Anthony M. Lowman

Poly(ethylene glycol)-containing hydrogels in drug delivery

The use of hydrogels as carriers for protein delivery has been a subject of significant recent research. In our recent work, we have shown that diffusion controlled delivery of proteins from hydrogels containing poly(ethylene glycol) (PEG) can be possible and controlled by the three-dimensional structure. In addition, a number of these hydrogel carriers are mucoadhesive and can be used for protein delivery. PEG star polymer gels have also been prepared by gamma-irradiation and have been used for protein delivery with and without molecular imprinting. The presence of a large number of functional groups in a small volume makes these polymers important for use in biological and pharmaceutical applications. PEG star polymer hydrogels were synthesized using gamma-irradiation and were characterized using swelling techniques. Equilibrium swelling studies were conducted to investigate the effects of molecular weight, number of star arms, concentration, and radiation dose.

Elucidation of the mechanism of incorporation of insulin in controlled release systems based on complexation polymers

The objective of this study was to investigate the insulin incorporation and release properties of poly(methacrylic acid-g-ethylene glycol) P(MAA-g-EG) microparticles as a function of copolymer composition. These microparticles exhibited unique pH-responsive characteristics in which interpolymer complexes were formed in acidic media and dissociated in neutral/basic environments. The microparticles containing equimolar amounts of MAA and PEG were capable of efficient insulin loading using equilibrium partitioning (>90%). Additionally, insulin release from the gel was significantly retarded in acidic media while rapid release occurred under neutral/basic conditions. In contrast, as the amount of MAA of the polymer was increased, the entrapment efficiency of insulin within the gel greatly reduced and the insulin was readily released from the polymer network in the acidic and neutral/basic media. In addition, in order to evaluate the potential application of the microparticles to other drugs, theophylline, vancomycin, fluorescein-isothiocyanate-labeled dextrans (FITC-Ds) with average molecular weights of 4400 (FITC-D-4), 12,000 (FITC-D-10) and 19,500 (FITC-D-20) were utilized as model hydrophilic drugs. The incorporation profiles showed that the uptake of theophylline and vancomycin to the microparticles was lower than that of insulin. Additionally, polymer microparticles loaded with theophylline and vancomycin exhibited pH-sensitive release behavior, however, the oscillatory behavior is less pronounced than those of insulin. The values of drug incorporation ratio showed that the microparticles were capable of incorporating almost 90% of insulin and 15% of vancomycin from solution. On the other hand, the other hydrophilic drugs showed very low incorporation efficiency to the microparticles. These data suggest that gels containing equimolar amounts of MAA:EG have the potential to be used as an oral carrier of peptide drugs, especially for insulin.

Uptake and release of budesonide from mucoadhesive, pH-sensitive copolymers and their application to nasal delivery

Microparticles of novel, bioadhesive graft copolymers of polymethacrylic acid and polyethylene glycol (P(MAA-g-EG)) were prepared. The aims of this study were to investigate the uptake and release kinetics of budesonide from P(MAA-g-EG) in vitro as well as the pharmacokinetics following nasal administration of the polymer contained budesonide. The loading of budesonide into the pH-sensitive polymers was examined using various ethanol solutions. Ethanol was required for drug solubilization but hindered hydrogel swelling at pH 7.2. Maximum loading of the drug in the polymer was obtained using 25% ethanol solutions. The release of budesonide from the polymer swollen in 25% ethanol solutions obeyed classical Fickian release behavior after an initial rapid drug burst. For nasal administration of budesonide-containing P(MAA-g-EG) the plasma concentration of budesonide was kept constant following a peak concentration of the drug approximately 45 min after administration.

Evaluation of porous networks of poly(2-hydroxyethyl methacrylate) as interfacial drug delivery devices

Long-term implantable drug delivery devices are desirable to achieve rapid and reliable delivery of bioactive substances to the body. The limitation of most implantable devices is the resulting chronic inflammatory response and fibrous encapsulation of the implant, which prevents effective drug delivery for prolonged periods. One method of overcoming this problem is the addition of an intermediary that could prevent capsule formation. Biocompatible materials with interconnected pore structures greater than 8-10 microm have been shown to support the ingrowth and maintenance of vascularized tissue. In this investigation, we evaluate the efficacy of using porous hydrogel sponges for the tissue interface in an implantable drug delivery device. Porous networks of poly(2-hydroxyethyl methacrylate) (PHEMA) were synthesized using a thermally initiated free-radical solution polymerization. To characterize the microstructure of the PHEMA networks, scanning electron microscopy and mercury porosimetry were used. By altering the solvent fraction in the reaction mixture, PHEMA sponges were synthesized with interconnected pores ranging in size from from 6 to 15 microm with porosities of 55% to 87%. Following the in vitro evaluation, sponges were attached to the distal end of a 20-gauge catheter tubing, and implanted subcutaneously and intraperitoneally. After 5 months implantation, insulin was infused into the devices from external pumps and rapid insulin absorption was observed in conjunction with dramatic lowering of blood glucose levels. From histological evaluation of explanted devices, we observed highly vascularized tissue surrounding the mesenteric implants. These results indicate that it may be possible to use PHEMA sponges for a tissue intermediary for long-term implantable drug delivery devices.

Molecular analysis of interpolymer complexation in graft copolymer networks

Interpolymer complexation is associated with the formation of hydrogen bonds between pendent and/or grafted groups of a single polymer or copolymers. The degree of complexation can be described theoretically in terms of complexation/decomplexation equilibrium characteristics. Graft copolymer networks of poly(methacrylic acid-g-ethylene glycol) were prepared by free radical solution polymerization of methacrylic acid and poly(ethylene glycol) monomethacrylate. The ensuing gels exhibited drastic network structural changes in response to environmental pH changes due to the formation/dissociation of interpolymer complexes. The molecular level degree of complexation was determined as a function of copolymer composition, PEG graft chain molecular weight and environmental pH. The largest degrees of complexation were observed in gels containing nearly equimolar amounts of the monomeric units and the longest molecular weight poly(ethylene glycol) grafts. These trends were predicted by the equilibrium complexation theory and were in good agreement with the experimental data.

A novel method for the direct fabrication of growth factor-loaded microspheres within porous nondegradable hydrogels: Controlled release for cartilage tissue engineering

Because of similar mechanical properties to native cartilage, synthetic hydrogels based on poly(vinyl alcohol) (PVA) have been proposed for replacement of damaged articular cartilage, but they suffer from a complete lack of integration with surrounding tissue. In this study, insulin-like growth factor-1 (IGF-1), an important growth factor in cartilage regeneration, was encapsulated in degradable poly(lactic-co-glycolic acid) (PLGA) microparticles embedded in the PVA hydrogels in a single step based on a double emulsion. The release of IGF-1 from these hydrogels was sustained over 6 weeks in vitro. Poly(glycolic acid) (PGA) fiber scaffolds were wrapped around the hydrogels, seeded with chondrocytes, and implanted subcutaneously in athymic mice. The release of IGF-1 enhanced cartilage formation in the layers surrounding the hydrogels, in terms of the content of extracellular matrix components and mechanical properties, and increased integration between the cartilage layers and the hydrogels, according to gross observation of the cross-sections and histology. The compressive modulus of the cartilage-hydrogel constructs without IGF-1 was 0.07±0.02MPa, compared to 0.17-0.2MPa for hydrogels that contained IGF-1. The biochemical and mechanical markers of cartilage formation were not different between the low and high concentrations of IGF-1, despite an order of magnitude difference in concentration. This study shows that the sustained release of IGF-1 can enhance tissue formation and points to a possible strategy for effecting integration with surrounding tissue.

Mechanical evaluation of poly(vinyl alcohol)-based fibrous composites as biomaterials for meniscal tissue replacement

In this study, poly(vinyl alcohol) (PVA) hydrogels were reinforced with ultrahigh molecular weight polyethylene (UHMWPE) and PP fibers and evaluated as potential nondegradable meniscal replacements. An investigation of hydrogel and composite mechanical properties indicates that fiber-reinforced PVA hydrogels could replicate the unique anisotropic modulus distribution present in the native meniscus; the most commonly damaged orthopedic tissue. More specifically, fibrous reinforcement successfully increased the tensile modulus of the biomaterial from 0.23±0.02MPa without any reinforcement to 258.1±40.1MPa at 29vol.% UHMWPE. Additionally, the molecular weight between cross-links, bound water and the microstructure of the PVA hydrogels were evaluated as a function of freeze-thaw cycles and polymer concentration to lend insight into the processes occurring during synthesis. These results suggest the presence of multiple mechanisms as causes for increasing hydrogel modulus with freeze-thaw cycling, including hydrogen bonding between amorphous and/or crystalline regions, and the formation of highly concentrated regions of mostly amorphous PVA chains. It is possible that the formation of regions with highly concentrated amounts of PVA increases the load-bearing ability of the hydrogels.

In vitro analysis of PNIPAAm–PEG, a novel, injectable scaffold for spinal cord repair

Nervous tissue engineering in combination with other therapeutic strategies is an emerging trend for the treatment of different CNS disorders and injuries. We propose to use poly(N-isopropylacrylamide)-co-poly(ethylene glycol) (PNIPAAm-PEG) as a minimally invasive, injectable scaffold platform for the repair of spinal cord injury (SCI). The scaffold allows cell attachment, and provides mechanical support and a sustained release of neurotrophins. In order to use PNIPAAm-PEG as an injectable scaffold for treatment of SCI, it must maintain its mass and volume over time in physiological conditions. To provide mechanical support at the injury site, it is also critical that the engineered scaffold matches the compressive modulus of the native neuronal tissue. This study focused on studying the ability of the scaffold to release bioactive neurotrophins and matching the material properties to those of the native neuronal tissue. We found that the release of both BDNF and NT-3 was sustained for up to 4 weeks, with a minimal burst exhibited for both neurotrophins. The bioactivity of the released NT-3 and BDNF was confirmed after 4 weeks. In addition, our results show that the PNIPAAm-PEG scaffold can be designed to match the desired mechanical properties of the native neuronal tissue, with a compressive modulus in the 3-5 kPa range. The scaffold was also compatible with bone marrow stromal cells, allowing their survival and attachment for up to 31 days. These results indicate that PNIPAAm-PEG is a promising multifunctional scaffold for the treatment of SCI.

The role of crystallization and phase separation in the formation of physically cross-linked PVA hydrogels

The biocompatibility, processing ease, and mechanical properties of freeze-thawed poly(vinyl alcohol) (PVA)-based hydrogels have encouraged significant research toward developing this material for various biomedical applications. Crystallization that occurs during the freeze-thawing process is cited in the literature as the primary mechanism responsible for the resultant mechanical properties. Further analysis, however, shows the presence of two unique mechanisms that contribute to PVAs mechanical properties. During freeze–thaw cycling water freezes causing phase separation, which facilitates crystallization. The impact of phase separation during freeze–thaw cycling was investigated by comparing freeze-thawed and aged PVA hydrogels. Aged hydrogels were not prepared by freezing and, therefore, did not exhibit significant phase separation. The amount of phase separation was discerned using optical microscopy in the hydrated state. Crystallinity and mechanical properties were also evaluated as a function of the number of cycles (for freeze-thawed gels) and aging time (for aged gels). For freeze-thawed hydrogels, crystallinity deviated significantly from the trend observed in compressive modulus, indicating that crystallinity was not the only factor determining the hydrogels mechanical properties. Phase separation was found to occur during freeze–thaw cycling independently of crystallization, especially at later freeze–thaw cycles (after the third). The trends observed for both crystallinity and modulus for aged hydrogels, however, were in better agreement with each other. Further evaluation of the mechanical properties of aged and freeze-thawed hydrogels with similar crystallinities indicated that freeze-thawed hydrogels have significantly higher modulus values (p < 0.05). As a result, phase separation, independently of crystallization, was determined to have a significant effect on gelation during freeze–thaw cycling. In particular, PVA-rich regions that are formed during phase separation, without additional cross-linking, are believed to have a significant effect on the resultant mechanical properties.

Biomimetic Materials And Design: Biointerfacial Strategies, Tissue Engineering And Targeted Drug Delivery

Biointerfacial strategies: the use of supported thin films of peptide amphiphiles as model systems of the extracellular matrix to study the effects of structure-function phenomena on cell adhesion engineering of integrin-specific biomimetic surfacesto control cell adhesion and function. Tissue engineering: biomaterials - synthetic and engineering strategies scaffolds for directing cellular responses and tissue formation. Targeted drug delivery: micropatterning biomimetic materials for bioadhesionand drug delivery bioinspired engineering of intelligent drug delivery systems and protein-polymer conjugates.