We have designed series of monodispersed nanohydrogels (NHGs) with sizes ranging from 20 to 400 nm, generated from mixtures of N-isopropylacrylamide, di-block (hydrophilic-hydrophobic), and tri-block (hydrophobic-hydrophilic-hydrophobic) copolymer acrylamide macro-monomers. When the monomers are mixed at high temperature they collapse into well-defined self-assemblies, which can be further polymerized leading to cross-linked NHGs with sizes matching the intermediate self-assemblies. The size of the self-assemblies can be tuned/calibrated by combining different ratios of the starting monomeric mixtures at high temperature. Herein, we defined the concept of” phantom monomers” which are the closest structure that mimic a selected monomer but lacks the active function for polymerization. The phantom monomer co-formulated with other monomers will be present in the intermediate self-assemblies due to its similarity with one of the active monomers. However, upon polymerization, the phantom monomer, lacking of a polymerizable function, will be excluded and a new NHGs will be generated. The comparative analyses of our previously obtained standard NHGs (G1) with those obtained here using phantom monomers (G2 and G3) not only put in evidence the self-assembly mediated mechanism but also prove the generation of new monodispersed NHG’s with improved drug-loading properties. As proof of concept, the NHGs were loaded with doxorubicin (DOX) and tested in cells. Results indicate that the drug-loading of the NHGs increases from G1 to G3 and while using the same drug concentration, different size NHGs affect differently the treated cells and disclose a different activity and localization as compared to free DOX by confocal microscopy.

我们设计了一系列单分散纳米水凝胶（NHG），其尺寸范围为20至400 nm，由N-异丙基丙烯酰胺，二嵌段（亲水-疏水）和三嵌段（疏水-亲水-疏水）共聚物丙烯酰胺的混合物生成-单体。当单体在高温下混合时，它们会塌陷成定义明确的自组装体，然后可以进一步聚合，从而生成尺寸与中间自组装体相匹配的交联NHG。可以通过组合高温下起始单体混合物的不同比例来调整/校准自组装体的大小。在此，我们定义了“幻影单体”的概念，它是模仿所选单体但缺乏聚合活性功能的最接近的结构。与其他单体共同配制的幻象单体将由于其与一种活性单体的相似性而存在于中间自组装物中。然而，在聚合时，缺乏可聚合功能的幻影单体将被排除，并会生成新的NHG。对我们先前获得的标准NHG（G1）与此处使用幻影单体（G2和G3）进行的比较分析，不仅证明了自组装介导的机理，而且证明了新的单分散NHG的产生，具有改善的载药量。作为概念验证，NHG装有阿霉素（DOX），并在细胞中进行了测试。结果表明，NHGs的载药量从G1增加到G3，并且使用相同的药物浓度，