Rapid Probing of Biological Surfaces with a Sparse-Matrix Peptide Library

Finding unique peptides to target specific biological surfaces is crucial to basic research and technology development, though methods based on biological arrays or large libraries limit the speed and ease with which these necessary compounds can be found. We reasoned that because biological surfaces, such as cell surfaces, mineralized tissues, and various extracellular matrices have unique molecular compositions, they present unique physicochemical signatures to the surrounding medium which could be probed by peptides with appropriately corresponding physicochemical properties. To test this hypothesis, a naïve pilot library of 36 peptides, varying in their hydrophobicity and charge, was arranged in a two-dimensional matrix and screened against various biological surfaces. While the number of peptides in the matrix library was very small, we obtained “hits” against all biological surfaces probed. Sequence refinement of the “hits” led to peptides with markedly higher specificity and binding activity against screened biological surfaces. Genetic studies revealed that peptide binding to bacteria was mediated, at least in some cases, by specific cell-surface molecules, while examination of human tooth sections showed that this method can be used to derive peptides with highly specific binding to human tissue

SPECIFIC AND NONSPECIFIC INTERACTIONS IN BACTERIAL ADHESION TO SOLID SUBSTRATA

Based on a literature review, a hypothesis is forwarded on the mechanism of initial bacterial adhesion to solid substrata, which accounts both for the role of specific microscopic surface components as well as for the role of non-specific macroscopic surface properties (surface free energy, zeta potential or hydrophobicity). Three distinct regions in the adhesion process are suggested in which at large and intermediate separation distances adhesion is mediated by the macroscopic surface properties as surface free energy and surface charge, respectively. At small separation distances specific short-range interactions can occur, leading to a strong and irreversible bonding, provided the water film present in between the interaction surfaces can be removed. A major role of hydrophobic groups, supposed to be associated with bacterial surface appendages is suggested to be its dehydrating capacity, enabling the removal of the vicinal water film yielding small areas of direct contact between protruberant parts of the cell surface and the substratum

EFFECT OF ZETA-POTENTIAL AND SURFACE-ENERGY ON BACTERIAL ADHESION TO UNCOATED AND SALIVA-COATED HUMAN-ENAMEL AND DENTIN

Physicochemical surface characteristics of early plaque-forming bacteria and of human tooth surfaces were measured to establish their role in bacterial adhesion to intact dental tissue slabs. In addition, the influence of an experimental salivary pellicle was evaluated. Strains of S. mutans, S. sanguis, S. salivarius, A. viscosus, and A. odontolyticus showed relatively high surface free energies (range, 99-128 mJ.m-2) and carried a negative surface charge, at both physiological (μ = 0.057) and low (μ = 0.020) ionic strengths of the medium. Very large differences in hydrophobicity were detected when the hexadecane adsorption test was used for measurement. Powdered enamel and dentin were also negatively charged at low ionic strength but were slightly positively charged in the physiological buffer. The surface free energy of enamel and dentin increased upon saliva coating, whereas the surface charge was always negative. The adhesion experiments showed: (1) large differences in the binding of various bacteria to the same surface; (2) an up to 20-fold difference in the binding of the same bacterium to different surfaces, although the binding of some strains was relatively independent of the type of surface or presence of a salivary pellicle; (3) a significant decrease in adhesion when the ionic strength of the medium was lowered, due to increased electrostatic repulsion (however, the adhesion of some bacteria was independent of the ionic strength of the medium); (4) different time-dependent adherence kinetics, depending on both the bacteria and nature of the solid surface; and (5) a propensity for plaque streptococci to bind to uncoated dentin. </jats:p