CN115926187B - 一种混合金属有机框架及其制备方法与应用 - Google Patents
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Abstract
本发明涉及一种混合金属有机框架及其制备方法与应用,步骤包括:S1、将HAuCl4溶液与MHA溶液依次加入至超纯水中,搅拌第一搅拌时间后,得白色的Au(I)‑MHA络合物;加入过量的NaOH溶液使Au(I)‑MHA络合物溶解,并加入NaBH4溶液,搅拌第二搅拌时间后,得Au25(MHA)18团簇;S2、将Au25(MHA)18团簇与混合金属离子混合,超声第三超声时间后,即得混合金属有机框架;其中,混合金属离子包括:IVB族金属的四价离子以及具有抗菌性能的金属离子。本发明构建的铈(IV)‑AuNCs‑Cu(II)混合金属有机框架(Mixed‑Metal Organic Frameworks,MMOFs)中铈(IV)发挥类DNase酶效果进一步降解EPS的同时,协助金纳米团簇以及Cu(II)发挥优越的抗菌抗生物膜效果,且由于MOFs具有缓释降解的作用,局部浓度不会爆发式增加,进而减轻对正常细胞的毒害作用。
Description
技术领域
本发明涉及生物医药技术领域,尤其涉及一种混合金属有机框架及其制备方法与应用。
背景技术
内植物相关骨感染(Implant-related bone infection)是在内植物置入手术(例如:骨折内固定术、假体植入术等)后发生的感染,一直是困扰骨科临床治疗的难题之一。入侵的细菌会在骨组织和内固定表面形成生物膜,使感染迁延难治。细菌生物膜(bacterialbiofilms)为包裹在自身分泌的基质中的细菌群落,是细菌为保护自身而形成的一种独特生存模式,其能抵御宿主免疫攻击并降低对抗生素的敏感性,生物膜内的细菌与浮游菌相比,对抗生素的耐受力提高了1000倍。
目前有关内植物相关骨感染的治疗,传统方法以外科清创加抗生素治疗为主,随后有庆大霉素珠链和抗生素骨水泥或人工骨等,但效果均不尽满意。近年来随着多学科交叉研究的发展,目前针对生物膜开发的抗菌策略包括:抗生物膜药物、光热光动力、超声/磁调控等。
生物膜中的胞外DNA(extracellular DNA,eDNA)通过连接细菌和其他EPS组分在生物膜的形成和稳定中发挥关键作用,已成为破坏生物膜的潜在靶点;一旦eDNA破裂,生物膜则容易被清除。脱氧核糖核酸酶(DNase)能通过靶向和水解eDNA来分解生物膜。
目前临床上抗生素的滥用导致了细菌耐药的发生发展,且其对生物膜内低代谢菌而言作用有限。新型无机材料是近年来治疗感染的新思路,主要包括Ag、Cu等纳米抗菌材料。纳米银是目前应用最为广泛的人工纳米材料之一,由于其高效、广谱、作用持久和不易产生耐药性等优良的抗菌特性,目前越来越多的研究人员将其应用于涂层、纤维、塑料和杀虫剂等方面。
然而,DNase作为一种天然酶有几个缺点,比如:价格昂贵、稳定性差、难以负载和修饰;纳米银解析出的Ag+对哺乳动物细胞具有毒副作用,大大限制了其体内应用。大部分抗菌策略由于生物安全性、抗生物膜效率及机体复杂的内环境,目前尚无法应用于临床。
因此,亟需一种双功能协同系统,在没有抗生素的情况下,既能破坏生物膜细胞外多聚物结构,又能同时杀灭细菌,且毒副作用更小的混合金属有机框架及其制备方法与应用。
发明内容
本发明的目的是针对现有技术中的不足,提供一种混合金属有机框架及其制备方法与应用。
为实现上述目的,本发明采取的技术方案是:
本发明的第一方面是提供一种混合金属有机框架的制备方法,步骤包括:
S1、将HAuCl4溶液与6-巯基己酸(MHA)溶液依次加入至超纯水中,搅拌第一搅拌时间后,得白色的Au(I)-MHA络合物;加入过量的NaOH溶液使所述Au(I)-MHA络合物溶解,并加入NaBH4溶液,搅拌第二搅拌时间后,得Au25(MHA)18团簇;
S2、将所述Au25(MHA)18团簇与混合金属离子混合,超声第三超声时间后,即得所述混合金属有机框架;
其中,所述混合金属离子包括:IVB族金属的四价离子以及具有抗菌性能的金属离子。
优选地,所述HAuCl4溶液以及所述NaBH4溶液为新鲜配制的溶液。
优选地,所述HAuCl4溶液、所述MHA溶液以及所述NaBH4溶液的摩尔比为1:2:2.24。
优选地,所述第一搅拌时间为2min-10min。
优选地,所述第二搅拌时间为1h-6h。
优选地,所述Au25(MHA)18团簇与所述混合金属离子的摩尔比为1:20。
优选地,所述IVB族金属的四价离子包括:Ce4+。
优选地,所述具有抗菌性能的金属离子包括:Cu2+或Zn2+中的至少一种。
优选地,所述IVB族金属的四价离子与所述具有抗菌性能的金属离子的摩尔比为1:1。
优选地,所述第三超声时间为2min-10min。
本发明的第二方面是提供一种采用如前所述制备方法制得的混合金属有机框架。
本发明的第三方面是提供一种如前所述混合金属有机框架在制备抗耐药性细菌或/和抗生物膜感染药物中的应用。
本发明采用以上技术方案,与现有技术相比,具有如下技术效果:
路易斯酸性金属离子(例如:Zn(II)、Ce(IV)和Cu(II))的人工生物催化剂具有一定的类DNase和RNase活性;铈(Cerium,Ce)作为一种稀土元素,具有独特的理化性质;金纳米团簇(Gold nanoclusters,AuNCs)具有超小的尺寸(<2nm),一般由少则数个、多则上百个金原子组成,从分子构成上来看,金纳米团簇具有确定的分子式及分子结构,类似于有机金属化合物,所以金纳米团簇是无机-有机的混合化合物,金纳米团簇具有优异的抗菌活性,且虽然同为贵金属,与银释放离子产生毒性不同,金是惰性金属,高度稳定而不容易解离成离子,是人们公认的高度生物相容性材料,即使是尺寸降低至2nm的金纳米团簇,其稳定性和生物安全性也得到了验证;金属有机框架(Metal-Organic Frameworks,MOFs)是一种具有开放骨架的配位聚合物,由多齿桥接有机配体/团簇和金属阳离子自组装而成,含有潜在的空隙,MOFs的优势包括:结构和化学多样性、高负载能力以及生物降解性;
本发明构建的铈(IV)-AuNCs-Cu(II)混合金属有机框架(Mixed-Metal OrganicFrameworks,MMOFs)中铈(IV)发挥类DNase酶效果进一步降解EPS的同时,协助金纳米团簇以及Cu(II)发挥优越的抗菌抗生物膜效果,且由于MOFs具有缓释降解的作用,局部浓度不会爆发式增加,进而减轻对正常细胞的毒害作用。
附图说明
图1为实施例2中MMOFs体外抗菌性能测试中琼脂平板上细菌菌落生长情况的结果图;
图2为实施例2中MMOFs体外抗菌性能测试中扫描电镜结果图;
图3为实施例2中MMOFs作用于ESBL大肠杆菌膜时的破坏机制示意图。
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的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。
需要说明的是,在不冲突的情况下,本发明中的实施例及实施例中的特征可以相互组合。
下面结合附图和具体实施例对本发明作进一步说明,但不作为本发明的限定。
实施例1
本实施例提供一种混合金属有机框架及其制备方法,制备步骤包括:
S1、将1.25mL 20mM新鲜配制的HAuCl4溶液与5mL 10mM的MHA溶液依次加入至10mL的超纯水中,搅拌5min后,得白色的Au(I)-MHA络合物;加入1.5mL 1M的NaOH溶液使所述Au(I)-MHA络合物溶解,并加入0.5mL 112mM新鲜配制的NaBH4溶液,于室温下搅拌3h后,超滤管收集得Au25(MHA)18团簇;
S2、将1mL 2mg/mL的所述Au25(MHA)18团簇溶液转移至离心管中,并加入混合金属离子溶液(阳离子为等摩尔比的Ce4+与Cu2+,阴离子为Cl-),所述Au25(MHA)18团簇与所述混合金属离子的摩尔比为1:20,超声5min后,即得所述混合金属有机框架。
实施例2
本实施例提供如实施例1所述混合金属有机框架(即MMOFs)的体外抗菌性能测试。
以MRSA作为模式菌株,将处于生长对数期的MRSA悬液倍稀释至106CFU/mL备用;将PBS(对照组)和MMOFs(实验组)置于24孔板中,每组5个复孔,用微量加样器吸取1mL MRSA菌液加入24孔板中,置于37℃恒温箱中培养24h:去除未黏附细菌,然后置于离心管中超声震荡1min,稀释100倍后取100μL涂于培养板上,放入37℃恒温箱中培养24h,菌落计数并拍照,使用平板法计数,并以每毫升菌落形成单位(CFU/mL)表示;对于扫描电镜,细菌培养结束后,用移液器吸弃培养液并用PBS轻柔漂洗三次,各加入2mL 2.5%戊二醛于4℃固定过夜,再依次用PBS溶液轻柔漂洗,乙醇梯度脱水,每次15min,最后进行真空干燥、喷金、扫描电镜观察。
如图1-2所示,与无MMOFs包裹的对照组相比,实验组中细菌数量明显减少,参示图1,实验组琼脂平板上细菌菌落数量明显少于对照组,参示图2,实验组钛片表面的细菌数量更少。
综上所述,本发明构建的铈(IV)-AuNCs-Cu(II)混合金属有机框架(Mixed-MetalOrganic Frameworks,MMOFs)中铈(IV)发挥类DNase酶效果进一步降解EPS的同时,协助金纳米团簇以及Cu(II)发挥优越的抗菌抗生物膜效果,且由于MOFs具有缓释降解的作用,局部浓度不会爆发式增加,进而减轻对正常细胞的毒害作用。
以上所述仅为本发明较佳的实施例,并非因此限制本发明的实施方式及保护范围,对于本领域技术人员而言,应当能够意识到凡运用本发明说明书及图示内容所作出的等同替换和显而易见的变化所得到的方案,均应当包含在本发明的保护范围内。
Claims (7)
1.一种混合金属有机框架的制备方法,其特征在于,步骤包括:
S1、将HAuCl4溶液与6-巯基己酸溶液依次加入至超纯水中,搅拌第一搅拌时间后,得白色的Au(I)-MHA络合物;加入过量的NaOH溶液使所述Au(I)-MHA络合物溶解,并加入NaBH4溶液,搅拌第二搅拌时间后,得Au25(MHA)18团簇;
S2、将所述Au25(MHA)18团簇与混合金属离子混合,超声第三超声时间后,即得所述混合金属有机框架;
其中,所述Au25(MHA)18团簇与所述混合金属离子的摩尔比为1:20;
其中,所述混合金属离子为Ce4+以及Cu2+;
其中,所述Ce4+与所述Cu2+的摩尔比为1:1。
2.根据权利要求1所述的制备方法,其特征在于,所述HAuCl4溶液以及所述NaBH4溶液为新鲜配制的溶液。
3.根据权利要求1所述的制备方法,其特征在于,所述HAuCl4溶液、所述MHA溶液以及所述NaBH4溶液的摩尔比为1:2:2.24。
4.根据权利要求1所述的制备方法,其特征在于,所述第一搅拌时间为2min-10min;所述第二搅拌时间为1h-6h。
5.根据权利要求1所述的制备方法,其特征在于,所述第三超声时间为2min-10min。
6.一种采用如权利要求1-5任一项所述制备方法制得的混合金属有机框架。
7.一种如权利要求6所述混合金属有机框架在制备抗耐药性细菌或/和抗生物膜感染药物中的应用。
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