Atomic Force Microscopy Study of Human Tooth Enamel Surfaces

Human enamel features from individual crystals up to prisms were observed by atomic force microscopy (AFM). Low magnification images of vestibular tooth surfaces show the existence of enamel prisms appearing as deep holes. Individual, parallel enamel crystals show lateral faces elongated and formed by the (100) planes of hydroxyapatite (HA). Height differences between (001) faces create the roughness of enamel surface. Individual (001) crystal faces can be observed clearly at higher magnification and show the characteristic hexagonal shape with 60° angles between (100) faces. This study confirms the applicability of AFM for studying biological hydroxyapatite crystals

Determination of structural parameters characterizing thin films by optical methods: A comparison between scanning angle reflectometry and optical waveguide lightmode spectroscopy

International audienceWe present a comparative study of the structural parameters characterizing thin macromolecular adsorbed films that are obtained from two optical techniques: optical waveguide lightmode spectroscopy ͑OWLS͒ and scanning angle reflectometry ͑SAR͒. We use polyelectrolyte multilayers and polyelectrolyte multilayers/protein films to perform this study. The comparison between the information obtained with the two methods is possible because the buildup of the polyelectrolyte multilayers is known to become substrate independent after the deposition of the first few polyelectrolyte layers. The analysis of the optical data requires usually to postulate a refractive index profile for the interface. Two profiles have been used: the homogeneous and isotropic monolayer and the bilayer profiles. When the refractive index profile of an adsorbed film is well approximated by a homogeneous and isotropic monolayer, as shown by using an analysis of the deposited films in terms of optical invariants, the two optical techniques lead to similar values for the film thickness and the optical mass. The situation is more complex in the case of the multilayers/protein films for which the calculated parameters can strongly depend upon the refractive index profile that is postulated to analyze the optical data. Whereas the optical mass and, to a lesser extent, the thickness seem fairly model independent for OWLS, they appear to be extremely sensitive to the model for SAR. For proteins deposited on top of the polyelectrolyte film, optical mass and protein thickness were found to be comparable when determined by OWLS and by SAR using the bilayer model. The data analysis of the SAR curves with the monolayer model leads to much larger and even physically unreasonable film thicknesses and optical masses. This was particularly noticeable for proteins having a large size ͑human serum albumin and fibrinogen͒, whereas both models lead to similar results for small sized proteins. By means of the different refractive index profiles, we show that great care must be taken in the physicochemical interpretation of the structural parameters determined by these optical techniques

Polyelectrolyte multilayer films with pegylated polypeptides as a new type of anti-microbial protection for biomaterials.

Adhesion of bacteria at the surface of implanted materials is the first step in microbial infection, leading to post-surgical complications. In order to reduce this adhesion, we show that poly(L-lysine)/poly(L-glutamic acid) (PLL/PGA) multilayers ending by several PLL/PGA-g-PEG bilayers can be used, PGA-g-PEG corresponding to PGA grafted by poly(ethylene glycol). Streaming potential and quartz crystal microbalance-dissipation measurements were used to characterize the buildup of these films. The multilayer films terminated by PGA and PGA-g-PEG were found to adsorb an extremely small amount of serum proteins as compared to a bare silica surface but the PGA ending films do not reduce bacterial adhesion. On the other hand, the adhesion of Escherichia coli bacteria is reduced by 72% on films ending by one (PLL/PGA-g-PEG) bilayer and by 92% for films ending by three (PLL/PGA-g-PEG) bilayers compared to bare substrate. Thus, our results show the ability of PGA-g-PEG to be inserted into multilayer films and to drastically reduce both protein adsorption and bacterial adhesion. This kind of anti-adhesive films represents a new and very simple method to coat any type of biomaterials for protection against bacterial adhesion and therefore limiting its pathological consequences.comparative studyevaluation studiesjournal articleresearch support, non-u.s. gov't2004 Mayimporte

Effect of hydrodynamic interactions on the distribution of adhering Brownian particles

Brownian dynamics simulations were used to study the adhesion of hard spheres on a solid surface by taking the hydrodynamic interactions into account. Special attention was paid to analyze the configuration of the assembly of adsorbed particles. These results were compared to configurations generated by the extensively studied random sequential adsorption (RSA) model. In our case the adsorption probability for a particle is almost uniform over the entire available surfae. This surprising result shows that RSA provides a good approximation to generate adsorbed particle configurations

Improvement of stability and cell adhesion properties of polyelectrolyte multilayer films by chemical cross-linking.

Poly(L-lysine)/hyaluronan (PLL/HA) films were chemically cross-linked with a water soluble carbodiimide (EDC) in combination with a N-hydroxysulfo-succinimide (NHS) to induce amide formation. Fourier transform infrared spectroscopy confirms the conversion of carboxylate and ammonium groups into amide bonds. Quartz crystal microbalance-dissipation reveals that the cross linking reaction is accompanied by a change in the viscoelastic properties of the films leading to more rigid films. After the cross-linking reaction, both positively and negatively ending films exhibit a negative zeta potential. It is shown by fluorescence recovery after photobleaching measured by confocal laser scanning microscopy that cross-linking dramatically reduces the diffusion of the PLL chains in the network. Cross linking also renders the films highly resistant to hyaluronidase, an enzyme that naturally degrades hyaluronan. Finally, the adhesion of chondrosarcoma cells on the films terminating either with PLL or HA is also investigated. Whereas the non cross-linked films are highly resistant to cell adhesion, the cells adhere and spread well on the cross-linked films.comparative studyjournal articleresearch support, non-u.s. gov't2004 Mar-Aprimporte

Collagen-based fibrillar multilayer films cross-linked by a natural agent.

Surface functionalization plays an important role in the design of biomedical implants, especially when layer forming cells, such as endothelial or epithelial cells, are needed. In this study, we define a novel nanoscale surface coating composed of collagen/alginate polyelectrolyte multilayers and cross-linked for stability with genipin. This buildup follows an exponential growth regime versus the number of deposition cycles with a distinct nanofibrillar structure that is not damaged by the cross-linking step. Stability and cell compatibility of the cross-linked coatings were studied with human umbilical vein endothelial cells. The surface coating can be covered by a monolayer of vascular endothelial cells within 5 days. Genipin cross-linking renders the surface more suitable for cell attachment and proliferation compared to glutaraldehyde (more conventional cross-linker) cross-linked surfaces, where cell clumps in dispersed areas were observed. In summary, it is possible with the defined system to build fibrillar structures with a nanoscale control of film thickness, which would be useful for in vivo applications such as inner lining of lumens for vascular and tracheal implants.journal articleresearch support, non-u.s. gov't2012 Jul 092012 06 13importe

Polysaccharide films built by simultaneous or alternate spray: a rapid way to engineer biomaterial surfaces.

We investigated polysaccharide films obtained by simultaneous and alternate spraying of a chitosan (CHI) solution as polycation and hyaluronic acid (HA), alginate (ALG), and chondroitin sulfate (CS) solutions as polyanions. For simultaneous spraying, the film thickness increases linearly with the cumulative spraying time and passes through a maximum for polyanion/CHI molar charge ratios lying between 0.6 and 1.2. The size of polyanion/CHI complexes formed in solution was compared with the simultaneously sprayed film growth rate as a function of the polyanion/CHI molar charge ratio. A good correlation was found. This suggests the importance of polyanion/polycation complexation in the simultaneous spraying process. Depending on the system, the film topography is either liquid-like or granular. Film biocompatibility was evaluated using human gingival fibroblasts. A small or no difference is observed in cell viability and adhesion between the two deposition processes. The CHI/HA system appears to be the best for cell adhesion inducing the clustering of CD44, a cell surface HA receptor, at the membrane of cells. Simultaneous or alternate spraying of CHI/HA appears thus to be a convenient and fast procedure for biomaterial surface modifications.journal articleresearch support, non-u.s. gov't2012 Jun 052012 05 23importe