Water treatment residual (WTR)-coated wood mulch for alleviation of toxic metals and phosphorus from polluted urban stormwater runoff.

Aluminum-based water treatment residual (WTR)-coated wood mulches were synthesized and tested for removal of heavy metals and phosphorus (P) in synthetic urban stormwater. WTRs are an industrial waste produced from coagulation in water treatment facilities, primarily composed of amorphous aluminum or iron hydroxides. Batch tests showed that the composite filter media could effectively adsorb 97% lead (Pb), 76% zinc (Zn), 81% copper (Cu) and 97% P from the synthetic stormwater (Pb = 100 µg/L, Zn = 800 µg/L, Cu = 100 µg/L, P = 2.30 mg/L, and pH = 7.0) within 120 min, due to the presence of aluminum hydroxides as an active adsorbent. The adsorption was a 2(nd)-order reaction with respect toward each pollutant. Column tests demonstrated that the WTR-coated mulches considerably alleviated the select pollutants under a continuous-flow condition over the entire filtration period. The effluent Pb, Zn, Cu, and P varied at 0.5-8.9%, 33.4-46.7%, 45.8-55.8%, and 6.4-51.9% of their respective initial concentrations with the increasing bed volume from 0 to 50. Synthetic precipitation leaching procedure (SPLP) and toxicity characteristic leaching procedure (TCLP) tests indicated that leached contaminants were all below the U.S. criteria, suggesting that the release of undesired chemicals under rainfall or landfilling conditions is not a concern during application. This study demonstrates that the WTR-coated mulches are a new, low-cost, and effective filter media for urban stormwater treatment. Equally important, this study provides a sustainable approach to beneficially reuse an industrial waste for environmental pollution control.

Facile co-assembly process to generate core-shell nanoparticles with functional protein corona.

A simple and robust protocol to maintain the structural feature of polymer-protein core-shell nanoparticles (PPCS-NPs) is developed based on the synergistic interactions between proteins and functional polymers. Using the self-assembly method, a broad range of proteins can be assembled to the selective water-insoluble polymers containing pyridine groups. The detailed analysis of the PPCS-NPs structure was conducted using FESEM and thin-sectioned TEM. The results illustrated that the protein molecules are located on the corona of the PPCS-NPs. While proteins are displacing between water and polymer to minimize the interfacial energy, the polymer offers a unique microenvironment to maintain protein structure and conformation. The proposed mechanism is based on a fine balance between hydrophobicity and hydrophilicity, as well as hydrogen bonding between proteins and polymer. The PPCS-NPs can serve as a scaffold to incorporate both glucose oxidase (GOX) and horseradish peroxidase (HRP) onto a single particle. Such a GOX-HRP bienzymatic system showed a ~20% increase in activity in comparison to the mixed free enzymes. Our method therefore provides a unique platform to preserve protein structure and conformation and can be extended to a number of biomolecules.

Mutant plant viruses with cell binding motifs provide differential adhesion strengths and morphologies.

The ability of Tobacco mosaic virus (TMV) to tolerate various amino acid insertions near its carboxy terminus is well-known. Typically these inserts are based on antigenic sequences for vaccine development with plant viruses as carriers. However, we determined that the structural symmetries and the size range of the viruses could also be modeled to mimic the extracellular matrix proteins by inserting cell-binding sequences to the virus coat protein. The extracellular matrix proteins play important roles in guiding cell adhesion, migration, proliferation, and stem cell differentiation. Previous studies with TMV demonstrated that the native and phosphate-modified virus particles enhanced stem cell differentiation toward bone-like tissues. Based on these studies, we sought to design and screen multiple genetically modified TMV mutants with reported cell adhesion sequences to expand the virus-based tools for cell studies. Here, we report the design of these mutants with cell binding amino acid motifs derived from several proteins, the stabilities of the mutants against proteases during purification and storage, and a simple and rapid functional assay to quantitatively determine adhesion strengths by centrifugal adhesion assay. Among the mutants, we found that cells on TMV expressing RGD motifs formed filopodial extensions with weaker attachment profiles, whereas the cells on TMV expressing collagen I mimetic sequence displayed little spreading but higher attachment strengths.

Electrospun fibrous scaffolds promote breast cancer cell alignment and epithelial-mesenchymal transition.

In this work we created electrospun fibrous scaffolds with random and aligned fiber orientations in order to mimic the three-dimensional structure of the natural extracellular matrix (ECM). The rigidity and topography of the ECM environment have been reported to alter cancer cell behavior. However, the complexity of the in vivo system makes it difficult to isolate and study such extracellular topographical cues that trigger cancer cells' response. Breast cancer cells were cultured on these fibrous scaffolds for 3-5 days. The cells showed elongated spindle-like morphology in the aligned fibers, whereas they maintained a mostly flat stellar shape in the random fibers. Gene expression profiling of these cells post seeding showed up-regulation of transforming growth factor ß-1 (TGFß-1) along with other mesenchymal biomarkers, suggesting that these cells undergo epithelial-mesenchymal transitions in response to the polymer scaffold. The results of this study indicate that the topographical cue may play a significant role in tumor progression.

Visualizing cell extracellular matrix (ECM) deposited by cells cultured on aligned bacteriophage M13 thin films.

Topographical features ranging from micro- to nanometers can affect cell orientation and migratory pathways, which are important factors in tissue engineering and tumor migration. In our previous study, a convective assembly of bacteriophage M13 resulted in thin films which could be used to control the alignment of cells. However, several questions regarding its underlying reasons to dictate cell alignment remained unanswered. Here, we further study the nanometer topographical features generated by the bacteriophage M13 crystalline film, which results in the alignment of the cells and extracellular matrix (ECM) proteins. Sequential imaging analyses at micro- and nanoscale levels of aligned cells and fibrillar matrix proteins were documented using scanning electron microscopy and immunofluorescence microscopy. As a result, we observed baby hamster kidney cells with higher degree of alignment on the ordered M13 substrates than NIH-3T3 fibroblasts, a difference which could be attributed to the intrinsic nature of the cells' production of ECM proteins. The results from this study provide a crucial insight into the topographical features of a biological thin film, which can be utilized to control the orientation of cells and surrounding ECM proteins.

Synthesis and electron microscopic analysis of the self-assembly of polymer and ferritin core-shell structures.

Self-assembly of poly(4-vinylpyridine) (P4VP) and ferritin produced a spherical core-shell structure, which was analyzed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In particular, for better understanding, the organization of such core-shell nanostructures, an optimal protocol for preparation of TEM thin sectioning of ferritin-P4VP composites, was developed. It entails fixing the ferritin-P4VP complex with 2.5% glutaraldehyde and infiltrating it with a mixture of acetone/resin while omitting the OsO(4) postfixation and ethanol dehydration steps of the conventional protocol. Using this method, a round-shaped thin section structure with unevenly distributed dark and white components was observed. The dark component from the thin section structure was determined to contain ferritin by energy-filtered TEM imaging and iron element mapping, whereas the white part was identified as P4VP by its energy dispersive X-ray spectroscopy (EDS) spectrum.

Synthesis and characterization of thermally responsive Pluronic F127-chitosan nanocapsules for controlled release and intracellular delivery of small molecules.

In this study, we synthesized empty core-shell structured nanocapsules of Pluronic F127 and chitosan and characterized the thermal responsiveness of the nanocapsules in size and wall-permeability. Moreover, we determined the feasibility of using the nanocapsules to encapsulate small molecules for temperature-controlled release and intracellular delivery. The nanocapsules are ∼37 nm at 37 °C and expand to ∼240 nm when cooled to 4 °C in aqueous solutions, exhibiting >200 times change in volume. Moreover, the permeability of the nanocapsule wall is high at 4 °C (when the nanocapsules are swollen), allowing free diffusion of small molecules (ethidium bromide, MW = 394.3 Da) across the wall, while at 37 °C (when the nanocapsules are swollen), the wall-permeability is so low that the small molecules can be effectively withheld in the nanocapsule for hours. As a result of their thermal responsiveness in size and wall-permeability, the nanocapsules are capable of encapsulating the small molecules for temperature-controlled release and intracellular delivery into the cytosol of both cancerous (MCF-7) and noncancerous (C3H10T1/2) mammalian cells. The cancerous cells were found to take up the nanocapsules much faster than the noncancerous cells during 45 min incubation at 37 °C. Moreover, toxicity of the nanocapsules as a delivery vehicle was found to be negligible. The Pluronic F127-chitosan nanocapsules should be very useful for encapsulating small therapeutic agents to treat diseases particularly when it is combined with cryotherapy where the process of cooling and heating between 37 °C and hypothermic temperatures is naturally done.

Comparative study of inhibition at multiple stages of amyloid-beta self-assembly provides mechanistic insight.

The "amyloid cascade hypothesis," linking self-assembly of the amyloid-beta protein (Abeta) to the pathogenesis of Alzheimer's disease, has led to the emergence of inhibition of Abeta self-assembly as a prime therapeutic strategy for this currently unpreventable and devastating disease. The complexity of Abeta self-assembly, which involves multiple reaction intermediates related by nonlinear and interconnected nucleation and growth mechanisms, provides multiple points for inhibitor intervention. Although a number of small-molecule inhibitors of Abeta self-assembly have been identified, little insight has been garnered concerning the point at which these inhibitors intervene within the Abeta assembly process. In the current study, a julolidine derivative is identified as an inhibitor of Abeta self-assembly. To gain insight into the mechanistic action of this inhibitor, the inhibition of fibril formation from monomeric protein is assessed quantitatively and compared with the inhibition of two distinct mechanisms of growth for soluble Abeta aggregation intermediates. This compound is observed to significantly inhibit soluble aggregate growth by lateral association while having little effect on soluble aggregate elongation via monomer addition. In addition, inhibition of soluble Abeta aggregate association exhibits an IC(50) with a somewhat lower stoichiometric ratio than the IC(50) determined for inhibition of fibril formation from monomeric Abeta. This quantitative comparison of inhibition within multiple Abeta self-assembly assays suggests that this compound binds the lateral surface of on-pathway intermediates exhibiting a range of sizes to prevent their association with other aggregates, which is required for further assembly into mature fibrils.

Controlled assembly of rodlike viruses with polymers.

A practical method to assemble rodlike tobacco mosaic virus and bateriophage M13 with polymers was developed, which afforded a 3D core-shell composite with morphological control.