Biocompatibility and internalization of molecularly imprinted nanoparticles

Molecularly imprinted polymers （MIP） are receiving increasing attention thanks to their robustness, stability, and inexpensive manufacture compared with their bio-analogues such as antibodies. The molecular imprinting process can be defined as the generation of molecular recognition sites in a synthetic polymer. The template-derived sites created within a polymeric matrix allow MIPs （often referred as plastic antibodies） to selectively recognize and bind to the target molecule. Therefore, MIPs can be used in sensors and in separation and diagnostics. Owing to their size and functional properties, MIP nanoparticles （NPs） can potentially be used in biomedicine, but comprehensive analysis of their interaction with cells and in vitro toxicological tests must be performed first. Herein, we report the synthesis of bare and core-shell imprinted NPs using an innovative solid-phase approach and the toxicological evaluation of such NPs in different cell lines （HaCaT, MEFs, HT1080, and macrophages）. We also evaluated the influence of the protein corona on particle stability, the internalization of NPs in cells, and the influence of various surface coatings. Studies on the metabolic effects of imprinted NPs on fibroblasts showed that bare MIPs do not alter cell metabolism, whereas some issues arise when specific particle coatings are used. Furthermore, in vitro cytokine release studies revealed that macrophages were not activated in the presence of the MIPs evaluated in this study. The results suggest that MIP NPs are biocompatible, paving the way for their in vivo application.

Development of a homogenous assay based on fluorescent imprinted nanoparticles for analysis of nitroaromatic compounds

Herein we describe the development of a homogeneous assay for the detection of 4-nitroaniline(4-NA)and 2,4-dinitroaniline(2,4-diNA).Thisassay relies on fluorescent molecularly imprinted nanoparticles(nano MIPs)which,upon interaction with the target analytes,generate areduction in fluorescence emission intensity(quenching).This is due to a resp on sive fluorescent monomer(N-2-propenyl-(5-dimethylamino)-1-naphthalene sulphonamide)employed in the manufacture of the nanoMIPs which,by virtue of the imprinting process,is capable of selectiveinteraction with the target an alyte,thus giving rise to a quenching effect.Selectivity experiments showed excellent recognition propertiestoward the target molecule.Under optimal conditions,the fluorescence intensity of these nano MIPs decreased as the concentration of theimprinted analyte increased from 10 nM to 2.71 pM.A linear relation between the negative logarithm of 4-NA or 2,4-diNA concentrations and the fluorescence intensity for both nanosystems was found(R^2=0.991 and R^2=0.9895),with excellent sensitivity(limit of detection(LOD)=7 and 6 nM,respectively).Furthermore,both nanosystems have been successfully applied for detection of 4-NA or 2,4-diNA in tap water,withrecoveries between 90%to 100.6%and 92%to 100.3%,respectively.Thanks to the versatility of the imprinting process,this nanosystemholds the potential for further development of several optical sensors for many other compounds.