CN116407676A - 一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法 - Google Patents
一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法 Download PDFInfo
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- CN116407676A CN116407676A CN202310465212.6A CN202310465212A CN116407676A CN 116407676 A CN116407676 A CN 116407676A CN 202310465212 A CN202310465212 A CN 202310465212A CN 116407676 A CN116407676 A CN 116407676A
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- hydrogel dressing
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Abstract
本发明公开了一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法。本发明以具有抗菌活性的羧甲基壳聚糖(CMCS)与聚乙烯醇(PVA)为水凝胶骨架,以多巴胺(DA)和硼砂为粘合剂和交联剂,采用原位合成和氧化自聚法,制备了具有多重动态可逆键的智能型CMCS/PVA/聚多巴胺(PDA)水凝胶敷料。以抗生素和植物源活性小分子等药物,在近红外光照射下不仅能够诱导聚多巴胺产生局部热疗作用,还可以通过时空调控药物释放能力,达到精准调控的目的。本方法制备的水凝胶敷料能够实现快速杀菌,同时拥有优异的生物生物相容性、高黏附性、光热稳定性和环境自适应性,能够用于急性和慢性等多种创面的保护和治疗。
Description
技术领域
本发明属于生物材料制备技术领域,涉及一种兼具光热与药物协同作用的高粘附性、快速自愈合、促进伤口愈合和生物相容性优异的多功能水凝胶敷料的制备方法。
背景技术
皮肤是人体与环境之间的主要物理屏障,皮肤损伤容易被细菌感染,体液流失并引起各种并发症,会影响人们的健康,甚至危及生命安全。伤口治疗在中国乃至全球范围内仍然是一个巨大的临床挑战,给每个国家的政府都带来了巨大的财政负担。尽管纱布、绷带和膜等传统伤口敷料已经开发并应用于诊所,但仍存在功能单一、更换过程中存在二次伤害并引发炎症等问题。在实际应用中,理想的敷料应满足诸如保持湿润环境、增强表皮迁移、促进血管生成、可粘附并轻松揭除、防止细菌感染和适用于不同部位等要求。水凝胶敷料因其丰富的3D结构和促进细胞水合作用进行气体和营养物质交换,为细胞生长、增值和迁移提供了一个潮湿的环境,加速伤口愈合。作为与生命组织最接近的生物功能材料,水凝胶可与人体组织直接接触,防止体外微生物感染,有效防止体液散失,并可传输氧气,促进伤口愈合。同时作为药物释放载体,对低分子溶质的良好透过性,也赋予了水凝胶药物传输系统的功能。水凝胶由于其优异的生化性能和力学性能,在伤口敷料领域已成为近几十年来最具竞争力的候选材料,并呈现出逐年增长的趋势,展现出了光明的前景。
利用物理和化学交联的方式设计水凝胶的结构并赋予其功能,实现集多种功能于一身的水凝胶敷料一直是人们不懈的追求。其中,近红外光热水凝胶敷料通过精确控制近红外光的辐射强度、时间和位置等达到精准调控杀菌的目的,成为近年来的研究热点。然而,开发一种既有光热与药物协同作用,又有高粘附性、超快速自愈性、促进伤口愈合和生物相容性等多功能水凝胶敷料,仍是一个巨大的挑战。目前研制的水凝胶敷料可填充伤口,不会与伤口形成粘连,但在更换时依旧会对伤者造成疼痛,尤其是对大面积创伤而言,更换伤口敷料需要很长时间,这会给病人造成极大的精神负担。同时,在使用时受到应力应变作用时易出现裂纹,破损和老化等现象,使凝胶的网络结构受损导致凝胶性能下降,限制了其应用。此外,在水凝胶敷料的设计中,大部分研究主要是考虑力学性能和生物相容性等,但对物理/化学协同治疗伤口的策略相对较少,并且其存在工艺比较复杂、成本较高、抗菌效果差和伤口愈合效果不佳等问题,限制了其进一步的使用。
发明内容
针对现有技术的不足,本发明主要是提供一种具有光热与药物协同作用的高粘附、快速自愈合、促进伤口愈合和生物相容性优异的多功能水凝胶敷料的制备方法。基于动态建构理论,以羧甲基壳聚糖(CMCS)和聚乙烯醇(PVA)为原料,以多巴胺(DA)为光热粘合剂和交联剂,以抗生素类和植物源活性小分子为模型药物,通过简单的原位合成和氧化自聚等方法制备了具有硼酸酯键、亚胺键和氢键等多重动态交联体系的载药CMCS/PVA/聚多巴胺(CPBD)水凝胶敷料。在近红外光照射下不仅能够诱导聚多巴胺产生局部热疗作用,还可以通过时空调控药物释放,达到精准调控的目的。同时,该水凝胶敷料还具有高粘附性、快速自愈合性、可拉伸性、光热稳定性、优异的抗菌能力、良好的生物相容性和促伤口愈合能力等性能,适用于急性和慢性等多种创面的保护和治疗。
本发明所述的一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法,其包括如下步骤:
(1)取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成一定浓度的PVA溶液。反应一段时间后,降至适宜温度,一定质量比的CMCS加入PVA溶液中,继续搅拌,形成透明的溶液;
(2)用一定浓度的氢氧化钠溶液对上述溶液的pH值进行调节,调至一定pH值后,加入不同量的DA,混合均匀,形成透明的溶液;
(3)最后,采用微流控技术将一定量的交联剂和适量的抗生素类和植物源活性小分子模型药物加入到上述体系中,保温反应一定时间,得到CPBD水凝胶敷料。
优选地,所述步骤(1)中PVA与CMCS的质量比为6:0~6:5,温度为30~65℃,机械搅拌速度为200~600r/min。
优选地,所述步骤(2)中氢氧化钠溶液的浓度为0.5~2.0mol/L,溶液的pH值为7.5~9.5,DA占PVA和CMCS总质量的3.0~20.0%。
优选地,所述步骤(2)中交联剂包括硼砂、硼酸和含有苯硼酸基团的4-乙烯基苯硼酸和苯硼酸等一种或几种以上,交联剂的浓度为0.01~0.2mol/L,总流速为0.6~1.5mL/min;抗生素类药物包括阿莫西林、盐酸环丙沙星、诺氟沙星、替卡西林、硫酸庆大霉素和妥布霉素等一种或几种以上,植物源活性小分子药物包括单宁酸、花青素、磷酸小檗碱等一种或几种以上;温度为40~70℃,反应时间为10~60min。
优选地,所述CPBD水凝胶敷料适用于抗感染治疗以及急性和慢性等多种创面的保护和治疗。
与现有技术相比,本发明的有益效果是:
(1)本发明的制备方法简单,便于操作,条件温和,解决了现有凝胶敷料中制备过程副产物较多,需精细操作的制约。
(2)利用硼酸酯键、亚胺键和氢键等多种动态共价键和动态非共价键获得快速自愈的CPBD水凝胶敷料,解决了使用时受应力-应变作用易出现裂纹、破损和老化等问题。凝胶切断后,在1min内可实现自我愈合,且在1min内的断裂-愈合的自愈合效率为80%,其与皮肤相匹配的力学性能,便于更换和固定,特别是适用于关节处和腋窝等特殊区域的急性和慢性创伤伤口。
(3)此发明制备的多功能CPBD水凝胶敷料具有丰富的三维网络结构,赋予其优异的吸液能力。在去离子水、生理盐水和磷酸盐缓冲液中的溶胀率分别为4215.00%、2057.48%和1203.20%。一方面能够吸附伤口浸出液、为细胞的增值和迁移以及新血管的生成提供支撑,另一方面水凝胶的多孔网络结构和CMCS中的正电荷具有止血功能。
(4)此发明制备的多功能CPBD水凝胶敷料具有优异的粘附性,可以在不同材质的表面进行粘附。在各种组织器官的表面具有较高的粘结强度,解决了与组织处固定困难等问题。
(5)此发明制备的多功能CPBD水凝胶敷料具有极佳的抗菌性能。结合CMCS的本征抗菌特性、药物和光热协同杀菌策略,与细菌共同培养7d时,抑菌直径均呈现先递增后保持不变,对大肠杆菌和金黄色葡萄球菌的杀菌率高达100%,展现出持久有效的抗菌能力。在近红外光照射下不仅能够诱导聚多巴胺产生局部热疗作用,还可以通过时空调控药物释放能力,达到精准调控的目的。
(6)此发明制备的多功能CPBD水凝胶敷料具有良好的生物相容性。CPBD水凝胶敷料的细胞存活率超过100%和溶血率均小于5%。动物实验表明,结合光热/药物协同治疗,CPBD水凝胶敷料将光热效应与抗生素等药物相结合,明显加速伤口愈合。在14d的治疗中,光热/药物协同治疗组的伤口愈合率达97.4%。
(7)此发明制备的多功能CPBD水凝胶敷料可以用于伤口愈合过程中的创伤愈合敷料、抗菌敷料、医用粘合剂和人工组织皮肤。
附图说明
图1CPBD水凝胶敷料的电子扫描电镜图;
图2CPBD水凝胶敷料的光热性能;
图3CPBD水凝胶敷料的对大肠杆菌和金黄色葡萄球菌的光热/药物协同抗菌效果;
具体实施方式
这些实施例仅限于说明本发明,但本发明并不局限与这些实施例。
实施案例1
取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成PVA溶液。反应1h后,降至30℃,形成透明的溶液;用0.5mol/L的氢氧化钠溶液加入到上述溶液中,使溶液的pH值为7.5。随后,将占PVA和CMCS总质量的2.0%的DA加入到上述溶液中,形成透明的溶液。最后,将0.01mol/L的硼酸,以0.6mL/min的总流速和单宁酸的浓度为0.5wt%加入到上述体系中,在温度为40℃的条件下反应0.5h,得到CPBD水凝胶敷料。
结果显示:CPBD水凝胶敷料呈现三维网络结构,且在去离子水、生理盐水和磷酸盐缓冲液中的溶胀率分别为472.00%、107.37%和83.67%。水凝胶切割后,30min能够实现完全自愈。粘结强度试验表明,CPBD水凝胶敷料易脱落,粘结强度较差。同时,CPBD水凝胶敷料对大肠杆菌和金黄色葡萄球菌有一定的抗菌作用,但抗菌效率仅为74.0%。
实施案例2
取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成PVA溶液。反应1h后,降至30℃,形成透明溶液;用0.5mol/L的氢氧化钠溶液加入到上述溶液中,使溶液的pH值为7.5。随后,将占PVA和CMCS总质量的10.0%的DA加入到上述溶液中,形成透明的溶液。最后,将0.01mol/L的硼酸,以0.6mL/min的总流速和单宁酸的浓度为0.5wt%加入到上述体系中,在温度为40℃的条件下反应0.5h,得到CPBD水凝胶敷料。
结果显示:CPBD水凝胶敷料呈现三维网络结构,且在去离子水、生理盐水和磷酸盐缓冲液中的溶胀率分别为861.46%、626.33%和774.19%。水凝胶切割后,15min能够实现完全自愈。粘结强度试验表明,CPBD水凝胶敷料对不同材质的基质的粘附作用较小,其中对猪皮的粘结强度为1.26KPa。同时,CPBD水凝胶敷料对大肠杆菌和金黄色葡萄球菌有一定的抗菌作用,但光热和药物协同抗菌效率仅为81.00%。
实施案例3
取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成PVA溶液。反应1h后,降至50℃,形成透明溶液;用0.5mol/L的氢氧化钠溶液加入到上述溶液中,使溶液的pH值为7.5。随后,将占PVA和CMCS总质量的10.0%的DA加入到上述溶液中,形成透明的溶液。最后,将0.01mol/L的硼酸,以0.6mL/min的总流速和单宁酸的浓度为0.5wt%加入到上述体系中,在温度为40℃的条件下反应0.5h,得到CPBD水凝胶敷料。
结果显示:CPBD水凝胶敷料呈现三维网络结构,且在去离子水、生理盐水和磷酸盐缓冲液中的溶胀率分别为817.45%、4.96.71%和345.67%。水凝胶切割后,18min能够实现完全自愈。粘结强度试验表明,CPBD水凝胶敷料对不同材质的基质具有一定的粘附作用,其中对猪皮的粘结强度为3.26KPa。同时,CPBD水凝胶敷料对大肠杆菌和金黄色葡萄球菌有一定的抗菌作用,但光热和药物协同抗菌效率仅为75.30%。
实施案例4
取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成PVA溶液。反应1h后,降至50℃,形成透明溶液;用0.5mol/L的氢氧化钠溶液加入到上述溶液中,使溶液的pH值为7.5。随后,将占PVA和CMCS总质量的10.0%的DA加入到上述溶液中,形成透明的溶液。最后,将0.1mol/L的硼砂,以0.6mL/min的总流速和阿莫西林的浓度为1.0wt%加入到上述体系中,在温度为40℃的条件下反应0.5h,得到CPBD水凝胶敷料。
结果显示:CPBD水凝胶敷料在去离子水、生理盐水和磷酸盐缓冲液中的溶胀率分别为547.87%、129.68%和201.37%。水凝胶切割后,11.9min能够实现完全自愈。粘结强度试验表明,CPBD水凝胶敷料对猪皮的粘结强度为3.46KPa。同时,CPBD水凝胶敷料对大肠杆菌和金黄色葡萄球菌有一定的抗菌作用,但光热和药物协同抗菌效率仅为88.02%。
实施案例5
取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成PVA溶液。反应1h后,降至40℃,形成透明溶液;用0.5mol/L的氢氧化钠溶液加入到上述溶液中,使溶液的pH值为7.5。随后,将占PVA和CMCS总质量的10.0%的DA加入到上述溶液中,形成透明的溶液。最后,将0.01mol/L的硼酸,以0.6mL/min的总流速和盐酸环丙沙星的浓度为1.0wt%加入到上述体系中,在温度为40℃的条件下反应0.5h,得到CPBD水凝胶敷料。
结果显示:CPBD水凝胶敷料呈现三维网络结构,且在去离子水、生理盐水和磷酸盐缓冲液中的溶胀率分别为805.68%、641.28%和701.41%。水凝胶切割后,13min能够实现完全自愈。粘结强度试验表明,CPBD水凝胶敷料对不同材质的基质具有一定的粘附作用,其中对猪皮的粘结强度为3.26KPa。同时,CPBD水凝胶敷料对大肠杆菌和金黄色葡萄球菌有一定的抗菌作用,但光热和药物协同抗菌效率仅为86.30%。
实施案例6
取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成PVA溶液。反应1h后,降至50℃,形成透明溶液;用0.5mol/L的氢氧化钠溶液加入到上述溶液中,使溶液的pH值为7.5。随后,将占PVA和CMCS总质量的10.0%的DA加入到上述溶液中,形成透明的溶液。最后,将0.1mol/L的硼砂,以0.6mL/min的总流速和盐酸环丙沙星的浓度为1.0wt%加入到上述体系中,在温度为40℃的条件下反应0.5h,得到CPBD水凝胶敷料。
结果显示:CPBD水凝胶敷料在去离子水、生理盐水和磷酸盐缓冲液中的溶胀率分别为1428.12%、613.11%和613.31%。水凝胶切割后,13min能够实现完全自愈。粘结强度试验表明,CPBD水凝胶敷料对猪皮的粘结强度为1.78KPa。同时,CPBD水凝胶敷料对大肠杆菌和金黄色葡萄球菌有一定的抗菌作用,但光热和药物协同抗菌效率仅为83.27%。
实施案例7
取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成PVA溶液。反应1h后,降至50℃,形成透明溶液;用0.5mol/L的氢氧化钠溶液加入到上述溶液中,使溶液的pH值为8.5。随后,将占PVA和CMCS总质量的10.0%的DA加入到上述溶液中,形成透明的溶液。最后,将0.1mol/L的硼砂,以0.6mL/min的总流速和盐酸环丙沙星的浓度为1.0wt%加入到上述体系中,在温度为40℃的条件下反应0.5h,得到CPBD水凝胶敷料。
结果显示:CPBD水凝胶敷料在去离子水、生理盐水和磷酸盐缓冲液中的溶胀率分别为1298.33%、531.76%和516.11%。水凝胶切割后,13min能够实现完全自愈。粘结强度试验表明,CPBD水凝胶敷料对猪皮的粘结强度为6.59KPa。同时,CPBD水凝胶敷料对大肠杆菌和金黄色葡萄球菌有一定的抗菌作用,但光热和药物协同抗菌效率仅为83.48%。
实施案例8
取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成PVA溶液。反应1h后,降至50℃,形成透明溶液;用0.5mol/L的氢氧化钠溶液加入到上述溶液中,使溶液的pH值为8.5。随后,将占PVA和CMCS总质量的10.0%的DA加入到上述溶液中,形成透明的溶液。最后,将0.1mol/L的硼酸,以1.3mL/min的总流速和盐酸环丙沙星的浓度为1.0wt%加入到上述体系中,在温度为40℃的条件下反应0.5h,得到CPBD水凝胶敷料。
结果显示:CPBD水凝胶敷料呈现三维网络结构,且在去离子水、生理盐水和磷酸盐缓冲液中的溶胀率分别为1361.46%、626.33%和774.19%。水凝胶切割后,15min能够实现完全自愈。粘结强度试验表明,CPBD水凝胶敷料对不同材质的基质具有一定的粘附作用,其中对猪皮的粘结强度为3.26KPa。同时,CPBD水凝胶敷料对大肠杆菌和金黄色葡萄球菌有一定的抗菌作用,但光热和药物协同抗菌效率仅为84.60%。
实施案例9
取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成PVA溶液。反应1h后,降至50℃,形成透明溶液;用0.5mol/L的氢氧化钠溶液加入到上述溶液中,使溶液的pH值为8.5。随后,将占PVA和CMCS总质量的20.0%的DA加入到上述溶液中,形成透明溶液。最后,将0.15mol/L的硼砂,以1.0mL/min的总流速和盐酸环丙沙星的浓度为1.5wt%加入到上述体系中,在温度为45℃的条件下反应1.0h,得到CPBD水凝胶敷料。
结果显示:CPBD水凝胶敷料呈现三维网络结构,且在去离子水、生理盐水和磷酸盐缓冲液中的溶胀率分别为4215.12%、2057.48%和1203.42%。水凝胶切割后,1min能够实现完全自愈,在1min内5次循环拉伸的自愈合效率约为80%。CPBD水凝胶敷料对金属、木头、玻璃、猪皮表面的黏结强度分别为11.02kPa、791.66kPa、67.65kPa和7.85kPa。在808nm近红外光照射下,CPBD水凝胶敷料的温度约40.8℃,五次交替照射后,水凝胶具有较好的光热稳定性。拉伸应力实验发现,它具有较好的拉伸性能,拉伸应力和断裂伸长率分别为39.48kPa和246.26%。此外,它还具有优异的光热抗菌能力(杀菌率近100%)、良好的生物相容性(细胞存活率大于100%和溶血率小于1.75%)和促进伤口愈合能力(光热和药物协同治疗时,大鼠的伤口愈合率为97.4%)。
Claims (8)
1.一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法,其特征在于,所述方法如下步骤进行:
(1)取PVA和去离子水加入三口烧瓶中,在95℃的温度下,形成一定浓度的PVA溶液。反应一段时间后,降至适宜温度,一定质量比的CMCS加入PVA溶液中,继续机械搅拌,形成均一透明溶液;
(2)用一定浓度的氢氧化钠溶液对上述溶液的pH值进行调节,调至一定pH值后,加入不同量的DA,混合均匀,形成透明的溶液;
(3)最后,采用微流控技术将一定量的交联剂和适量的抗生素和植物源活性药物加入到上述体系中,保温反应一定时间,得到CMCS/PVA/聚多巴胺(CPBD)水凝胶。
2.根据权利要求1所述的一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法,其特征在于所述步骤(1)中的PVA:CMCS的质量比为6:0~6:5,水浴加热温度为30~65℃,机械搅拌速度为200~600r/min。
3.根据权利要求1所述的一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法,其特征在于所述步骤(2)中的pH值进行调节,并加入不同质量比的DA,其中氢氧化钠溶液的浓度为0.5~2.0mol/L,溶液的pH值为7.5~9.5,DA占PVA和CMCS总质量的3.0~25.0%。
4.根据权利要求1所述的一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法,其特征在于所述步骤(3)中采用微流控技术将一定量的交联剂,其中交联剂包括硼砂、硼酸、苯硼酸和含有苯硼酸基团的4-乙烯基苯硼酸等一种或几种以上,交联剂的浓度为0.01~0.2mol/L,总流速为0.6~1.5mL/min。
5.根据权利要求1所述的一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法,其特征在于所述步骤(3)中适量的抗生素类和植物源活性药物加入到上述体系中,其中抗生素药物包括阿莫西林、盐酸环丙沙星、诺氟沙星、替卡西林、硫酸庆大霉素和妥布霉素等一种或几种以上;植物源活性药物包括单宁酸、花青素和磷酸小檗碱等一种或几种以上。
6.根据权利要求1所述的一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法,其特征在于所述步骤(3)中适量的抗生素和植物源活性药物的含量为0.5~2.0wt%。
7.根据权利要求1所述的一种兼具光热抗菌作用的超快速自愈合水凝胶敷料的制备方法,其特征在于所述步骤(3)中保温反应一定时间,温度为40~70℃,反应时间为0.5~2.0h。
8.一种如权利要求1~6任一所述方法制备的一种具有光热与药物协同作用的高粘附、超快速自愈合、促进伤口愈合和生物相容性的多功能水凝胶敷料,其制备方法简单,易于操作。通过构建硼酸酯键、亚胺键和氢键等多种动态可逆键,获得具有快速自愈合能力的CPBD水凝胶;引入DA制备具有光热响应和粘性的聚多巴胺(PDA),赋予其光热特性;在体系中加入抗生素类和植物源活性小分子模型药物,制备载药CPBD水凝胶。通过近红外光和药物的协同作用,达到时空调控药物释放的目的。制备的CPBD水凝胶敷料可以用于伤口愈合过程中的创伤愈合敷料、抗菌敷料、医用粘合剂和人工组织皮肤。
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CN117050336A (zh) * | 2023-07-13 | 2023-11-14 | 海南大学 | 聚乙烯醇/羧甲基壳聚糖/PDA@EGCG NPs水凝胶及其制备方法和应用 |
CN117050336B (zh) * | 2023-07-13 | 2024-05-24 | 海南大学 | 聚乙烯醇/羧甲基壳聚糖/PDA@EGCG NPs水凝胶及其制备方法和应用 |
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