Determination of the optical interference of iron oxide nanoparticles in fluorometric cytotoxicity assays.

Nanomaterials are known to exhibit unique interactions with light. Iron oxide nanoparticles (IONPs), composed of magnetite (black iron oxide) specifically, are known to be highly absorptive throughout the visible portion of the spectrum. We sought to investigate and overcome optical interference of IONPs in colorimetric, fluorometric and luminescence assays by introducing additional controls and determining the concentration-dependent contribution to optical artifacts which could confound, skew, or invalidate results. We tested the in vitro cytotoxicity of ∼8 nm spherical magnetite nanoparticles capped with alginate on a human lung carcinoma (A549) cell line for different exposure periods and at various concentrations. We observed significant interference with both the MTT reagent and the absorption at 590 nm, a concentration-dependent reduction in the luminescence, fluorescence at ∼490 nm (viability marker), and fluorescence at 530 nm (cytotoxicity marker). After introducing an additional correction, we obtained more accurate results, including a clear decrease in viability at 12-h post-treatment, with apparent near complete recovery after 24-h in addition to a dose-independent, time-dependent alteration in the cell proliferation rate. A small increase in cytotoxicity was noted at the 24-h timepoint at the two highest concentrations. According to our results, the MTT reagents appear to interact substantially with IONPs at concentrations above 0.1 mg/mL, therefore, this assay is not recommended for IONP cytotoxicity assessment at higher concentrations.

Templated synthesis enhances the cobalt adsorption capacity of a porous organic polymer.

Divalent transition metals such as Co(II) are important targets for removal from water sources, due to their potential toxicity as well as their high value. In this study, we found that a series of porous organic polymers based on amide-linked tetraphenylmethane units are effective Co(II) ion adsorbents in aqueous solution. To increase the density of Co(II) binding sites, we then developed a templated synthesis in which the branched, rigid monomers are pre-assembled around Co(II) ions prior to polymerization. After polymer formation, the Co(II) template ions are removed to yield a material rich in Co(II) binding sites. Ion adsorption isotherms show that the Co(II)-templated material has an ion adsorption capacity significantly greater than those of the non-templated materials, highlighting the utility of a templated synthetic route. SEM and TEM images show the morphology of the templated polymer to be dramatically different from the non-templated polymers and to be similar in size and shape to the Co(II)-monomer precursors, emphasizing the role of the template ions in directing the formation of the resulting polymer. This guest-templated approach requires no functionalization of the generic monomer and represents a promising synthetic route to high-capacity ion adsorbents for water purification and aqueous separations.

Hybrid Block Copolymer Synthesis by Merging Photoiniferter and Organocatalytic Ring-Opening Polymerizations.

Access to block copolymers from monomers that do not polymerize via a common mechanism requires initiation from a multifunctional species that allows orthogonal polymerization chemistries. We disclose a strategy to provide well-defined polyacrylamido-b-polyether block copolymers by a one-pot combination of photoiniferter polymerization and organocatalytic ring-opening polymerization (ROP) using a hydroxy-functionalized trithiocarbonate photoiniferter as the dual initiator at ambient temperature. Our results reveal good compatibility between the two polymerization systems and highlight that they can be performed in arbitrary order or simultaneously with good retention of the thiocarbonylthio functionality. We also demonstrate selective temporal control over the photoiniferter polymerization during concurrent ROP. We harnessed the efficiency of combining these polymerization systems to provide tailor-made block copolymers from chemically distinct monomers.

Block Copolymer Sequence Inversion through Photoiniferter Polymerization.

Block copolymers prepared by reversible addition-fragmentation chain transfer (RAFT) polymerization are often restricted to a specific comonomer blocking sequence that is dictated by intermediate radical stability and relative radical leaving group abilities. Techniques that provide alternative pathways for reinitiation of thiocarbonylthio-terminated polymers could allow access to block copolymer sequences currently unobtainable through the RAFT process. We report a method for preparing "inverted" block copolymers, whereby the traditional order of monomer addition has been reversed through the use of photoiniferter-mediated radical polymerization. Specifically, thiocarbonylthio photolysis of xanthate- and dithiocarbamate-functional macromolecular chain transfer agents (macro-CTAs) led to the direct formation of leaving group macroradicals otherwise unaffordable by an addition-fragmentation mechanism. We believe this method could provide a route to synthesize multiblock copolymers of synthetically challenging comonomer sequences.

Probing Membrane Hydration at the Interface of Self-Assembled Peptide Amphiphiles Using Electron Paramagnetic Resonance.

The relative hydrophilicity at the interface of a nanoparticle was measured utilizing electron paramagnetic resonance (EPR) spectroscopy. The supramolecular structure was assembled from spin-labeled peptide amphiphiles (PA) derived from N-carboxy anhydrides (NCA). Cyanuric chloride, or 2,4,6-trichloro-1,3,5-triazine (TCT), was used as a modular platform to synthesize the spin-labeled, lipid-mimetic macroinitiator used for the ring-opening polymerization of γ-benzyl-l-glutamic acid NCA to produce polyglutamate-b-dodecanethiol2. Through static and dynamic light scattering, as well as transmission electron microscopy, PAs with DP of 50 and 17 were shown to assemble into stable nanoparticles with an average hydrodynamic radius of 117 and 84 nm, respectively. Continuous wave EPR spectroscopy revealed that the mobility parameter (h-1/h0) and 2Aiso of the nitroxide radical increased with increasing pH, in concert with the deprotonation of the PE side chains and associated helix-coil transition. These results are consistent with an increase in the relative hydration and polarity at the nanoparticle interface, which would be dependent on the secondary structure of the polypeptide. This research suggests that a pH stimulus could be used to facilitate water diffusion through the membrane.

Synthetic upcycling of polyacrylates through organocatalyzed post-polymerization modification.

The direct transformation of commercially available commodity polyacrylates into value-added materials was achieved. We demonstrate how 1,5,7-triazabicyclo[4.4.0]dec-5-ene, serving as a nucleophilic catalyst, can be used to catalyze acyl substitution reactions of acrylic polymers in the presence of alcohol and amine nucleophiles. Furthermore, we found that organocatalytic transesterification exhibits high selectivity towards sterically unhindered esters, thus providing a new route towards site-selective acyl substitution of macromolecular materials. Combining this methodology with reversible-deactivation radical polymerization (RDRP) techniques such as reversible addition-fragmentation chain-transfer (RAFT) polymerization allowed for the precise functionalization of sterically-differentiated acrylic copolymers and polymeric chain ends. We envision this approach to expedite functional polymer synthesis and provide access to functional macromolecules prepared from inexpensive, hydrolytically-stable polymeric precursors.

Stimuli-responsive peptide-based ABA-triblock copolymers: unique morphology transitions with pH.

We report the synthesis and solution characterization of poly(L-lysine)-b-poly(propylene oxide)-b-poly(L-lysine) (KPK) triblock copolymers with high lysine weight fractions (>75 wt%). In contrast to PK diblock copolymers in this composition range, KPK triblock copolymers exhibit morphology transitions as a function of pH. Using a combination of light-scattering and microscopy techniques, we demonstrate spherical micelle-vesicle and spherical micelle-disk micelle transitions for different K fractions. We interpret these morphology changes in terms of the energy penalty associated with folding the core P block to form a spherical micelle in relation to the interfacial curvature associated with different charged states of the K block.