Amphiphilic multi-arm star-block copolymers for encapsulation of fragrance molecules

Novel amphiphilic multi-arm star-block copolymers with a hyperbranched core, a hydrophobic inner shell, and a hydrophilic outer shell have been prepared from a commercial hyperbranched polyester macroinitiator by ring-opening polymerization of -caprolactone, followed by atom transfer radical polymerization of tert-butyl acrylate (tBuA). Hydrolysis of the tert-butyl groups was then used to convert the poly(tBuA) blocks to poly(acrylic acid), resulting in stable amphiphilic core-shell structures with significantly higher degrees of functionality than reported so far in the literature. A strong correlation between the maximum concentration of selected hydrophobic guest molecules and the concentration of amphiphilic star-block copolymer in aqueous solution was observed by 1H NMR, demonstrating the capacity of these copolymers to encapsulate and disperse significant loadings (up to about 27 wt.-%) of volatile hydrophobic molecules such as fragrances in water.

Design of amphiphilic nanocapsules based on hyperbranched star-block copolymers for the encapsulation and the controlled release of olfactory compounds

Céline Ternat

Effective encapsulation of small, volatile, weakly water-soluble molecules such as flavors and fragrances is necessary to protect them from degradation, to increase their lifetime, and to improve their water dispersion and fixation depending on the substrate of interest. The aim of this project has been to improve encapsulation and release in order to optimize the performance of fragrance molecules in water-based household (detergents, softeners, etc.) and body care applications (shampoos, lotions, etc.), and fine perfumery applications. More specifically, the aim has been to investigate the effectiveness of "unimolecular micelles" based on amphiphilic multi-arm star-block copolymers with a hyperbranched core with more than 26 functional groups, a hydrophobic inner and a hydrophilic outer shell. These have been prepared from a commercial hyperbranched polyester macroinitiator (HBP) by ring-opening polymerization of ε-caprolactone, followed by the atom transfer radical polymerization (ATRP) of tert-butyl acrylate (tBuA). Hydrolysis of the tert-butyl groups has then been used to convert the poly(tBuA) blocks to poly(acrylic acid) (PAA), resulting in HBP-(PCL)p-(PAA)q pH-dependent amphiphilic star-block copolymers with good control of the molecular weight distribution. These were shown to form stable nanocapsules with a well defined core-shell architecture, as confirmed by thermal and microstructural characterization. The necessity of the core-shell architecture for the effective encapsulation of fragrance molecules in aqueous dispersion has been demonstrated by NMR (nuclear magnetic resonance). The extent of encapsulation reflects the dynamic equilibrium between the free molecules and the fragrance/polymer complex and is dependent on the octanol/water partition coefficient (logP) of the fragrance compounds, as demonstrated with a similar non ionic core-shell architecture HBP-(PBMA)37-(PPEGMA)39 composed of a hydrophobic poly(butyl methacrylate) (PBMA) core and a hydrophilic poly(polyethylene glycol) methyl ether methacrylate (PPEGMA) shell (provided by the Polymer Laboratory of the EPFL). The fragrance loadings in the polymer (HBP-(PCL)p-(PAA)q and HBP-(PBMA)37-(PPEGMA)39), which reached up to 30 wt% depending on the type of the fragrance molecule, were argued to be linked to their solubility in the hydrophobic core of the star-block copolymer. Moreover, under conditions representative of real applications (fine perfumery and softener applications) the star-block copolymers significantly extended the time over which the concentration of certain volatiles remained above the human olfactory threshold. Effective encapsulation and delayed release of small hydrophobic molecules was hence demonstrated to be possible with the present systems. The straightforward synthesis, tailorable chemistry and globular architecture of the star-block copolymers investigated here therefore offer promise for the development of relatively cheap encapsulants with affinities for specific components in fragrance packages, and release triggered by selected substrates according to their surface chemistry.

EPFL2007

Polymer Brushes as Substrates for Microarray Applications

Raphaël Barbey

The development of innovative bioanalytical technologies that enable the discovery of new therapeutics or the creation of systems for rapid, accurate and parallel analyses has gained an extraordinary interest in the biomedical sector in the last 10-15 years. Since proteins are the key-targets for such essential discoveries and developments, a particular emphasis has been placed on proteomics (study of proteins), including the investigation of protein structures, changes of protein expression upon treatment of cellular systems, the role of post-translational protein modifications on cell signaling events and the validation of selected proteins as disease-specific markers or pharmaceutical targets. These studies usually aim at using crude biological samples as the starting materials, which typically contain a large variety of proteins at concentrations covering a wide dynamic range and where the low abundant proteins are often the biologically most relevant. Therefore, the challenge is to create analytical tools which are able to cope with this large protein concentration range and which provide sufficiently high sensitivity to measure low abundant analytes in the presence of a high abundant protein matrix. The information that is expected from such technologies may soon exert a drastic change on the pace of medical research and considerably impact on the care of patients. As protein microarrays offer a powerful tool for the rapid, parallel analysis of complex samples and only require a minimal amount of reagents, they are currently one of the hot topics of life sciences, which strive to provide such analytical solutions. One of the current limiting factors in the field of protein microarrays for achieving the required analytical sensitivity is the amount of proteins that can be immobilized on the proteomic chip surface. Polymer brushes, which consist of an arrangement of polymer chains that are attached by one end to a substrate in a stretched, well-defined chain conformation, can increase the surface area available for protein binding, and thus act as a new category of 3D protein microarray substrates. With the advent of surface-initiated controlled polymerization techniques, extremely precise control over the thickness, composition, architecture and grafting density can be achieved, which allow to finely tune the properties of the polymer brush. In addition, these polymer chains may contain, natively or after specific post-polymerization reaction of their side chains, a high density of functional groups that can react with proteins. The different approaches available for the preparation of such surface-tethered polymer chains, the strategies allowing control over their architecture and properties as well as examples of applications related to protein binding are presented in Chapter 1. This Thesis describes the use of polymer brushes to create a new type of 3D proteomic chips, with the ultimate goal to improve the protein binding capacity compared to existing chip surface, without altering the accessibility of the detection reagents or increasing the intrinsic fluorescence of the background. The objective of this research work is to prepare polymer brush-based substrates that can outperform the sensitivity (i.e. increase the signal-to-noise ratio) that is obtained with a commercial microarray system, which is used as a benchmark in this Thesis. In this regard, functional polymer brushes containing epoxide groups that can react with the nucleophilic moieties of proteins have been chosen as promising candidates. In a preliminary set of experiments, the potential of glycidyl methacrylate-containing brushes to bind (bio)molecules via a post-polymerization modification reaction under mild conditions, i.e. at room temperature in aqueous medium, has been studied (Chapter 2). To this end, poly(glycidyl methacrylate) brushes, as well as several poly(glycidyl methacrylate)-co-poly(2-(diethylamino)ethyl methacrylate) copolymer brushes of different composition have been prepared by surface-initiated atom transfer radical (co)polymerization. The kinetics of the post-polymerization modification reaction of these (co)polymer brushes were followed using various model primary amines, as well as proteins. The introduction of tertiary amine groups, incorporated in the polymer brushes such as 2-(diethylamino)ethyl methacrylate units, was demonstrated not only to enhance the rate of the epoxide ring-opening reaction but also to induce higher protein binding capacities as compared to poly(glycidyl methacrylate) homopolymer brushes. The improved loading capacities of the (co)polymer brushes were evaluated in Chapter 3 under conditions that more closely resemble real microarray applications, such as the use of automated nanospotters to deposit proteins and fluorescence readers to detect protein immobilization. In this regard, commercially-available proteomic chips were coated with the epoxide-containing (co)polymer brushes and the spotting conditions as well as the proteins were varied. These studies revealed that the loading capacity depends on the pH and on the investigated protein, but also that the (co)polymer brush-coated substrates show far higher loading capacities than the standard, unmodified proteomic chips. This investigation also ascertained the supremacy of brushes containing 2-(diethylamino)ethyl methacrylate units compared to homopolymer brushes, as well as the superiority of thick (co)polymer brushes in comparison to their thinner equivalents. These (co)polymer brushes were subsequently used as platforms for real, although simplified, protein assays. The modified proteomic chips were spotted with unlabeled proteins that were further recognized with detection antibodies via either a one-step or a two-step approach. The clear benefits observed for copolymer brushes as compared to the existing technology under these realistic assay conditions nicely support the fact that the interior volume of the brush remains accessible for the detection reagents. In conclusion, the combination of the high loading capacity and the remaining accessibility for the detection partners make these (co)polymer brushes, and particularly the poly(glycidyl methacrylate)-co-poly(2-(diethylamino)ethyl methacrylate) brushes, excellent candidates for the improvement of the sensitivity in protein microarray applications.

EPFL2010

Temperature-Controlled Masking/Unmasking of Cell-Adhesive Cues with Poly(ethylene glycol) Methacrylate Based Brushes

Solenne Desseaux, Harm-Anton Klok

Thin, thermoresponsive polymer coatings that allow to reversibly modulate cell adhesion and detachment are attractive substrates for cell sheet engineering. Usually, this is accomplished by applying thin poly(N-isopropylacrylamide) (PNIPAM) coatings, which allow cell adhesion via nonspecific interactions above the collapse temperature (TT) of the surface-attached polymer and cell detachment upon cooling below TT. This Article presents an alternative, thermoresponsive polymer platform that is based on 2-(2-methoxyethoxy)ethyl methacrylate (MEO2MA) containing copolymer brushes prepared via surface-initiated atom transfer radical polymerization (SI-ATRP). These brushes are interesting as they gradually collapse and dehydrate upon increasing the temperature from 10 to 40 degrees C, yet resist nonspecific adhesion of cells over this entire temperature window. The MEO2MA based brushes presented here were modified via a two-step postpolymerization modification protocol to introduce cell-adhesive RGD containing peptide ligands. The possibility to reversibly control the swelling and collapse of these brush films by varying temperature allows to modulate the effectively available surface concentration of these cell-adhesive cues and thus provides a way to mask/unmask their biological activity. As a first proof of concept, this Article demonstrates that these MEO2MA brush copolymer films enable integrin-mediated adhesion of 3T3 fibroblasts at 37 degrees C and allow release of these cells by cooling to 23 degrees C. The use of cell-adhesive ligands, which can be thermoreversibly masked/unmasked, is attractive as it enables the use of serum-free cell culture conditions. This is advantageous since it avoids possible concerns regarding eventual toxicity and immunological side effects of serum proteins and also provides opportunities to select for particular cell types and for enhanced control over cell stimulation and differentiation.

American Chemical Society2014