From Unimolecular Template to Silver Nanocrystal Clusters: An Effective Strategy to Balance Antibacterial Activity and Cytotoxicity

Silver nanomaterials have attracted a great deal of interest due to their broad-spectrum antimicrobial activity. However, it is still challenging to balance the high antibacterial efficiency with low damage to biological cells of silver nanostructures, especially when the diameter decreases to less than 10 nm. Here, we developed a new type of Ag nanohybrid material via a unimolecular micelle template method, which presents amazing antibacterial activities and almost noncytotoxicity. First, water-soluble multiarm star-shaped brushlike copolymer alpha-CD-g-[(PEO40-g-PAA(50))-b-PEO5](18) was precisely synthesized and its micelle behavior in different solvents was revealed. Then, nanocrystal clusters assembled by Ag grains (Ag@Template NCs) were prepared through an in situ redox route using the unimolecular micelle of alpha-CD-g-[(PEO40-g-PAA(50))-b-PEO5](18) as the soft template, AgNO3 as a precursor, and tetrabutylammonium borohydride (TBAB) as the reducing agent. The overall size of the achieved Ag@Template NCs is controlled by the template structure at around 40 nm (D-h in DMF), and the size of the Ag grain can be easily regulated from similar to 1 to similar to 5 nm by adjusting the feeding ratio of AgNO3/acrylic acid (AA) units in the template from 1:10 to 1:1. Benefitting from the structural design of the template, all Ag@Template NCs prepared here exhibit excellent dispersibility and chemical stability in different aqueous environments (neutral, pH = 5.5, and 0.9% NaCl physiological saline solution), which play a crucial role in the long-term storage and potential application in a complex physiological environment. The antibacterial and cytotoxicity tests indicate that Ag@Template NCs display much better performance than Ag nanoparticles (Ag NPs), which have a comparable overall size of similar to 25 nm. The inhibitory capability of Ag@Template NCs to bacteria strongly depends on the grain size. Specifically, the Ag@Template-1 NC assembled by the smallest grains (1.6 +/- 0.3 nm) presents the best antibacterial activity. For E. coli (-), the MIC value is as low as 5 mu g/mL (0.36 mu g/mL of Ag), while for S. aureus (+), the value is around 10 mu g/mL (0.72 mu g/mL of Ag). The survival rate of L02 cells and lactate dehydrogenase assay together illustrate the low cytotoxicity possessed by the prepared Ag@Template NCs. Therefore, the proposed Ag@Template NC structure successfully resolves the high reactivity, instability, and fast oxidation issues of the ultrasmall Ag nanoparticles, and integrates high antibacterial efficiency and nontoxicity to biological cells into one platform, which implies its broad potential application in biomedicine.

Automated summary

Silver nanomaterials have shown broad-spectrum antimicrobial activity, with the challenge of balancing high antibacterial efficiency and low damage to biological cells. A new type of Ag nanohybrid material was developed using a unimolecular micelle template method, demonstrating excellent antibacterial activities and almost no cytotoxicity. This structure successfully addresses the reactivity, instability, and fast oxidation issues of ultrasmall silver nanoparticles, integrating high antibacterial efficiency and nontoxicity into one platform for potential biomedical applications.