CN112295008B - 一种具有抗炎止血功能的生物活性敷料 - Google Patents

一种具有抗炎止血功能的生物活性敷料 Download PDF

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CN112295008B
CN112295008B CN202011249079.3A CN202011249079A CN112295008B CN 112295008 B CN112295008 B CN 112295008B CN 202011249079 A CN202011249079 A CN 202011249079A CN 112295008 B CN112295008 B CN 112295008B
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parts
dressing
chitosan
salicylic acid
hemostatic
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CN112295008A (zh
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罗宇仙
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Guangzhou Tuwei Kechuang Biotechnology Co ltd
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Guangzhou Tuwei Kechuang Biotechnology Co ltd
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Abstract

本发明公开了一种具有抗炎止血功能的生物活性敷料及其制备方法。所述敷料包括如壳聚糖5.0‑10.0份、海藻酸钠5.0‑10.0份、止血药物5.0‑15.0份、水杨酸1.0‑3.0份和β‑环糊精1.0‑10.0份。所述敷料实现水杨酸的可控释放，降低水杨酸对皮肤的刺激和毒副作用；另外，本发明通过止血药物、薄荷醇和水杨酸三者之间的协同作用实现了快速止血和伤口愈合的效果，同时原料丰富，制作成本低，可广泛应用于小型出血性伤口创面的处理。

Description

一种具有抗炎止血功能的生物活性敷料
技术领域
本发明属于医用材料技术领域，具体涉及一种抗炎止血功能的生物活性敷料、制备方法及其应用。
背景技术
诸如棉质纱布、无纺布类的传统伤口敷料在过去得到了广泛的应用，然而由于其容易粘在伤口部位而容易造成在更换或去除敷料时伤口部位的二次创伤，以及较差的屏障作用而容易导致伤口的病菌感染，且对伤口创面无明显的愈合作用。因此，急需一种对外部病菌起到有效阻隔作用，同时具有理想的伤口愈合、抗菌性效果的敷料。
壳聚糖基敷料是一类天然的生物大分子材料，其具有诸多优良的生物材料性能，因而近年作为敷料在医疗领域得到了不断的应用。专利CN201810694061.0公开了一种壳聚糖生物大分子，其通过将壳聚糖和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)活化剂和多酚类化合物反应，得到多酚和温敏性化合物接枝改性的壳聚糖，实现了敷料的湿粘附及温度响应性；专利CN104013990A公开了一种具有儿茶酚基结构的壳聚糖生物医学材料，通过将儿茶酚基结构接枝到壳聚糖从而提高了壳聚糖的性能，改善组织的亲和性，从而使止血海绵具有很好的止血效果；CN110772659A公开了一种石墨烯接枝改性的壳聚糖医用敷料，其具有特殊的理疗聚合功能，特别是物理抗菌和远红外辐射促进伤口方面；CN105107008B公开了一种羟丁基壳聚糖/氧化海藻酸钠/纳米银复合敷料，其中所述的凝胶敷料快速吸收渗液，又具有良好的舒适性。
然而，现有的壳聚糖基敷料多是通过壳聚糖改性来提高敷料的性能。由于壳聚糖分子间或分子内的氢键的大量形成，造成壳聚糖分子较差的溶解度，这极大地限制了壳聚糖基材料对水难溶性药物分子的有效装载且相应地导致药效不显著问题，从而限制了壳聚糖基材料在药用敷料领域中的广泛应用。
目前，利用环糊精来包合各种水难溶性药物分子形成环糊精包合物，然后再将包合物添加到壳聚糖基载体材料中，进而改善水难溶性药物分子在壳聚糖基敷料中的有效装载量是最近研究较多的一种途径。然而，包合物也存在各种各样的问题，如药物分子从包合物中的释放速率相比原药物存在过快或过慢的问题。
水杨酸是一种脂溶性的有机酸，由于其原料易得、制备简单以及其优异的杀菌、防腐、消炎、消肿、止痒等功能而被广泛地用于制备各种皮肤用化妆品或药品中。然而，水杨酸难溶于性水，这大大限制水杨酸在各种水性制剂、特别是水凝胶制剂中的广泛应用。
为了克服现有技术的不足，在本发明中，通过选择壳聚糖、水杨酸的环糊精包合物、药物成分及添加剂，合理的配比各种组分，实现了水杨酸在壳聚糖基凝胶敷料中的有效装载和可控释放，从而显著地缩短了止血时间且加快伤口出血部位的愈合。
发明内容
本发明的一个目的在于解决水杨酸在壳聚糖基水性凝胶中的有效装载的问题，同时实现了水杨酸通过水凝胶中的可控释放。
本发明的另一个目的在于通过壳聚糖、海藻酸钠、保湿剂、止血药物、薄荷醇和水杨酸之间的配合和协同作用，实现了快速止血、止痒、无残留、促进出血创面的愈合，减少了水杨酸的刺激性，并提供了一种具有良好的舒适性、易贴易揭的药用止血敷料。
本发明的目的通过以下技术方案实现：
一种具有抗炎止血功能的生物活性敷料，包括如下质量份数的原料：壳聚糖5.0-10.0份、海藻酸钠5.0-10.0份、止血药物5.0-15.0份、水杨酸1.0-3.0份和β-环糊精1.0-10.0份。
具体地，一种具有抗炎止血功能的生物活性敷料，包括如下质量份数的原料：
壳聚糖5.0-10.0份，如5.0份、6.0份、7.0份、.08份、9.0份、10.0份或者它们之间任意范围内的任意值；
海藻酸钠5.0-10.0份，如5.0份、6.0份、7.0份、8.0份、9.0份、10.0份或者它们之间任意范围内的任意值；
止血药物5.0-15.0份，如5.0份、6.0份、7.0份、8.0份、9.0份、10.0份、11.0份、12.0份、13.0份、14.0份、15.0份或者它们之间任意范围内的任意值；
水杨酸1.0-3.0份，如1.0份、1.5份、2.0份、2.5份、3.0份或者它们之间任意范围内的任意值；和
β-环糊精1.0-10.0份，如1.0份、2.0份、3.0份、4.0份、5.0份、6.0份、7.0份、8.0份、9.0份、10.0份或者它们之间任意范围内的任意值。
其中，所述敷料还包括薄荷醇，优选地，所述敷料中所述薄荷醇的质量份数为1.0-5.0份，如1.0份、1.5份、2.0份、2.5份、3.0份、3.5份、4.0份、4.5份、5.0份或者它们之间任意范围内的任意值。
其中，所述敷料还包括交联剂和保湿剂。
优选地，所述敷料包括交联剂0.5-2.0份，如0.5份、0.6份、0.7份、0.8份、0.9份、1.0份、1.2份、1.4份、1.6份、1.8份、2.0份或者它们之间任意范围内的任意值。
优选地，所述敷料包括保湿剂3.0-8.0份，如3.0份、3.5份、4.0份、4.5份、5.0份、5.5份、6.0份、6.5份、7.0份、7.5份、8.0份或者它们之间任意范围内的任意值。
其中，所述壳聚糖为水溶性的低分子量壳聚糖，且所述壳聚糖的分子量为5000-100000道尔顿。
其中，所述壳聚糖为改性壳聚糖，优选地所述壳聚糖选自羟甲基壳聚糖、羟乙基壳聚糖、羟丙基壳聚糖和季铵化壳聚糖中的一种或多种，优选地所述壳聚糖为羟甲基壳聚糖，如购自青岛汇智生物工程有限公司的羟甲基壳聚糖产品。
其中，所述止血药物选自三七、艾叶、白及、槐花、侧柏叶、大蓟和茜草中的一种或多种。
其中，所述止血药物是研制粉末或提取物的形式。优选地，所述提取物为采用现有技术已知的水提取浓缩技术制得的去离子水提取物。
其中，所述交联剂选自氯化钙、氯化钠、氯化钾和碳酸钙中的一种或多种，优选地所述交联剂为氯化钙。
其中，所述保湿剂选自甘油、丙二醇、甘露醇和山梨醇中的一种或多种，优选地所述保湿剂为甘油。
其中，所述止血药物：薄荷醇：水杨酸的质量份数比为(6-9)：(2-5)：(1-3)，优选地，所述止血药物：薄荷醇：水杨酸的质量份数比为8：3：2。在本发明中，发明人预料不到地发现在薄荷醇和水杨酸共存的情况下，水凝胶的抑菌效果明显优于仅存在水杨酸的效果。原因可能在于薄荷醇是萜类渗透促进剂，其能够增加药物从水溶性基质向角质层的传递。在本发明中，薄荷醇明显地促进水杨酸向皮肤皮层内的渗透，进而实现了水杨酸在可控释放的快速发挥作用。
另外，申请人还注意到，氯化钙除了具有交联大分子材料壳聚糖和海藻酸钠的作用外，还在一定程度上与水凝胶的吸湿性有关。在壳聚糖和海藻酸钠的浓度一定时，随着氯化钙浓度的增加，水凝胶的吸湿性逐渐减小，但当氯化钙的浓度过低时，形成的凝胶的强度则较差，凝胶强度较低。而随着甘油浓度的增加，水凝胶的吸湿性逐渐增大，但其强度相应地下降。特别地，当氯化钙和甘油之间的质量份数比为3:10时，实现最佳的止血性能和吸湿性的组合效果。
本发明还提供了上述生物活性敷料的制备方法，包括如下步骤：
a)将壳聚糖、海藻酸钠、止血药物、交联剂和保湿剂加入到去离子水中，4-6℃充分搅拌溶解得到混合液；
b)将β-环糊精和水杨酸添加至去离子水中，超声处理3-5分钟，60-80℃加热搅拌1-1.5小时使水杨酸和β-环糊精充分溶解，混匀得到包合物溶液，冷冻干燥得到包合物粉末；
c)将步骤b)制得的包合物粉末添加到a)制得的溶液中，搅拌均匀，静置除泡；和
d)将步骤c)制得的溶液铺膜，冷冻干燥，紫外消毒，得到凝胶敷料。
优选地，在步骤b)中将β-环糊精和水杨酸以1：3的质量比加入到去离子水中。
在步骤d)中，所述包合物粉末以最终凝胶敷料中的水杨酸含量不超过2％的量加入所述a)的溶液中。
可选地，在步骤a)中加入薄荷醇。
本发明还提供了所述生物活性敷料在制备用于治疗出血性创伤的药物中的用途。
本发明的有益效果体现在：
(1)本发明通过β-环糊精与水杨酸之间形成包合物，有效地提高水杨酸作为杀菌防腐剂在敷料载体中的有效装载量。同时，由于环糊精包合，实现了水杨酸在水凝胶敷料中的受控释放，克服了水杨酸的稳定性差和对皮肤刺激性大的缺陷，延长了水杨酸对皮肤创口的作用时间和效果。
(2)本发明充分利用了大分子生物材料壳聚糖、海藻酸钠与止血药物成分、杀菌剂、交联剂和保湿剂各自的功效以及它们之间的协同作用，使得所制备的敷料具有快速止血、杀菌止痒、抑菌性能好等优点。另外，本发明的生物活性敷料对人体无任何副作用，使用安全，易贴易揭，对皮肤刺激性极小。
(3)发明人还预料不到地发现，敷料中薄荷醇的存在，和水杨酸产生了协同效应，比单独使用水杨酸能够加速对皮肤皮层内的渗透作用，从而刺激皮肤角质层在较短的时间内做出应激反应，从而促进皮肤出血性创口的快速止血和愈合。
具体实施方式
下面将结合具体实施例对本发明的技术方案进行清楚、完整地描述，显然，所描述的实施例仅是出于举例的目的，而不是全部。基于本发明中的实施例，本领域普通技术人员将更好地理解和掌握本发明所要求保护的技术方案及其实现的技术效果。
(一)生物活性敷料的制备
实施例1生物活性敷料1的制备
首先，按原料的质量份数计，将羟甲基壳聚糖8.0份、海藻酸钠7.0份、艾叶提取物8.0份、薄荷醇3.0份、氯化钙1.5份和甘油5.0份加入到适量去离子水中，4℃下以150-200rpm在振荡机上充分搅拌溶解得到混合液A；将β-环糊精6.0份和水杨酸2.0份添加至适量去离子水中，超声处理3-5分钟，70℃加热搅拌1.5小时使水杨酸和β-环糊精充分溶解，混匀，得到包合物溶液，冷冻干燥得到包合物粉末；将所得到的包合物粉末加入到混合液A中，以150-200rpm在振荡机上充分搅拌均匀得到混合液B；将混合液B铺膜，在冷冻干燥机中冷冻干燥24小时，紫外杀菌，得到水凝胶敷料。
实施例2生物活性敷料2的制备
除了不添加薄荷醇外，其余的成分含量以及操作条件与实施例1相同。
实施例3生物活性敷料3的制备
除了将实施例1中的壳聚糖的质量份比换成5.0份、海藻酸钠的质量份比换成10.0份和薄荷醇的质量份比换成1.0份，其余的成分含量以及操作条件都不变。
实施例4生物活性敷料4的制备
除了将实施例1中的壳聚糖的质量份比换成10.0份、海藻酸钠的质量份比换成5.0份和薄荷醇的质量份比换成5.0份，其余的成分含量以及操作条件都不变。
实施例5生物活性敷料5的制备
除了将实施例1中的薄荷醇的质量份比换成7.0份，其余的成分含量以及操作条件都不变。
实施例6生物活性敷料6的制备
除了将实施例1中的艾叶提取物的质量份比换成12.0份，其余的成分含量以及操作条件都不变。
实施例7生物活性敷料7的制备
除了将实施例1中的水杨酸的质量份比换成5.0份，其余的成分含量以及操作条件都不变。
实施例8生物活性敷料8的制备
除了将实施例1中的氯化钙的质量份比换成0份和甘油的质量份数比换成8.0份外，其余的成分含量以及操作条件都不变。
实施例9生物活性敷料9的制备
除了将实施例1中的氯化钙的质量份比换成2.0份和甘油的质量份数比换成0份外，其余的成分含量以及操作条件都不变。
实施例10生物活性敷料10的制备
除了将实施例1中的氯化钙的质量份比换成2.0份和甘油的质量份数比换成8.0份外，其余的成分含量以及操作条件都不变。
对比例1
除了不添加海藻酸钠且将实施例1中的壳聚糖的质量份比换成15.0份外，其余的成分含量以及操作条件都不变。
对比例2
除了将实施例1中的艾叶提取物的质量份比换成3.0份外，其余的成分含量以及操作条件与实施例1相同。
对比例3
除了将实施例1中的水杨酸的质量份比换成0.5份外，其余的成分含量以及操作条件与实施例1相同。
上述各个实施例和对比例的组分质量份数配比，具体如表1中所示：
表1
Figure BDA0002771003450000071
(二)测试指标
1、吸水性：分别取实施例1-10和对比例1-3的样品少量，称重W1，在去离子水中浸泡24h后，取出，用滤纸吸干表面水分，再次对凝胶称重W2，通过公式：吸水率(％)＝水凝胶增重的部分(W2-W1)/水凝胶初始重量(W1)X100，计算各个样品的吸水率。
2、止血性能
选取健康的体重为2.0-2.5kg的新西兰白兔为实验动物，随机分组，乙醚吸入麻醉，制作肝脏创伤出血模型。造成肝敞开型伤口，出血5-6s后，于出血部位施用各个实施例和对比例的生物活性敷料，记录止血时间，观察止血效果。
3、抑菌性能
采用抑菌圈法测定各个敷料的抑菌性能。将金黄色葡萄球菌(Staphylococcusaureus)活化，进行斜面接种，于37℃培养约18-24h，活化1-2代，用4ml无菌生理盐水冲洗斜面上的菌液。取菌种量相当的菌液均匀地涂布在平板培养基上，将直径0.6cm的圆形滤纸片置于培养基中，用牛津杯取各个实施例和对比例样品的水凝胶敷料，置于滤纸片上，每个样品三次重复，于37℃培养箱中培养约24h，测量抑菌圈的直径大小。
对本发明的实施例1-10和对比例1-3所制备的生物活性敷料进行吸水性能、止血性能和金黄色葡萄糖球菌抑菌性能测试，结果如下表2所示：
表2
Figure BDA0002771003450000081
Figure BDA0002771003450000091
上述结果表明，对于凝胶的吸湿性能，尽管壳聚糖和海藻酸钠均为吸附水分的主要载体基质，但是其他辅料成分也显著影响凝胶的吸水性能。在本实验中，发明人注意到，不存在薄荷醇的处理的样品的吸水率显著地低于存在薄荷醇的处理，然而过量的薄荷醇在一定程度上抑制了壳聚糖和海藻酸钠之间形成的三维立体交联网络结构的吸水溶胀性能，进而限制凝胶的吸水量。另外，氯化钙和甘油也显著影响凝胶的吸湿性能。
对于止血性能，当敷料中不存在止血药物时(对比例2)，尽管壳聚糖和海藻酸钠材料也具有止血效果，但创口的止血时间显著地大于其他处理的止血时间。其中，与不含水杨酸的处理(对比例3)相比，实施例1的处理的止血时间显著地更短，这表明尽管水杨酸不直接具有止血作用，但是其本身固有的抗菌消炎功效在一定程度上刺激皮肤局部细胞在短时间内做出应激反应，进而协同止血药物一起促使大量的凝血因子和大量的纤维蛋白生成，促使血管收缩而止血。另外，与不含薄荷醇的处理(实施例2)相比，薄荷醇在一定程度上促进止血药物和水杨酸的经皮渗透，进而使得在单位时间内更高剂量的止血成分和水杨酸到达创口部位，刺激创口部分的细胞更快速地释放各种有利于凝血的因子而止血；但是过量的薄荷醇(实施例5)并没有更好地实现细胞中凝血因子的快速释放，其原因有待于进一步研究。
对于抑菌性能，水杨酸对于各个凝胶样品中的抑菌性能起着决定性的作用。其中，水杨酸和止血药物的缺乏(对比例2和3)，则显著地影响了凝胶的抑菌效果。此外，即使水杨酸存在，但是薄荷醇不存在，或者水杨酸和薄荷醇之间的比例也影响水凝胶的抑菌特性。发明人意外地发现，当交联剂与保湿剂的比例为3:10时，水凝胶的止血和抑菌性能都明显地优于其他处理。
从以上分析可以看出，本发明的生物活性敷料的功能是多种组分的协同互作的结果。
本发明的内容不限于以上所列举的各个实施例，本领域普通技术人员应知晓，在不偏离本发明的精神和实质的情况下，通过阅读本发明的说明书可对本发明的技术特征进行各种替代、修改和组合，所述各种替代、修改和组合后的技术方案均为本发明的权利要求的保护范围所涵盖。

Claims (5)

1.一种具有抗炎止血功能的生物活性敷料，其特征在于：所述敷料包括如下质量份数的原料：壳聚糖5.0-10.0份、海藻酸钠5.0-10.0份、止血药物5.0-15.0份、水杨酸1.0-3.0份和β-环糊精1.0-10.0份；
所述敷料还包括薄荷醇1-5份、交联剂0.5-2.0份和保湿剂3-8份；
其中，所述止血药物：薄荷醇：水杨酸的质量份数比为(6-9)：(2-5)：(1-3)；
所述止血药物选自三七、艾叶、白及、槐花、侧柏叶、大蓟和茜草中的一种或多种。
2.根据权利要求1所述的生物活性敷料，其特征在于，所述壳聚糖选自羟甲基壳聚糖、羟乙基壳聚糖、羟丙基壳聚糖和季铵化壳聚糖中的一种或多种。
3.根据权利要求1所述的生物活性敷料，其特征在于，所述交联剂为氯化钙；和所述保湿剂选自甘油、丙二醇、甘露醇和山梨醇中的一种或多种。
4.一种权利要求1-3中任一项所述的生物活性敷料的制备方法，包括如下步骤：
a)将壳聚糖、海藻酸钠、薄荷醇、止血药物、交联剂和保湿剂加入到去离子水中，4-6℃充分搅拌溶解得到混合液；
b)将β-环糊精和水杨酸添加至去离子水中，超声处理3-5分钟，60-80℃加热搅拌1-1.5小时使水杨酸和β-环糊精充分溶解，得到包合物溶液，冷冻干燥得到包合物粉末；
c)将步骤b)制得的包合物粉末添加到a)制得的溶液中，搅拌均匀，静置除泡；和
d)将步骤c)制得的溶液铺膜，冷冻干燥，紫外消毒，得到凝胶敷料。
5.根据权利要求1-3中任一项所述的生物活性敷料或者通过权利要求4所述的制备方法制备的生物活性敷料在制备用于治疗出血性创伤的药物中的用途。
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