Carole C. Perry

Chemistry of aqueous silica nanoparticle surfaces and the mechanism of selective peptide adsorption

Control over selective recognition of biomolecules on inorganic nanoparticles is a major challenge for the synthesis of new catalysts, functional carriers for therapeutics, and assembly of renewable biobased materials. We found low sequence similarity among sequences of peptides strongly attracted to amorphous silica nanoparticles of various size (15-450 nm) using combinatorial phage display methods. Characterization of the surface by acid base titrations and zeta potential measurements revealed that the acidity of the silica particles increased with larger particle size, corresponding to between 5% and 20% ionization of silanol groups at pH 7. The wide range of surface ionization results in the attraction of increasingly basic peptides to increasingly acidic nanoparticles, along with major changes in the aqueous interfacial layer as seen in molecular dynamics simulation. We identified the mechanism of peptide adsorption using binding assays, zeta potential measurements, IR spectra, and molecular simulations of the purified peptides (without phage) in contact with uniformly sized silica particles. Positively charged peptides are strongly attracted to anionic silica surfaces by ion pairing of protonated N-termini, Lys side chains, and Arg side chains with negatively charged siloxide groups. Further, attraction of the peptides to the surface involves hydrogen bonds between polar groups in the peptide with silanol and siloxide groups on the silica surface, as well as ion-dipole, dipole-dipole, and van-der-Waals interactions. Electrostatic attraction between peptides and particle surfaces is supported by neutralization of zeta potentials, an inverse correlation between the required peptide concentration for measurable adsorption and the peptide pI, and proximity of cationic groups to the surface in the computation. The importance of hydrogen bonds and polar interactions is supported by adsorption of noncationic peptides containing Ser, His, and Asp residues, including the formation of multilayers. We also demonstrate tuning of interfacial interactions using mutant peptides with an excellent correlation between adsorption measurements, zeta potentials, computed adsorption energies, and the proposed binding mechanism. Follow-on questions about the relation between peptide adsorption on silica nanoparticles and mineralization of silica from peptide-stabilized precursors are raised.

Biosilicification: the role of the organic matrix in structure control.

Abstract. Silicon (although never in the elemental form) is present in all living organisms and is required for the production of structural materials in single-celled organisms through to higher plants and animals. Hydrated amorphous silica is a mineral of infinite functionality and yet it is formed into structures with microscopic and macroscopic form. Research into the mechanisms controlling the process have highlighted proteins and proteoglycans as possible control molecules. Such molecules are suggested to play a critical role in the catalysis of silica polycondensation reactions and in structure direction. This article reviews information on silica form and function, silica condensation chemistry, the role of macromolecules in structure control and in vitro studies of silica formation using biomolecules extracted from biological silicas. An understanding of the mechanisms by which biological organisms regulate mineral formation will assist in our understanding of the essentiality of silicon to life processes and in the generation of new materials with specific form and function for industrial application in the 21st century.

Antibiotic mediated synthesis of gold nanoparticles with potent antimicrobial activity and their application in antimicrobial coatings

We report a one-pot synthesis of spherical gold nanoparticles (52–22 nm) and their capping with cefaclor, a second-generation antibiotic, without use of other chemicals. The differently sized gold nanoparticles were fabricated by controlling the rate of reduction of gold ions in aqueous solution by varying the reaction temperature (20–70 °C). The primary amine group of cefaclor acted as both the reducing and capping agent for the synthesis of gold nanoparticles leaving the β-lactam ring of cefaclor available for activity against microbes. Antimicrobial testing showed that cefaclor reduced gold nanoparticles have potent antimicrobial activity against both Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria as compared to cefaclor or gold nanoparticles alone. The minimum inhibition concentrations (MICs) of cefaclor reduced gold nanoparticles were 10 µg mL−1 and 100 µg mL−1 for S. aureus and E. coli respectively. The cefaclor reduced gold nanoparticles were further coated onto poly(ethyleneimine) (PEI) modified glass surfaces to obtain antimicrobial coatings suitable for biomedical applications and were tested against E. coli as an exemplar of activity. The antimicrobial coatings were very robust under adverse conditions (pH 3 and 10), inhibited the growth of E. coli on their surfaces, and could be used many times with retained activity. Results from a combined spectroscopic (FTIR) and microscopic study (AFM) suggest that the action of these novel particles is through the combined action of cefaclor inhibiting the synthesis of the peptidoglycan layer and gold nanoparticles generating “holes” in bacterial cell walls thereby increasing the permeability of the cell wall, resulting in the leakage of cell contents and eventually cell death.

The use of high aspect ratio photoresist (SU-8) for super-hydrophobic pattern prototyping

In this work we present a reliable technique for the production of large areas of high aspect-ratio patterns and describe their use as model super-hydrophobic systems. The high thickness and straight sidewalls possible with SU-8 were used to generate dense patterns of small pillars. These photoresist patterns could be used directly, without the need for micromoulding. A method is given allowing resist thickness to be varied over a wide range and a bottom antireflective layer was used to simplify patterning on reflective substrates. This patterning technique allows rapid testing of wetting theories, as pattern size and depth can be varied simply and samples can be produced in sufficient numbers for laboratory use. We show how the static contact angle of water varies with pattern height for one sample-pattern and how static and dynamic contact angles vary with dimension using high aspect-ratio patterns.

Peptide Adsorption on Silica Nanoparticles: Evidence of Hydrophobic Interactions

Molecular recognition and interactions at the interface between biomolecules and inorganic materials determine important phenomena such as protein adsorption, cell adhesion to biomaterials, or the selective response of biosensors. Events occurring at the biomolecule-inorganic interface, despite their importance, are still poorly understood, thus limiting control of interfacial properties and response. In this contribution, using well-characterized silica nanoparticles and a series of peptides having heterogeneous physicochemical properties (S1: KLPGWSG, S2: AFILPTG, and S3: LDHSLHS) identified from biopanning against the same particles, we identify the driving forces that govern peptide-silica binding. Binding isotherms obtained by fluorimetric assay under different pH conditions allowed us to demonstrate the impact of binding environment (pH) on adsorption behavior of a given peptide-surface silica nanoparticle. Our experimental data suggest a multistep adsorption mechanism leading to the formation of multilayers on silica, in which the prevailing interactions (i.e., electrostatic or hydrophobic/hydrogen bonding) and their relative contribution to the binding event are governed by the identity of the peptide itself, the substrates surface functionality (hydrophilic or hydrophobic), and the peptide bulk concentration and solution bulk pH. Our studies show how it is possible to modulate peptide uptake on silica, or in fact on any particle, by changing either the surface properties or, more simply, the binding environment. In addition, the data reveal an intrinsic bias toward positively charged sequences in the elution conditions used in the biopanning protocol with much information about strong binder sequence diversity being lost during panning.

Plastron properties of a superhydrophobic surface

Most insects and spiders drown when submerged during flooding or tidal inundation, but some are able to survive and others can remain submerged indefinitely without harm. Many achieve this by natural adaptations to their surface morphology to trap films of air, creating plastrons which fix the water-vapor interface and provide an incompressible oxygen-carbon dioxide exchange surface. Here the authors demonstrate how the surface of an extremely water-repellent foam mimics this mechanism of underwater respiration and allows direct extraction of oxygen from aerated water. The biomimetic principle demonstrated can be applied to a wide variety of man-made superhydrophobic materials.

Silicification: The Processes by Which Organisms Capture and Mineralize Silica

Silicification is widespread in the biological world and occurs in bacteria, single-celled protists, plants, invertebrates and vertebrates. Minerals formed in the biological environment often show unusual physical properties (e.g., strength, degree of hydration) and often have structures that exhibit order on many length scales. The minerals are formed from an environment that is undersaturated with respect to silicon and under conditions of around neutral pH and low temperature ca. 4–40°C. Formation of the mineral may occur intra- or extra-cellularly and specific biochemical locations for mineral deposition that include lipids, proteins and carbohydrates are known. The significance of the cellular machinery cannot be over emphasized and it is with advances in experimental techniques (cell biology and materials characterization) and advances in understanding (including the ability to design laboratory experiments to mimic the biological environment) that much progress has been made in the field in recent years. In most cases the formation of the mineral phase is linked to cellular processes and if we understand this process the knowledge so gained could be used to good effect in designing new materials for biomedical, optical and other applications. The study of living organisms could result in wealth generation/creation. It should be noted that although significant advances have been made in the last ten years, new questions have arisen and there are many areas requiring exploration. This contribution will place emphasis on the systems for which most is known, namely sponges and diatoms, however, it should be borne in mind that many other organisms from single-celled species such as choanoflagellates (Mann et al. 1982) and radiolaria through to higher plants and molluscs such as the limpet (Mann et al. 1986) make use of silica and form species-specific structures that show structural organization on several length scales. Examples of some of the silicified structures observed …

From biosilicification to tailored materials: Optimizing hydrophobic domains and resistance to protonation of polyamines

Considerable research has been directed toward identifying the mechanisms involved in biosilicification to understand and possibly mimic the process for the production of superior silica-based materials while simultaneously minimizing pollution and energy costs. Molecules isolated from diatoms and, most recently sponges, thought to be key to this process contain polyamines with a propylamine backbone and variable levels of methylation. In a chemical approach to understanding the role of amine (especially propylamine) structures in silicification we have explored three key structural features: (i) the degree of polymerization, (ii) the level of amine methylation, and (iii) the size of the amine chain spacers. In this article, we show that there are two factors critical to their function: the ability of the amines to produce microemulsions and the presence of charged and uncharged amine groups within a molecule, with the latter feature helping to catalyze silicic acid condensation by a proton donor/acceptor mechanism. The understanding of amine–silicate interactions obtained from this study has enabled the controlled preparation of hollow and nonporous siliceous materials under mild conditions (circumneutral pH, room temperature, and in all aqueous systems) possibly compatible with the conditions used by biosystems. The “rules” identified from our study were further used predictively to modulate the activity of a given amine. We believe that the outcomes of the present contribution will form the basis for an approach to controlling the growth of inorganic materials by using tailor-made organic molecules.

An overview of the fundamentals of the chemistry of silica with relevance to biosilicification and technological advances

Biomineral formation is widespread in nature, and occurs in bacteria, single‐celled protists, plants, invertebrates, and vertebrates. Minerals formed in the biological environment often show unusual physical properties (e.g. strength, degree of hydration) and often have structures that exhibit order on many length scales. Biosilica, found in single‐celled organisms through to higher plants and primitive animals (sponges), is formed from an environment that is undersaturated with respect to silicon, and under conditions of approximately neutral pH and relatively low temperatures of 4−40 °C compared to those used industrially. Formation of the mineral may occur intracellularly or extracellularly, and specific biochemical locations for mineral deposition that include lipids, proteins and carbohydrates are known. In most cases, the formation of the mineral phase is linked to cellular processes, an understanding of which could lead to the design of new materials for biomedical, optical and other applications. In this contribution, we describe the aqueous chemistry of silica, from uncondensed monomers through to colloidal particles and 3D structures, that is relevant to the environment from which the biomineral forms. We then describe the chemistry of silica formation from alkoxides such as tetraethoxysilane, as this and other silanes have been used to study the chemistry of silica formation using silicatein, and such precursors are often used in the preparation of silicas for technological applications. The focus of this article is on the methods, experimental and computational, by which the process of silica formation can be studied, with an emphasis on speciation.

Osteoinductive silk-silica composite biomaterials for bone regeneration.

Osteoinductive and biodegradable composite biomaterials for bone regeneration were prepared by combining silk fibroin with silica particles. The influence of these composite systems on osteogenesis was evaluated with human mesenchymal stem cells (hMSCs) subjected to osteogenic differentiation. hMSCs adhered, proliferated, and differentiated towards osteogenic lineages on silk/silica films. The addition of the silica to the silk films influenced gene expression leading to upregulation of bone sialoprotein (BSP) and collagen type 1 (Col 1) osteogenic markers. Evidence for early bone formation in the form of collagen fibers and apatite nodules was obtained on the silk/silica films. Collagen fibers were closely associated with apatite deposits and overall collagen content was higher for the silica containing samples. Also, smaller sized silica particles (24 nm-2 μm) with large surface area facilitated silica biodegradation in vitro through particle dissolution, leading to ∼5-fold decrease in silica content over 10 weeks. These results indicate the suitability of silk/silica composite system towards bone regeneration, where degradation/remodeling rates of the organic and inorganic components can be controlled.