明的水凝胶敷料通过在水凝胶中添加牛角瓜纤维（CGF）及载药丝素蛋白微球（SFMP）以实现分散应力，牛角瓜纤维（CGF）及载药丝素蛋白微球（SFMP）能够显著提高水凝胶的力学性能；此外，搭载黄连素等抗菌药物的丝素蛋白微球缓释效果好，与水凝胶结合使敷料具备更佳的抗炎抗菌促愈合功能。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作介绍。
图1是未搭载黄连素（berberine，Ber）的丝素蛋白纳微米颗粒（SFMP）（a）与搭载黄连素的丝素蛋白微球Ber@SFMP（b）的扫描电子显微镜（SEM）图。
图2是所制备的Ber@SFMP对大肠杆菌（E.coil）及金黄色葡萄球菌（S.aureus）的抑制效果图，C组是以普通棉布作为对照组的细菌生长情况。
图3为Ber@SFMP中Ber随时间变化的累计释放速率。
图4为海藻酸钠（Sodium alginate，SA）水凝胶的形貌（a）与Ber@SFMP与CGF增强SA水凝胶的形貌（b），增强后水凝胶形貌发生明显变化。
图5为SA水凝胶在分别负载2 mg Ber@SFMP（SA-2Ber@SFMP）、3 mg CGF（SA-3CGF）、负载3mg Ber@SFMP（SA-3Ber@SFMP）以及1mg CGF与2 mg Ber@SFMP（SA-2Ber@SFMP-1CGF）都负载的情况下的压缩强力测试结果图。
具体实施方式
需要说明的是，在不冲突的情况下，本申请中的实施例及实施例中的特征可以相互组合。
为使本申请实施例的目的、技术方案和优点更加清楚，下面将结合本申请实施例中的附图，对本申请实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例是本申请的一部分实施例，而不是全部的实施例。基于本申请中的实施例，本领域普通技术人员在没有做出创造性劳动的前提下所获得的所有其他实施例，都属于本申请保护的范围。
实施例1 丝素蛋白（silk fibroin，SF）溶液的制备
将蚕丝浸泡在碳酸钠溶液 (0.02 mol/L)中，质量体积比为1:400，煮沸30 min后，用去离子水洗涤三次去除残余丝胶，晾干过夜；
将脱胶后的蚕丝置于溴化锂（LiBr）溶液 (9.3 mol/L，蚕丝与LiBr溶液的质量体积比为1:4) 浸泡，60ºC孵育4 h；
将溶解的SF转移到透析袋中，在去离子水中透析2天，透析后，将SF液离心2次(9000 rpm，15 min)；然后收集上清液，得到浓度7 wt%的SF溶液。
实施例2制备搭载黄连素（berberine，Ber）的丝素蛋白微球（Ber@SFMP）。
将实施例1制备的SF溶液中加入去离子水稀释至浓度为1 wt%，再与磷酸钾溶液（1.25M，pH=8.5）以体积比为1:5的比例混合，在25ºC下，以500 rpm的速度磁力搅拌5 min，混合均匀后置于4 ºC冰箱2 h，随后在室温下静置24 h；将静置后的混合溶液以12000 rpm的速度离心10 min，倒出上清液，沉淀加入去离子水以35%功率超声3 min，进行重悬，在同样的离心速度和超声下用去离子水清洗3次，最后重悬得到丝素蛋白纳米颗粒悬浮液，进行冻干，得到丝素蛋白纳微米颗粒粉末（SFMP）。
使用去离子水分别配置2 mg/mL的Ber和SF纳微米颗粒溶液，超声分散两者，按质量比1:1将两溶液混合均匀，均匀混合后，在25ºC以500 rpm 的速度进行搅拌，避光过夜。最后，使用微量离心机在室温条件下以12000 rpm 的速度离心10 min，收集上清液待测，沉淀加入1 mL去离子水分散均匀进行重悬，重复以上操作3次，将上清液保存待后续测试，将重悬后的样品加去离子水以35%功率超声3 min后进行冻干，得到黄连素负载的丝素蛋白微球Ber@SFMP。
实施例3 制备牛角瓜纤维（Calotropis Gigantea fiber，CGF）基丝素蛋白水凝胶敷料
取100 mL去离子水于烧杯中，再加入6 g海藻酸钠（Sodium alginate，SA）粉末，室温下机械搅拌30 min，等到SA完全溶解后，真空12小时除去气泡，得到SA溶液，记为溶液A。
用去离子水制得10 mL浓度0.1 mg/mL牛角瓜纤维分散液，再加入2 mg Ber@SFMP制备Ber@SFMP-CGF溶液，记为溶液B。
取5 g CaCl2粉末于100 mL去离子水中，搅拌30 min得到CaCl2溶液。
将溶液A与溶液B以10/10的体积混合，再喷洒CaCl2溶液进行交联后，静置，得到牛角瓜纤维基丝素蛋白水凝胶敷料。
对比例1本对比例与实施例3相比，将2 mg Ber@SFMP加入10 mL去离子水中，得到溶液B。
对比例2本对比例与实施例3相比，用去离子水制得10 mL浓度0.3mg/mL牛角瓜纤维分散液，即为溶液B。
对比例3本对比例与实施例3相比，将3 mg Ber@SFMP加入10 mL去离子水中，得到溶液B。
效果例1对实施例2制备得到的Ber@SFMP与没有经过黄连素负载的丝素蛋白微球的电位性质进行验证，如表1所述，微球载药后（即Ber@SFMP）的电位变高，这是由于带正电的黄连素水溶液与带负电的SF溶液发生电荷中和，粒径的变化也论证了微球表面进行了载药，载药率可达42.36%；此外，从图1未搭载黄连素（Ber）的丝素蛋白纳微米颗粒（SFMP）（a）与Ber@SFMP（b）的扫描电子显微镜（SEM）对比图可以看出，载药后的Ber@SFMP相比未载药的SFMP粒径增加，表面形貌发生改变，聚集了黄连素颗粒。
表1载药微球性能表征
效果例2按照GB/T 20944.3-2008对所制备的Ber@SFMP以0.1 mg/mL的浓度置于PBS中进行24 h震荡，对大肠杆菌（E.coil）及金黄色葡萄球菌（S.aureus）的抑制效果进行表征，C 组是以普通棉布作为对照组的细菌生长情况，结果：Ber@SFMP对大肠杆菌和金黄色葡萄球菌的抑制率分别为51.70±1.55%与99.51±0.23%，满足国标规定的抑菌效果要求（＞50%）。
效果例3为探究Ber@SFMP 中 Ber 的突释及缓释效果（这关系着其用于伤口敷料抗菌抗炎等的效果），分别取定量的 Ber@SFMP 悬浮于1 mmol/L PBS（pH 7.4）溶液中，然后转移分装至10 mL的离心管（浓度为1 mg/mL），离心管密闭放置在摇床上，并将摇床置于37ºC烘箱中，特定时间点（1、3、6、9、12、24、48、72、96、120、144 小时）取出离心管，离心管离心换液后继续放置烘箱中摇床上振荡，上清液（释放液）用紫外分光光度计检测吸光度，以PBS的吸光度作为基线，并根据 Ber 溶于1 mmol/L PBS 紫外分光吸光度与药物浓度的标准曲线计算出释放液中 Ber 的含量，并以此计算 Ber 的累计释放速率。从图3可以看出，Ber@SFMP缓释效果优秀，当以1 mg/mL 在1 mmol/L PBS（pH 7.4）进行缓释测试时，前6 h可释放18.81%，满足敷料的抑菌效果要求。
效果例4图5 为SA水凝胶在分别负载2 mg Ber@SFMP（SA-2Ber@SFMP，即对比例1制备的牛角瓜纤维基丝素蛋白水凝胶敷料）、3 mg CGF（SA-3CGF，即对比例2制备的牛角瓜纤维基丝素蛋白水凝胶敷料）、3 mg Ber@SFMP（SA-3Ber@SFMP，即对比例3制备的牛角瓜纤维基丝素蛋白水凝胶敷料）以及1mg CGF与2 mg Ber@SFMP（SA-2Ber@SFMP-1CGF，即实施例3制备的牛角瓜纤维基丝素蛋白水凝胶敷料）都负载的情况下的压缩强力测试情况，可看出4种负载方式对SA水凝胶均有增强，但Ber@SFMP和CGF 都存在的情况下，水凝胶的抵抗压缩变形的能力明显最强。
以上述依据本申请的理想实施例为启示，通过上述的说明内容，相关工作人员完全可以在不偏离本项申请技术思想的范围内，进行多样的变更以及修改。本项申请的技术性范围并不局限于说明书上的内容，必须要根据权利要求范围来确定其技术性范围。
Claims (9)
1.一种牛角瓜纤维基水凝胶敷料，其特征在于，组分包括牛角瓜纤维、海藻酸钠以及载药丝素蛋白微球，所述牛角瓜纤维基水凝胶敷料由所述组分经交联得到。
2.根据权利要求1所述的牛角瓜纤维基水凝胶敷料，其特征在于，所述载药丝素蛋白微球为搭载抗菌药物的丝素蛋白微球，所述抗菌药物为黄连素、壳聚糖、甲壳质、芦荟、黄芩或金银花中的一种或多种。
3.根据权利要求1或2所述的一种牛角瓜纤维基水凝胶敷料，其特征在于，所述牛角瓜纤维基水凝胶敷料还包括交联剂氯化钙。
4.一种权利要求1-3任一项所述的牛角瓜纤维基水凝胶敷料的制备方法，其特征在于，将牛角瓜纤维、海藻酸钠以及载药丝素蛋白微球经交联制备得到。
5.根据权利要求4所述的牛角瓜纤维基水凝胶敷料的制备方法，其特征在于，先将牛角瓜纤维与载药丝素蛋白微球分散在水中形成分散液，再与海藻酸钠溶液混合，通过交联反应制备得到牛角瓜纤维基水凝胶敷料。
6.根据权利要求4或5所述的牛角瓜纤维基水凝胶敷料的制备方法，其特征在于，先将牛角瓜纤维与载药丝素蛋白微球分散在水中形成分散液，再与海藻酸钠溶液混合，加入交联剂氯化钙，通过交联反应制备得到牛角瓜纤维基水凝胶敷料。
7.根据权利要求4或5所述的牛角瓜纤维基水凝胶敷料的制备方法，其特征在于，所述载药丝素蛋白微球由抗菌药物与丝素蛋白纳微米颗粒反应得到。
8.根据权利要求7所述的牛角瓜纤维基水凝胶敷料的制备方法，其特征在于，所述丝素蛋白纳微米颗粒的制备方法包括如下步骤：将丝素蛋白溶液与磷酸盐溶液混合反应，经冷藏、静置、离心，将沉淀加入去离子水后超声，洗涤、最后重悬、冻干，得到丝素蛋白纳微米颗粒。
9.权利要求1-3任一项所述的牛角瓜纤维基水凝胶敷料或根据权利要求4-8任一项权利要求所述的方法制备得到的牛角瓜纤维基水凝胶敷料在制备皮肤创面修复材料中的用途。
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| CN104327283A (zh) * | 2014-10-16 | 2015-02-04 | 苏州经贸职业技术学院 | 一种复合丝素蛋白水凝胶及其制备方法、应用 |
| CN105854068A (zh) * | 2015-01-21 | 2016-08-17 | 天津开发区金衫包装制品有限公司 | 一种含中药抗菌成分黄连素的海藻酸钠纳米纤维医用敷料及其制备方法 |
| CN107261196A (zh) * | 2017-05-16 | 2017-10-20 | 苏州大学 | 一种抗菌丝素蛋白材料及其制备方法 |
| CN109836595A (zh) * | 2019-01-28 | 2019-06-04 | 北京航空航天大学 | 一种仿生软骨掺杂有序磁性纳米短纤维拼接双层水凝胶的制备方法 |
| CN110003502A (zh) * | 2019-04-09 | 2019-07-12 | 西安培华学院 | 一种丝素蛋白纳米微球及其制备方法 |
| CN110917400A (zh) * | 2019-12-05 | 2020-03-27 | 中山大学 | 一种纳米杂化丝素蛋白水凝胶及其制备方法和应用 |
| CN111101250A (zh) * | 2020-01-11 | 2020-05-05 | 南通纺织丝绸产业技术研究院 | 一种多功能牛角瓜纤维混纺纱线的制备方法 |
| CN111803697A (zh) * | 2020-07-16 | 2020-10-23 | 太原理工大学 | 一种载药海藻酸钠/明胶复合水凝胶型创可贴的制备方法 |
| CN112957521A (zh) * | 2021-04-01 | 2021-06-15 | 浙江理工大学 | 一种载青蒿素脂质体的海藻酸盐-丝素蛋白复合水凝胶的制备方法 |
| CN113736026A (zh) * | 2021-09-14 | 2021-12-03 | 山东盛和纺织股份有限公司 | 一种改性牛角瓜纤维及其制备方法、改性牛角瓜纤维/es纤维热风非织造布、高吸湿口罩 |
| CN114668883A (zh) * | 2022-04-21 | 2022-06-28 | 安徽大学 | 一种姜黄素负载海藻酸钠/聚乙烯醇/丝素蛋白复合支架的制备方法 |
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