Solute transport analysis in pH-responsive,complexing hydrogels of poly(methacrylic acid-g-ethylene glycol)

We report on the preparation and properties of hydrogels of poly(methacrylic acid-g-ethylene glycol) that exhibit pH-responsive swelling behavior due to the reversible formation/dissociation of interpolymer complexes. Because of their nature, these materials may be useful in drug delivery applications. In this work, we studied the diffusional behavior of three solutes of varying molecular size in the complexing hydrogels as a function of solution pH. The ability of these gels to control the solute diffusion rates was strongly dependent on the molecular size of the solute and the environmental pH. The diffusion coefficients for solutes were calculated as a function of pH and were lower in acidic than neutral or basic media due to the formation of interpolymer complexes in the gels. However, the ratio of the solute radius to the network mesh size also was a significant factor in the overall behavior of these gels. The diffusion coefficient of the smallest solute, proxyphylline, studied only changed by a factor of five between the complexed and uncomplexed state. However, for the largest solute, FITC-dextran, which has a molecular radius ten times greater than proxyphylline, the diffusion coefficients of the drugs in complexed and uncomplexed gels varied by almost two orders of magnitude. These results are explained in terms of mesh size characteristics of the gels.     

Drug transport mechanisms and release kinetics from molecularly designed poly(acrylic acid-g-ethylene glycol) hydrogels

Controlled drug release devices of pH-sensitive, complexing poly(acrylic acid-g-ethylene glycol) (P(AA-g-EG)) hydrogels were prepared by free radical solution UV polymerization. The effects of hydrogel composition, polymerization conditions and surrounding environment on theophylline release kinetics and drug transport mechanisms were evaluated in these P(AA-g-EG) polymer networks. Release studies indicated a dependence of the theophylline release kinetics and diffusion coefficients on the hydrogel structure, polymerization conditions and pH of the environment. The theophylline transport mechanism was studied by fitting experimental data to five different model equations and calculating the corresponding parameters. The Akaike information criterion was also considered to elucidate the best-fit equation. Results indicated that in most release cases, the drug release mechanism was anomalous (non-Fickian). This indicates that such systems may, under certain conditions, provide release characteristics approaching zero-order release. The pH of the dissolution medium appeared to have a strong effect on the drug transport mechanism. At more basic pH values, Case II transport was observed, indicating a drug release mechanism highly influenced by macromolecular chain relaxation. The results obtained in this research work lead us to the conclusion that P(AA-g-EG) hydrogels can be successfully used as drug delivery systems. Their versatility to be designed with specifically tuned release properties renders these biomaterials promising pharmaceutical carriers for therapeutic agents.     

Fibrinolytic behaviour of streptokinase-immobilized poly(methacrylic acid-g-ethylene oxide)

Streptokinase was immobilized on poly(methacrylic acid-g-ethylene oxide) surfaces by means of a modified coupling reagent method. The surfaces were activated with a carbodiimide to form an O-acyl isourea derivative, which, upon addition of the enzyme, condensed to form the corresponding amide. The quantity of enzyme immobilized on the surface increased as the reaction time with carbodiimide and streptokinase increased. The fibrinolytic activity of the immobilized enzyme systems was measured by a clot lysis assay. The dynamics of fibrin clot lysis was investigated for a period of up to 500 min. The lysis reaction mechanism was found to approximate a first-order fibrin degradation. In addition, the kinetic rate constant was found to increase with increasing immobilized enzyme content.     

Investigation of translational motion of poly(ethylene glycol) macromolecules in poly(methacrylic acid) hydrogels

The translational mobility of linear macromolecules of poly(ethylene glycol) (PEG) within a weakly cross-linked poly(methacrylic acid) (PMAA) hydrogel was investigated by means of the pulse field gradient (PFG) NMR method in order to reveal the effect of PMAA/PEG complex formation. It was found that inside the collapsed gel a fraction of the PEG molecules has self-diffusion characteristics like those of the network chains. This suggests the formation of an interpolymer complex, as a result of which some linear molecules acquired the dynamic properties of the network chains. Another fraction of the PEG macromolecules inside the collapsed gel enjoyed free diffusion, for they were not included in the complex with PMAA. In contrast, within the swollen gel (at concentrations of PEG higher than 5 wt.-%) the self-diffusion coefficient of all PEG molecules was independent of the diffusion time, which indicates an absence of the interpolymer complex (or at least that its lifetime is negligibly short).     

Effects of sterilization on poly(ethylene glycol) hydrogels

The past few decades have witnessed a dramatic increase in the development of polymeric biomaterials. These biomaterials have to undergo a sterilization procedure before implantation. However, many sterilization procedures have been shown to profoundly affect polymer properties. Poly(ethylene glycol) hydrogels have gained increasing importance in the controlled delivery of therapeutics and in tissue engineering. We evaluated the effect of ethylene oxide (EtO), hydrogen peroxide (H(2)O(2)), and gamma sterilization of poly(ethylene glycol) hydrogels on properties relevant to controlled drug delivery and tissue engineering. We observed that the release of cyclosporine (CyA) (an immunosuppressive drug that is effective in combating tissue rejection following organ transplantation) was significantly affected by the type of sterilization. However, that was not the case with rhodamine B, a dye. Hence, the drug release characteristics were observed to be dependent not only on the sterilization procedure but also on the type of agent that needs to be delivered. In addition, differences in the swelling ratios for the sterilized and unsterilized hydrogels were statistically significant for 1:1 crosslinked hydrogels derived from the 8000 MW polymer. Significant differences were also observed for gamma sterilization for 1:1 crosslinked hydrogels derived from the 3350 MW polymer and also the 2:1 crosslinked hydrogels derived from the 8000 MW polymer. Atomic force microscopy (AFM) studies revealed that the roughness parameter for the unsterilized and EtO-sterilized PEG hydrogels remained similar. However, a statistically significant reduction of the roughness parameter was observed for the H(2)O(2) and gamma-sterilized samples. Electron spin resonance (ESR) studies on the unsterilized and the sterilized samples revealed the presence of the peroxy and the triphenyl methyl carbon radical in the samples. The gamma and the H(2)O(2)-sterilized samples were observed to have a much higher concentration of the radical pecies when compared with the EtO and the unsterilized samples.     

Cell adhesion on poly(propylene fumarate-co-ethylene glycol) hydrogels

We synthesized poly(propylene fumarate-co-ethylene glycol) block copolymers [P(PF-co-EG)] that were crosslinked to form hydrogels and investigated the effect of copolymer composition on cell adhesion to the hydrogels. These copolymers were water soluble when the molar ratio of ethylene glycol repeating unit to propylene fumarate repeating unit was higher than 4.4. The water content of swollen hydrogels increased from 29 to 63% and the water contact angle decreased from 38 to 21 degrees as the molar ratio increased from 0.6 to 4.4. No significant change in either property was observed for ratios higher than 4.4. In a cell adhesion assay under serum-free conditions, the number of adherent platelets and smooth muscle cells decreased from 21 to 2% and from 78 to 20% of the initial seeding density, respectively, as the molar ratio increased from 0.6 to 7.8. Adherent smooth muscle cells did not spread on the hydrogels of the compositions tested. Adherent platelets did not show any filopodia. These results suggest that the hydrophilicity of P(PF-co-EG) hydrogels is one of the factors affecting cell adhesion, and that copolymer modification may be required for enhancing cell adhesion for an application involving the copolymers as in situ crosslinkable cell carriers.     

Cartilage-like mechanical properties of poly (ethylene glycol)-diacrylate hydrogels

Hydrogels prepared from poly-(ethylene glycol) (PEG) have been used in a variety of studies of cartilage tissue engineering. Such hydrogels may also be useful as a tunable mechanical material for cartilage repair. Previous studies have characterized the chemical and mechanical properties of PEG-based hydrogels, as modulated by precursor molecular weight and concentration. Cartilage mechanical properties vary substantially, with maturation, with depth from the articular surface, in health and disease, and in compression and tension. We hypothesized that PEG hydrogels could mimic a broad range of the compressive and tensile mechanical properties of articular cartilage. The objective of this study was to characterize the mechanical properties of PEG hydrogels over a broad range and with reference to articular cartilage. In particular, we assessed the effects of PEG precursor molecular weight (508?Da, 3.4?kDa, 6?kDa, and 10?kDa) and concentration (10-40%) on swelling property, equilibrium confined compressive modulus (H(A0)), compressive dynamic stiffness, and hydraulic permeability (k(p0)) of PEG hydrogels in static/dynamic confined compression tests, and equilibrium tensile modulus (E(ten)) in tension tests. As molecular weight of PEG decreased and concentration increased, hydrogels exhibited a decrease in swelling ratio (31.5-2.2), an increase in H(A0) (0.01-2.46?MPa) and E(ten) (0.02-3.5?MPa), an increase in dynamic compressive stiffness (0.055-42.9?MPa), and a decrease in k(p0) (1.2?×?10(-15) to 8.5?×?10(-15)?m(2)/(Pa?s)). The frequency-dependence of dynamic compressive stiffness amplitude and phase, as well as the strain-dependence of permeability, were typical of the time- and strain-dependent mechanical behavior of articular cartilage. H(A0) and E(ten) were positively correlated with the final PEG concentration, accounting for swelling. These results indicate that PEG hydrogels can be prepared to mimic many of the static and dynamic mechanical properties of articular cartilage.     

Binding specificity of alpha-bilirubin-imprinted poly(methacrylic acid-co-ethylene glycol dimethylacrylate) toward alpha-bilirubin

alpha-Bilirubin is an important index to determine the liver's functions. Poly(methacrylic acid-co-ethylene glycol dimethylacrylate) (poly(MAA-co-EGDMA)) imprinted with alpha-bilirubin was proposed and shown to be able to bind alpha-bilirubin specifically. The extraction condition was also discussed. Polymers prepared by imprinting bilirubin in poly(MAA-co-EGDMA) and in poly(beta-cyclodextrin-co-EGDMA) were compared. In this work, binding specificity of molecularly imprinted polymer (MIP) toward the target template, alpha-bilirubin, is discussed. Adsorption profile of alpha-bilirubin by bilirubin-imprinted poly(MAA-co-EGDMA) was measured as a function of time, from where the equilibrium could be determined. Two hours was determined to be the proper time for adsorption. Biliverdin as a rather similar analog compound of bilirubin was chosen for comparison of binding specificity in this study. Progesterone as well as testosterone was also chosen for study because they both co-exist in serum with bilirubin and might interfere with binding capacity of imprinted poly(MAA-co-EGDMA) toward alpha-bilirubin. Specificity of this polymer for bilirubin was thus confirmed by tasks carried out in mixture solutions comprised of compounds above. It is true that biliverdin contained in the binary mixture did affect the binding capacity of bilirubin. Nevertheless, polymer's binding specificity for bilirubin was essentially sufficient for recognition of alpha-bilirubin in the presence of other compounds. How MIP identified the target template molecule, alpha-bilirubin, is also elucidated.     

Osteogenic potential of poly(ethylene glycol)-poly(dimethylsiloxane) hybrid hydrogels

Growth factors have been shown to be potent mediators of osteogenesis. However, their use in tissue-engineered scaffolds not only can be costly but also can induce undesired responses in surrounding tissues. Thus, the ability to specifically induce osteogenic differentiation in the absence of exogenous growth factors through manipulation of scaffold material properties would be desirable for bone regeneration. Previous research indicates that addition of inorganic or hydrophobic components to organic, hydrophilic scaffolds can enhance multipotent stem cell (MSC) osteogenesis. However, the combined impact of scaffold inorganic content and hydrophobicity on MSC behavior has not been systematically explored, particularly in three-dimensional (3D) culture systems. The aim of the present study was therefore to examine the effects of simultaneous increases in scaffold hydrophobicity and inorganic content on MSC osteogenic fate decisions in a 3D culture environment toward the development of intrinsically osteoinductive scaffolds. Mouse 10T? MSCs were encapsulated in a series of novel scaffolds composed of varying levels of hydrophobic, inorganic poly(dimethylsiloxane) (PDMS) and hydrophilic, organic poly(ethylene glycol) (PEG). After 21 days of culture, increased levels of osteoblast markers, runx2 and osteocalcin, were observed in scaffolds with increased PDMS content. Bone extracellular matrix (ECM) molecules, collagen I and calcium phosphate, were also elevated in formulations with higher PDMS:PEG ratios. Importantly, this osteogenic response appeared to be specific in that markers for chondrocytic, smooth muscle cell, and adipocytic lineages were not similarly affected by variations in scaffold PDMS content. As anticipated, the increase in scaffold hydrophobicity accompanying increasing PDMS levels was associated with elevated scaffold serum protein adsorption. Thus, scaffold inorganic content combined with alterations in adsorbed serum proteins may underlie the observed cell behavior.     

Modulation of the pH-responsive properties of poly(L-lysine iso-phthalamide) grafted with a poly(ethylene glycol) analogue

A pH responsive pseudopeptide, poly(L-lysine iso-phthalamide), has been modified with a hydrophilic poly(ethylene glycol) analogue, Jeffamine M-1000 and the effect of grafting ratio on the pH responsive behaviour of the grafted polymers in aqueous solution investigated using fluorescence and 1H NMR spectroscopy. It was demonstrated that at below 35.1 wt% grafting, the modified polymers retained the pH-driven conformational transition of the parent polymer from an expanded structure at high degrees of ionisation to a compact hydrophobically stabilised structure at low degrees of ionisation. The onset of pH response and the pH range over which the conformational transition occurred varied significantly with degree of grafting. At Jeffamine M-1000 ratios in excess of 48.0 wt%, the graft polymer existed in a micellular form over the whole pH studied. Potential applications in drug delivery of both the linear and micellular forms are discussed.     

Spatially controlled bacterial adhesion using surface-patterned poly(ethylene glycol) hydrogels

We constructed surface-patterned hydrogels using low-energy focused electron beams to locally crosslink poly(ethylene glycol) (PEG) thin films on silanized glass substrates. Derived from electron-beam lithography, this technique was used to create patterned hydrogels with well-defined spatial positions and degrees of swelling. We found that cells of the bacterium Staphylococcus epidermidis adhered to and grew on the silanized glass substrates. These cells did not, however, adhere to surfaces covered by high-swelling lightly crosslinked PEG hydrogels. This finding is consistent with the cell-repulsiveness generally attributed to PEGylated surfaces. In contrast, S. epidermidis cells did adhere to surfaces covered by low-swelling highly crosslinked hydrogels. By creating precise patterns of repulsive hydrogels combined with adhesive hydrogels or with exposed glass substrate, we were able to spatially control the adhesion of S. epidermidis. Significantly, adhesive areas small enough to trap single bacterial cells could be fabricated. The results suggest that the lateral confinement imposed by cell-repulsive hydrogels hindered the cell proliferation and development into larger bacterial colonies.     

Biodegradable and thermoreversible hydrogels of poly(ethylene glycol)-poly(epsilon-caprolactone-co-glycolide)-poly(ethylene glycol) aqueous solutions

The aqueous solutions of triblock copolymers of poly(ethylene glycol)-poly(epsilon-caprolactone-co-glycolide)-poly(ethylene glycol) [PEG-P(CL-GA)-PEG] undergoing sol-gel transition as the temperature increases from 20 to 60 degrees C were successfully prepared. The thermogelling block copolymers were synthesized by subtle control of the hydrophilic/hydrophobic balance and the chain microstructures. The amphiphilic block copolymer formed micelles in aqueous solution, and the micelle aggregated as the temperature increased. The sol-gel transition of the copolymer aqueous solutions was studied focusing on the structure-property relationship. GA was incorporated into the polymer chain to prevent crystallization of PCL component and increase the polymer degradation. It is expected to be a promising long-term delivery system for pH-sensitive drugs, proteins, and genes.     

Transparent, elastomeric and tough hydrogels from poly(ethylene glycol) and silicate nanoparticles

The structures and mechanical properties of both physically and covalently cross-linked nanocomposite hydrogels made from poly(ethylene glycol) (PEG) and silicate nanoparticles (Laponite RD) are investigated. Injectable nanocomposite precursor solutions can be covalently cross-linked via photopolymerization. The resulting hydrogels are transparent and have interconnected pores, high elongation and toughness. These properties depend on the hydrogel composition, polymer-nanoparticle interactions and degree of cross-linking (both physical and covalent). Covalent cross-linking of polymer chains leads to the formation of an elastic network, whereas physical cross-linking between nanoparticles and polymer chains induces viscoelastic properties. At high deformations covalent bonds may be broken but physical bonds rebuild and to some extent self-heal the overall network structure. Addition of silicate also enhances the bioactivity and adhesiveness of the hydrogel as these materials stick to soft tissue as well as to hard surfaces. In addition, MC3T3-E1 mouse preosteoblast cells readily adhere and spread on nanocomposite hydrogel surfaces. Collectively, the combinations of properties such as elasticity, stiffness, interconnected network, adhesiveness to surfaces and bio-adhesion to cells provide inspiration and opportunities to engineer mechanically strong and elastic tissue matrixes for orthopedic, craniofacial and dental applications.     

Photopolymerized poly(ethylene glycol)/poly(L-lysine) hydrogels for the delivery of neural progenitor cells

Neural progenitor cells (NPCs) have shown promise in a number of models of disease and injury, but for these cells to be safe and effective, they must be directed to differentiate appropriately following transplantation. We have developed a photopolymerized hydrogel composed of macromers of poly(ethylene glycol) (PEG) bound to poly(L-lysine) (PLL) that supports NPC survival and directs differentiation. Green fluorescent protein (GFP) positive NPCs were encapsulated in these gels and demonstrated survival up to 17 days. When encapsulated in the gels at a photoinitiator concentration of 5.0 mg/ml, few NPCs (0.5 +/- 0.25%) demonstrated apoptosis. Furthermore, 55 +/- 6% of the NPCs cultured within the gels in epidermal growth factor (EGF) containing media differentiated into a mature neuronal cell type (neurofilament 200 positive) while the remainder 44 +/- 8% were undifferentiated (nestin positive). A small percentage, 1 +/- 0.4%, expressed the astrocytic marker glial acidic fibrilary protein. Photopolymerized PEG/PLL gels promote the survival and direct the differentiation of NPCs, making this system a promising delivery vehicle for NPCs in the treatment of injuries and diseases of the central nervous system.     

Synthesis, characterization and drug release from three-arm poly(epsilon-caprolactone) maleic acid/poly(ethylene glycol) diacrylate hydrogels

A biodegradable polymer network hydrogel with both hydrophobic and hydrophilic components was synthesized and characterized. The hydrophobic and hydrophilic components were a three-arm poly(epsilon-caprolactone) maleic acid (PGCL-Ma, as the hydrophobic constituent) and poly(ethylene glycol) diacrylate macromer (PEGDA, as a hydrophilic constituent), respectively. These two polymers were chemically photo-crosslinked to generate a three-dimensional network structure, which were characterized by FT-IR, DSC and SEM. The swelling property of the networks was studied in phosphate-buffered saline (PBS, pH 7.4). The results of this study showed that a wide-range swelling property was obtained by changing the composition ratio of PGCL-Ma to PEGDA. The in vitro release of bovine serum albumin (BSA) from these hydrogels as a function of the PEGDA to PGCL-Ma composition ratio and incubation time was examined and we found that the incorporation of PEGDA into PGCL-Ma increased the initial burst release of BSA. As the PEGDA component increased, the rate of formation of a loose three-dimensional (3D) network structure increased; consequently, the sustained rate and extent of BSA release increased. We suggest that the release of BSA was controlled by both diffusion of BSA through swelling of the hydrophilic phase during an early stage and degradation of the hydrophobic phase during a late stage; and that the relative magnitude of diffusion versus degradation controlled release depended on composition ratio and immersion time.     

Biodegradable DNA-enabled poly(ethylene glycol) hydrogels prepared by copper-free click chemistry

Significant research has focused on investigating the potential of hydrogels in various applications and, in particular, in medicine. Specifically, hydrogels that are biodegradable lend promise to many therapeutic and biosensing applications. Endonucleases are critical for mechanisms of DNA repair. However, they are also known to be overexpressed in cancer and to be present in wounds with bacterial contamination. In this work, we set out to demonstrate the preparation of DNA-enabled hydrogels that could be degraded by nucleases. Specifically, hydrogels were prepared through the reaction of dibenzocyclooctyne-functionalized multi-arm poly(ethylene glycol) with azide-functionalized single-stranded DNA in aqueous solutions via copper-free click chemistry. Through the use of this method, biodegradable hydrogels were formed at room temperature in buffered saline solutions that mimic physiological conditions, avoiding possible harmful effects associated with other polymerization techniques that can be detrimental to cells or other bioactive molecules. The degradation of these DNA-cross-linked hydrogels upon exposure to the model endonucleases Benzonase^® and DNase I was studied. In addition, the ability of the hydrogels to act as depots for encapsulation and nuclease-controlled release of a model protein was demonstrated. This model has the potential to be tailored and expanded upon for use in a variety of applications where mild hydrogel preparation techniques and controlled material degradation are necessary including in drug delivery and wound healing systems.     

Photopolymerized poly(ethylene glycol)/poly(L-lysine) hydrogels for the delivery of neural progenitor cells

Neural progenitor cells (NPCs) have shown promise in a number of models of disease and injury, but for these cells to be safe and effective, they must be directed to differentiate appropriately following transplantation. We have developed a photopolymerized hydrogel composed of macromers of poly(ethylene glycol) (PEG) bound to poly(L-lysine) (PLL) that supports NPC survival and directs differentiation. Green fluorescent protein (GFP) positive NPCs were encapsulated in these gels and demonstrated survival up to 17 days. When encapsulated in the gels at a photoinitiator concentration of 5.0 mg/ml, few NPCs (0.5 ± 0.25%) demonstrated apoptosis. Furthermore, 55 ± 6% of the NPCs cultured within the gels in epidermal growth factor (EGF) containing media differentiated into a mature neuronal cell type (neurofilament 200 positive) while the remainder 44 ± 8% were undifferentiated (nestin positive). A small percentage, 1 ± 0.4%, expressed the astrocytic marker glial acidic fibrilary protein. Photopolymerized PEG/PLL gels promote the survival and direct the differentiation of NPCs, making this system a promising delivery vehicle for NPCs in the treatment of injuries and diseases of the central nervous system.     

Conformational transitions in poly(methacrylic acid) gel/poly(ethylene glycol) complexes. Effect of the gel cross-linking density

We present results on the effect of the cross-linking density on the collapse of poly-(methacrylic acid) gels induced by the interaction with linear poly(ethylene glycol) and on their subsequent reswelling (reentrant transition). The abruptness and the amplitude of both transitions increase with decreasing cross-linking density of the gel. A special feature of the reswelling transition for the PMAA gel/PEG system is the existence of a wide range of PEG concentrations where both collapsed and swollen states of the gel are stable. The conditions for the existence of such a region are easier realized for slightly cross-linked gels. The corresponding theoretical explanation is given. The swelling degree of the collapsed gels increases with increasing cross-linking density, which is explained by the presence of a certain amount of hydrophilic PEG loops in highly cross-linked gels.     

Preparation and characterization of poly(ethylene glycol) hydrogels cross-linked by hydrolyzable polyrotaxane

PEG hydrogels cross-linked by a hydrolyzable polyrotaxane were prepared and their hydrolytic erosion characterized in terms of supramolecular dissociation of the polyrotaxane. The hydrolyzable polyrotaxane, in which many alpha-cyclodextrins (alpha-CDs) are threaded onto a poly(ethylene glycol) (PEG) chain capped with L-phenylalanine via ester linkages, was used as a multifunctional cross-linker: the PEG network was covalently bound to hydroxyl groups of alpha-CDs in the polyrotaxane. The contact angle and water content of the hydrogels were varied with the polyrotaxane content in the feed. In vitro hydrolysis study revealed that the time to reach complete gel erosion was shortened by increasing the polyrotaxane content in the feed in relation to the decreased number of chemical cross-links between PEG and alpha-CDs in the polyrotaxane. The hydrogel degradation in a physiological condition was found to be followed by bulk mechanism. These findings suggest that changing the preparative conditions such as polyrotaxane content will make it possible to control programmed gel erosion for tissue engineering.