Nano- and sub-micron porous polyelectrolyte multilayer assemblies: biomimetic surfaces for human corneal epithelial cells.

In vivo, corneal epithelial cells adhere on basement membranes that exhibit porosity on the nanoscale with the diameters of pores and fibers ranging from 20 to 200 nm. Polyelectrolyte multilayers with porosity ranging from the nano to the microscale were assembled to mimic the pore sizes of corneal membranes in vivo. The average pore diameter was found to be 100 nm and 600 nm for the nanoporous and sub-micron porous films respectively. In this study, a purely physical feature, specifically, porosity, provided cues to human corneal epithelial cells. Porous surfaces that exhibited either 100 nm or 600 nm pore diameters supported corneal cell adhesion, however, nanoscale porosity significantly enhanced corneal epithelial cellular response. Corneal epithelial cell proliferation and migration speeds were significantly higher on nanoporous topographies. The actin cytoskeletal organization was well defined and vinculin focal adhesions were found in cells presented with a nanoscale environment. These trends prevailed for fibronectin-coated surfaces as well suggesting that for human corneal epithelial cells, the physical environment plays a defining role in guiding cell behavior.

Polyelectrolyte multilayers with intrinsic antimicrobial functionality: the importance of mobile polycations.

Cationic contact-killing is an important strategy for creating antimicrobial surfaces that prevent viable bacteria attachment. Recent studies have shown that highly swollen, compliant surfaces resist bacterial attachment and a sufficient density of mobile cationic charge can effectively disrupt bacterial cell membranes. Polyelectrolyte multilayers (PEMs), a popular coating system for surface modification, have been used to kill bacteria through the incorporation of contact-killing or leaching biocides. In this work, we show that manipulation of multilayer assembly and postassembly conditions (e.g., pH) to expose mobile cationic charge can create antimicrobial PEMs without the addition of specific biocidal species. As a model system, we explored PEMs comprising poly(allylamine hydrochloride) (PAH) and poly(sodium 4-styrene sulfonate) (SPS) assembled at high pH and subsequently immersed in low pH solutions. This system undergoes a reversible pH-dependent swelling transition, and we demonstrate that antimicrobial functionality at physiological pH conditions can be turned on and off with suitable pH treatment. In both airborne and waterborne bacteria assays, the viability of two strains of Gram positive Staphylococcus epidermidis (S. epidermidis), one biofilm forming and one nonbiofilm forming, and two strains of Gram negative Escherichia coli (E. coli) was effectively reduced on SPS/PAH multilayers displaying accessible cationic charge. To generalize our results, the pH assembly conditions of PEMs comprising poly(acrylic acid) (PAA) and PAH were also modified to introduce antibacterial capabilities.

Substrata mechanical stiffness can regulate adhesion of viable bacteria.

The competing mechanisms that regulate adhesion of bacteria to surfaces and subsequent biofilm formation remain unclear, though nearly all studies have focused on the role of physical and chemical properties of the material surface. Given the large monetary and health costs of medical-device colonization and hospital-acquired infections due to bacteria, there is considerable interest in better understanding of material properties that can limit bacterial adhesion and viability. Here we employ weak polyelectrolyte multilayer (PEM) thin films comprised of poly(allylamine) hydrochloride (PAH) and poly(acrylic acid) (PAA), assembled over a range of conditions, to explore the physicochemical and mechanical characteristics of material surfaces controlling adhesion of Staphylococcus epidermidis bacteria and subsequent colony growth. Although it is increasingly appreciated that eukaryotic cells possess subcellular structures and biomolecular pathways to sense and respond to local chemomechanical environments, much less is known about mechanoselective adhesion of prokaryotes such as these bacteria. We find that adhesion of viable S. epidermidis correlates positively with the stiffness of these polymeric substrata, independently of the roughness, interaction energy, and charge density of these materials. Quantitatively similar trends observed for wild-type and actin analogue mutant Escherichia coli suggest that these results are not confined to only specific bacterial strains, shapes, or cell envelope types. These results indicate the plausibility of mechanoselective adhesion mechanisms in prokaryotes and suggest that mechanical stiffness of substrata materials represents an additional parameter that can regulate adhesion of and subsequent colonization by viable bacteria.

Biochemical functionalization of polymeric cell substrata can alter mechanical compliance.

Biochemical functionalization of surfaces is an increasingly utilized mechanism to promote or inhibit adhesion of cells. To promote mammalian cell adhesion, one common functionalization approach is surface conjugation of adhesion peptide sequences such as Arg-Gly-Asp (RGD), a ligand of transmembrane integrin molecules. It is generally assumed that such functionalization does not alter the local mechanical properties of the functionalized surface, as is important to interpretations of macromolecular mechanotransduction in cells. Here, we examine this assumption systematically, through nanomechanical measurement of the nominal elastic modulus of polymer multilayer films of nanoscale thickness, functionalized with RGD through different processing routes. We find that the method of biochemical functionalization can significantly alter mechanical compliance of polymeric substrata such as weak polyelectrolyte multilayers (PEMs), increasingly utilized materials for such studies. In particular, immersed adsorption of intermediate functionalization reagents significantly decreases compliance of the PEMs considered herein, whereas polymer-on-polymer stamping of these same reagents does not alter compliance of weak PEMs. This finding points to the potential unintended alteration of mechanical properties via surface functionalization and also suggests functionalization methods by which chemical and mechanical properties of cell substrata can be controlled independently.