Surface-initiated atom-transfer radical polymerization (SI-ATRP) of bactericidal polymer brushes on poly(lactic acid) surfaces.

We have modified the surface of poly(lactic acid) (PLA) by bromination in the presence of N-bromosuccinimide (NBS) under UV irradiation. This new approach to impart functionality to the surface does not effect the bulk of the material. Brominated PLA surfaces served as initiators for atom-transfer radical polymerization (SI-ATRP) of 2-(methacryloyloxy)ethyl]trimethylammonium chloride, a quaternary ammonium methacrylate (QMA). Grafting of poly(QMA) brushes rendered PLA films hydrophilic and these films displayed a three-order of magnitude increase in antimicrobial efficacy against Gram-negative bacteria such as Escherichia coli as compared to unmodified PLA. The two-step strategy described here to modify PLA surface represents a useful route to modified PLA materials for biomedical and antimicrobial packaging applications.

Azide-Substituted Polylactide: A Biodegradable Substrate for Antimicrobial Materials via Click Chemistry Attachment of Quaternary Ammonium Groups.

Polylactide (PL) co-polymers substituted with pendant azide groups (azido-PL) were synthesized by the nucleophilic conjugate addition of 3-azido-1-propanethiol to a co-polymer of PL containing α,ß-unsaturated ester units, poly(lactide-co-methylene glycolide) (ene-PL) that is obtained from the base-promoted dehydrochlorination of poly(lactide-co-chlorolactide) (chloro-PL). Alternatively, azido-PL was prepared by the treatment of chloro-PL with 3-azido-1-propanethiol without isolation of the ene-PL intermediate. The azido-PL was functionalized by copper-catalyzed [3 + 2] cycloaddition reactions with four alkynes: propargyl 4-methoxybenzoate, N,N,N-trimethyl-N-propargylammonium bromide, N,N-dimethyl-N-octyl-N-propargylammonium bromide, and N,N,N-trioctyl-N-propargylammonium bromide. Polymer adducts with N,N,N-trioctyl-N-propargylammonium bromide displayed potent antimicrobial activity both in suspension and as a polymer film.

Peptide-functionalized poly[oligo(ethylene glycol) methacrylate] brushes on dopamine-coated stainless steel for controlled cell adhesion.

The modification of the surface of surgical implants with cell adhesion ligands has emerged as a promising approach to improve biomaterial-host interactions. However, these approaches are limited by the non-specific adsorption of biomolecules and uncontrolled presentation of desired bioactive ligands on implant surfaces. This leads to sub-optimal integration with host tissue and delayed healing. Here we present a strategy to grow non-fouling polymer brushes of oligo(ethylene glycol) methacrylate by atom transfer radical polymerization from dopamine-functionalized clinical grade 316 stainless steel. These brushes prevent non-specific adsorption of proteins and attachment of cells. Subsequently, the brushes can be modified with covalently tethered adhesive peptides that provide controlled cell adhesion. This approach may therefore have broad application to promote bone growth and improvements in osseointegration. STATEMENT OF SIGNIFICANCE: Stainless steel (SS) implants are widely used clinically for orthopaedic, spinal, dental and cardiovascular applications. However, non-specific adsorption of biomolecules onto implant surfaces results in sub-optimal integration with host tissue. To allow controlled cell-SS interactions, we have developed a strategy to grow non-fouling polymer brushes that prevent protein adsorption and cell adhesion and can be subsequently functionalized with adhesive peptides to direct cell adhesion and signaling. This approach has broad application to improve osseointegration onto stainless steel implants in bone repair.

Competition Between π-π and C-H/π Interactions: A Comparison of the Structural and Electronic Properties of Alkoxy-Substituted 1,8-Bis((propyloxyphenyl)ethynyl)naphthalenes.

The structural and electronic consequences of π-π and C-H/π interactions in two alkoxy-substituted 1,8-bis- ((propyloxyphenyl)ethynyl)naphthalenes are explored by using X-ray crystallography and electronic structure computations. The crystal structure of analogue 4, bearing an alkoxy side chain in the 4-position of each of the phenyl rings, adopts a π-stacked geometry, whereas analogue 8, bearing alkoxy groups at both the 2- and the 5-positions of each ring, has a geometry in which the rings are splayed away from a π-stacked arrangement. Symmetry-adapted perturbation theory analysis was performed on the two analogues to evaluate the interactions between the phenylethynyl arms in each molecule in terms of electrostatic, steric, polarization, and London dispersion components. The computations support the expectation that the π-stacked geometry of the alkoxyphenyl units in 4 is simply a consequence of maximizing π-π interactions. However, the splayed geometry of 8 results from a more subtle competition between different noncovalent interactions: this geometry provides a favorable anti-alignment of C-O bond dipoles, and two C-H/π interactions in which hydrogen atoms of the alkyl side chains interact favorably with the π electrons of the other phenyl ring. These favorable interactions overcome competing π-π interactions to give rise to a geometry in which the phenylethynyl substituents are in an offset, unstacked arrangement.

Closely stacked oligo(phenylene ethynylene)s: effect of π-stacking on the electronic properties of conjugated chromophores.

In this work, a bicyclo[4.4.1]undecane scaffold is used to hold oligo(phenylene ethynylene) units in a cofacially stacked arrangement along the entire length of the conjugated units. We study the impact that the resulting strong interchain interactions have on the photophysical properties. The length of the individual oligomer branches was varied from three to five rings to investigate the effect of conjugation on the electronic properties of the stacked segments. Absorption and fluorescence spectra were recorded and compared to those of the corresponding unstacked analogues. Time-dependent density functional theory calculations were carried out and helped to rationalize the low-energy features present in the fluorescence spectra of the stacked systems. The calculations indicate that the low-energy emissions are due to the presence of excimer-like states. The stronger intensity of the low-energy fluorescence band observed in the five-ring stacked system compared to the three-ring analogue is attributed to the smaller activation barrier that separates the local intrachain state and the excimer-like state in the former compound.

Multitiered 2D pi-stacked conjugated polymers based on pseudo-geminal disubstituted [2.2]paracyclophane.

Interchain interactions between pi-systems have a strong effect on the electronic structure of conjugated organic materials. This influence has previously been explored by the spectroscopic and electrochemical characterization of molecules in which pairs of conjugated oligomers are held in a stacked fashion by attachment to a rigid scaffold. We have prepared a new polymer which uses a pseudo-geminal disubstituted [2.2]paracyclophane scaffold to hold 1,4-bis(phenylethynyl)-2,5-dialkoxybenzene (PE(3)) chromophores in a pi-stacked fashion over their entire length and in an extended multitier arrangement. Solutions of this new polymer display a Stokes shift of 171 nm, compared to just ca. 30 nm for previous models in which only the terminal phenyl rings of the PE(3) chromophore are held in a stacked arrangement. This suggests that interchain interactions of pi-systems over their entire length in a multitier assembly provides for relaxation of the excited state to a stable "phane" electronic state which is responsible for emission. This stabilization is not available in the stacked dimer or other regioisomers of the polymer which possess lesser degrees of overlap. Thus, the architecture of the soluble polymer mimics that of segments of conjugated polymers in semiconducting thin films and will provide a platform for the exploration of the nature of charge carriers and excitons in these important materials.

Multivalent integrin-specific ligands enhance tissue healing and biomaterial integration.

Engineered biointerfaces covered with biomimetic motifs, including short bioadhesive ligands, are a promising material-based strategy for tissue repair in regenerative medicine. Potentially useful coating molecules are ligands for the integrins, major extracellular matrix receptors that require both ligand binding and nanoscale clustering for maximal signaling efficiency. We prepared coatings consisting of well-defined multimer constructs with a precise number of recombinant fragments of fibronectin (monomer, dimer, tetramer, and pentamer) to assess how nanoscale ligand clustering affects integrin binding, stem cell responses, tissue healing, and biomaterial integration. Clinical-grade titanium was grafted with polymer brushes that presented monomers, dimers, trimers, or pentamers of the alpha(5)beta(1) integrin-specific fibronectin III (7 to 10) domain (FNIII(7-10)). Coatings consisting of trimers and pentamers enhanced integrin-mediated adhesion in vitro, osteogenic signaling, and differentiation in human mesenchymal stem cells more than did surfaces presenting monomers and dimers. Furthermore, ligand clustering promoted bone formation and functional integration of the implant into bone in rat tibiae. This study establishes that a material-based strategy in which implants are coated with clustered bioadhesive ligands can promote robust implant-tissue integration.

Saccharide polymer brushes to control protein and cell adhesion to titanium.

Attaining control over the surface chemistry of titanium is critical to its use in medical implants, especially to address complications such as infection and loosening of implants over time, which still present significant challenges. The surface-initiated atom transfer radical polymerization (SI-ATRP) of a saccharide-substituted methacrylate, 2-gluconamidoethyl methacrylate (GAMA), affords dense polymer brushes that resist protein adsorption and cell adhesion. We further tailored the nature of the surfaces by covalent attachment of an adhesion peptide to afford control over cell adhesion. Whereas unmodified poly(GAMA) brushes prevent cell adhesion, brushes with a tethered GFOGER-containing peptide sequence promote the deposition of confluent well-spread cells. The presentation of adhesion proteins on a robust bioresistive background in this fashion constitutes a versatile approach to the development of new biomaterials.

Polymer brushes and self-assembled monolayers: Versatile platforms to control cell adhesion to biomaterials (Review).

This review focuses on the surface modification of substrates with self-assembled monolayers (SAMs) and polymer brushes to tailor interactions with biological systems and to thereby enhance their performance in bioapplications. Surface modification of biomedical implants promotes improved biocompatibility and enhanced implant integration with the host. While SAMs of alkanethiols on gold substrates successfully prevent nonspecific protein adsorption in vitro and can further be modified to tether ligands to control in vitro cell adhesion, extracellular matrix assembly, and cellular differentiation, this model system suffers from lack of stability in vivo. To overcome this limitation, highly tuned polymer brushes have been used as more robust coatings on a greater variety of biologically relevant substrates, including titanium, the current orthopedic clinical standard. In order to improve implant-bone integration, the authors modified titanium implants with a robust SAM on which surface-initiated atom transfer radical polymerization was performed, yielding oligo(ethylene glycol) methacrylate brushes. These brushes afforded the ability to tether bioactive ligands, which effectively promoted bone cell differentiation in vitro and supported significantly better in vivo functional implant integration.

Synthesis and modification of functional poly(lactide) copolymers: toward biofunctional materials.

A polylactide copolymer with pendant benzyloxy groups has been synthesized by the copolymerization of a benzyl-ether substituted monomer with lactide. Debenzylation of the polymer to provide pendant hydroxyl groups followed by modification with succinic anhydride affords the corresponding carboxylic acid functionalized copolymer that is amenable to standard carbodiimide coupling conditions to attach amine-containing biological molecules. An amino-substituted biotin derivative was coupled to the carboxyl functional groups of copolymer films as proof-of-concept. In a demonstration of the function of these new materials, an RGD-containing peptide sequence was tethered to copolymer films at various densities and was shown to enhance the adhesion of epithelial cells. This strategy provides the opportunity for the attachment of a variety of ligands, allowing for the fabrication of a versatile class of biodegradable, biocompatible materials.

The effect of integrin-specific bioactive coatings on tissue healing and implant osseointegration.

Implant osseointegration, defined as bone apposition and functional fixation, is a requisite for clinical success in orthopaedic and dental applications, many of which are restricted by implant loosening. Modification of implants to present bioactive motifs such as the RGD cell-adhesive sequence from fibronectin (FN) represents a promising approach in regenerative medicine. However, these biomimetic strategies have yielded only marginal enhancements in tissue healing in vivo. In this study, clinical-grade titanium implants were grafted with a non-fouling oligo(ethylene glycol)-substituted polymer coating functionalized with controlled densities of ligands of varying specificity for target integrin receptors. Biomaterials presenting the alpha5beta1-integrin-specific FN fragment FNIII 7-10 enhanced osteoblastic differentiation in bone marrow stromal cells compared to unmodified titanium and RGD-presenting surfaces. Importantly, FNIII 7-10-functionalized titanium significantly improved functional implant osseointegration compared to RGD-functionalized and unmodified titanium in vivo. This study demonstrates that bioactive coatings that promote integrin binding specificity regulate marrow-derived progenitor osteoblastic differentiation and enhance healing responses and functional integration of biomedical implants. This work identifies an innovative strategy for the rational design of biomaterials for regenerative medicine.

Stacked conjugated oligomers as molecular models to examine interchain interactions in conjugated materials.

Previous studies of the redox states of linear conjugated oligomers as models for polarons and bipolarons in conjugated polymers do not fully address the influence of intermolecular interactions on the electronic structure of conjugated systems in the solid state. Fusion of oligothiophenes onto a bicyclo[4.4.1]undecane core holds the conjugated oligomers in a permanent cofacial stack. One- and two-electron oxidation of the stacked oligomers affords mono(radical cation)s and dications that serve as models for polarons and bipolarons in p-doped conjugated polymers and demonstrates the effect of pi-stacking on the electronic structure of these species.

Functional lactide monomers: methodology and polymerization.

Side-chain-functionalized lactide analogues have been synthesized from commercially available amino acids and polymerized using stannous octoate as a catalyst. The synthetic strategy presented allows for the incorporation of any protected amino acid for the preparation of functionalized diastereomerically pure lactide monomers. The resulting functionalized cyclic monomers can be homopolymerized and copolymerized with lactides and then quantitatively deprotected forming new functional poly(lactide)-based materials. This strategy allows for the introduction of functional groups along a poly(lactide) (PLA) backbone that after deprotection can be viewed as chemical handles for further functionalization of PLA, yielding improved biomaterials for a variety of applications.

Switchable gratings by spatially periodic alignment of liquid crystals via patterned photopolymerization.

Spatially periodic patterning of the anchoring condition of a nematic liquid crystal (NLC) within a polymer matrix via a patterned photopolymerization affords a novel and facile method to prepare electrically switchable diffraction gratings. UV irradiation through a photomask of two comonomers, with opposite tendencies to align the NLC and also with different reactivity ratios, leads to definition of areas with either homeotropic or planar alignment of the NLC. Photopolymerization-induced diffusion of the monomers accounts for the spatial distribution of the concentration of these monomers. The resulting diffraction gratings are switchable under low electric fields and possess structural stability offered by the polymer matrix.

Integrin binding specificity regulates biomaterial surface chemistry effects on cell differentiation.

Biomaterial surface chemistry has profound consequences on cellular and host responses, but the underlying molecular mechanisms remain poorly understood. Using self-assembled monolayers as model biomaterial surfaces presenting well defined chemistries, we demonstrate that surface chemistry modulates osteoblastic differentiation and matrix mineralization independently from alterations in cell proliferation. Surfaces were precoated with equal densities of fibronectin (FN), and surface chemistry modulated FN structure to alter integrin adhesion receptor binding. OH- and NH(2)-terminated surfaces up-regulated osteoblast-specific gene expression, alkaline phosphatase enzymatic activity, and matrix mineralization compared with surfaces presenting COOH and CH(3) groups. These surface chemistry-dependent differences in cell differentiation were controlled by binding of specific integrins to adsorbed FN. Function-perturbing antibodies against the central cell binding domain of FN completely inhibited matrix mineralization. Furthermore, blocking antibodies against beta(1) integrin inhibited matrix mineralization on the OH and NH(2) surfaces, whereas function-perturbing antibodies specific for beta(3) integrin increased mineralization on the COOH substrate. These results establish surface-dependent differences in integrin binding as a mechanism regulating differential cellular responses to biomaterial surfaces. This mechanism could be exploited to engineer materials that control integrin binding specificity to elicit desired cellular activities to enhance the integration of biomaterials and improve the performance of biotechnological culture supports.

Control of anchoring of nematic fluids at polymer surfaces created by in situ photopolymerization.

In situ photopolymerization of alkyl acrylate monomers in the presence of a nematic fluid provides a cellular matrix of liquid crystalline droplets in which the chemical structure of the encapsulating polymer exerts control over the alignment (anchoring) of the liquid crystalline molecules. Control is obtained by variation of the alkyl side chains and through copolymerization of two dissimilar monofunctional acrylates. For example, among a series of poly(methylheptyl acrylate)s, the 1-methylheptyl analogue prefers planar anchoring of a nematic (TL205) over the temperature range studied. However, the polymers of other methylheptyl side chains display a homeotropic-to-planar anchoring thermal transition temperature similar to that of the n-heptyl analogue. Copolymerization of two monofunctional acrylates with opposing tendencies of aligning liquid crystal leads to tunability of anchoring behavior over a wide temperature range. The broad anchoring transitions we observed provide a way of achieving highly tilted anchoring.

Surface chemistry modulates focal adhesion composition and signaling through changes in integrin binding.

Biomaterial surface properties influence protein adsorption and elicit diverse cellular responses in biomedical and biotechnological applications. However, the molecular mechanisms directing cellular activities remain poorly understood. Using a model system with well-defined chemistries (CH3, OH, COOH, NH2) and a fixed density of the single adhesive ligand fibronectin, we investigated the effects of surface chemistry on focal adhesion assembly and signaling. Surface chemistry strongly modulated integrin binding and specificity--alpha5beta1 integrin binding affinity followed the pattern OH>NH2=COOH>CH3, while integrin alphaVbeta3 displayed the relationship COOH>NH2>>OH=CH3. Immunostaining and biochemical analyses revealed that surface chemistry modulates the structure and molecular composition of cell-matrix adhesions as well as focal adhesion kinase (FAK) signaling. The neutral hydrophilic OH functionality supported the highest levels of recruitment of talin, alpha-actinin, paxillin, and tyrosine-phosphorylated proteins to adhesive structures. The positively charged NH2 and negatively charged COOH surfaces exhibited intermediate levels of recruitment of focal adhesion components, while the hydrophobic CH3 substrate displayed the lowest levels. These patterns in focal adhesion assembly correlated well with integrin alpha5beta1 binding. Phosphorylation of specific tyrosine residues in FAK also showed differential sensitivity to surface chemistry. Finally, surface chemistry-dependent differences in adhesive interactions modulated osteoblastic differentiation. These differences in focal adhesion assembly and signaling provide a potential mechanism for the diverse cellular responses elicited by different material properties.

Surface chemistry modulates fibronectin conformation and directs integrin binding and specificity to control cell adhesion.

Integrin-mediated cell adhesion to proteins adsorbed onto synthetic surfaces anchors cells and triggers signals that direct cell function. In the case of fibronectin (Fn), adsorption onto substrates of varying properties alters its conformation/structure and its ability to support cell adhesion. In the present study, self-assembled monolayers (SAMs) of alkanethiols on gold were used as model surfaces to investigate the effects of surface chemistry on Fn adsorption, integrin binding, and cell adhesion. SAMs presenting terminal CH(3), OH, COOH, and NH(2) functionalities modulated adsorbed Fn conformation as determined through differences in the binding affinities of monoclonal antibodies raised against the central cell-binding domain (OH > COOH = NH(2) > CH(3)). Binding of alpha(5)beta(1) integrin to adsorbed Fn was controlled by SAM surface chemistry in a manner consistent with antibody binding (OH > COOH = NH(2) > CH(3)), whereas alpha(V) integrin binding followed the trend: COOH >> OH = NH(2) = CH(3), demonstrating alpha(5)beta(1) integrin specificity for Fn adsorbed onto the NH(2) and OH SAMs. Cell adhesion strength to Fn-coated SAMs correlated with alpha(5)beta(1) integrin binding (OH > COOH = NH(2) > CH(3)), and experiments with function-perturbing antibodies demonstrated that this receptor provides the dominant adhesion mechanism in this cell model. This work establishes an experimental framework to analyze adhesive mechanisms controlling cell-surface interactions and provides a general strategy of surface-directed control of adsorbed protein activity to manipulate cell function in biomaterial and biotechnological applications.

Influence of pi-stacking on the redox properties of oligothiophenes: (alpha-alkyloligo-thienyl)para[2.2]cyclophanes.

[structure: see text] Oligothienyl-substituted paracyclophanes bearing alpha-thienyl substituents, which block polymerization at these sites, undergo oxidation to form stable radical cations and dications. Splitting of the first voltammetric oxidation wave of stacked dimers into two single-electron processes corresponding to formation of the stacked mono(radical cation) and the stacked bis(radical cation) illustrates the influence of pi-stacking on the behavior of conjugated chains, which serve as models for stacked polarons in conjugated polymers.