Joseph M. DeSimone

Strategies in the design of nanoparticles for therapeutic applications

Engineered nanoparticles have the potential to revolutionize the diagnosis and treatment of many diseases; for example, by allowing the targeted delivery of a drug to particular subsets of cells. However, so far, such nanoparticles have not proved capable of surmounting all of the biological barriers required to achieve this goal. Nevertheless, advances in nanoparticle engineering, as well as advances in understanding the importance of nanoparticle characteristics such as size, shape and surface properties for biological interactions, are creating new opportunities for the development of nanoparticles for therapeutic applications. This Review focuses on recent progress important for the rational design of such nanoparticles and discusses the challenges to realizing the potential of nanoparticles.

The effect of particle design on cellular internalization pathways

The interaction of particles with cells is known to be strongly influenced by particle size, but little is known about the interdependent role that size, shape, and surface chemistry have on cellular internalization and intracellular trafficking. We report on the internalization of specially designed, monodisperse hydrogel particles into HeLa cells as a function of size, shape, and surface charge. We employ a top-down particle fabrication technique called PRINT that is able to generate uniform populations of organic micro- and nanoparticles with complete control of size, shape, and surface chemistry. Evidence of particle internalization was obtained by using conventional biological techniques and transmission electron microscopy. These findings suggest that HeLa cells readily internalize nonspherical particles with dimensions as large as 3 μm by using several different mechanisms of endocytosis. Moreover, it was found that rod-like particles enjoy an appreciable advantage when it comes to internalization rates, reminiscent of the advantage that many rod-like bacteria have for internalization in nonphagocytic cells.

Synthesis of fluoropolymers in supercritical carbon dioxide

Fluoropolymers are used in many technologically demanding applications because of their balance of high-performance properties. A significant impediment to the synthesis of variants of commercially available amorphous fluoropolymers is their general insolubility in most solvents except chlorofluorocarbons (CFCs). The environmental concerns about CFCs can be circumvented by preparing these technologically important materials in supercritical fluids. The homogeneous solution polymerization of highly fluorinated acrylic monomers can be achieved in supercritical carbon dioxide by using free radical methods. In addition, detailed decomposition rates and efficiency factors were measured for azobisisobutyronitrile in supercritical carbon dioxide and were compared to those obtained with conventional liquid solvents.

Continuous liquid interface production of 3D objects

Fast, continuous, 3D printing Although three-dimensional (3D) printing is now possible using relatively small and low-cost machines, it is still a fairly slow process. This is because 3D printers require a series of steps to cure, replenish, and reposition themselves for each additive cycle. Tumbleston et al. devised a process to effectively grow solid structures out of a liquid bath. The key to the process is the creation of an oxygen-containing “dead zone” between the solid part and the liquid precursor where solidification cannot occur. The precursor liquid is then renewed by the upward movement of the growing solid part. This approach made structures tens of centimeters in size that could contain features with a resolution below 100 µm. Science, this issue p. 1349 Solid parts are elevated from a liquid resin pool at a speed of hundreds of millimeters per hour. Additive manufacturing processes such as 3D printing use time-consuming, stepwise layer-by-layer approaches to object fabrication. We demonstrate the continuous generation of monolithic polymeric parts up to tens of centimeters in size with feature resolution below 100 micrometers. Continuous liquid interface production is achieved with an oxygen-permeable window below the ultraviolet image projection plane, which creates a “dead zone” (persistent liquid interface) where photopolymerization is inhibited between the window and the polymerizing part. We delineate critical control parameters and show that complex solid parts can be drawn out of the resin at rates of hundreds of millimeters per hour. These print speeds allow parts to be produced in minutes instead of hours.

Dispersion Polymerizations in Supercritical Carbon Dioxide

Conventional heterogeneous dispersion polymerizations of unsaturated monomers are performed in either aqueous or organic dispersing media with the addition of interfacially active agents to stabilize the colloidal dispersion that forms. Successful stabilization of the polymer colloid during polymerization results in the formation of high molar mass polymers with high rates of polymerization. An environmentally responsible alternative to aqueous and organic dispersing media for heterogeneous dispersion polymerizations is described in which supercritical carbon dioxide (CO2) is used in conjunction with molecularly engineered free radical initiators and amphipathic molecules that are specifically designed to be interfacially active in CO2. Conventional lipophilic monomers, exemplified by methyl methacrylate, can be quantitatively (>90 percent) polymerized heterogeneously to very high degrees of polymerization (>3000) in supercritical CO2 in the presence of an added stabilizer to form kinetically stable dispersions that result in micrometer-sized particles with a narrow size distribution.

Using mechanobiological mimicry of red blood cells to extend circulation times of hydrogel microparticles

It has long been hypothesized that elastic modulus governs the biodistribution and circulation times of particles and cells in blood; however, this notion has never been rigorously tested. We synthesized hydrogel microparticles with tunable elasticity in the physiological range, which resemble red blood cells in size and shape, and tested their behavior in vivo. Decreasing the modulus of these particles altered their biodistribution properties, allowing them to bypass several organs, such as the lung, that entrapped their more rigid counterparts, resulting in increasingly longer circulation times well past those of conventional microparticles. An 8-fold decrease in hydrogel modulus correlated to a greater than 30-fold increase in the elimination phase half-life for these particles. These results demonstrate a critical design parameter for hydrogel microparticles.

PEGylated PRINT nanoparticles: The impact of PEG density on protein binding, macrophage association, biodistribution, and pharmacokinetics

In this account, we varied PEGylation density on the surface of hydrogel PRINT nanoparticles and systematically observed the effects on protein adsorption, macrophage uptake, and circulation time. Interestingly, the density of PEGylation necessary to promote a long-circulating particle was dramatically less than what has been previously reported. Overall, our methodology provides a rapid screening technique to predict particle behavior in vivo and our results deliver further insight to what PEG density is necessary to facilitate long-circulation.

More Effective Nanomedicines through Particle Design

Nanomedicine is an emerging field that applies concepts in nanotechnology to develop novel diagnostics and therapies. Physical and chemical properties of particles, including size, shape, modulus, surface charge and surface chemistry, play an important role in determining particle-cell interactions, cellular trafficking mechanisms, biodistribution, and pharmacokinetics. This discussion focuses on both nanoparticles and microparticles since microparticles can also provide many insights for the development of drug carriers and possess advantages over nanoparticles in certain applications. This review covers recent major advancement in the nanomedicine field and also highlights studies using the PRINT technology.

Extraction of a hydrophilic compound from water into liquid CO2 using dendritic surfactants

Dendrimers are well defined, highly branched polymers that adopt a roughly spherical, globular shape in solution. Their cores are relatively loosely packed and can trap guest molecules, and by appropriate functionalization of the branch tips the macromolecules can act as unimolecular micelle-like entities. Here we show that dendrimers with a fluorinated shell are soluble in liquid carbon dioxide and can transport CO2-insoluble molecules into this solvent within their cores. Specifically, we demonstrate the extraction of a polar ionic dye, methyl orange, from water into CO2 using these fluorinated dendrimers. This observation suggests possible uses of such macromolecules for the remediation of contaminated water, the extraction of pharmaceutical products from fermentation vessels, the selective encapsulation of drugs for targeted delivery and the transport of reagents for chemical reactions (such as polymerization) in liquid and supercritical CO2 solvents.

Design of Nonionic Surfactants for Supercritical Carbon Dioxide

Interfacially active block copolymer amphiphiles have been synthesized and their self-assembly into micelles in supercritical carbon dioxide (CO2) has been demonstrated with small-angle neutron scattering (SANS). These materials establish the design criteria for molecularly engineered surfactants that can stabilize and disperse otherwise insoluble matter into a CO2 continuous phase. Polystyrene-b-poly(1,1-dihydroperfluorooctyl acrylate) copolymers self-assembled into polydisperse core-shell-type micelles as a result of the disparate solubility characteristics of the different block segments in CO2. These nonionic surfactants for CO2 were shown by SANS to be capable of emulsifying up to 20 percent by weight of a CO2-insoluble hydrocarbon into CO2. This result demonstrates the efficacy of surfactant-modified CO2 in reducing the large volumes of organic and halogenated solvent waste streams released into our environment by solvent-intensive manufacturing and process industries.