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CN114652854B - 一种双锁mof材料及其制备和应用 - Google Patents

一种双锁mof材料及其制备和应用 Download PDF

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CN114652854B
CN114652854B CN202210208303.7A CN202210208303A CN114652854B CN 114652854 B CN114652854 B CN 114652854B CN 202210208303 A CN202210208303 A CN 202210208303A CN 114652854 B CN114652854 B CN 114652854B
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王英文
张盾
戚鹏
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Abstract

本发明属于纳米材料和生物医学领域，具体涉及一种双锁MOF材料及其制备和应用。双锁MOF材料为以ZIF‑90作为核，其外部包裹酸敏感水凝胶，形成壳核结构的带有双锁响应的双锁MOF材料。本发明制备的双锁MOF材料具有优异的酸和ATP同时刺激响应释放性能。本发明制备的双锁MOF具有良好的稳定性、生物相容性，在药物传递、疾病治疗方面具有广阔的应用前景。

Description

一种双锁MOF材料及其制备和应用
技术领域
本发明属于纳米材料和生物医学领域，具体涉及一种双锁MOF材料及其制备和应用。
背景技术
细菌在表面的定植和随后的生物被膜的形成是关键的全球问题之一。生物膜是胞外多糖基质中微生物的复杂群落，不可逆地粘附在材料表面，导致生物污染和伤口感染。目前，对抗生物膜最常用的策略是使用抗菌药物，然而，由于缺乏对生物膜的靶向性和选择性，使用过量的药物导致了耐药细菌的出现和脱靶毒性的增加。
基于纳米载体的抗菌素传递系统因其能够克服非选择性积累所引起的毒性并提高抗菌素的生物利用度而受到广泛关注。利用刺激响应表面修饰纳米载体，可以实现选择性靶向货物释放。然而，由于人体内部环境复杂，单刺激纳米载体有时不能有效识别生物膜区域。例如，当酸性pH值作为生物膜的指示值时，酸性细胞器或其他外部酸刺激也可能触发货物释放，从而增加脱靶毒性和潜在的安全问题。因此，进一步提高对生物膜的靶向效率对于精准医学治疗和成像至关重要，现阶段纳米载体涉及广泛的纳米材料，如二氧化硅纳米粒子、碳纳米管、金纳米粒子和量子、脂质体和聚合物，但都只对单一刺激有响应，尚未发现对两种刺激连续响应的纳米载体，而pH/ATP双锁纳米载体为解决这一问题提供了一个有前景的方法。
发明内容
本发明的目的在于针对目前现有的不能有效识别生物膜区域的问题，提供一种双锁MOF材料及其制备和应用。
为实现上述目的，本发明采用的技术方案为：
一种双锁MOF材料，双锁MOF材料为以ZIF-90作为核，其外部包裹酸敏感水凝胶，形成壳核结构的带有双锁响应的双锁MOF材料。
所述水凝胶外壳为聚丙烯酸，氯化钙，碳酸钠按摩尔比为1:1:1混合，而后常温(22-25℃)反应2-2.5小时。
所述双锁MOF材料内核为以2-甲醛咪唑和醋酸锌常温反应生成的ZIF-90。
所述ZIF-90为2-甲醛咪唑和醋酸锌按摩尔比为10:1混合，而后常温(22-25℃)反应30-35分钟。
一种双锁MOF材料的制备方法，将2-甲醛咪唑和醋酸锌常温下反应获得ZIF-90，而后将其与聚丙烯酸和氯化钙混合，获得酸敏感水凝胶为外壳内部包裹ZIF-90的壳核结构的带有双锁相应的双锁MOF材料。
所述ZIF-90与聚丙烯酸和氯化钙的物料关系比例为1:2:2-1:2.5:2.5。
一种双锁MOF材料的应用，所述材料在药物传递中的应用。进一步，所述材料在药物传递和肿瘤治疗中的应用。
本发明以2-甲醛咪唑和醋酸锌常温反应生成MOF材料，将荧光染料包裹在MOF中，通过将聚丙烯酸和氯化钙反应生成的酸刺激响应水凝胶包裹在MOF表面，形成双锁MOF材料，发现制备的双锁MOF材料具有优异的酸和ATP连续刺激响应释放性能。其中，所述ZIF-90内包裹荧光染料Rho 6G，以证实双锁MOF材料的刺激响应性能。
本发明的有益效果在于：
本发明以2-甲醛咪唑和醋酸锌常温反应生成MOF材料内核，将荧光染料包裹在MOF内核中，通过将聚丙烯酸和氯化钙反应生成的酸刺激响应水凝胶包裹在MOF表面，形成双锁MOF材料，发现制备的双锁MOF材料具有优异的酸和ATP连续刺激响应释放性能。在模拟生物膜微环境中，双锁MOF材料展现良好的刺激响应性，本发明制备的双锁MOF纳米载体由于其特殊的壳核结构，具有酸响应、ATP响应的连续刺激响应的性能，且具有良好的稳定性，选择性，不易被复杂环境干扰。在药物传递、疾病治疗方面具有广阔的应用前景。
本发明双锁MOF在双刺激响应释放方面具有巨大的潜力，进而可用于药物传递和生物膜根除。
附图说明
图1为本发明实施例1提供的双锁MOF的TEM图，其中，A为罗丹明6G@ZIF-90、B为罗丹明6G@ZIF-90@ACC/PAA；
图2为本发明实施例1提供的双锁MOF在刺激响应的TEM图，其中，A为双锁MOF在pH＝4.0溶液中反应15min、B为双锁MOF在pH＝4.0，1mM ATP溶液中反应15min；
图3为本发明实施例提供的双锁MOF不同pH处理时的荧光值变化图；
图4为本发明实施例提供的酸性环境下，不同浓度ATP处理时双锁MOF的荧光值变化图；
图5为不同组装方法下，纳米载体的荧光值变化图。
具体实施方式
以下结合实例对本发明的具体实施方式做进一步说明，应当指出的是，此处所描述的具体实施方式只是为了说明和解释本发明，并不局限于本发明。
本发明通过形成特定的壳核结构的纳米双锁MOF，该纳米载体具有酸响应、ATP响应的连续刺激响应的性能，且具有良好的稳定性，选择性，不易被复杂环境干扰。
实施例1：
双锁MOF的制备：
1)将48.05mg 2-咪唑甲醛(0.5M)、4.79mg罗丹明6G(1mM)溶于10.0mL DMF中，搅拌15min，然后缓慢加入含有0.05M醋酸锌的5.0mL DMF。然后依次用DMF、超纯水、乙醇洗涤，离心(10000rpm,5min)。最后，将产品分散在超纯水中，得到罗丹明6G@ZIF-90原液(100mg mL-1)。(参见图1A)。
2)将114mg聚丙烯酸(PAA,0.1M),111mg氯化钙(0.1M)和500μL罗丹明6G@ZIF-90(100mg mL-1)添加到10.0mL超纯水中搅拌1h。然后，将10.0mL 0.1M Na2CO3溶液快速加入并搅拌1h。离心所得产品并用超纯水洗净(10000rpm,10分钟)，再分散在超纯水中得到罗丹明6G@ZIF-90@ACC/PAA。(参见图1B)。
通过透射电子显微镜(TEM)表征制备的罗丹明6G@ZIF-90和罗丹明6G@ZIF-90@ACC/PAA的形貌。如图1A和图1B所示，罗丹明6G@ZIF-90和罗丹明6G@ZIF-90@ACC/PAA都是均匀的纳米颗粒。罗丹明6G@ZIF-90纳米颗粒表面粗糙，尺寸均匀，平均直径110-150nm。聚丙烯酸/氯化钙包裹后，罗丹明6G@ZIF-90@ACC/PAA材料的尺寸比罗丹明6G@ZIF-90变大，约为150-200nm。
实施例2：
双锁MOF的刺激响应性：
取5μL上述实施例制备的罗丹明6G@ZIF-90@ACC/PAA(100mg mL-1)置于3mL PBS(pH＝4.0)溶液中孵育15分钟，通过TEM观察其形态变化(参见图2A)。取5μL制备的罗丹明6G@ZIF-90@ACC/PAA(100mg mL-1)置于3mL PBS(pH＝4.0)溶液中，并加入5μL ATP(1M)溶液孵育15分钟，通过TEM观察其形态变化(参见图2B)。
如图2A和图2B所示，酸处理后的罗丹明6G@ZIF-90@ACC/PAA聚丙烯酸/氯化钙水凝胶溶解，罗丹明6G@ZIF-90保持完整，且尺寸与图1A相同，酸和ATP共同处理后，聚丙烯酸/氯化钙水凝胶溶解且罗丹明6G@ZIF-90颗粒裂解失去完整形态，证明了双锁MOF的刺激响应性。
实施例3：
不同pH时双锁MOF的刺激响应性：
将50μL罗丹明6G@ZIF-90@ACC/PAA(1mg mL-1)分别加入240μL不同pH的PBS缓冲溶液中，再加入10μL ATP(1mM)溶液，震荡反应60min，测量溶液在激发波长为525nm，发射波长552nm时的荧光强度。由图3可知，双锁MOF纳米材料的荧光值随时间延长而增加，约30分钟可达到释放平衡，此外，不同pH环境下，纳米颗粒荧光变化有较大区别，随着环境体系酸性增加，荧光分子释放逐渐增多，体现出良好的pH刺激响应性。
实施例4：
不同ATP浓度时双锁MOF的刺激响应性：
将50μL罗丹明6G@ZIF-90@ACC/PAA(1mg mL-1)加入240μL pH分别4.0和6.0的PBS缓冲溶液中，再加入10μL不同浓度的ATP溶液，震荡反应60min，测量溶液在激发波长为525nm，发射波长552nm时的荧光强度。由图4可知，在酸性条件下，荧光值随ATP浓度的增加而增加，且在ATP浓度为1μM时体系已产生较明显的荧光响应，证明罗丹明6G@ZIF-90@ACC/PAA纳米颗粒有灵敏的ATP响应性。
实施例5：
不同组装方法制备纳米载体的荧光响应性：
1)将48.05mg 2-咪唑甲醛(0.5M)、4.79mg罗丹明6G(1mM)溶于10.0mL DMF中，搅拌15min，然后缓慢加入含有0.05M醋酸锌的5.0mL DMF。然后依次超纯水洗涤，将产品分散在超纯水中，得到罗丹明6G@ZIF-90原液(100mg mL-1)。将114mg聚丙烯酸(PAA,0.1M),111mg氯化钙(0.1M)和500μL罗丹明6G@ZIF-90(100mg mL-1)添加到10.0mL超纯水中搅拌1h。然后，将10.0mL 0.1M Na2CO3溶液快速加入并搅拌1h。离心所得产品并用超纯水洗净，此为制备方法1，即实施例1制备所得材料。
2)将114mg聚丙烯酸(PAA,0.1M),111mg氯化钙(0.1M)和4.79mg罗丹明6G(1mM)添加到10.0mL超纯水中搅拌1h。然后，将10.0mL 0.1M Na2CO3溶液快速加入并搅拌1h。离心所得产品并用超纯水洗净，得到原液(100mg mL-1)。将48.05mg 2-咪唑甲醛(0.5M)、50μL原液(100mg mL-1)溶于10.0mL DMF中，搅拌15min，然后缓慢加入含有0.05M醋酸锌的5.0mL DMF。然后依次超纯水洗涤，将产品分散在超纯水中，此为制备方法2。
分别取5μL上述实施例制备纳米材料置于3mL PBS(pH＝4.0)溶液中，并加入5μLATP(1M)溶液孵育15分钟，观察其荧光变化，并放置14天再次观察荧光数值。结果表明仅制备方法1的组装方式可以合成具有双刺激的目标纳米载体，且该纳米载体具有良好的稳定性，浸泡14天后荧光值没有显著变化。

Claims (3)

1.一种双锁MOF材料，其特征在于：双锁MOF材料为以ZIF-90作为核，其外部包裹酸敏感水凝胶，形成壳核结构的带有双锁响应的双锁MOF材料；
具体制备方法为：
1)将48.05mg 2-咪唑甲醛、4.79mg罗丹明6G溶于10.0mL DMF中，搅拌15min，然后缓慢加入含有0.05M醋酸锌的5.0mL DMF；然后依次用DMF、超纯水、乙醇洗涤，10000rpm离心5min；最后，将产品分散在超纯水中，得到罗丹明6G@ZIF-90原液；
2)将114mg聚丙烯酸，111mg氯化钙和500μL罗丹明6G@ZIF-90添加到10.0mL超纯水中搅拌1h；然后，将10.0mL 0.1M Na2CO3溶液快速加入并搅拌1h；10000rpm离心10min所得产品并用超纯水洗净，再分散在超纯水中得到罗丹明6G@ZIF-90@ACC/PAA。
2.按权利要求1所述的双锁MOF材料的制备方法，其特征在于：
1)将48.05mg 2-咪唑甲醛、4.79mg罗丹明6G溶于10.0mL DMF中，搅拌15min，然后缓慢加入含有0.05M醋酸锌的5.0mL DMF；然后依次用DMF、超纯水、乙醇洗涤，10000rpm离心5min；最后，将产品分散在超纯水中，得到罗丹明6G@ZIF-90原液；
2)将114mg聚丙烯酸，111mg氯化钙和500μL罗丹明6G@ZIF-90添加到10.0mL超纯水中搅拌1h；然后，将10.0mL 0.1M Na2CO3溶液快速加入并搅拌1h；10000rpm离心10min所得产品并用超纯水洗净，再分散在超纯水中得到罗丹明6G@ZIF-90@ACC/PAA。
3.一种权利要求1所述双锁MOF材料的应用，其特征在于：所述材料在药物传递中作为载体的应用。
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