Renee V. Goreham

Antibacterial surfaces by adsorptive binding of polyvinyl-sulphonate-stabilized silver nanoparticles.

This paper presents a novel and facile method for the generation of efficient antibacterial coatings which can be applied to practically any type of substrate. Silver nanoparticles were stabilized with an adsorbed surface layer of polyvinyl sulphonate (PVS). This steric layer provided excellent colloidal stability, preventing aggregation over periods of months. PVS-coated silver nanoparticles were bound onto amine-containing surfaces, here produced by deposition of an allylamine plasma polymer thin film onto various substrates. SEM imaging showed no aggregation upon surface binding of the nanoparticles; they were well dispersed on amine surfaces. Such nanoparticle-coated surfaces were found to be effective in preventing attachment of Staphylococcus epidermidis bacteria and also in preventing biofilm formation. Combined with the ability of plasma polymerization to apply the thin polymeric binding layer onto a wide range of materials, this method appears promising for the fabrication of a wide range of infection-resistant biomedical devices.

pH-tunable gradients of wettability and surface potential

Smart materials that can sense and respond to changes in the environment are of interest in numerous and diverse applications. In this paper, we report gradient surfaces where wettability and surface potential respond to changes in the pH. The gradients are produced by controlling the concentration of amine and carboxyl acid groups across the surface. The response of surface wettability to pH changes was studied by water contact angle measurements. The potential across the surface was determined by atomic force microscopy-based surface force measurements. These studies showed that at low pH the surface potential changes from “no charge” at the acid end to a positive charge at the amine end. At high pH the surface potential changed from negative at the acid end to “no charge” at the amine side. At an intermediate pH the charge across the surface changes from negative at the acid end to positive at the amine end. Potential applications include separation or guidance of charged entities such as particles, proteins or bacteria.

Surface Bound Amine Functional Group Density Influences Embryonic Stem Cell Maintenance

Gradient surfaces are highly effective tools to screen and optimize cell- surface interactions. Here, the response of embryonic stem (ES) cell colonies to plasma polymer gradient surfaces is investigated. Surface chemistry ranged from pure allylamine (AA) plasma polymer on one end of the gradient to pure octadiene (OD) plasma polymer on the other end. Optimal surface chemistry conditions for retention of pluripotency were identified. Expression of the stem cell markers alkaline phosphatase (AP) and Oct4 varied with the position of the ES cell colonies across the OD-AA plasma polymer gradient. Both markers were more strongly retained on the OD plasma polymer rich regions of the gradients. The observed variation of expression across the plasma polymer gradient increased with duration of stem cell culture. While maximum cell adhesion to the gradient substrate occurred at a nitrogen- to-carbon (N/C ratio) of approximately 0.1, Oct4 and AP expression was best retained at an N/C ratio < 0.04. Stem cell marker expression correlated with colony size and morphology: more compact, multilayered colonies with prominent F-actin staining arose as the N/C ratio decreased. Disruption of actin polymerization using Y-27632 ROCK inhibitor resulted in a collapse of the multilayer colony structure into monolayers with limited cell-cell contact. A corresponding decrease in expression of AP and Oct4 was observed. Oct4 expression along with 3D colony morphology was partially rescued on the OD plasma polymer rich regions of the gradient.

Versatile gradients of chemistry, bound ligands and nanoparticles on alumina nanopore arrays

Nanoporous alumina (PA) arrays produced by self-ordering growth, using electrochemical anodization, have been extensively explored for potential applications based upon the unique thermal, mechanical and structural properties, and high surface-to-volume ratio of these materials. However, the potential applications and functionality of these materials may be further extended by molecular-level engineering of the surface of the pore rims. In this paper we present a method for the generation of chemical gradients on the surface of PA arrays based upon plasma co-polymerization of two monomers. We further extend these chemical gradients, which are also gradients of surface charge, to those of bound ligands and number density gradients of nanoparticles. The latter represent a highly exotic new class of materials, comprising aligned PA, capped by gold nanoparticles around the rim of the pores. Gradients of chemistry, ligands and nanoparticles generated by our method retain the porous structure of the substrate, which is important in applications that take advantage of the inherent properties of these materials. This method can be readily extended to other porous materials.

Subtle changes in surface chemistry affect embryoid body cell differentiation: lessons learnt from surface-bound amine density gradients.

Advanced approaches to direct the differentiation of embryonic stem cells are highly sought after. The surface-bound chemical gradient format is a powerful screening approach that can be deployed to study changes in stem cell behavior as a function of subtle changes in surface chemistry. Here, we investigate the spontaneous differentiation of cells derived from differentiating mouse embryoid body (mEB) cells into endoderm, mesoderm, and ectoderm following culture on surface-bound gradients of chemical functional groups in the absence of differentiation-biasing bioactive factors. Gradients were created using a diffusion-controlled plasma polymerization technique. The generated coating ranged from hydrophobic 1,7-octadiene (OD) plasma polymer at one end of the gradient to a more hydrophilic allylamine (AA) plasma polymer on the opposite end. The gradient surface was divided into seven equal regions of progressively increasing AA plasma polymer content and mEB cell response within these regions was compared. Cells adhered preferentially to the central regions of the gradient; however, cell proliferation increased toward AA-plasma-polymer-rich end of the gradient. Variation in the expression of germ layer markers was noted across the gradient surface. High AA:OD plasma polymer ratios triggered cell differentiation toward both mesoderm and ectoderm. Expression of tissue-specific markers, in particular, KRT18, AFP, and TNNT2, was strikingly responsive to subtle changes in surface chemistry, exhibiting vastly different expression levels between adjacent regions. Our results suggest that the surface-bound gradient platform is well suited to screening surface chemistries for use in the field of stem cell technologies and regenerative medicine.

Particle number density gradient samples for nanoparticle metrology with atomic force microscopy

Atomic force microscopy (AFM) can provide a link in the traceability chain between dimensional measurement techniques for nanoparticles, such as dynamic light scattering and differential centrifugal sedimentation, and the realization of the definition of the SI metre. Despite the size of nanoparticles being well within the resolution range of typical AFMs, the accurate measurement of nanoparticles with AFM presents a number of challenges. One of these challenges is the number density of particles deposited on substrates for AFM imaging and measurement. If the number density is too low, it is difficult to obtain adequate measurement statistics, whereas a number density that is too high can result in particle agglomeration on the substrate and make it difficult to image sufficient substrate area to obtain a reliable reference for height measurements. We present imaging and measurement results of 16 nm gold nanoparticles deposited on a substrate functionalized to produce a surface with a number density gradient across the sample. This substrate functionalization shows great potential for achieving reliable and efficient nanoparticle metrology with AFM.