Influence of Ionic Strength and Specific Ion Effects on Polyelectrolyte Multilayer Films with pH-Responsive Behavior.

Layer-by-layer assembled multilayer films have shown great potential for different applications owing to their responsive behavior. Herein, we systematically investigated the effects of composition, salt concentration, and ion specificity on the pH responsiveness of covalently crosslinked chitosan and alginate dialdehyde multilayer films. The changes in film swelling were measured using ellipsometry from low (0.01 mM) to high (3 M) salt (NaCl or NaSCN) concentrations at pH 3, 6, and 9. The swelling responses to increasing ionic strength matched the swelling responses observed for polyzwitterionic and weak monocomponent polyelectrolyte films and depended on the multilayer composition, pH, and ion specificity. Finally, we used the ellipsometric data to demonstrate that the pH responsiveness of such multilayer films, as measured using a quartz crystal microbalance with dissipation monitoring, strongly depends on the ionic condition under which the responses were measured. We thus show that erroneous conclusions about the pH responsiveness of polyelectrolyte multilayer films can be easily obtained if the ionic environment of the application does not closely resemble the ionic condition under which the pH responsiveness is tested.

Self-Cross-Linkable Chitosan-Alginate Complexes Inspired by Mussel Glue Chemistry.

In this study, mussel-inspired chemistry, based on catechol-amine reactions, was adopted to develop self-cross-linkable chitosan-alginate (Chi-Alg) complexes. To do so, the biopolymers were each substituted with ∼20% catechol groups (ChiC and AlgC), and then four complex combinations (Chi-Alg, ChiC-Alg, Chi-AlgC, ChiC-AlgC) were prepared at the surface and in bulk solution. Based on QCM-D and lap shear adhesion tests, the complex with catechol only on Chi (ChiC-Alg) did not show a significant variation from the control complex (Chi-Alg). Conversely, the complexes with catechol on alginate (Chi-AlgC and ChiC-AlgC) rendered a self-cross-linking property and enhanced cohesive properties.

Ion-Specific Antipolyelectrolyte Effect on the Swelling Behavior of Polyzwitterionic Layers.

In this study, we systematically investigate the interactions between mobile ions generated from added salts and immobile charges within a sulfobetaine-based polyzwitterionic film in the presence of five salts (KCl, KBr, KSCN, LiCl, and CsCl). The sulfobetaine groups contain quaternary alkylammonium and sulfonate groups, giving the positive and negative charges. The swelling of the zwitterionic film in the presence of different salts is compared with the swelling behavior of a polycationic or polyanionic film containing the same charged groups. For such a comparative study, we design cross-linked terpolymer films with similar thicknesses, cross-link densities, and charge fractions, but with varying charged moieties. While the addition of salt in general leads to a collapse of both cationic and anionic films, the presence of specific types of mobile anions (Cl-, Br-, and SCN-) considerably influences the swelling behavior of polycationic films. We attribute this observation to a different degree of ion-pair formations between the different types of anionic counterions and the immobile cationic quaternary alkylammonium groups in the films where highly polarizable counterions such as SCN- lead to a high degree of ion pairing and less polarizable counterions, such as Cl-, cause a low degree of ion pairing. Conversely, we do not observe any substantial effect of varying the type of cationic counterions (K+, Li+, and Cs+), which we assign to the lack of ion pairing between the weakly polarizable cations and the immobile anionic sulfonate groups in the films. In addition, we observe that the zwitterionic films swell with increasing ionic strength and the degree of swelling is anion dependent, which is in agreement with previous reports on the "antipolyelectrolyte effect". Herein, we explain this ion-specific swelling behavior with the different cation and anion abilities to form ion pairs with quaternary alkylammonium and sulfonate in the sulfobetaine groups.

Fibrin formation and fractal organization at cationic, anionic, and zwitterionic polymer coated interfaces.

Biomaterial-associated thrombosis remains a persistent challenge whenever medical devices are inserted in blood vessels. The issue is principally addressed by the development of antithrombogenic coatings that prevent the formation of blood clots, e.g. by limiting adsorption of fibrin - the core protein network of a clot. Charged polymers (i.e. polyelectrolytes and zwitterionic polymers) show potential as coating materials for medical devices, and we here investigate these polymer coatings in the context of biomaterial-associated thrombosis. Our findings indicate that fibrin polymerization can yield a surface-dependent distribution of fractal-like branched structures and amorphous aggregates, with surface-induced fibrin formation dominating for anionic polymer interfaces and recruitment of bulk-formed fibrin dominating for cationic polymer interfaces. In addition, we identify coatings containing zwitterionic sulfobetaine groups as promising candidates for antithrombogenic biomaterials.

Enhancing the sweat resistance of sunscreens.

BACKGROUND: While sunbathing of performing outdoor sport activities, sunscreens are important for protection of uncovered skin against ultraviolet (UV) radiation. However, perspiration negatively affects the performance of a sunscreen film by weakening its substantivity and uniformity through the activation of two mechanisms, namely sunscreen wash-off and sunscreen redistribution. MATERIAL AND METHODS: We used a perspiring skin simulator to investigate the effect of sunscreen formulation on its efficiency upon sweating. Specifically, we modified the sunscreen formulation by incorporating a hydrophobic film former and adding water-absorbing particles. Sunscreen performance before and after perspiration is assessed by in vitro sun protection factor measurements, direct detection of changes in the sunscreen distribution using UV reflectance imaging, and by coherent anti-Stokes Raman scattering (CARS) microscopy for microscopic characterization of the UV filter relocation. RESULTS: The results show that incorporating a hydrophobic film former can decrease sunscreen wash-off due to sweating, while an excessive amount of film former might negatively affect the sunscreen distribution. The addition of water-absorbing particles, on the other hand, had either a negative or positive impact on the sunscreen substantivity, depending on the particle properties. While the addition of large water-absorbing particles appeared to increase sunscreen redistribution, smaller particles that could form a gel-like structure upon contact with water, appeared to change sunscreen wetting and sweat droplet spreading, thereby decreasing sunscreen wash-off and sunscreen redistribution. CONCLUSIONS: We find that using a combination of hydrophobic film formers, which increase water resistance, and small water-absorbing particles, which change the wetting behavior, can make sunscreen formulations more sweat-resistant and less runny.

Parameterization of the optical constants of polydopamine films for spectroscopic ellipsometry studies.

Bio-inspired polydopamine coatings offer vast possibilities for surface modification of materials. The thickness of such nanometric coatings is usually estimated based on ellipsometry measurements. However, the complex light-absorbing nature of polydopamine is often overlooked when analyzing such data, which can result in inaccurate estimations of the coating thickness as well as the optical properties. In this study, we prepared and characterized three polydopamine coatings where the film thickness and surface roughness are systematically varied. For each case, we developed suitable optical models and showed how an inappropriate optical model can provide inaccurate estimates of the coating properties. AFM height profiles were obtained from scratched areas of each sample to verify the thickness values estimated by ellipsometry. The results confirm that polydopamine coatings, depending on the oxidation conditions, can possess different structural and optical properties, and thus require unique optical models for the ellipsometry analysis.

Water vapor permeation through topical films on a moisture-releasing skin Model.

BACKGROUND: Covering the skin by topical films affects the skin hydration and transepidermal water loss (TEWL). In vivo studies to investigate the water vapor permeation through topical films are complicated, expensive, ethically not preferred, and time- and labor-consuming. The objective of this study was to introduce an in vitro and subject-independent alternative evaluation method to predict the breathability of topical formulations. METHODS: In this study, we developed an in vitro setup to simulate the TEWL values of human skin and investigated the breathability of five polymeric film formers used in topical formulations. Furthermore, a comparative in vivo TEWL study was performed on ten human volunteers with defined areas of skin covered with films of two selected polymers possessing different barrier properties. RESULTS: By employing the in vitro setup, a vinylpyrrolidone/acrylates/lauryl methacrylate copolymer was determined to form the most breathable film, whereas acrylates/octylacrylamide copolymer and shellac films showed the highest barrier properties. The in vivo TEWL study demonstrated the same relative barrier properties for the acrylates/octylacrylamide and polyurethane-64 films, despite a more complex driving force for water vapor permeation due to moisture accumulation on the covered skin surfaces. CONCLUSION: We obtained a good correlation between the in vitro and in vivo results, demonstrating that our model can categorize different polymeric film formers based on their breathability when applied to human skin. This information can aid in selecting suitable film-forming polymers for topical formulations with either breathable or occluding functionalities.

In vitro skin model for characterization of sunscreen substantivity upon perspiration.

OBJECTIVE: The resistance of sunscreens to the loss of ultraviolet (UV) protection upon perspiration is important for their practical efficacy. However, this topic is largely overlooked in evaluations of sunscreen substantivity due to the relatively few well-established protocols compared to those for water resistance and mechanical wear. METHODS: In an attempt to achieve a better fundamental understanding of sunscreen behaviour in response to sweat exposure, we have developed a perspiring skin simulator, containing a substrate surface that mimics sweating human skin. Using this perspiring skin simulator, we evaluated sunscreen performance upon perspiration by in vitro sun protection factor (SPF) measurements, optical microscopy, ultraviolet (UV) reflectance imaging and coherent anti-Stokes Raman scattering (CARS) microscopy. RESULTS AND CONCLUSION: Results indicated that perspiration reduced sunscreen efficiency through two mechanisms, namely sunscreen wash-off (impairing the film thickness) and sunscreen redistribution (impairing the film uniformity). Further, we investigated how the sweat rate affected these mechanisms and how sunscreen application dose influenced UV protection upon perspiration. As expected, higher sweat rates led to a large loss of UV protection, while a larger application dose led to larger amounts of sunscreen being washed-off and redistributed but also provided higher UV protection before and after sweating.

OBJECTIF: La résistance des écrans solaires à la perte de protection contre les ultraviolets (UV) à cause de la transpiration est importante quant à leur efficacité pratique. Cependant, ce point est généralement négligé dans les évaluations de la substantivité des écrans solaires en raison du nombre relativement faible de protocoles bien établis, en comparaison avec ceux pour la résistance à l'eau et l'usure mécanique. MÉTHODES: Dans le but de parvenir à une meilleure compréhension fondamentale du comportement des écrans solaires en cas d'exposition à la sueur, nous avons développé un simulateur de peau transpirante, dont la surface de substrat imite la transpiration de la peau humaine. À l'aide de ce simulateur, nous avons évalué les performances des écrans solaires lors de la transpiration par des mesures in vitro du facteur de protection solaire (FPS), par microscopie optique, par imagerie de la réflectance ultraviolette (UV) et par microscopie cohérente de diffusion Raman anti-Stokes (coherent anti-Stokes Raman scattering, CARS). RÉSULTATS ET CONCLUSION: Les résultats ont montré que la transpiration réduisait l'efficacité de l'écran solaire en raison de deux mécanismes, à savoir le lavage de l'écran solaire (altération de l'épaisseur du film) et la redistribution de l'écran solaire (altération de l'uniformité du film). De plus, nous avons étudié comment le taux de transpiration affectait ces mécanismes et comment la dose d'application d'écran solaire influençait la protection UV en cas de transpiration. Comme l'on pouvait s'y attendre, des taux de sueur plus élevés ont entraîné une perte importante de protection contre les UV, tandis qu'une dose d'application plus importante a conduit à des quantités plus importantes d'écran solaire lavé et redistribué, mais a également fourni une protection contre les UV plus élevée avant et après la transpiration.

Swelling Behavior, Interaction, and Electrostatic Properties of Chitosan/Alginate Dialdehyde Multilayer Films with Different Outermost Layer.

In this study, self-cross-linked chitosan/alginate dialdehyde multilayer films, capped with either alginate dialdehyde (6 layers) or chitosan (7 layers), were fabricated using the layer-by-layer method. The disruption of the electrostatic equilibrium when exposing the fabricated layers to acidic and alkaline conditions causes swelling within the film and independently in the outermost layer, showing dependence on the ionic strength. Spectroscopic ellipsometry and quartz crystal microbalance with dissipation monitoring were employed to examine the swelling behavior. Atomic force microscopy colloidal probe measurements were conducted to assess the surface forces between the multilayer films at different pH and ionic strengths. Finally, the electrostatic properties of the multilayer films were examined at different pH and ionic strengths using zeta potential measurements. The results suggest that stimuli-responsiveness and overall swelling behavior of the polysaccharide multilayer films significantly depend on the outermost layer, an effect that should expectedly become more pronounced the thinner the film becomes.

How Dissociation of Carboxylic Acid Groups in a Weak Polyelectrolyte Brush Depend on Their Distance from the Substrate.

A weak polyelectrolyte brush is composed of a layer of polyacids or polybases grafted by one end of their chains to a substrate surface. For such brush layers immersed in an aqueous solution, the dissociation behavior of the acidic or basic groups and the structural and physical properties of the brush layer will thus be strongly dependent on the environmental conditions. For a polyacid brush layer consisting of, e.g., poly(acrylic acid), this means that the chains in the brush layer will be charged at high pH and uncharged at low pH. However, theoretical scaling laws not only foresee the structural changes occurring in response to the pH-induced dissociation behavior but also how the dissociation behavior of the brush layer depends on the ionic strength of the aqueous solution and the density of acidic groups within the brush layer. We have herein employed spectroscopic ellipsometry and a quartz crystal microbalance with dissipation monitoring (QCM-D) to experimentally evaluate the theoretically predicted dissociation and structural behavior of PAA brushes. Spectroscopic ellipsometry allows us to study the brush thickness as a function of pH and ionic strength, while QCM-D gives us an opportunity to investigate the swelling behavior of PAA brushes at various penetration depths of propagating acoustic waves. Our studies show that the dissociation degree of the carboxylic acid groups in a PAA brush increases with increasing distance from the substrate. Moreover, the ionic strength enhances carboxylic acid dissociation, such that a higher ionic strength leads to a narrower distribution and higher average dissociation degree. In conclusion, our results provide an experimental verification of the theoretically predicted gradient in the degree of dissociation of the acid groups in weak polyacid brush layers and shows that at a pH value equal to approximately the average pKa value of the brush, the state of the acid groups varies from being almost uncharged to almost fully dissociated depending on the ionic strength and vertical position in the brush.

Water transport in polymer composites through swelling-induced networks of hydrogel particles.

Water diffusion in polymer composites is not only affected by the chemical nature of the materials but also by their internal structures. To enable the design of polymer composites with controlled diffusion kinetics, we investigate the effect of hydrogel particle networks on the water transport. The composites in this study comprise hydrogel particles based on sodium poly(acrylic acid), which are incorporated at different concentrations into a soft and sticky polymer matrix. Through the use of X-ray micro computed tomography, the internal structure of the polymer composites is examined and the interparticle distances are calculated. The structure of the composites is then related to the water diffusion kinetics upon exposure to saline solution as well as humid air. Even though the hydrogel particles are isolated and the interparticle distances are in the order of several micrometers, a sudden increase in the water diffusion kinetics is observed above a critical concentration. Due to the low water permeability of the matrix, such a change in the water diffusion kinetics is indicative of network formation. During hydration, swelling enables the hydrogels to overcome the interparticle distances and form a network for water transport.

A novel starch-binding laccase from the wheat pathogen Zymoseptoria tritici highlights the functional diversity of ascomycete laccases.

BACKGROUND: Laccases are multicopper oxidases, which are assigned into auxiliary activity family 1 (AA1) in the CAZy database. These enzymes, catalyzing the oxidation of phenolic and nonphenolic substrates coupled to reduction of O2 to H2O, are increasingly attractive as eco-friendly oxidation biocatalysts. Basidiomycota laccases are well characterized due to their potential in de-lignification of lignocellulose. By contrast, insight into the biochemical diversity of Ascomycota counterparts from saprophytes and plant pathogens is scarce. RESULTS: Here, we report the properties of the laccase from the major wheat pathogen Zymoseptoria tritici (ZtrLac1A), distinguished from common plant fungal pathogens by an apoplastic infection strategy. We demonstrate that ZtrLac1A is appended to a functional starch-binding module and displays an activity signature disfavoring relatively apolar phenolic redox mediators as compared to the related biochemically characterized laccases. By contrast, the redox potential of ZtrLac1A (370 mV vs. SHE) is similar to ascomycetes counterparts. The atypical specificity is consistent with distinctive sequence substitutions and insertions in loops flanking the T1 site and the enzyme C-terminus compared to characterized laccases. CONCLUSIONS: ZtrLac1A is the first reported modular laccase appended to a functional starch-specific carbohydrate binding module of family 20 (CBM20). The distinct specificity profile of ZtrLac1A correlates to structural differences in the active site region compared to previously described ascomycetes homologues. These differences are also highlighted by the clustering of the sequence of ZtrLac1A in a distinct clade populated predominantly by plant pathogens in the phylogenetic tree of AA1 laccases. The possible role of these laccases in vivo merits further investigations. These findings expand our toolbox of laccases for green oxidation and highlight the binding functionality of CBM-appended laccases as versatile immobilization tags.

Salt-Induced Control of the Grafting Density in Poly(ethylene glycol) Brush Layers by a Grafting-to Approach.

In this work, a method to obtain control of the grafting density during the formation of polymer brush layers by the grafting-to method of thiolated poly(ethylene glycol) onto gold is presented. The grafting density of the polymer chains was adjusted by adding Na2SO4 in concentrations between 0.2 and 0.9 M to the aqueous polymer solution during the grafting process. The obtained grafting densities ranged from 0.26 to 1.60 chains nm-2, as determined by surface plasmon resonance. The kinetics of the grafting process were studied in situ by a quartz crystal microbalance with dissipation, and a mushroom to brush conformational transition was observed when the polymer was grafted in the presence of Na2SO4. The transition from mushroom to brush was only observed for long periods of grafting, highlighting the importance of time to obtain high grafting densities. Finally, the prepared brush layer with the highest grafting density showed high resistance to the adsorption of bovine serum albumin, while layers with a lower grafting density showed only limited resistance.

Surface Forces between Highly Charged Cationic Polyelectrolytes Adsorbed to Silica: How Control of pH and the Adsorbed Amount Determines the Net Surface Charge.

Atomic force microscopy (AFM) and quartz crystal microbalance with dissipation (QCM-D) were employed to investigate the pH-dependent adsorption of poly(diallyldimethylammonium chloride) (polyDADMAC) to silica surfaces as well as the surface forces between these layers. It was found that polyDADMAC adopted a relatively flat conformation when adsorbed to a silica surface and that the adsorbed amount increased with increasing pH. From the surface force measurements it is evident that the surface undergoes a charge reversal upon saturation with polyDADMAC, at the three different investigated pH values, and that some degree of charge regulation of the silica surface takes place during the adsorption process. Finally, the overcharging phenomenon is discussed in terms of a geometrical mismatch due to the different average spacing between the surface charges on the silica surface and the size of the DADMAC monomer.

Hofmeister Effect on PNIPAM in Bulk and at an Interface: Surface Partitioning of Weakly Hydrated Anions.

The effect of sodium fluoride, sodium trichloroacetate, and sodium thiocyanate on the stability and conformation of poly(N-isopropylacrylamide), in bulk solution and at the gold-aqueous interface, is investigated by differential scanning calorimetry, dynamic light scattering, quartz crystal microbalance, and atomic force microscopy. The results indicate a surface partitioning of the weakly hydrated anions, i.e., thiocyanate and trichloroacetate, and the findings are discussed in terms of anion-induced electrostatic stabilization. Although attractive polymer-ion interactions are suggested for thiocyanate and trichloroacetate, a salting-out effect is found for sodium trichloroacetate. This apparent contradiction is explained by a combination of previously suggested mechanisms for the salting-out effect by weakly hydrated anions.

Electrical Double-Layer and Ion Bridging Forces between Symmetric and Asymmetric Charged Surfaces in the Presence of Mono- and Divalent Ions.

An atomic force microscope, employing the colloidal probe technique, was used to study the interactions between six different combinations of silane-functionalized silica surfaces in NaCl and CaCl2 solutions. The surfaces consisted of monolayers of the apolar trimethoxy(octyl)silane, the positively charged (3-aminopropyl)trimethoxysilane, and the negatively charged (3-mercaptopropyl)trimethoxysilane. The interactions between the three symmetric systems, as well as between the three asymmetric combinations of surfaces, were measured and compared to calculated electrical double-layer forces. The results demonstrated that the long-range interactions between the surfaces in all cases were dominated by double-layer forces, while short-range interactions, including adhesion, were dominated by ion bridging forces in the cases where both interaction surfaces favored adsorption of calcium ions. The study thus also demonstrates how surface force studies in mono- and divalent salt solutions can be used as an analytical tool for probing specific functional groups on heterogeneous surfaces.

Hofmeister effect of salt mixtures on thermo-responsive poly(propylene oxide).

The Hofmeister series is a classification of ions regarding their ability to stabilize or destabilize aqueous solutions of proteins, polymers and other molecules which are partly miscible with water. In this study, we employ differential scanning calorimetry to investigate how the stability of aqueous solutions of poly(propylene oxide) is affected by mixtures of ions with different location in the Hofmeister series. Our results show that the Hofmeister effects of pure salt species are not always linearly additive and that the relative effect of some ions can be reversed depending on the composition of the salt mixture as well as by the absolute and relative concentration of the different species. We suggest that these results can lead to a better understanding of the potential role of the Hofmeister effect in regulation of biological processes, which does always take place in salt mixtures rather than solutions containing just single salt species.

Direct measurement of colloidal interactions between polyaniline surfaces in a UV-curable coating formulation: the effect of surface hydrophilicity/hydrophobicity and resin composition.

The interactions between polyaniline particles and polyaniline surfaces in polyester acrylate resin mixed with 1,6-hexanediol diacrylate monomer have been investigated using contact angle measurements and the atomic force microscopy colloidal probe technique. Polyaniline with different characteristics (hydrophilic and hydrophobic) were synthesized directly on spherical polystyrene particles of 10 µm in diameter. Surface forces were measured between core/shell structured polystyrene/polyaniline particles (and a pure polystyrene particle as reference) mounted on an atomic force microscope cantilever and a pressed pellet of either hydrophilic or hydrophobic polyaniline powders, in resins of various polymer:monomer ratios. A short-range purely repulsive interaction was observed between hydrophilic polyaniline (doped with phosphoric acid) surfaces in polyester acrylate resin. In contrast, interactions between hydrophobic polyaniline (doped with n-decyl phosphonic acid) were dominated by attractive forces, suggesting less compatibility and higher tendency for aggregation of these particles in liquid polyester acrylate compared to hydrophilic polyaniline. Both observations are in agreement with the conclusions from the interfacial energy studies performed by contact angle measurements.

Surface grafted chitosan gels. Part II. Gel formation and characterization.

Responsive biomaterial hydrogels attract significant attention due to their biocompatibility and degradability. In order to make chitosan based gels, we first graft one layer of chitosan to silica, and then build a chitosan/poly(acrylic acid) multilayer using the layer-by-layer approach. After cross-linking the chitosan present in the polyelectrolyte multilayer, poly(acrylic acid) is partly removed by exposing the multilayer structure to a concentrated carbonate buffer solution at a high pH, leaving a surface-grafted cross-linked gel. Chemical cross-linking enhances the gel stability against detachment and decomposition. The chemical reaction between gluteraldehyde, the cross-linking agent, and chitosan was followed in situ using total internal reflection Raman (TIRR) spectroscopy, which provided a molecular insight into the complex reaction mechanism, as well as the means to quantify the cross-linking density. The amount of poly(acrylic acid) trapped inside the surface grafted films was found to decrease with decreasing cross-linking density, as confirmed in situ using TIRR, and ex situ by Fourier transform infrared (FTIR) measurements on dried films. The responsiveness of the chitosan-based gels with respect to pH changes was probed by quartz crystal microbalance with dissipation (QCM-D) and TIRR. Highly cross-linked gels show a small and fully reversible behavior when the solution pH is switched between pH 2.7 and 5.7. In contrast, low cross-linked gels are more responsive to pH changes, but the response is fully reversible only after the first exposure to the acidic solution, once an internal restructuring of the gel has taken place. Two distinct pKa's for both chitosan and poly(acrylic acid), were determined for the cross-linked structure using TIRR. They are associated with populations of chargeable groups displaying either a bulk like dissociation behavior or forming ionic complexes inside the hydrogel film.

Surface grafted chitosan gels. Part I. Molecular insight into the formation of chitosan and poly(acrylic acid) multilayers.

Composite polyelectrolyte multilayers of chitosan and low molecular weight poly(acrylic acid) (PAA) have been assembled by sequential adsorption as a first step toward building a surface anchored chitosan gel. Silane chemistry was used to graft the first chitosan layer to prevent film detachment and decomposition. The assembly process is characterized by nonlinear growth behavior, with different adsorption kinetics for chitosan and PAA. In situ analysis of the multilayer by means of surface sensitive total internal reflection Raman (TIRR) spectroscopy, combined with target factor analysis of the spectra, provided information regarding composition, including water content, and ionization state of weak acidic and basic groups present in the thin composite film. Low molecular weight PAA, mainly in its protonated form, diffuses into and out of the composite film during adsorption and rinsing steps. The higher molecular weight chitosan shows a similar behavior, although to a much lower extent. Our data demonstrate that the charged monomeric units of chitosan are mainly compensated by carboxylate ions from PAA. Furthermore, the morphology and mechanical properties of the multilayers were investigated in situ using atomic force microscopy operating in PeakForce tapping mode. The multilayer consists of islands that grow in lateral dimension and height during the build-up process, leading to close to exponentially increasing roughness with deposition number. Both diffusion in and out of at least one of the two components (PAA) and the island-like morphology contribute to the nonlinear growth of chitosan/PAA multilayers.