Richard A. Gemeinhart

Matrix metalloproteases: Underutilized targets for drug delivery

Pathophysiological molecules in the extracellular environment offer excellent targets that can be exploited for designing drug targeting systems. Matrix metalloproteases (MMPs) are a family of extracellular proteolytic enzymes that are characterized by their overexpression or overactivity in several pathologies. Over the last two decades, the MMP literature reveals heightened interest in the research involving MMP biology, pathology and targeting. This review describes various strategies that have been designed to utilize MMPs for targeting therapeutic entities. Key factors that need to be considered in the successful design of such systems have been identified based on the analyses of these strategies. Development of targeted drug delivery using MMPs has been steadily pursued; however, drug delivery efforts using these targets need to be intensified and focused to realize the clinical application of the fast developing fundamental MMP research.

Cellular Fate of a Modular DNA Delivery System Mediated by Silica Nanoparticles

Development of efficient molecular medicines, including gene therapeutics, RNA therapeutics, and DNA vaccines, depends on efficient means of transfer of DNA or RNA into the cell. Potential problems, including toxicity and immunogenicity, surrounding viral methods of DNA delivery have necessitated the use of nonviral, synthetic carriers. To better design synthetic carriers, or transfection reagents, the modular design of viruses has inspired a modular approach to DNA and RNA delivery. Each modular component can be designed to circumvent each of the many barriers. The modular approach will allow modification of individual components for a specific application. By utilizing a dense silica nanoparticle to form a ternary complex, transfection efficiency of a DNA‐transfection reagent complex was increased by a factor of approximately 10 by concentrating the DNA at the surface of cells. Surface modification of the silica nanoparticles allowed determination of the cellular uptake mechanism with only minor alteration of transfection efficiency. Nanoparticles are internalized by an endosome‐lysosomal route followed by perinuclear accumulation. The modification mechanism confirms that surface modification of the modular system can allow specific moieties to be incorporated into the modular system without significant alteration of the transfection efficiency. By showing that the modular system based upon concentration of DNA at the level of the cell can be used to increase transfection efficiency, we have shown that further modification of the system may better target DNA delivery and overcome other barriers of DNA expression.

pH-sensitivity of fast responsive superporous hydrogels

Stimuli-sensitive hydrogels (or smart hydrogels) are hydrogels that swell or shrink in response to small changes in environmental conditions in which they are placed. While the extent of swelling or shrinking may be large, the kinetics of such changes is slow, since the diffusion of water into and out of the hydrogel is a slow process. To obtain fast responses, we have prepared superporous hydrogels (SPHs) that can swell or shrink extremely fast regardless of their dimensions. The swelling and shrinking are orders of magnitude faster than expected for a nonporous hydrogel of the same dimensions. Water molecules are taken up into the SPHs by capillary forces, and this makes water uptake much faster than diffusion. The swelling ratio of the poly(acrylamide- co -acrylic acid) (p(AM-co-AA)) SPHs was dependent on the pH and ionic strength of the medium. The effect of pH was most pronounced and the effect of ionic strength was observed at all pH values. SPHs made at pH around 5 showed transient maximum swelling when exposed to pH 1.2 medium due to the transient low hydrogen ion concentration inside the swelling SPHs. The p(AM-co-AA) SPHs showed repeated swelling and shrinking by alternating the medium pH between 1.2 and 7.5, and the changes in swelling ratio was quite fast occurring in a matter of a minute. This fast sensitivity may make the stimuli sensitive hydrogels useful in many applications not previously possible. These materials can be used for applications where a single-piece hydrogel is more advantageous than hydrogel microparticulates.

In Vitro Evaluation of Macroporous Hydrogels to Facilitate Stem Cell Infiltration, Growth, and Mineralization

Hydrogels have gained acceptance as biomaterials in a wide range of applications, including pharmaceutical formulations, drug delivery, and tissue sealants. However, exploiting the potential of hydrogels as scaffolds for cell transplantation, tissue engineering, and regenerative medicine still remains a challenge due to, in part, scaffold design limitations. Here, we describe a highly interconnected, macroporous poly(ethylene glycol) diacrylate hydrogel scaffold, with pores ranging from 100 to 600 microm. The scaffold exhibits rapid cell uptake and cell seeding without the need of any external force or device with high incorporation efficiency. When human mesenchymal stem cells are seeded within the porous scaffolds, the scaffolds were found to promote long-term stem cell viability, and on exposure to osteogenic medium, elicit an mineralization response as evaluated by an increased alkaline phosphatase activity (per cell) and calcium and phosphate content within the constructs. The atomic composition of the mineralized matrix was further determined by energy dispersive spectroscopy and found to be similar to calcium-deficient hydroxyapatite, the amorphous biological precursor of bone. The macroporous design of the hydrogel appears advantageous over similar porous hydrogel scaffolds with respect to ease of synthesis, ease of stem cell seeding, and its ability to support long-term stem cell survival and possible differentiation.

Understanding the adsorption mechanism of chitosan onto poly(lactide-co-glycolide) particles

Polyelectrolyte-coated nanoparticles or microparticles interact with bioactive molecules (peptides, proteins or nucleic acids) and have been proposed as delivery systems for these molecules. However, the mechanism of adsorption of polyelectrolyte onto particles remains unsolved. In this study, cationic poly(lactide-co-glycolide) (PLGA) nanoparticles were fabricated by adsorption of various concentrations of a biodegradable polysaccharide, chitosan (0-2.4g/L), using oil-in-water emulsion and solvent evaporation techniques. The particle diameter, zeta-potential, and chitosan adsorption of chitosan-coated PLGA nanoparticles confirmed the increase of polyelectrolyte adsorption. Five adsorption isotherm models (Langmuir, Freundlich, Halsey, Henderson, and Smith) were applied to the experimental data in order to better understand the mechanism of adsorption. Both particle diameter and chitosan adsorption increased with chitosan concentration during adsorption. A good correlation was obtained between PLGA-chitosan nanoparticle size and adsorbed chitosan on the surface, suggesting that the increased particle size was primarily due to the increased chitosan adsorption. The zeta-potential of chitosan-coated PLGA nanoparticles was positive and increased with chitosan adsorbed until a maximum value (+55mV) was reached at approximately 0.4-0.6g/L; PLGA nanoparticles had a negative zeta-potential (-20mV) prior to chitosan adsorption. Chitosan adsorption on PLGA nanoparticles followed a multilayer adsorption behavior, although the Langmuir monolayer equation held at low concentrations of chitosan. The underlying reasons for adsorption of chitosan on PLGA nanoparticles were thought to be the cationic nature of chitosan, high surface energy and microporous non-uniform surface of PLGA nanoparticles.

Pore structure of superporous hydrogels

Hydrogels with a fast swelling property have been synthesized using a gas blowing technique. Since those hydrogels possess interconnected pores of which diameters are in the order of a few hundred micrometers, they are called “superporous hydrogels” (SPHs). The fast swelling of SPHs in aqueous solution is due to the absorption of water by capillary pressure through interconnected pores (i.e. open channels). Because of the importance of pore structures on the fast swelling property, effects of surface porosity on the swelling kinetics were examined. The surface chemistry of the polymerization mold made of glass was varied using various silanes, and the surface morphology of the synthesized SPHs was examined by scanning electron microscopy. The porosity was measured using mercury porosimetry. Despite differences in surface morphology and surface porosity of SPHs, the swelling kinetics were not changed significantly. The internal pore structures remained the same as the surface porosity changed. The study indicates that the swelling of SPHs is predominantly determined by the internal pore structures, and small differences in the surface porosity do not alter the overall swelling kinetics of SPHs. Copyright

Mechanistic Examination of Protein Release from Polymer Nanofibers

Therapeutic proteins have emerged as a significant class of pharmaceutical agents over the past several decades. The potency, rapid elimination, and systemic side effects have prompted the need of spatiotemporally controlled release for proteins maybe more than any other active therapeutic molecules. This work examines the release of two model protein compounds, bovine serum albumin (BSA) and an anti-integrin antibody (AI), from electrospun polycaprolactone (PCL) nanofiber mats. The anti-integrin antibody was chosen as a model of antibody therapy; in particular, anti-integrin antibodies are a promising class of therapeutic molecules for cancer and angiogenic diseases. The release kinetics were studied experimentally and interpreted in the framework of a recently published theory of desorption-limited drug release from nondegrading--or very slowly degrading--fibers. The results are consistent with a protein release mechanism dominated by desorption from the polymer surface, while the polycaprolactone nanofibers are not degrading at an appreciable rate.

Hybrid Nanocrystals: Achieving Concurrent Therapeutic and Bioimaging Functionalities toward Solid Tumors

Bioimaging and therapeutic agents accumulated in ectopic tumors following intravenous administration of hybrid nanocrystals to tumor-bearing mice. Solid, nanosized paclitaxel crystals physically incorporated fluorescent molecules throughout the crystal lattice and retained fluorescent properties in the solid state. Hybrid nanocrystals were significantly localized in solid tumors and remained in the tumor for several days. An anticancer effect is expected of these hybrid nanocrystals.

Developing nanocrystals for cancer treatment.

Nanocrystals are carrier-free solid drug particles that are sized in the nanometer range and have crystalline characteristics. Due to high drug loading (as high as 100%) - free of organic solvents or solubilizing chemicals - nanocrystals have become attractive in the field of drug delivery for cancer treatment. Top-down and bottom-up approaches have been developed for preparing anticancer nanocrystals. In this review, preparation methods and in vivo performance of anticancer nanocrystals are discussed first, followed by an introduction of hybrid nanocrystals in cancer theranostics.

Effect of compression on fast swelling of poly(acrylamide-co-acrylic acid) superporous hydrogels.

Superporous hydrogels (SPHs) swell to a large size in a very short time. In many applications it is preferred to compress SPHs to reduce the overall dimension in the dried state. The effects of compression on the swelling property of SPHs were examined. The swelling property of the compressed SPHs was dependent on the orientation of the SPHs during compression. If SPHs were compressed in an orientated manner so that they retained interconnected porous structure, they were able to swell to near equilibrium within 10 min of immersion in aqueous fluids. If SPHs were compressed in a manner that did not retain the open pore structure, the swelling rate was greatly reduced. The results showed that the SPHs could be compressed without significant sacrifice of the fast swelling property if compressed in a proper orientation. Because pores were formed owing to the generation of gas which rose from bottom to the top of the container, the compression parallel to the pore formation resulted in preservation of the pore structure, and thus fast swelling property. The ability to compress SPHs, maintaining the fast swelling property, is expected to be useful in various applications including development of gastric retention devices for oral drug delivery.