Innovation in nanomedicine through materials nanoarchitectonics.

Materials nanoarchitectonics has led to important innovations in the design and construction of systems in nanoelectronics, nanomachinery, and energy conversion. Recent publications point to the fact that the same approach may be applied successfully to other fields. In this Perspective, we define the key features of materials nanoarchitectonics and examine how they can be used to address current challenges in nanomedicine, placing emphasis on colloidal agents for therapeutic and diagnostic applications.

Novel micellar CB2 receptor agonist with anti-inflammatory action for treating corneal alkali burns in a mouse model.

Introduction: Moderate corneal alkali burns such as those sustained from accidental exposure to household chemicals are treated with topical corticosteroids. Side effects include increased intraocular pressure and slowing of wound healing. Here, we compare the effects of a cannabinoid receptor 2 (CB2r) agonist, TA-A001, that is involved in wound healing with that of the corticosteroid, prednisolone. Methods: TA-A001 was encapsulated with a polymeric micelle comprising polyvinylpyrrolidone: polylactide block copolymers referred to as SmartCelle™ to allow delivery of the very hydrophobic drug. Mouse corneas were given moderate alkali burns. Different doses of TA-A001 of 0.125%, 0.25% and 0.5% were used to treat the burns in comparison to the corticosteroid, prednisolone. Results: TA-A001 at 0.25% and 0.5% allowed for faster wound closure. However, the higher 0.5% dose also induced unwanted neovascularization. By comparison, burned corneas treated with prednisolone showed slower healing as well as disorganization of the cornea. Although 0.25% TA-A001 appeared to produce the most-optimal responses, this dose resulted in marked expression of the macrophage chemoattractant protein, MCP-1. However, there was also an increase in CD163 positive stained M2 anti-inflammatory macrophages in the TA-A001 corneas. TA-A001 treated corneas showed the presence of sensory nerve fibers throughout the corneal epithelium including the superficial cell layers as did Substance P staining. Discussion: We found that TA-A001 at the 0.25% doses was able to modulate inflammation resulting from a moderate alkali burn to the cornea. With more extensive testing, TA-A001 might prove to be a potential alternative to corticosteroids for treating alkali burns or other causes of corneal inflammation.

Tuning the properties and functions of 17ß-estradiol-polysaccharide conjugates in thin films: impact of sample history.

In addition to its role in the regulation of sex-related processes, 17ß-estradiol (E2) participates in the prevention and treatment of cardiovascular diseases via nongenomic pathways mediated by estrogen receptors (ER-α) located in the cell membrane. To achieve specific nongenomic activity of E2, we linked E2 (4.4 mol %) to chitosan-phosphorylcholine (CH-PC) (20 mol % PC). Injections of ER-α solutions (5 to 100 nmol L(-1)) over rehydrated CH-PC-E2 thin films led to permanent adsorption of ER-α to the film surface, as detected by quartz crystal microbalance with dissipation (QCM-D). However, ER-α did not bind onto CH-PC-E2 films formed in situ and never dried. X-ray photoelectron spectroscopy (XPS) analysis of spin-cast CH-PC-E2 films revealed significant E2 enrichment of the topmost section of the film, attributed to the preferential migration of E2 toward the film/air interface upon drying. Mechanical analysis of CH-PC-E2 films in the frequency domain probed by QCM-D indicated that rehydrated films behave as an entangled network with junction points formed by self-assembly of hydrophobic E2 moieties and by ion pairing among PC groups, whereas films formed in situ are entangled polymer solutions with temporary junctions. The structural analysis presented offers useful guidelines for the study of amphiphilic biomacromolecules designed for therapeutic use as thin films.

Probing the stability of biocompatible sodium hyaluronate/chitosan nanocoatings against changes in salinity and pH.

The stability of polyelectrolyte multilayer assemblies was investigated with emphasis on the effects of solution ionic strength, pH, and polymer molecular weight on the film thickness and surface topography. The multilayers consisting of two polysaccharides, the polyanion sodium hyaluronate (HA) and the polycation chitosan (CH) were studied using surface plasmon resonance (SPR) spectroscopy, impedance quartz crystal microbalance (QCM), and atomic force microscopy (AFM). SPR/QCM experiments show that coatings consisting of four HA/CH bilayers assembled at pH 4.5 in the presence of 0.15 M NaCl are stable in NaCl solutions of concentration less than 0.8 M. These multilayers are stable when placed in contact with aqueous solutions ranging in pH from 3.5 to 9. The molecular weight of the polysaccharides has only a marginal effect on the stability of the films in the range explored here (HA: Mn = 360,000 or 31,000 g/mol; CH: Mn = 160,000 or 30,000 g/mol). AFM imaging reveals that different mechanisms may account for the multilayers stability versus salt and pH treatments. While increasing the ionic strength induces reorganization of the surface topography from isolated spherical islets to elongated worm-like features, changes in pH have no appreciable effects on the coating topography prior to complete disintegration.

Phosphorylcholine-modified chitosan films as effective promoters of cell aggregation: correlation between the films properties and cellular response.

This study describes chitosan-phosphorylcholine (CH-PC) films able to support the formation of cell aggregates (spheroids), which are important for tissue engineering and pharmacological studies. The surface topography, charge, thickness, and rheology of CH-PC thin films were characterized by AFM, zeta-potential measurements, SPR spectroscopy, and QCM-D measurements. The CH-PC films are highly hydrated gels, independently of the level of PC incorporation (15-40 mol-% PC/glucosamine units). QCM-D studies established that the amount of fibrinogen adsorbed on CH-PC films decreased with increasing PC content. CH-PC surfaces underwent a transition from moderately cell-adhesive (CH-PC15) to non-adhesive (CH-PC40). Optical micrographs of HUVEC and MCF-7 cell lines cultured on CH-PC surfaces showed that they form spheroids on CH-PC25 and CH-PC40 films.

Self-assembled nanogels of cholesteryl-modified polysaccharides: effect of the polysaccharide structure on their association characteristics in the dilute and semidilute regimes.

The assembly of cholesteryl derivatives of the highly branched polysaccharide mannan Mw = (5.2 x 104 g/mol) in dilute aqueous solution was investigated by 1H nuclear magnetic resonance (NMR) spectroscopy, size-exclusion chromatography coupled with multiangle laser scattering (SEC-MALLS), dynamic light scattering (DLS), atomic force microscopy (AFM), fluorescence quenching, and fluorescence depolarization measurements. In the dilute regime, cholesteryl-bearing mannans (CHM) containing approximately 1 cholesteryl group per 100 mannopyranose units formed nanogels with a hydrodynamic radius (RH) of approximately 20 nm containing approximately 8 macromolecules held together via hydrophobic nanodomains consisting of approximately 9 cholesteryl groups. Their density Phih ( approximately 0.02) was significantly lower than the density ( approximately 0.16) of nanogels formed by a cholesteryl derivative of the linear polysaccharide pullulan (CHP) of identical molar mass and level of cholesteryl substitution. In the semidilute regime, CHM nanogels formed a macrogel network for concentrations higher than 12.5% w/w, whereas CHP nanogels underwent macrogelation only above a threshold concentration of 8.0% w/w, as revealed by oscillatory and steady-shear viscosity measurements. The differences in the solution properties of CHM and CHP reflect differences in their assembly on the molecular level, in particular, the size and number of hydrophobic nanodomains and the hydration level. They are attributed to differences in the mobility of the cholesteryl groups which, itself, can be traced to the fact that in CHM the cholesteryl groups are predominantly linked to short oligomannopyranose branches, whereas in CHP they are linked to the polymer main chain. Our study provides a novel means to nanoengineer polysaccharide nanogels which may find unique biotechnological applications.

Construction of viscoelastic biocompatible films via the layer-by-layer assembly of hyaluronan and phosphorylcholine-modified chitosan.

Films of hyaluronan (HA) and a phosphorylcholine-modified chitosan (PC-CH) were constructed by the polyelectrolyte multilayer (PEM) deposition technique and their buildup in 0.15 M NaCl was followed by atomic force microscopy, surface plasmon resonance spectroscopy (SPR), and dissipative quartz crystal microbalance (QCM). The HA/PC-CH films were stable over a wide pH range (3.0-12.0), exhibiting a stronger resistance against alkaline conditions as compared to HA/CH films. The loss and storage moduli, G' and G", of the films throughout the growth of eight bilayer assemblies were derived from an impedance analysis of the QCM data recorded in situ. Both G' and G" values were one order of magnitude lower than the moduli of HA/CH films. The fluid gel-like characteristics of HA/PC-CH multilayers were attributed to their high water content (50 wt %), which was estimated by comparing the surface coverage values derived from SPR and QCM measurements. Given the versatility of the PEM methodology, HA/PC-CH films are attractive tools for developing biocompatible surface coatings of controlled mechanical properties.

Unified model of association-induced lower critical solution temperature phase separation and its application to solutions of telechelic poly(ethylene oxide) and of telechelic poly(N-isopropylacrylamide) in water.

The authors present a model describing the coexistence of hydrophobic association and phase separation with lower critical solution temperature (LCST) in aqueous solutions of polymers carrying short hydrophobic chains at both chain ends (telechelic associating polymers). The LCST of these solutions is found to decrease along the sol/gel transition curve as a result of both end-chain association (association-induced phase separation) and direct hydrophobic interaction of the end chains with water. The authors relate the magnitude of the LCST decrease to a hydration cooperativity parameter sigma. The LCST decreases substantially (approximately 100 K) in the case of random hydration (sigma=1), whereas only a small shift (approximately 5-10 K) occurs in the case of cooperative hydration (sigma=0.3). The molecular weight dependence of the LCST drop is studied in detail in each case. The results are compared with experimental observations of the cloud points of telechelic poly(ethylene oxide) solutions, in which random hydration predominates, and of telechelic poly(N-isopropylacrylamide) solutions, in which cooperative hydration prevails.

Interactions in water of alkyl and perfluoroalkyl surfactants with fluorocarbon- and hydrocarbon-modified poly(N-isopropylacrylamides).

Fluorescence spectroscopy and isothermal titration calorimetry (ITC) have been used to study the interactions in water at 25 degrees C of two anionic surfactants--sodium dodecyl sulfate (SDS) and sodium perfluorononanoate (SPFN)--with various pyrene-labeled hydrophobically modified poly(N-isopropylacrylamides) (HM-PNIPAM) grafted at random with small amounts of fluorocarbon chains (1H,1H-perfluorooctyl, CH2C7F15); (PNIPAM-F), or (n-octadecyl, C18H37) (PNIPAM-HPy) or both (PNIPAM-F/HPy). In aqueous solution, the copolymers form micellar structures consisting of a loose corona of hydrated poly(N-isopropylacrylamide) chains and a hydrophobic core rich in hydrocarbon or fluorocarbon groups. From fluorescence studies based on changes in the ratio of pyrene excimer to monomer emission intensity, it has been established (1) that mixed SDS/C18H37 clusters form along the polymer chain upon addition of SDS to either PNIPAM-HPy or PNIPAM-F/HPy and (2) that SPFN does not interact with the hydrocarbon-rich microdomains of the polymeric micelles. The conclusions were corroborated by ITC experiments, which yield the overall enthalpy change associated with polymer/surfactant interactions. They provided strong evidence (1) that SDS molecules adsorb along the PNIPAM main chain but do not mix with the fluorocarbon-rich microdomains of PNIPAM-F or PNIPAM-F/HPy and (2) that SPFN associates with the perfluorocarbon substituents of PNIPAM-F and PNIPAM-F/HPy but has a poor affinity for the polymer chain.

Effect of molecular weight on the exponential growth and morphology of hyaluronan/chitosan multilayers: a surface plasmon resonance spectroscopy and atomic force microscopy investigation.

The layer-by-layer growth of multilayer assemblies of two polysaccharides, the polyanion hyaluronan (HA) and the polycation chitosan (CH), was investigated using atomic force microscopy (AFM) and surface plasmon resonance (SPR) spectroscopy, with primary emphasis on the effect of the polysaccharide molecular weights on the film thickness and surface morphology. The HA/CH multilayers exhibit an exponential increase of the optical film thickness with the number of deposited bilayers. We show that the multilayer thickness at a given stage depends on the size of both CH, the diffusing polyelectrolyte, and HA, the non-diffusing species. Assemblies (12 bilayers) of high molecular weight polysaccharides (HA, 360,000; CH, 160,000) were twice as thick (approximately 900 nm vs approximately 450 nm) as those obtained with low molecular weight polymers (HA, 30,000; CH, 31,000), as assessed by AFM scratch tests. The exponential growth rate is the same for the high and low molecular weight pairs; the larger film thicknesses observed by SPR and by AFM arising from an earlier onset of the steep exponential growth phase in the case of the high molecular weight pair. In all cases, isolated islets form during the deposition of the first CH layer onto the underlying HA. Upon further film growth, individual islets coalesce into larger vermiculate features. The transition from distinct islands to vermiculate structures depends on the molecular weights of the polysaccharides and the lower molecular weight construct presents larger worm-like surface domains than the high molecular weight pair.

Delivery platform for hydrophobic drugs: prodrug approach combined with self-assembled multilayers.

We report the design of a platform for the delivery of hydrophobic drugs via a macromolecular prodrug approach combined with LbL-assembled polyelectrolyte multilayers. A hyaluronan ester prodrug of the chemotherapeutic drug paclitaxel has been synthesized. Conjugation of the drug to hyaluronan through a labile succinate ester did not inhibit its activity. Using quartz crystal microbalance, atomic force microscopy, and UV spectroscopy, we have shown that the presence of the hydrophobic paclitaxel moieties does not prohibit the layer-by-layer construction of the multilayers. Release of the drug from the paclitaxel-loaded multilayers upon hydrolysis of the ester linkage resulted in a drastic cell death. Application of this delivery platform to substrates such as colloids, biomedical implants, or vascular tissues may lead to new therapeutic strategies.