CN114588524B - 一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法 - Google Patents
一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法 Download PDFInfo
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Abstract
本发明涉及一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法,包括如下步骤:S1:取20μL‑30μL的咪唑修饰石墨烯量子点(IMZ‑GQDs)溶液(30mg/mL)滴加在PDMS模板表面,置于真空干燥箱内;S2:取出后刮除表面多余溶液,再次真空干燥;S3:取约25μL的PVA/PVP混合溶液滴加在PDMS模板表面,置于真空干燥箱内;S4:取出后刮除表面多余溶液及气泡;S5:室温下干燥24h后脱模,并保存于干燥器内。本发明的有益效果为:IMZ‑GQDs MNs将抗菌性纳米材料与微针技术相结合,用于安全、高效、可控的眼部给药,能有效突破眼表解剖屏障同时溶解释放出IMZ‑GQDs破坏细菌内膜完整性、降低膜电位,最终导致细菌死亡,以达到治疗细菌性角膜炎的目的。
Description
技术领域
本发明涉及医疗技术领域,特别涉及一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法。
背景技术
石墨烯量子点(GQDs)具有光稳定性高、溶解性好、易于修饰和毒性低的优势,多通过与过氧化氢(H2O2)协同作用或光动力(PDT)杀菌,然而这些额外刺激的加入可能不适用于眼表等敏感组织。
带正电荷的纳米抗菌材料近年来颇受关注。咪唑(IMZ)作为自然界最常见的五元杂环化合物之一,是组氨酸的组成部分,参与体内多种酶促反应。咪唑环可经烷基化反应成为带正电荷的阳离子,同时具有良好的稳定性和安全性,广泛应用于制药领域,用于治疗微生物感染、癌症、阿茨海默症等。
此外,纳米材料对眼部的有效给药仍存在挑战,滴眼液/眼膏形式难以突破解剖屏障达到有效治疗浓度,提高应用浓度则可能增加细胞毒性;植入眼内的药物递送系统可实现药物的可控释放,但需手术干预,存在风险。因此,临床亟需一种更加安全、高效、可控的眼部给药方式。
发明内容
针对现有技术存在的不足,本发明的目的在于提供一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法,以解决上述问题。
本发明的技术方案是这样实现的:一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法,包括如下步骤:
S1:取20μL-30μL的咪唑修饰石墨烯量子点(IMZ-GQDs)溶液(30mg/mL)滴加在PDMS模板表面,置于真空干燥箱内,使溶液进入针尖并压出空气;
S2:取出后刮除表面多余溶液,再次真空干燥,使溶液干燥浓缩至尖端;
S3:取约25μL的PVA/PVP混合溶液滴加在PDMS模板表面,置于真空干燥箱内,使溶液进入针尖并压出空气;
S4:取出后刮除表面多余溶液及气泡;
S5:室温下干燥24h后脱模,获得的咪唑修饰石墨烯量子点的可溶性微针贴片(IMZ-GQDs MNs)保存于干燥器内。
通过采用上述技术方案,负载咪唑修饰石墨烯量子点(IMZ-GQDs)的可溶性微针贴片(IMZ-GQDs MNs)结合了微针技术与抗菌纳米材料两种新兴技术的优点,用于安全、高效、可控的眼部给药,这种微针贴片可以很好地贴附于角膜表面,减少因泪液代谢造成的药物损失,能有效突破眼表解剖屏障,穿透泪膜和角膜上皮层,在角膜基质层溶解释放出IMZ-GQDs;IMZ-GQDs可通过咪唑阳离子特性破坏细菌内膜完整性、降低膜电位,表现出对于革兰阳性菌的显著抗菌效果;IMZ-GQDs MNs可安全应用于眼部且对兔眼金葡菌性角膜炎的疗效显著,可为细菌性角膜炎以及其他眼病的治疗提供参考。
本发明进一步设置为:所述步骤S3和S4中的PVA/PVP混合溶液通过如下步骤制得:
a:称取一定量聚乙烯醇(PVA,Mw=31000Da)固体加入去离子水中,使用加热磁力搅拌器(110℃,900rpm)磁力搅拌3h,获得15%(w/v)的均质透明PVA溶液,静置去除气泡;
b:称取一定量聚乙烯基吡咯烷酮(PVP,K-30)固体溶于去离子水中,室温搅拌均匀,获得10%(w/v)的均质透明PVP溶液;
c:将15%(w/v)PVA溶液与10%(w/v)PVP溶液按3:2的比例混匀,获得均质透明的PVA/PVP混合溶液,去除气泡,待用。
通过采用上述技术方案,PVA/PVP混合溶液能有效的作为IMZ-GQDs的基底,并使IMZ-GQDs更容易脱模。
本发明进一步设置为:所述步骤S1、S2、S3中的真空干燥箱参数均设为-0.08MPa,20℃。
通过采用上述技术方案,能有效的使溶液进入PDMS模板内,并压出空气,使其成型效果更好。
本发明进一步设置为:所述步骤S1、S2、S3中在真空干燥箱内维持10min。
通过采用上述技术方案,能有效的压出PDMS模板内的空气,使其IMZ-GQDs MNs的成型效果更好。
本发明进一步设置为:所述步骤S1中取25μL的IMZ-GQDs溶液(30mg/mL)。
通过采用上述技术方案,因为IMZ-GQDs体外抗菌的最佳浓度为100μg/mL,而健康人眼角膜体积约为60μL,可得出IMZ-GQDs单眼的有效用量应为6μg,所以25μL的IMZ-GQDs溶液(30mg/mL)会更加接近IMZ-GQDs单眼的有效用量。
本发明进一步设置为:所述步骤S4中刮除表面多余溶液及气泡后还需另外滴加25μL的PVA/PVP混合溶液作为基底。
通过采用上述技术方案,PDMS模板内的IMZ-GQDs更容易取出脱模,还能使IMZ-GQDs的使用更加的方便。
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简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动性的前提下,还可以根据这些附图获得其他的附图。
图1中的(A)为在IMZ-GQDs(100μg/mL)不同作用时间下金黄色葡萄球菌的存活率(*:P<0.05,**:P<0.01,***:P<0.001,n=3)及相应菌落图片;(B)为活死染色:(a-c)对照组和(d-f)IMZ-GQDs(100μg/mL,3h)处理的金黄色葡萄球菌,标尺为50μm;(C)为SEM图片:(a)对照组和(b)IMZ-GQDs(100μg/mL,3h)处理的金黄色葡萄球菌;
图2为本发明中IMZ-GQDs MNs的制备流程示意图;
图3为本发明中Blank MNs与IMZ-GQDs MNs的:(a)俯视位,(b)横截面光学显微镜图,(c)SEM图片;
图4为经不同处理的兔眼角膜:(a)裂隙灯显微镜图片,(b)钴蓝光下荧光素钠染色图片。
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
如图1-图4所示,本发明公开了一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法,包括如下步骤:
S1:取20μL-30μL的咪唑修饰石墨烯量子点(IMZ-GQDs)溶液(30mg/mL)滴加在PDMS模板表面,置于真空干燥箱内,使溶液进入针尖并压出空气;
S2:取出后刮除表面多余溶液,再次真空干燥,使溶液干燥浓缩至尖端;
S3:取约25μL的PVA/PVP混合溶液滴加在PDMS模板表面,置于真空干燥箱内,使溶液进入针尖并压出空气;
S4:取出后刮除表面多余溶液及气泡;
S5:室温下干燥24h后脱模,获得的咪唑修饰石墨烯量子点的可溶性微针贴片(IMZ-GQDs MNs)保存于干燥器内。
通过采用上述技术方案,负载咪唑修饰石墨烯量子点(IMZ-GQDs)的可溶性微针贴片(IMZ-GQDs MNs)结合了微针技术与抗菌纳米材料两种新兴技术的优点,用于安全、高效、可控的眼部给药,这种微针贴片可以很好地贴附于角膜表面,减少因泪液代谢造成的药物损失,能有效突破眼表解剖屏障,穿透泪膜和角膜上皮层,在角膜基质层溶解释放出IMZ-GQDs;IMZ-GQDs可通过咪唑阳离子特性破坏细菌内膜完整性、降低膜电位,表现出对于革兰阳性菌的显著抗菌效果;IMZ-GQDs MNs可安全应用于眼部且对兔眼金葡菌性角膜炎的疗效显著,可为细菌性角膜炎以及其他眼病的治疗提供参考。
在本发明实施例中,所述步骤S3和S4中的PVA/PVP混合溶液通过如下步骤制得:
a:称取一定量聚乙烯醇(PVA,Mw=31000Da)固体加入去离子水中,使用加热磁力搅拌器(110℃,900rpm)磁力搅拌3h,获得15%(w/v)的均质透明PVA溶液,静置去除气泡;
b:称取一定量聚乙烯基吡咯烷酮(PVP,K-30)固体溶于去离子水中,室温搅拌均匀,获得10%(w/v)的均质透明PVP溶液;
c:将15%(w/v)PVA溶液与10%(w/v)PVP溶液按3:2的比例混匀,获得均质透明的PVA/PVP混合溶液,去除气泡,待用。
通过采用上述技术方案,PVA/PVP混合溶液能有效的作为IMZ-GQDs的基底,并使IMZ-GQDs更容易脱模。
在本发明实施例中,所述步骤S1、S2、S3中的真空干燥箱参数均设为-0.08MPa,20℃。
通过采用上述技术方案,能有效的使溶液进入PDMS模板内,并压出空气,使其成型效果更好。
在本发明实施例中,所述步骤S1、S2、S3中在真空干燥箱内维持10min。
通过采用上述技术方案,能有效的压出PDMS模板内的空气,使其IMZ-GQDs MNs的成型效果更好。
在本发明实施例中,所述步骤S1中取25μL的IMZ-GQDs溶液(30mg/mL)。
通过采用上述技术方案,因为IMZ-GQDs体外抗菌的最佳浓度为100μg/mL,而健康人眼角膜体积约为60μL,可得出IMZ-GQDs单眼的有效用量应为6μg,所以25μL的IMZ-GQDs溶液(30mg/mL)会更加接近IMZ-GQDs单眼的有效用量。
在本发明实施例中,所述步骤S4中刮除表面多余溶液及气泡后还需另外滴加25μL的PVA/PVP混合溶液作为基底。
通过采用上述技术方案,PDMS模板内的IMZ-GQDs更容易取出脱模,还能使IMZ-GQDs的使用更加的方便。
以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (6)
1.一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法,其特征在于,包括如下步骤:
S1:取20μL-30μL的咪唑修饰石墨烯量子点(IMZ-GQDs)溶液(30mg/mL)滴加在PDMS模板表面,置于真空干燥箱内,使溶液进入针尖并压出空气;
S2:取出后刮除表面多余溶液,再次真空干燥,使溶液干燥浓缩至尖端;
S3:取约25μL的PVA/PVP混合溶液滴加在PDMS模板表面,置于真空干燥箱内,使溶液进入针尖并压出空气;
S4:取出后刮除表面多余溶液及气泡;
S5:室温下干燥24h后脱模,获得的咪唑修饰石墨烯量子点的可溶性微针贴片(IMZ-GQDs MNs)保存于干燥器内。
2.根据权利要求1所述的一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法,其特征在于,所述步骤S3和S4中的PVA/PVP混合溶液通过如下步骤制得:
a:称取一定量聚乙烯醇(PVA,Mw=31000Da)固体加入去离子水中,使用加热磁力搅拌器(110℃,900rpm)磁力搅拌3h,获得15%(w/v)的均质透明PVA溶液,静置去除气泡;
b:称取一定量聚乙烯基吡咯烷酮(PVP,K-30)固体溶于去离子水中,室温搅拌均匀,获得10%(w/v)的均质透明PVP溶液;
c:将15%(w/v)PVA溶液与10%(w/v)PVP溶液按3:2的比例混匀,获得均质透明的PVA/PVP混合溶液,去除气泡,待用。
3.根据权利要求1所述的一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法,其特征在于,所述步骤S1、S2、S3中的真空干燥箱参数均设为-0.08MPa,20℃。
4.根据权利要求1所述的一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法,其特征在于,所述步骤S1、S2、S3中在真空干燥箱内维持10min。
5.根据权利要求1所述的一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法,其特征在于,所述步骤S1中取25μL的IMZ-GQDs溶液(30mg/mL)。
6.根据权利要求1所述的一种用于治疗细菌性角膜炎的基于石墨烯量子点构建的可溶性微针贴片的制备方法,其特征在于,所述步骤S4中刮除表面多余溶液及气泡后还需另外滴加25μL的PVA/PVP混合溶液作为基底。
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