CN115025278A - 一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料及其制备方法 - Google Patents
一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料及其制备方法 Download PDFInfo
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- CN115025278A CN115025278A CN202210752415.9A CN202210752415A CN115025278A CN 115025278 A CN115025278 A CN 115025278A CN 202210752415 A CN202210752415 A CN 202210752415A CN 115025278 A CN115025278 A CN 115025278A
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- Prior art keywords
- houttuynia cordata
- honey
- carboxylated chitosan
- preparation
- hydrogel dressing
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- Medicinal Preparation (AREA)
Abstract
本发明提供了一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料及其制备方法，属于医用水凝胶敷料技术领域。该水凝胶敷料包含以下组分：羧化壳聚糖1.5～7.5wt％，羧甲基纤维素2～8wt％，鱼腥草粉0.5～1.5wt％，蜂蜜1～3wt％，余量为水。本发明通过将羧化壳聚糖溶于超纯水中，加入羧甲基纤维素溶解，再加入鱼腥草粉和蜂蜜，混匀后超声处理，然后静置消除气泡，得到该水凝胶敷料。该制备方法工艺简单，制备条件温和，原料均为自然界丰富易得的生物材料，制备的水凝胶敷料具有优异的生物相容性和降解性，以及良好的抗菌消炎、止血镇痛、吸收伤口渗液、促进伤口愈合及组织修复再生的功能。
Description
技术领域
本发明属于医用水凝胶敷料技术领域，具体涉及一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料及其制备方法。
背景技术
临床上处理患者伤口时需要在去除坏死组织后在伤口上覆盖敷料，以防止细菌侵入和控制水分损失。传统敷料(如纱布、棉垫等)虽然来源广泛、质地柔软、成本低，但其止血效果不令人满意，没有保湿作用，对创面的愈合也没有促进作用，肉芽组织容易长入纱布网眼中，导致粘连结痂，而且敷料浸透时易导致外源性感染，更换时易造成二次创伤。相比于传统敷料，水凝胶是一些高聚物或共聚物吸收大量水分后形成的溶胀交联半固体，具有良好的柔韧性和更好的生物相容性，其制作的敷料吸收液体性能好，能够为创面创造一个利于组织再生的湿润环境和低氧环境，而且水凝胶的滑弹状态可有效地避免伤口粘连造成二次伤害。因此，水凝胶敷料在医用敷料领域有更广阔的应用前景。
目前，水凝胶类型有明胶水凝胶、壳聚糖水凝胶、海藻酸水凝胶、透明质酸水凝胶、葡聚糖水凝胶、共聚物水凝胶、MOFs水凝胶等。其中，壳聚糖水凝胶的主要原料是天然可降解多糖壳聚糖及其水溶性衍生物，不仅具有优良的生物相容性和可降解性，而且还具有抗菌、防腐、止血和促进伤口愈合等特殊功能。
中国专利CN111035801B公开了“银纳米团簇基壳聚糖水凝胶敷料及其制备方法和应用”，其采用戊二醛化学交联壳聚糖并加入纳米银粒子，所制备敷料能够有效避免伤口感染，对于伤口愈合具有积极作用；但使用的化学交联剂戊二醛有一定毒性且对人的皮肤粘膜有刺激性，无机纳米银粒子的成本较高且与有机物存在分散相容问题。中国专利CN103480034B公开了“辐照交联壳聚糖/明胶/聚乙烯醇水凝胶敷料及其制备方法和应用”，采用高能电子束或γ光子照射进行辐照交联制备的水凝胶敷料可应用于外伤止血、消炎、烫伤、手术后伤口愈合等领域；但辐射交联方法会使聚合物发生降解，分子量变小，交联和降解同时发生，从而在体系中引入杂质，造成后处理困难，从而使其应用受到限制。中国专利CN109731121B公开了“一种含有介孔二氧化硅的纤维素和壳聚糖复合敷料的制备方法”，通过共混物理交联制备的敷料具备较高的拉伸强度，具有多孔结构，透气性、保湿性良好，具有长效杀菌抑菌作用；但制备过程较为复杂，制备条件不易控制，且添加了具有一定副作用的抗菌药、消炎药及伤口愈合药等西药。中国专利CN102764447B公开了“水凝胶敷料及其制备工艺”，通过将羟乙基纤维素、丙二醇、水以及羧甲基壳聚糖或壳聚糖季铵盐共混制备了一种生物相容性及降解性较好的水凝胶；但该水凝胶敷料功能比较单一，在处理伤口时主要起杀菌功能。
鉴于上述壳聚糖水凝胶技术方案中存在的问题，需要开发一种生物相容性与降解性良好，同时具有消炎、抗菌、止血、吸收伤口渗液、促进伤口愈合且无毒性的功能性生物医用伤口敷料。
发明内容
本发明的目的在于针对现有壳聚糖水凝胶敷料存在的不足，提供一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料及其制备方法，该羧化壳聚糖水凝胶敷料制备工艺简单，生产成本低廉，将羧化壳聚糖与鱼腥草和蜂蜜相结合，使得该水凝胶敷料不仅具有良好的生物相容性和可降解性，还具有良好的抗菌消炎、止血镇痛、吸收伤口渗液、促进伤口愈合及组织修复再生等功能。
为了实现上述目的，本发明采用如下技术方案：
一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料，以质量百分比计，所述水凝胶敷料包含以下组分：羧化壳聚糖1.5％～7.5％，羧甲基纤维素2％～8％，鱼腥草粉0.5％～1.5％，蜂蜜1％～3％，余量为水。
本发明中选用由壳聚糖经羧化改性得到的羧化壳聚糖，不仅易溶于水，还具有优良的吸湿、保湿、调理等功能，以及良好的抑菌、防腐、止血和促进伤口愈合等功能；所使用的羧甲基纤维素溶入水后，具有增稠、保湿、成膜、黏接、乳化及悬浮等作用；所使用的鱼腥草粉末是天然而又安全的抗生素，其内含癸酰乙醛、月桂醛、α-蒎烯等抗菌有效成分，使其具有抗菌、抗病毒、消炎止血、增强机体免疫力等作用；所使用的蜂蜜含有果糖、葡萄糖、酶类、维生素、微量元素等，使其具有抗菌消炎、促进伤口愈合及促进组织再生的修复能力。
为确保羧化壳聚糖在水中溶解性更好，所述羧化壳聚糖的羧化度≥60％，且1wt％水溶液的粘度为10～80mpa·s；所述羧甲基纤维素作为增稠剂，其分子量为52～70万，且2wt％水溶液的粘度为1500～4500mpa·s；所述蜂蜜具有促进组织再生的能力，所含果糖和葡萄糖质量含量≥60％。
本发明还提供了上述含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料的制备方法，包括以下步骤：
S1、将所述羧化壳聚糖溶于超纯水中得到羧化壳聚糖溶液；
S2、将所述羧甲基纤维素溶于所述羧化壳聚糖溶液中，得到混合溶液；
S3、将所述混合溶液加热至38～42℃，再加入所述鱼腥草粉和蜂蜜，充分混匀，得悬浮液；
S4、将所述悬浮液超声处理，室温下静置消除气泡，即得所述含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料。
优选的，步骤S3中将所述混合溶液加热至40℃。
优选的，步骤S3中所述鱼腥草粉的制备方法如下：将鱼腥草择去根须后用水清洗干净，然后将洗好的鱼腥草切成小段并烘干，再粉碎、过筛网，得所述鱼腥草粉末；筛网目数≥200目。
进一步优选的，将洗好的鱼腥草切成小段，置于80℃下干燥48h。
优选的，步骤S3中充分混匀是指在100～150rpm搅拌速度下搅拌1～3h。
优选的，步骤S4中超声处理是指以400～800W的功率超声20～60min。
优选的，室温下静置消除气泡的时间为20～40h。
与现有技术相比，本发明具有以下有益效果：
(1)本发明提供的含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料中，羧化壳聚糖、羧甲基纤维素、鱼腥草粉及蜂蜜均为天然有机生物质材料，它们不仅具有良好的生物相容性和生物安全性，而且各组分之间通过官能团的静电作用、氢键结合以及分子链的缠结等结合形式进行物理交联，形成的凝胶具有热可逆性和良好的生物降解性。
(2)本发明的制备工艺简单，制备条件温和，制备过程无高能光源辐照，无交联剂、引发剂及其他任何有毒有害物质的添加，产品纯净无杂质、安全无毒、环境友好；此外，该水凝胶敷料制备使用的各种原料自然界丰富易得，因此生产成本低廉，经济效益高。
(3)本发明通过各组分之间的协同作用，使获得的复合水凝胶敷料的作用效果显著优于单独使用其中某一组分时的效果，从而成为同时具有抗菌消炎、止血镇痛、吸收伤口渗液、促进伤口愈合及组织修复再生等多功能的医用敷料，对烧伤或烫伤、外科手术后的伤口和外伤、溃疡等的治疗具有很好的效果。
具体实施方式
以下结合实施例对本发明技术方案进行清楚、完整的描述，显然，所描述的实施例仅仅是本发明的一部分实施例，而不是全部的实施例。基于本发明的实施例，本领域普通技术人员在没有做出创造性劳动的前提下所得到的所有其它实施例，都属于本发明所保护的范围。本领域技术人员依据以下实施方式所作的任何等效变换或替代，均属于本发明的保护范围之内。
以下各实施例中，除筛网目数不同外，鱼腥草粉均按如下方法制备：
将鱼腥草择去根须后用水将杂质清洗干净，然后将洗好的鱼腥草切成小段并置于80℃烘箱中干燥48h，再将干燥好的鱼腥草通过植物粉碎机粉碎成粉末，过筛网，筛网目数≥200目，得所述鱼腥草粉。
实施例1
本实施例提供了一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料，以质量百分比计，包含以下组分：羧化壳聚糖1.5％，羧甲基纤维素8％，鱼腥草粉1.5％，蜂蜜1％，余量为水；其中羧化壳聚糖的羧化度为60％，1wt％羧化壳聚糖水溶液的粘度为80mpa·s；羧甲基纤维素的分子量为70万，2wt％羧甲基纤维素水溶液的粘度为4500mpa·s；蜂蜜中果糖含量为32wt％，葡萄糖含量为28wt％，果糖和葡萄糖总含量为60wt％；
所述水凝胶敷料的制备方法包括以下步骤：
S1、称取羧化壳聚糖，加入超纯水中，搅拌至全部溶解，得羧化壳聚糖溶液；
S2、称取羧甲基纤维素，加入步骤S1的羧化壳聚糖溶液中，搅拌至全部溶解；
S3、将步骤S2得到的溶液加热至42℃，加入粒度为200目的鱼腥草粉和蜂蜜，以150rpm速度搅拌1h；
S4、将步骤S3得到的溶液在700W功率下超声分散30min，然后在室温下静置40h消除气泡，得到含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料；
对照组11：与实施例1不同之处在于没有步骤S3，其他均相同；
对照组12：与实施例1不同之处在于步骤S3中只加入鱼腥草粉末而不加入蜂蜜，其他均相同；
对照组13：与实施例1不同之处在于步骤S3中只加入蜂蜜而不加入鱼腥草粉末，其他均相同。
按照如下方法对实施例1和对照组11～13制备的水凝胶敷料进行抗菌试验：
将实施例1和对照组11～13制得的水凝胶敷料分别按照1％的质量比溶于琼脂培养基(主要成分包含：10g/L蛋白胨，3g/L牛肉膏，5g/L氯化钠，18g/L琼脂，于121℃高压灭菌15min)，然后用移液枪分别吸取市购的大肠杆菌、金黄色葡萄球菌和白色念珠菌悬液各0.1mL，并用涂布棒分别使三种细菌悬液均匀分布在营养琼脂培养基上，最后置于37℃培养箱中培养48h后计算抑菌率，实验结果如表1所示：
表1水凝胶敷料对不同细菌的抑菌率
| 实施例1 | 对照组11 | 对照组12 | 对照组13 | |
| 大肠杆菌抑菌率 | 99.99％ | 62.35％ | 87.78％ | 85.41％ |
| 金黄色葡萄球菌抑菌率 | 100.00％ | 64.68％ | 90.52％ | 87.34％ |
| 白色念珠菌抑菌率 | 99.98％ | 60.76％ | 86.46％ | 84.68％ |
从表1中可以看出，实施例1和对照组11～13制备的水凝胶敷料均具有抗菌活性。对照组11作为空白组(未加鱼腥草粉末和蜂蜜)，其抑菌活性来源于水溶性羧化壳聚糖的抑菌性；只添加了鱼腥草粉末的对照组12和只添加了蜂蜜的对照组13的抑菌率均高于对照组11，说明羧化壳聚糖水凝胶中加入鱼腥草或蜂蜜后对其抗菌效果具有促进作用；实施例1制备的水凝胶敷料与三个对照组相比，表现出了优异的抑菌效果，说明羧化壳聚糖、鱼腥草粉末、蜂蜜之间产生了相互有益的协同效应，因此，本发明制备的含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料可为伤口提供一个清洁无菌的愈合环境。
实施例2
本实施例提供了一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料，以质量百分比计，包含以下组分：羧化壳聚糖3.5％，羧甲基纤维素2％，鱼腥草粉1.0％，蜂蜜3％，余量为水；其中羧化壳聚糖的羧化度为80％，1wt％羧化壳聚糖水溶液的粘度为40mpa·s；羧甲基纤维素的分子量为52万，2wt％羧甲基纤维素水溶液的粘度为1500mpa·s；蜂蜜中果糖含量为40wt％，葡萄糖含量为35wt％，果糖和葡萄糖总含量为75wt％；
所述水凝胶敷料的制备方法与实施例1基本相同，不同之处在于，步骤S3中加热至40℃，鱼腥草粉的粒度为300目，以130rpm速度搅拌1.5h；步骤S4中在400W功率下超声60min，在室温下静置25h消除气泡。
对照组21：与实施例2不同之处在于没有步骤S3，其他均相同；
对照组22：与实施例2不同之处在于步骤S3中只加入鱼腥草粉末而不加入蜂蜜，其他均相同；
对照组23：与实施例2不同之处在于步骤S3中只加入蜂蜜而不加入鱼腥草粉末，其他均相同。
按照如下方法对实施例2和对照组21～23制备的水凝胶敷料进行止血试验：
将小白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，有血液溢出后立即进行止血处理。其中，实施例2和对照组21～23分别采用各自制备的水凝胶敷料进行止血处理；空白组I不进行任何处理，空白组II只用传统医用纱布包扎，空白组III采用云南白药处理；每组小白鼠各3只。根据分组记录止血平均时间，实验结果如表2所示：
表2不同止血方式所需止血时间
从表2可以看出，实施例2、对照组22、对照组23制备的水凝胶敷料的止血时间都比云南白药的短，尤其是实施例2的水凝胶敷料的止血速度比云南白药提高53.9％，而且本发明制备的水凝胶敷料止血后还可以继续使用，但是云南白药粉止血后需要清理掉，否则会对伤口愈合产生不利影响。实施例2制备的水凝胶敷料的止血效果显著优于对照组21～23，止血时间只有传统医用纱布止血时间的13.5％；这主要是得益于羧化壳聚糖、鱼腥草和蜂蜜之间的协同作用，使得实施例2制备的水凝胶敷料的凝血效果很好，使创伤表面的流血量迅速减少，从而在较短时间内达到止血目的。
实施例3
本实施例提供了一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料，以质量百分比计，包含以下组分：羧化壳聚糖7.5％，羧甲基纤维素4％，鱼腥草粉0.5％，蜂蜜1.5％，余量为水；其中羧化壳聚糖的羧化度为70％，1wt％羧化壳聚糖水溶液的粘度为60mpa·s；羧甲基纤维素的分子量为58万，2wt％羧甲基纤维素水溶液的粘度为2500mpa·s；蜂蜜中果糖含量为42wt％，葡萄糖含量为38wt％，果糖和葡萄糖总含量为80wt％；
所述水凝胶敷料的制备方法与实施例1基本相同，不同之处在于，步骤S3中加热至38℃，鱼腥草粉的粒度为270目，以100rpm速度搅拌3h；步骤S4中在800W功率下超声20min，在室温下静置20h消除气泡。
对照组31：与实施例3不同之处在于没有步骤S3，其他均相同；
对照组32：与实施例3不同之处在于步骤S3中只加入鱼腥草粉末而不加入蜂蜜，其他均相同；
对照组33：与实施例3不同之处在于步骤S3中只加入蜂蜜而不加入鱼腥草粉末，其他均相同。
按照如下方法对实施例3和对照组31～33制备的水凝胶敷料进行伤口愈合试验：
将麻醉后的小白鼠臀部被毛剔除，然后用75℃电烙铁置于剃毛皮肤处10s，得到直径为1cm的圆形烫伤创面，然后取3cm×3cm的水凝胶敷料覆盖在创面上，每隔2天更换一次敷料，直至创口完全愈合(临床上认为创面愈合率达到95％即可认为完全愈合)，期间观察敷料与创面的粘合情况及创面的愈合情况。空白组采用传统医用纱布覆盖创面。实验结果如表3所示：
表3不同水凝胶敷料伤口愈合试验结果
从表3可以看出，实施例3和对照组31～33制备的水凝胶敷料用于处理烫伤创面时的伤口处愈合时间均短于传统医用纱布组，新生肉芽组织生成后具有弹性且无脂肪液化现象发生，痊愈后创面也较为平整；而空白组(使用传统医用纱布)的伤口愈合时间长，且每次更换纱布时都会黏连，从而导致创口恢复慢，伤口愈合后表面轻微发红且有明显疤痕。实施例3、对照组32、对照组33的水凝胶敷料变成薄膜的时间以及伤口愈合时间均比对照组31的短，实施例3的水凝胶敷料变成薄膜的时间和伤口愈合时间又都比对照组32、对照组33的更短；这说明实施例3中在羧化壳聚糖水凝胶中同时加入鱼腥草和蜂蜜后，通过三者之间的协同作用，使得本发明制备的含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料对于外部创伤所致的伤口具有优异的伤口渗液吸收能力、促进伤口愈合及组织修复再生的能力。
实施例4
本实施例提供了一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料，以质量百分比计，包含以下组分：羧化壳聚糖5.5％，羧甲基纤维素6％，鱼腥草粉1.2％，蜂蜜2.4％，余量为水；其中羧化壳聚糖的羧化度为65％，1wt％羧化壳聚糖水溶液的粘度为70mpa·s；羧甲基纤维素的分子量为65万，2wt％羧甲基纤维素水溶液的粘度为4000mpa·s；蜂蜜中果糖含量为40wt％，葡萄糖含量为35wt％，果糖和葡萄糖总含量为75wt％；
所述水凝胶敷料的制备方法与实施例1基本相同，不同之处在于，步骤S3中加热至40℃，鱼腥草粉的粒度为400目，以120rpm速度搅拌2h；步骤S4中在600W功率下超声40min，在室温下静置30h消除气泡。
对照组41：与实施例4不同之处在于没有步骤S3，其他均相同；
对照组42：与实施例4不同之处在于步骤S3中只加入鱼腥草粉末而不加入蜂蜜，其他均相同；
对照组43：与实施例4不同之处在于步骤S3中只加入蜂蜜而不加入鱼腥草粉末，其他均相同。
按照实施例1中的抗菌试验方法、实施例2中的止血试验方法、实施例3中的伤口愈合试验方法分别对实施例4和对照组41～43进行试验，试验结果见表4：
表4不同水凝胶敷料的应用效果测试结果
从表4可以看出，实施例4在羧化壳聚糖水凝胶中同时加入鱼腥草和蜂蜜后，与不添加鱼腥草和蜂蜜的对照组41、只添加了鱼腥草的对照组42和只添加了蜂蜜的对照组43相比，不仅表现出了优异的抑菌效果，而且止血时间、水凝胶敷料变成薄膜时间、伤口处愈合时间均得到显著缩短，伤口愈合情况表现良好。这是因为在羧化壳聚糖水凝胶中同时加入鱼腥草和蜂蜜后，羧化壳聚糖继续发挥其抑菌、防腐、止血和促进伤口愈合等功能；而富含抗菌有效成分的鱼腥草在抗菌、抗病毒、消炎止血、增强机体免疫力等方面发挥加强作用，尤其是在止血镇痛方面加强作用效果更明显；而富含果糖、葡萄糖、酶类、维生素、微量元素等的蜂蜜则在抗菌消炎、促进伤口愈合及促进组织再生方面发挥加强作用，尤其是在吸收伤口渗液、促进伤口愈合及促进组织修复再生等方面的加强作用效果更明显。通过羧化壳聚糖、鱼腥草和蜂蜜三种生物质原料之间的协同效应，使得本发明制备的含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料不仅具有优异的生物相容性及降解性，还同时具有抗菌消炎、止血镇痛、吸收伤口渗液、促进伤口愈合及组织修复再生等特点，是一种对烧伤或烫伤、外科手术后的伤口或外伤、溃疡等有治疗效果的功能性生物医用伤口敷料。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明的保护范围。对于任何熟悉本领域的技术人员来说，本发明可以有各种更改和变化。任何依据本发明申请保护范围及说明书内容所作的简单的等效变化和修饰，均应包含在本发明的保护范围之内。
Claims (9)
1.一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料，其特征在于，以质量百分比计，所述水凝胶敷料包含以下组分：羧化壳聚糖1.5％～7.5％，羧甲基纤维素2％～8％，鱼腥草粉0.5％～1.5％，蜂蜜1％～3％，余量为水。
2.根据权利要求1所述的一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料，其特征在于，所述羧化壳聚糖的羧化度≥60％，且1wt％水溶液的粘度为10～80mpa·s；所述羧甲基纤维素的分子量为52～70万，且2wt％水溶液的粘度为1500～4500mpa·s；所述蜂蜜中果糖和葡萄糖质量含量≥60％。
3.权利要求1或2所述的一种含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料的制备方法，其特征在于，包括以下步骤：
S1、将所述羧化壳聚糖溶于超纯水中得到羧化壳聚糖溶液；
S2、将所述羧甲基纤维素溶于所述羧化壳聚糖溶液中，得到混合溶液；
S3、将所述混合溶液加热至38～42℃，再加入所述鱼腥草粉和蜂蜜，充分混匀，得悬浮液；
S4、将所述悬浮液超声处理，室温下静置消除气泡，即得所述含鱼腥草和蜂蜜的羧化壳聚糖水凝胶敷料。
4.根据权利要求3所述的制备方法，其特征在于，步骤S3中将所述混合溶液加热至40℃。
5.根据权利要求3所述的制备方法，其特征在于，步骤S3中所述鱼腥草粉的制备方法如下：将鱼腥草择去根须后用水清洗干净，然后将洗好的鱼腥草切成小段并烘干，再粉碎、过筛网，得所述鱼腥草粉；筛网目数≥200目。
6.根据权利要求5所述的制备方法，其特征在于，将洗好的鱼腥草切成小段，置于80℃下干燥48h。
7.根据权利要求3所述的制备方法，其特征在于，步骤S3中充分混匀是指在100～150rpm搅拌速度下搅拌1～3h。
8.根据权利要求3所述的制备方法，其特征在于，步骤S4中超声处理是指以400～800W的功率超声20～60min。
9.根据权利要求3所述的制备方法，其特征在于，室温下静置消除气泡的时间为20～40h。
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