CN113274543A - 一种新型黏附抗菌温度耐受功能水凝胶制备方法及应用 - Google Patents

一种新型黏附抗菌温度耐受功能水凝胶制备方法及应用 Download PDF

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CN113274543A
CN113274543A CN202110548138.5A CN202110548138A CN113274543A CN 113274543 A CN113274543 A CN 113274543A CN 202110548138 A CN202110548138 A CN 202110548138A CN 113274543 A CN113274543 A CN 113274543A
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antibacterial
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林权
刘厚
赵月
冯钰斌
杨柏
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Jilin University
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Abstract

本发明提供了一种新型黏附抗菌温度耐受功能水凝胶制备方法及应用。采用抗菌高分子与明胶以及丙烯酰胺进行反应制备多功能水凝胶，具有优良的黏附性和力学性能，7天以上持久的抗菌性，在比较宽的温度范围(‑20℃–60℃)内保持各种优良性能，特别是具有宽温度范围下的抗菌性能，能够长期保湿。用于不同环境下急/慢性创面的敷料以及组织再生辅助材料。

Description

一种新型黏附抗菌温度耐受功能水凝胶制备方法及应用
技术领域
本发明属于高分子材料技术领域，具体涉及一种新型黏附、抗菌、温度耐受功能水凝胶的制备及其在糖尿病足伤口敷料方面的应用。
背景技术
糖尿病足是糖尿病患者中最常见的并发症之一，它会增加患者发病，感染，非创伤性截肢，甚至早期死亡的风险。由于高糖的伤口环境，致使糖尿病足成为一种极易被细菌感染的慢性伤口疾病，严重影响了患者的身心健康，也带来了沉重的社会经济负担。在临床治疗中，伤口敷料逐渐成为促进糖尿病足伤口修复的主要策略手段。传统的糖尿病足伤口敷料以静电纺丝纤维、脱脂棉、天然或者合成的纱布(绷带)等干敷料为主，其存在着黏连伤口，愈合慢，易感染以及易结痂等问题。近年来，以水凝胶为主的湿敷料在促进糖尿病足伤口愈合方面展现出极大的潜力，并引起了人们广泛的研究兴趣。水凝胶是一类具有三维立体的高分子网状结构的材料，其多孔结构能够吸收创面渗出液，高的水含量也能保持伤口的湿润性，更好的促进伤口愈合。为了满足更高的需求，水凝胶敷料被赋予了抗菌，黏附，载药等多种功能，能够通过密闭伤口，抑菌等作用来加快糖尿病足伤口的愈合速度。
考虑到处于热带、极地地区等拥有极端温度的地区国家，以及长时间处于野外、边疆等极端环境中的患者，一种温度耐受且能长久保湿的多功能水凝胶敷料是迫切需要的。甘油是一种生物相容性良好的黏稠状液体，在水凝胶体系中加入适量的甘油可以降低水凝胶的凝固点，从而达到抗冻的效果，并且甘油分子可以与水分子以及水凝胶的其余成分之间形成大量氢键，从而极大地减缓水分子在高温条件下的挥发，使水凝胶敷料能够长久保湿。这就使得水凝胶敷料能够在很宽的温度范围(-20℃–60℃)内仍然保持其原有的黏附，抗菌等性质，从而有效的促进糖尿病足伤口的愈合。这也拓展了水凝胶敷料的使用范围，为制备更加多功能的，更具适应性的水凝胶敷料提供了思路。
发明内容
本发明的目的是提供一种新型黏附、抗菌、温度耐受水凝胶的制备及其在糖尿病足伤口敷料方面的应用。
该方法首先将一定比例的抗菌高分子、明胶、丙烯酰胺、亚甲基双丙烯酰胺溶解到甘油/水的二元混合溶剂中，混合均匀后采用热聚合的方法形成半贯穿的凝胶网络结构，之后放到4℃冰箱中进一步反应，获得明胶形成的三螺旋结构和聚丙烯酰胺交联结构的双网络结构水凝胶。水凝胶表面丰富的官能团赋予了其良好的黏附性；抗菌高分子的加入，不仅使水凝胶拥有了持久的抗菌性，而且其与聚丙烯酰胺形成超分子弱相互作用构筑互穿网络；另外，本发明的水凝胶组成成分构建了H键、范德华力等相互作用，锁住水分子，获得在宽温度范围-20℃–60℃能保持生物相容性好，无毒和优异的特征机械性能、抗菌性能等各种性能。通过细胞毒性试验，动物试验等进一步证明了该抗菌性水凝胶可以作为伤口敷料用于糖尿病足的伤口修复。结合以上特点，这种新型黏附、抗菌、温度耐受性多功能水凝胶有很大的潜力作为一种新型伤口敷料在糖尿病足等慢性伤口，以及组织工程等生物医学领域大展身手。
本发明所述的新型黏附、抗菌、温度耐受水凝胶的制备方法，其具体步骤如下：
(1)制备水/甘油的体积比为4:1～3:2的混合溶液，取此混合溶液20mL置于圆底烧瓶中，再加入1.0–5.0g明胶，于60℃中连续搅拌30min，待成为均匀溶液后，缓慢加入1.0–4.0g抗菌高分子，继续搅拌20min。冷却至室温后，依次加入4.0–8.0g丙烯酰胺(AM)、4.0–6.0mg N,N’-亚甲基双丙烯酰胺(MBA)和80.0–120.0mg过硫酸铵(APS)，搅拌均匀。然后将得到的混合溶液倒入聚四氟乙烯模具中，在60℃下反应5h形成半贯穿网络水凝胶。
(2)将上述水凝胶冷却后放置于4℃冰箱中反应30–60分钟后取出，形成最终的多功能双网络水凝胶。
上述方法中，抗菌高分子可以是聚赖氨酸，季铵盐壳聚糖，聚六亚甲基胍盐酸盐等天然或者合成高分子抑菌剂。
上述方法中，如果选择聚赖氨酸作为抗菌高分子，其分子量限制在是4000–4300kDa。
本发明优点如下：1.本发明水凝胶具有优良的黏附性和力学性能，针对不同有机和无机表面均具有较强的黏附性和力学强度，可以作为伤口敷料便于固定与更换，也可以避免外力对伤口所造成的二次损伤；2.具有7天以上持久的抗菌性，可以保证材料长期发挥抗菌作用，并且抗菌剂无泄漏，不会对伤口造成二次伤害；3.水凝胶可在比较宽的温度范围(-20℃–60℃)内保持各种优良性能，特别是具有宽温度范围下的抗菌性能，拓宽了伤口敷料的实际使用场景；4.水凝胶长时间含水率可以达到60％以上，并且能够长期保湿，有助于伤口的愈合；5.水凝胶冻干后的扫描电镜图显示其为疏松多孔结构，便于吸收伤口渗液，也可以用于药物以及生物活性分子的储存与释放；6.本发明水凝胶原料来源广，易保存，生物相容性好，有利于产品的商品化；7.本发明水凝胶制备操作过程简便，绿色无污染；8.本发明适用于不同环境下急/慢性创面的敷料以及组织再生辅助材料等，具有广阔的应用前景。
附图说明
图1：为实施例1所制备的多功能水凝胶敷料的相转变图。通过比较倾斜的试剂瓶中物质的前后变化我们可以看出，水凝胶前驱体溶液为淡黄色透明液体(图A)，在加入APS聚合，以及4℃冷却后变为淡黄色的凝胶(图B)。
图2：为实施例1所制备的多功能水凝胶敷料冻干后的扫描电镜图。从图中我们可以清楚的看到水凝胶内部疏松多孔的结构，利于药物的搭载与释放，以及物质的交换。
图3：为实施方案1所制备的多功能水凝胶敷料与不同材料的黏附图。我们可以看到所制备的水凝胶可以黏附到不同物质表面，具有较强的黏附性。
图4：为实施方案1所制备的多功能水凝胶敷料的体外抗菌实验图。图中百分数为加入抗菌高分子的质量分数，可以看到随着加入抗菌高分子的浓度变大，抑菌环也越来越大，表明本发明的水凝胶有很好的抗菌性能。
图5：为实施方案1所制备的多功能水凝胶敷料修复小鼠糖尿病足伤口图。从图中我们可以看到在经过18天的伤口修复后，对照组小鼠伤口并无太大改善，没有完全愈合；而水凝胶组小鼠伤口已完全愈合，说明我们的水凝胶伤口敷料可以有效的促进糖尿病足伤口的愈合。
图6:为实施方案1所制备的多功能水凝胶敷料在不同温度下的体外抗菌实验结果。针对对照组和G-PAGL水凝胶组的大肠杆菌分别在4，37和45℃下的生长曲线图。结果表明本发明的水凝胶在比较宽的温度范围都有优良的抗菌性能。
具体实施方式
实施例1：
(1)将1.0明胶加入到一个含有20mL的去离子水/甘油(体积比7:3)混合溶液的圆底烧瓶中，在60℃中连续搅拌半小时，待明胶完全溶解后，再缓慢加入2.0g聚赖氨酸(ε-PL，分子量4000–4300kDa)，继续搅拌20分钟。冷却至室温后，依次加入6.0g丙烯酰胺(AM)单体、6.0mg N,N’-亚甲基双丙烯酰胺(MBA)和120.0mg过硫酸铵(APS)，搅拌均匀。然后将得到的混合溶液倒入模具中，在60℃下反应5h形成半贯穿网络水凝胶。
(2)将上述水凝胶放于4℃冰箱中反应60分钟后，形成最终的多功能双网络水凝胶。
实施例2：
(1)将1.0明胶加入到一个含有20mL的去离子水/甘油(体积比4:1)混合溶液的圆底烧瓶中，在60℃中连续搅拌半小时，待明胶完全溶解后，再缓慢加入2.0g季铵盐壳聚糖，继续搅拌20分钟。冷却至室温后，依次加入6.0g丙烯酰胺(AM)单体、6.0mg N,N’-亚甲基双丙烯酰胺(MBA)和120.0mg过硫酸铵(APS)，搅拌均匀。然后将得到的混合溶液倒入模具中，在60℃下反应5h形成半贯穿网络水凝胶。
(2)将上述水凝胶放于4℃冰箱中反应60分钟后，形成最终的多功能双网络水凝胶。
实施例3：
(1)将1.0明胶加入到一个含有20mL的去离子水/甘油(体积比4:1)混合溶液的圆底烧瓶中，在60℃中连续搅拌半小时，待明胶完全溶解后，再缓慢加入2.0g聚六亚甲基胍盐酸盐，继续搅拌20分钟。冷却至室温后，依次加入6.0g丙烯酰胺(AM)单体、6.0mg N,N’-亚甲基双丙烯酰胺(MBA)和120.0mg过硫酸铵(APS)，搅拌均匀。然后将得到的混合溶液倒入模具中，在60℃下反应5h形成半贯穿网络水凝胶。
(2)将上述水凝胶放于4℃冰箱中反应60分钟后，形成最终的多功能双网络水凝胶。

Claims (5)

1.一种新型黏附抗菌温度耐受功能水凝胶的制备方法，其特征在于：具体步骤如下：
(1)制备水/甘油的体积比为4:1～3:2的混合溶液，取此混合溶液20mL置于圆底烧瓶中，再加入1.0–5.0g明胶，于60℃中连续搅拌30min，待成为均匀溶液后，缓慢加入1.0–4.0g抗菌高分子，继续搅拌20min；冷却至室温后，依次加入4.0–8.0g丙烯酰胺(AM)、4.0–6.0mgN,N’-亚甲基双丙烯酰胺(MBA)和80.0–120.0mg过硫酸铵(APS)，搅拌均匀；然后将得到的混合溶液倒入聚四氟乙烯模具中，在60℃下反应5h形成半贯穿网络水凝胶；
(2)将上述水凝胶冷却后放置于4℃冰箱中反应30–60分钟后取出，形成最终的多功能双网络水凝胶。
2.如权利要求1所述的一种新型黏附抗菌温度耐受功能水凝胶的制备方法，其特征在于：步骤(1)中所述的抗菌高分子可以是聚赖氨酸，季铵盐壳聚糖，聚六亚甲基胍盐酸盐等天然或者合成高分子抑菌剂。
3.如权利要求1所述的一种新型黏附抗菌温度耐受功能水凝胶的制备方法，其特征在于：步骤(1)中所述的如果选择聚赖氨酸作为抗菌高分子，其分子量限制在是4000–4300kDa。
4.权利要求1-3所述的任意一种新型黏附抗菌温度耐受功能水凝胶的制备方法制备的水凝胶，其特征在于：水凝胶具有优良的黏附性、持久的抗菌性，可以在宽温度范围-20℃–60℃能保持生物相容性好、无毒和优异的特征机械性等各种性能。
5.权利要求1-3所述的任意一种新型黏附抗菌温度耐受功能水凝胶的制备方法制备的水凝胶，其特征在于：水凝胶可以作为敷料在糖尿病足伤口修复方面应用。
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