Smart Polymeric Nanocarriers of Met-enkephalin.

This study describes a novel approach to polymeric nanocarriers of the therapeutic peptide met-enkephalin based on the aggregation of thermoresponsive polymers. Thermoresponsive bioconjugate poly((di(ethylene glycol) monomethyl ether methacrylate)-ran-(oligo(ethylene glycol) monomethyl ether methacrylate) is synthesized by AGET ATRP using modified met-enkephalin as a macroinitiator. The abrupt heating of bioconjugate water solution leads to the self-assembly of bioconjugate chains and the formation of mesoglobules of controlled sizes. Mesoglobules formed by bioconjugates are stabilized by coating with cross-linked two-layer shell via nucleated radical polymerization of N-isopropylacrylamide using a degradable cross-linker. The targeting peptide RGD, containing the fluorescence marker carboxyfluorescein, is linked to a nanocarrier during the formation of the outer shell layer. In the presence of glutathione, the whole shell is completely degradable and the met-enkephalin conjugate is released. It is anticipated that precisely engineered nanoparticles protecting their cargo will emerge as the next-generation platform for cancer therapy and many other biomedical applications.

Transfer of fibroblast sheets cultured on thermoresponsive dishes with membranes.

In cell or tissue engineering, it is essential to develop a support for cell-to-cell adhesion, which leads to the generation of cell sheets connected by extracellular matrix. Such supports must be hydrophobic and should result in a detachable cell sheet. A thermoresponsive support that enables the cultured cell sheet to detach using only a change in temperature could be an interesting alternative in regenerative medicine. The aim of this study was to evaluate plates covered with thermoresponsive polymers as supports for the formation of fibroblast sheets and to develop a damage-free procedure for cell sheet transfer with the use of membranes as transfer tools. Human skin fibroblasts were seeded on supports coated with a thermoresponsive polymer: commercial UpCell™ dishes (NUNC™) coated with thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) and dishes coated with thermoresponsive poly(tri(ethylene glycol) monoethyl ether methacrylate) (P(TEGMA-EE)). Confluent fibroblast sheets were effectively cultured and harvested from both commercial PNIPAM-coated dishes and laboratory P(TEGMA-EE)-coated dishes. To transfer a detached cell sheet, two membranes, Immobilon-P(®) and SUPRATHEL(®), were examined. The use of SUPRATHEL for relocating the cell sheets opens a new possibility for the clinical treatment of wounds. This study established the background for implementing thermoresponsive supports for transplanting in vitro cultured fibroblasts.

Nonviral Plasmid DNA Carriers Based on N,N'-Dimethylaminoethyl Methacrylate and Di(ethylene glycol) Methyl Ether Methacrylate Star Copolymers.

Star polymers with random and block copolymer arms made of cationic N,N'-dimethylaminoethyl methacrylate (DMAEMA) and nonionic di(ethylene glycol) methyl ether methacrylate (DEGMA) were synthesized via atom transfer radical polymerization (ATRP) and used for the delivery of plasmid DNA in gene therapy. All stars were able to form polyplexes with plasmid DNA. The structure and size of the polyplexes were precisely determined using light scattering and cryo-TEM microscopy. The hydrodynamic radius of a complex of DNA with star was dependent on the architecture of the star arms, the DEGMA content and the number of amino groups in the star compared to the number of phosphate groups of the nucleic acid (N/P ratio). The smallest polyplexes (Rh90°âˆ¼50 nm) with positive zeta potentials (∼15 mV) were formed of stars with N/P=6. The introduction of DEGMA into the star structure caused a decrease of polyplex cytotoxicity in comparison to DMAEMA homopolymer stars. The overall transfection efficiency using HT-1080 cells showed that the studied systems are prospective gene delivery agents. The most promising results were obtained for stars with random copolymer arms of high DEGMA content.

Controlling the Crystallinity of Thermoresponsive Poly(2-oxazoline)-Based Nanolayers to Cell Adhesion and Detachment.

Semicrystalline, thermoresponsive poly(2-isopropyl-2-oxazoline) (PIPOx) layers covalently bonded to glass or silica wafers were obtained via the surface-termination of the living polymer chains. Polymer solutions in acetonitrile were exposed to 50 °C for various time periods and were poured onto the functionalized solid wafers. Fibrillar crystallites formed in polymerization solutions settled down onto the wafers next to the amorphous polymer. The amount of crystallites adsorbed on thermoresponsive polymer layers depended on the annealing time of the PIPOx solution. The wettability of PIPOx layers decreased with the increasing amount of crystallites. The higher content of crystallites weakened the temperature response of the layer, as evidenced by the philicity and thickness measurements. Semicrystalline thermoresponsive PIPOx layers were used as biomaterials for human dermal fibroblasts (HDFs) culture and detachment. The presence of crystallites on the PIPOx layers promoted the proliferation of HDFs. Changes in the physicochemical properties of the layer, caused by the temperature response of the polymer, led to the change in the cells shape from a spindle-like to an ellipsoidal shape, which resulted in their detachment. A supporting membrane was used to assist the detachment of the cells from PIPOx biosurfaces and to prevent the rolling of the sheet.

Relevance of the poly(ethylene glycol) linkers in peptide surfaces for proteases assays.

Poly(ethylene glycol)s (PEGs) with different lengths were used as linkers during the preparation of peptide surfaces for protease detection. In the first approach, the PEG monolayers were prepared using a "grafting to" method on 3-aminopropyltrietoxysilane (APTES)-modified silicon wafers. Protected peptides with a fluorescent marker were synthesized by Fmoc solid phase synthesis. The protected peptide structures enabled their site-specific immobilization onto the PEG surfaces. Alternatively, the PEG-peptide surface was obtained by immobilizing a PEG-peptide conjugate directly onto the modified silicon wafer. The surfaces (composition, grafting density, hydrophilicity, and roughness) were characterized by time-of-flight-secondary ion mass spectrometry (ToF-SIMS), X-ray photoelectron spectroscopy (XPS), contact angle (CA), and atomic force microscopy (AFM). Introducing the PEG linker between the peptide and surface increased their resistance toward nonspecific protein adsorption. The peptide surfaces were examined as analytical platforms to study the action of trypsin as a representative protease. The products of the enzymatic hydrolysis were analyzed by fluorescence spectroscopy, electrospray ionization-mass spectrometry (ESI-MS), and ToF-SIMS. Conclusions about the optimal length of the PEG linker for the analytical application of PEG-peptide surfaces were drawn. This work demonstrates an effective synthetic procedure to obtain PEG-peptide surfaces as attractive platforms for the development of peptide microarrays.

Polymeric nanoparticle engineering: from temperature-responsive polymer mesoglobules to gene delivery systems.

A novel approach for the preparation of nano- and microcapsules in aqueous solutions by using thermoresponsive polymer (TRP) templates (mesoglobules) is described. The method comprised three steps: formation of mesoglobules, coating the templates by seeded radical copolymerization, followed by core dissolution and core removal upon cooling. When mesoglobule entraps biomacromolecules during the process of their formation, it makes it possible to load a controlled amount of bioactive compounds without covalent attachment. Special attention is paid to the mesoglobule dissolution upon cooling, as well as their loading efficiency. Details on the outer shell formation and the possibilities for targeting ligands incorporation and control of the shell porosity are discussed. Finally, the seeded radical copolymerization was used for covering DNA complexes with cationic copolymers bearing TRP blocks. This Review is an attempt to convince researchers of the promising perspectives for using mesoglobules as potential reservoirs, carriers, and transferring agents for biologically active substances.

Poly(2-substituted-2-oxazoline) surfaces for dermal fibroblasts adhesion and detachment.

The thermoresponsive surfaces of brush structure (linear polymer chains tethered on the surface) based on poly(2-isopropyl-2-oxazoline)s and copolymers of 2-ethyl-2-oxazoline and 2-nonyl-2-oxazoline were obtained using the grafting-to method. The living oxazoline (co)polymers have been synthesized by cationic ring-opening polymerization and subsequently terminated by the reactive amine groups present on the surface. The changes in the surface morphology, philicity and thickness occurring during surface modification were monitored via atomic force microscopy, contact angle and ellipsometry. The thickness of the (co)poly(2-substituted-2-oxazoline) layers ranged from 4 to 11 nm depending on the molar mass of immobilized polymer and reversibly varied with the temperature changes. This confirmed thermoresponsive properties of obtained surfaces. The obtained polymer surfaces were used as a support for dermal fibroblast culture and detachment. The fibroblasts' adhesion and proliferation on the polymer surfaces were observed when the culture temperature was above the cloud point temperature of the immobilized polymer. Lowering the temperature resulted in the detachment of the dermal fibroblast sheets from the polymer layers, which makes these surfaces suitable for the treatment of wounds and in skin tissue engineering.

Synthesis and characterisation of PEG-peptide surfaces for proteolytic enzyme detection.

Peptide surfaces were obtained by the covalent immobilisation of fluorescently labelled pentapeptides carboxyfluorescein-glycine-arginine-methionine-leucine-glycine, either directly or through a poly(ethylene glycol) (PEG) linker on modified silicon wafers. Each step during the preparation of the peptide surfaces was confirmed by several surface characterisation techniques. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) and X-ray photoelectron spectroscopy were used to determine the surface composition, the wafers philicity was measured by contact angle and atomic force microscopy was used to investigate the surface morphology. Exposure of the peptide surfaces to trypsin resulted in the release of a fluorescently labelled peptide product, which allowed the kinetics of the enzymatic reaction to be followed with the aid of fluorescence spectroscopy. The electrospray ionisation mass spectrometry analysis of the post-digestion solution confirmed that the pentapeptides attached to the solid support undergo specific trypsin hydrolysis at the C-terminus of the arginine residues. Detailed surface analyses before and after the enzyme action was performed using ToF-SIMS. Because of the limited accessibility of the short peptide directly attached to the surface, a quantitative yield of enzymatic hydrolysis was observed only in case when the peptide was bound through the PEG linker. The insertion of the PEG linker increased the number of immobilised peptides and the rate of enzymatic digestion which consequently improved the quality of the enzyme assays. The described approach may be used for different peptide sequences designed for other proteases.

Thermal Behaviour of Common Thermoresponsive Polymers in Phosphate Buffer and in Its Salt Solutions.

Thermoresponsive polymers are a promising material for drug nanocarrier preparation, which makes the study of their aggregation in physiological conditions very important. In this paper, the thermal behaviour of the thermoresponsive polymers poly(N-isopropylacrylamide), poly(2-isopropyl-2-oxazoline-co-2-n-propyl-2-oxazoline) and poly[(2-hydroxyethyl methacrylate)-co-oligo(ethylene glycol) methyl ether methacrylate] were studied in phosphate buffer (PBS) and solutions of its salts in concentration as in PBS. The thermal response of the polymers was measured using UV-Vis and dynamic light scattering (DLS). The salts shifted the cloud point temperature (TCP) of the (co)polymers to higher values compared to the TCP of aqueous polymer solutions. In PBS and NaCl solutions, all polymers exhibited an unexpected and previously unreported transmittance profile. During heating, an additional aggregation of polymers appeared above the TCP accompanied by the formation of a precipitate. In monosodium phosphate solutions and pure water, the studied polymers showed lower critical solution temperature (LCST-type) behaviour. DLS measurements showed that a salt influenced the size of the resulting polymer particles. The sizes and stability of particles depended on the heating rate. In PBS and NaCl solutions, the size of particles in the dispersion decreased above 60 °C, and the precipitate appeared on the bottom of the cuvette. The additional aggregation of polymer and its falling out of solution may hinder the removal of carriers from the body and has to be taken into account when preparing nanocarriers.

Micellization of Polystyrene-b-Polyglycidol in Dioxane and Water/Dioxane Solutions.

In this work, the self-assembly of a series of amphiphilic polystyrene-b-polyglycidol (PS-b-PGL) diblock copolymers in dioxane and dioxane/water mixtures is presented. The PS-b-PGL have an average degree of polymerization (DP) of PS block equal to 29 units and varied degrees of polymerization for the glycidol segments with DPs of 13, 42, 69 and 117. In dioxane, amphiphilic diblock copolymers form micelles with the hydrophilic PGL placed in the core. Critical micelle concentration (CMC) was determined based on the intensity of scattered light vs. concentration. The micelle size was measured by dynamic light scattering and transmission electron microscopy. Also, the behaviour of the copolymer was studied in water/dioxane solutions by following the changes of scattered light intensity with the addition of water to the system. Critical water content (CWC) of the studied systems decreased as the initial PS-b-PGL concentration in dioxane increased. This process was accompanied by a decrease in the size of aggregate formed. For a given initial copolymer concentration, the size of copolymer aggregates decreased linearly with increasing the length of the PGL block.

Selective Partial Hydrolysis of 2-isopropyl-2-oxazoline Copolymers towards Decreasing the Ability to Crystallize.

Poly(2-isopropyl-2-oxazoline) (PiPrOx) is readily prone to crystallization both in solid and from solutions. This feature is detrimental for certain applications. Here, we examine whether the presence of unsubstituted ethyleneimine (EI) units, a gradient distributed within a polymer chain composed of 2-isopropyl-2-oxazoline (iPrOx) and 2-methyl-2-oxazoline (MOx) units, decreases the ability to crystallize the copolymer and affects thermal properties compared to the homopolymer of iPrOx. We assumed that the separation of stiff iPrOx units by the more flexible EI will affect the spatial arrangements of the ordered chains, slightly plasticize and, as a result, decrease their ability to crystallize. The selective hydrolysis of gradient iPrOx and 2-methyl-2-oxazoline (MOx) copolymers, carried out under mild conditions, led to iPrOx/MOx/EI copolymers. To the best of our knowledge, the selective hydrolysis of these copolymers has never been carried out before. Their thermal properties and crystallization abilities, both in a solid state and from an aqueous solution, were analyzed. Based on the analysis of polymer charge and cytotoxicity studies, the potential use of the copolymers obtained was indicated in some biological systems.

Thermoresponsive Nanogels of Modified Poly((di(ethylene glycol) methyl ether methacrylate)-co-(2-aminoethyl methacrylate))s.

A series of copolymers of di(ethylene glycol) methyl ether methacrylate (D) and 2-aminoethyl methacrylate (A) (P(D-co-A)) with variable ratios of comonomers were synthesized using atom transfer radical polymerization. Then, the amino groups of obtained copolymers were modified to clickable azide or prop-2-yn-1-yl carbamate groups. A thermoresponsive copolymers were obtained with the value of cloud point temperature (TCP) dependent on the type and number of functional groups in the copolymer and on the concentration of solutions. For P(D-co-A) copolymers, the TCP increased with increasing content of 2-aminoethyl methacrylate comonomer. The presence of azide and prop-2-yn-1-yl carbamate groups caused the changes of TCP of modified copolymers. All studied copolymers in dilute aqueous solutions aggregated above TCP to nanoparticles with sizes dependent on the solution concentration, heating procedures, and types and numbers of functional groups present in a copolymer chain. The presence of hydrophilic elements in the chain and the increase in the copolymer concentration led to the enlargement of the particle sizes. Aggregates were crosslinked using click reaction between an azide and prop-2-yn-1-yl carbamate groups that led to stable thermoresponsive nanogels. A systematic study of the behavior of copolymers allowed the determination of the chains useful for possible application in drug delivery.

Processing of (Co)Poly(2-oxazoline)s by Electrospinning and Extrusion from Melt and the Postprocessing Properties of the (Co)Polymers.

Poly(2-oxazoline) (POx) matrices in the form of non-woven fibrous mats and three-dimensional moulds were obtained by electrospinning and fused deposition modelling (FDM), respectively. To obtain these materials, poly(2-isopropyl-2-oxazoline) (PiPrOx) and gradient copolymers of 2-isopropyl- with 2-n-propyl-2-oxazoline (P(iPrOx-nPrOx)), with relatively low molar masses and low dispersity values, were processed. The conditions for the electrospinning of POx were optimised for both water and the organic solvent. Also, the FDM conditions for the fabrication of POx multi-layer moulds of cylindrical or cubical shape were optimised. The properties of the POx after electrospinning and extrusion from melt were determined. The molar mass of all (co)poly(2-oxazoline)s did not change after electrospinning. Also, FDM did not influence the molar masses of the (co)polymers; however, the long processing of the material caused degradation and an increase in molar mass dispersity. The thermal properties changed significantly after processing of POx what was monitored by increase in enthalpy of exo- and endothermic peaks in differential scanning calorimetry (DSC) curve. The influence of the processing conditions on the structure and properties of the final material were evaluated having in a mind their potential application as scaffolds.

Amniotic Stem Cells Cultured on Thermoresponsive Polymers Allow Obtaining a Full Cell Sheet.

Amniotic stem cells promote adhesion and migration of epithelial cells. Obtaining a full sheet containing amniotic stem cells seems to be the best solution for the treatment of burn wounds. The main advantage of this method is obtaining a full sheet of cells by lowering the temperature below the transition temperature, which does not affect extracellular matrix. The purpose of this work was to produce a skin substitute-a full sheet consisting of amniotic mesenchymal stem cells-and compare with well-known fibroblast sheet. Amniotic membrane cells revealed better tendency to full sheet detachment than fibroblasts. Confluence after 24 hours was always higher on polymer-coated dishes than on normal polypropylene dishes. Also viability was higher than on the control culture dish, while the number of apoptotic cells was always highest on polypropylene (control). Ile-Lys-Val-ala-Val (IKVAV) 0.28 addition to poly (poly [ethylene glycol] ethyl methacrylate) (PTEGMA) caused best cell confluence and highest percentage of cells in mitosis phase of cell cycle, but also worst cell detachment was observed in both cell types on PTEGMA IKVAV 0.28. Viability of cells transferred in cell sheet form onto a new culture dish was higher than when detached as suspension enzymatically. Additionally, percentage of apoptotic cells transferred in cell sheet form onto a new culture dish was always lower than when detached as suspension enzymatically. Culturing of PTEGMA, PTEGMA IKVAV 0.28 and PTEGMA IKVAV 0.14 have a stimulating effect on number of cells in mitosis in amniotic cell culture even after cell sheet transfer onto a new dish, whereas such effect with fibroblast was not observed.

Poly(2-oxazoline) Matrices with Temperature-Dependent Solubility-Interactions with Water and Use for Cell Culture.

In this work, we studied the stability of matrices with temperature-dependent solubility and their interactions with water at physiological temperature for their application in cell culture in vitro. Gradient copolymers of 2-isopropyl- with 2-n-propyl-2-oxazoline (P(iPrOx-nPrOx)) were used to prepare the matrices. The comonomer ratio during polymerization was chosen such that the cloud point temperature (TCP) of the copolymer was below 37 °C while the glass transition (Tg) was above 37 °C. The role of the support for matrices in the context of their stability in aqueous solution was examined. Therefore, matrices in the form of both self-supported bulk polymer materials (fibrillar mats and molds) and polymer films supported on the silica slides were examined. All of the matrices remained undissolved when incubated in water at a temperature above TCP. For the self-supported mats and molds, we observed the loss of shape stability, but, in the case of films supported on silica slides, only slight changes in morphology were observed. For a more in-depth investigation of the origin of the shape deformation of self-supported matrices, we analyzed the wettability, thickness, and water uptake of films on silica support because the matrices remained undeformed under these conditions. It was found that, above the TCP of P(iPrOx-nPrOx), the wettability of the films decreased, but at the same time the films absorbed water and swelled. We examined how this specific behavior of the supported films influenced the culture of fibroblasts. The temperature-dependent solubility of the matrices and the possibility of noninvasive cell separation were also examined.

Thermoresponsive P(HEMA-co-OEGMA) copolymers: synthesis, characteristics and solution behavior.

Random, thermoresponsive copolymers of 2-hydroxyethyl methacrylate (HEMA) and oligo(ethylene glycol) methyl ether methacrylate M n = 300 (OEGMA) were synthesized via atom transfer radical polymerization (ATRP) in a DMSO/H2O solvent mixture. Reactivity ratios were determined by the extended Kelen-Tudos method and found to be close to 1. Studies confirmed the randomness of the obtained copolymers. The thermoresponsiveness in water and in phosphate buffer (PBS) solutions and the influence of copolymer composition and solution concentration on the cloud point temperature (T cp) were investigated. Phase transitions in water solutions were reversible and narrow. The response of P(HEMA-co-OEGMA) to temperature could be adjusted in the range from 66.5 °C to 21.5 °C by changing the HEMA content. In PBS solutions, significant differences in the heating/cooling cycle were observed for all investigated concentrations. The presence of kosmotropic salts in PBS decreased the T cp value and caused thermal aggregation of chains to form a macroscopic aggregate at temperatures above the T cp.

Fibroblast and keratinocyte crosstalk: the effect of a poly(tri[ethylene glycol] ethyl ether methacrylate) thermoresponsive surface on short-term co-culture.

The treatment of difficult-to-treat wounds can be challenging. Although a number of approaches have been investigated, the healing process may be slow and unsatisfactory. An alternative approach is the use of a continuous sheet of skin cells applied over a wound which may improve cell implantation and patient recovery. To analyse the gene expression profile of fibroblast/keratinocyte co-culture on poly(tri[ethylene glycol] ethyl ether methacrylate) (P[TEGMA-EE]), a thermoresponsive biocompatible surface. Cultures were grown for 72 hours as a continuous layer on P(TEGMA-EE). Assays for genotoxicity, cell morphology, and fluorescence-assisted flow cytometry were performed to exclude adverse effects. A gene expression profile related to the extracellular matrix was investigated by microarray analysis. For fibroblast monocultures and fibroblast/keratinocyte co-cultures maintained for 72 hours on P(TEGMA-EE), no change in morphology or specific surface markers, or DNA damage (comet assay) was observed, relative to control surface. Moreover, no detrimental impact was ascertained based on microarray analysis. In response to lowered temperature, the detachment of a continuous cell layer sheet from the thermoresponsive surface was observed. When gene expression was compared between fibroblasts cultured alone and co-cultured with keratinocytes on P(TEGMA-EE), 10 genes were shown to be differentially expressed. Of these genes, six were significantly differentially expressed between cultures grown on P(TEGMA-EE) and human skin samples. Our results indicate that P(TEGMA-EE) is fully biocompatible and is therefore a suitable surface for successful preparation and recovery of two-layered fibroblast/keratinocyte co-culture as a continuous sheet of cells.

Photocrosslinking of Polyglycidol and Its Derivative: Route to Thermoresponsive Hydrogels.

Hydrogels of biologically well-tolerated, high-molar-mass polyglycidol (PGl) and its thermoresponsive derivative poly(glycidol-co-ethyl glycidyl carbamate) have been obtained by direct UV crosslinking in the solid state. Polymers with molar masses up to 1.45 × 106 g mol-1 were crosslinked in the presence of benzophenone or (4-benzoylbenzyl)trimethylammonium chloride as photosensitizers. The photosensitizer concentration was varied from 2 to 10 wt%. The influence of polymer composition and photosensitizer type and amount on the crosslinking efficiency, swelling and temperature behavior of the obtained hydrogels was investigated. The photocrosslinking of PGl and poly(glycidol-co-ethyl glycidyl carbamate) led to hydrogels with swelling degrees up to 1700%. The swelling degrees of the hydrogels decreased with the increase of the environmental temperature indicating the thermoresponsive nature of gels. The swelling of obtained gels can be controlled by varying the composition of the copolymer precursor and by the network density.

Synthesis, Characterization and Cytotoxicity of Novel Thermoresponsive Star Copolymers of N,N'-Dimethylaminoethyl Methacrylate and Hydroxyl-Bearing Oligo(Ethylene Glycol) Methacrylate.

Novel, nontoxic star copolymers of N,N-dimethylaminoethyl methacrylate (DMAEMA) and hydroxyl-bearing oligo(ethylene glycol) methacrylate (OEGMA-OH) were synthesized via atom transfer radical polymerization (ATRP) using hyperbranched poly(arylene oxindole) as the macroinitiator. Stars with molar masses from 100,000 g/mol to 257,000 g/mol and with various amounts of OEGMA-OH in the arms were prepared. As these polymers can find applications, e.g., as carriers of nucleic acids, drugs or antibacterial or antifouling agents, in this work, much attention has been devoted to exploring their solution behavior and their stimuli-responsive properties. The behavior of the stars was studied in aqueous solutions under various pH and temperature conditions, as well as in PBS buffer, in Dulbecco's modified Eagle's medium (DMEM) and in organic solvents for comparison. The results indicated that increasing the content of hydrophilic OEGMA-OH units in the arms up to 10 mol% increased the cloud point temperature. For the stars with an OEGMA-OH content of 10 mol%, the thermo- and pH-responsivity was switched off. Since cytotoxicity experiments have shown that the obtained stars are less toxic than homopolymer DMAEMA stars, the presented studies confirmed that the prepared polymers are great candidates for the design of various nanosystems for biomedical applications.

Degradable polymeric nanoparticles by aggregation of thermoresponsive polymers and "click" chemistry.

This study describes a novel approach to the preparation of crosslinked polymeric nanoparticles of controlled sizes that can be degraded under basic conditions. For this purpose thermoresponsive copolymers containing azide and alkyne functions were obtained by ATRP of di(ethylene glycol) monomethyl ether methacrylate (D) and 2-aminoethyl methacrylate (A) followed by post polymerization modification. The amino groups of A were reacted with propargyl chloroformate or 2-azido-1,3-dimethylimidazolinium hexafluorophosphate, which led to two types of copolymers. Increasing the temperature of aqueous solutions of the mixed copolymers caused their aggregation into spherical nanoparticles composed of both types of chains. Their dimensions could be controlled by changing the concentration and heating rate of the solutions. Covalent stabilization of aggregated chains was performed by a "click" reaction between the azide and alkyne groups. Due to the presence of a carbamate bond the nanoparticles undergo pH dependent degradation under mild basic conditions. The proposed procedure opens a route to new carriers for the controlled release of active species.