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The tensile properties of alginate hydrogels.

Alginate hydrogels are currently being employed and explored for a broad range of medical applications including cell encapsulation, drug delivery, and tissue engineering. In these capacities, knowledge of the mechanical and material properties of the hydrogels and the properties that govern and influence them is necessary to adequately design and effectively use these systems. Although much is known about the mechanical properties of alginate in compression and shear, little is known about the tensile characteristics. Thus, an extensive tensile assessment of alginate hydrogels was completed as a function of alginate type, formulation, gelling conditions, incubation, and strain rate. In general, the initial tensile behavior and properties of alginate hydrogels were highly dependent on the choice of the alginate polymer and how it was processed. Specifically, high guluronic acid containing alginate polymers yielded stronger, more ductile hydrogels than high mannuronic acid containing alginates. The ultimate stress, ultimate strain, and tensile modulus were decreased by increased phosphate concentrations, solution reconstitution with phosphate buffered saline instead of culture media, and peptide modification. Incubation of hydrogels for at least 7 days diminished many of the initial tensile property differences associated with formulation and gelling conditions. Overall, by controlling the specific alginate polymer and processing methods, a wide range of tensile properties are available from these hydrogels.

Alginates↗

Crosslinked hyaluronic acid hydrogels: a strategy to functionalize and pattern.

The physiological activity of hyaluronic acid (HA) polymers and oligomers makes it a promising material for a variety of applications. The development of HA-hydrogel scaffolds with improved mechanical stability against degradation and biochemical functionality may enhance their application to tissue engineering. In this report, a crosslinking strategy targeting the alcohol groups via a poly(ethylene glycol) diepoxide crosslinker was investigated for the generation of degradable HA hydrogels. To provide support for cell adhesion in vitro, collagen was incorporated into the HA solution prior to the crosslinking process. The hydrogels have a continuous exterior and a porous interior, with pore diameters ranging from 6 to 9 microm. HA and HA-collagen hydrogels degrade in the presence of hyaluronidase and collagenase enzymes, indicating that the chemical modification does not prevent biodegradation. Complete degradation of the hydrogels occurred within 14 days in hyaluronidase (100 U/ml) and 3 days in collagenase (66 U/ml). Pattern transfer was employed to introduce a surface topography onto the hydrogel, which was able to orient cell growth. Furthermore, the hydrogels could be functionalized with the biomolecule neutravidin by incorporation of biotin along the HA backbone. This biotinylation approach may allow attachment of bioactive molecules that are conjugated to avidin.

Absorbable Implants↗

Osteogenic differentiation of rat bone marrow stromal cells cultured on Arg-Gly-Asp modified hydrogels without dexamethasone and beta-glycerol phosphate.

In this study, we investigated the effect of signaling peptides incorporated into oligo(poly(ethylene glycol) fumarate) (OPF) hydrogels on in vitro differentiation and mineralization of marrow stromal cells (MSCs) cultured in media without soluble osteogenic supplements (dexamethasone and beta-glycerol phosphate). When MSCs were cultured for 16 days on OPF hydrogels modified with Arg-Gly-Asp (RGD) containing peptides, the normalized cell number was dependent on the peptide concentration between days 0 and 5 and reached comparable values at day 10 regardless of the concentration. The alkaline phosphatase (ALP) activity of MSCs on the peptide-modified OPF hydrogels was also concentration-dependent: ALP activity showed peaks on day 10 or day 13 on OPF hydrogels modified with 2.0 and 1.0 micromol peptide/g, which were significantly greater than those on the OPF hydrogels modified with 0.1 micromol peptides/g or no peptide. A characteristic marker of osteoblastic differentiation, osteopontin (OPN), was detected for all the test groups. However, OPN secretion between days 0 and 10 was significantly higher on the peptide modified hydrogels compared to that on tissue culture-treated polystyrene. Taken together, the results indicate that the presence of signaling peptide allows for a favorable microenvironment for MSCs to differentiate into osteoblasts and produce mineralized matrix, although the soluble factors may further enhance calcium deposition. These findings further support the usefulness of OPF hydrogels as scaffolds for guided bone regeneration, and represent an initial step in exploring the complex relationship between soluble and insoluble factors in osteogenic differentiation on biodegradable materials.

Absorbable Implants↗

A novel pH- and ionic-strength-sensitive carboxy methyl dextran hydrogel.

A fast and simple method for the preparation of pH-sensitive hydrogel membranes for drug delivery and tissue engineering applications has been developed using carbodiimide chemistry. The hydrogels were formed by the intermolecular cross-linking of carboxymethyl dextran (CM-dextran) using 1-ethyl-(3-3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). Infrared spectra of the hydrogels suggest the formation of ester bonds between the hydroxyl and carboxyl groups in the CM-dextran. The porosity of the hydrogels produced, as shown by protein diffusion, increases in response to changes in the pH and the ionic strength of the external medium. The results show pH-dependent swelling behaviour arising from the acidic pedant groups in the polymer network. The diffusion of the protein lysozyme through the hydrogel membranes increased with increases in both pH (5.0-9.0) and ionic strength. The effect of changes of pH and ionic strength on the hydrogel's permeability was shown to be reversible. Scanning electron microscopy of these hydrogels showed that pH-dependent changes in permeability are mirrored by morphological changes in gel structure.

Dextrans↗

Synthesis, properties and controlled release behaviors of hydrogel networks using cyclodextrin as pendant groups.

Based on inclusion character of beta-cyclodextrin (beta-CD) with drug molecule and low glass transition temperature of poly(2-hydroxyethyl acrylate) (PHEA), a series of hydrogels with different compositions were synthesized by the copolymerization of a monovinyl cyclodexrin monomer with 2-hydroxyethyl acrylate (HEA). The structure and properties of the hydrogels were characterized by FTIR, DSC, TGA and swelling measurements. It is found that swelling ratios of these beta-CD hydrogels can keep a relative stability in the range of pH from 1.4 to 7.4, and are not sensitive to change in NaCl concentration. Using drug N-acety-5-methoxytryptamine (melatonin, MEL) as a model molecule, the controlled drug release behaviors of these hydrogels were investigated. The results indicate that the diffusion and permeation of MEL from the hydrogels may be a dominant factor for its release. Owing to the formation of MEL/beta-CD retarding diffusion rate of MEL, a sustained release of MEL from hydrogel with high content of beta-CD can be obtained compared with hydrogel PHEA without beta-CD.

Absorption↗

Evaluation of an in situ forming hydrogel wound dressing based on oxidized alginate and gelatin.

Wound dressings that can be formed in situ offer several advantages over the use of preformed dressings such as conformability without wrinkling or fluting in the wound bed, ease of application and improved patient compliance and comfort. Here we describe such an in situ forming hydrogel wound dressing from gelatin, oxidized alginate and borax. Periodate oxidized alginate rapidly cross-links proteins such as gelatin in the presence of borax to give in situ forming hydrogels that are both non-toxic and biodegradable. The composite matrix has the haemostatic effect of gelatin, the wound healing-promoting feature of alginate and the antiseptic property of borax to make it a potential wound dressing material. The hydrogel was found to have a fluid uptake of 90% of its weight which would prevent the wound bed from accumulation of exudates. The water vapour transmission rate (WVTR) of the hydrogel was found to be 2686+/-124 g/m2/day indicating that the hydrogel can maintain a moist environment over wound bed in moderate to heavily exuding wound which would enhance epithelial cell migration during the healing process. The wound healing efficacy of hydrogel was evaluated in experimental full thickness wounds using a rat model which demonstrated that within 2 weeks, the wound covered with gel was completely filled with new epithelium without any significant adverse reactions. These in situ forming hydrogels fulfil many critical elements desirable in a wound dressing material.

Alginates↗

The effect of ethylene glycol methacrylate phosphate in PEG hydrogels on mineralization and viability of encapsulated hMSCs.

Owing to their resistance to protein adsorption, poly(ethylene glycol) (PEG) hydrogels are unable to support the adhesion of human mesenchymal stem cells (hMSCs). Human MSCs attach to various substrates through cell adhesion proteins, and osteopontin is an important cell adhesion protein for both MSCs and osteoblasts found in bone. As such, we hypothesized that cell adhesion to a PEG hydrogel could be dramatically improved by actively promoting the formation of a mineral phase throughout the hydrogel. This was accomplished using the photoreactive, phosphate-containing molecule ethylene glycol methacrylate phosphate (EGMP). When EGMP was incorporated into the PEG hydrogel, it resulted in the formation of a bone-like mineral phase, as verified using compositional analysis and X-ray diffraction. Furthermore, cell viability of gel-encapsulated hMSCs was increased in the presence of EGMP from 15% in the absence of EGMP to 97% in the presence of 50 mm EGMP. This improvement in cell viability was thought to be due to the ability of mineralized hydrogels to sequester cell-secreted osteopontin. It was found that EGMP-containing PEGDA hydrogels are able to sequester the important cell adhesion protein osteopontin, and these hydrogels promoted hMSC adhesion and spreading.

Calcium Phosphates↗

Examination of the effects of poly(N-vinylpyrrolidinone) hydrogels in direct and indirect contact with cells.

Poly(N-vinylpyrrolidinone) (PNVP) has been used in various biomedical applications for many years. This study explores two PNVP hydrogels for their biocompatibility with skin cells and their ability to support the growth of skin cells in direct and indirect contact with the cells. Two crosslinked PNVP's were investigated, one crosslinked with ethylene glycol dimethacrylate (EGDMA) and the other crosslinked with diethylene glycol bisallylcarbonate (DEGBAC). The different crosslinkers lead to hydrogels with different mechanical and slightly different biological properties. While neither hydrogel proved to be a suitable substrate for culturing cells (based on fibroblasts and a range of other cells), indirect contact with both showed them to be biocompatible and even stimulatory to fibroblasts. The P(NVP-co-DEGBAC) hydrogel stimulated fibroblast viability more reliably than the P(NVP-co-EGDMA) hydrogel when in indirect contact with cells. This effect was shown to be independent of the presence of foetal calf serum in the culture media, and could not be explained by any hydrogel breakdown products during the course of these experiments. Rather the phenomenon was observed to be the result of a dynamic interaction between the hydrogels and the cells.

Cells, Cultured↗

Synthesis of cell-adhesive dextran hydrogels and macroporous scaffolds.

Dextran hydrogels have been previously investigated as drug delivery vehicles and more recently as macroporous scaffolds; however, the non-cell-adhesive nature of dextran has limited its utility for tissue engineering. To overcome this limitation, macroporous scaffolds of methacrylated dextran (Dex-MA) copolymerized with aminoethyl methacrylate (AEMA) were synthesized, thereby introducing primary amine groups for covalent immobilization of extracellular-matrix-derived peptides. The amino group density for hydrogels copolymerized with 0.5 wt% AEMA was found to be 36.1+/-0.4 micromol/cm(3) by elemental analysis. To further enhance cellular interaction, poly(Dex-MA-co-AEMA) hydrogels were modified with either CRGDS or a mixture of CDPGYIGSR and CQAASIKVAV (1:1, v/v) using sulfo-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC). The immobilized peptide concentration was determined using amino acid analysis at: 2.6+/-0.9 micromol/cm(3) for CRGDS-derived hydrogels and 2.2+/-0.3 micromol/cm(3) plus 1.9+/-0.2 micromol/cm(3) for CDPGYIGSR plus CQAASIKVAV-derived hydrogels, respectively. Cellular interactions of primary embryonic chick dorsal root ganglia (DRGs) were compared on the hydrogels. Cell adhesion and neurite outgrowth on poly(Dex-MA) increased with copolymerization of AEMA and further improved with peptide modification and significantly for CDPGYIGSR/CQAASIKVAV-derived poly(Dex-MA-co-AEMA) hydrogels. Moreover, DRGs penetrated within the first 600 microm of the scaffolds, thereby demonstrating the potential of this scaffold for guided cell and axonal regeneration in vivo.

Adhesives↗

Model development and numerical simulation of electric-stimulus-responsive hydrogels subject to an externally applied electric field.

Based on a multi-phasic mixture theory with consideration of ionic diffusion and convection, a multi-physic model, called the multi-effect-coupling electric-stimulus (MECe) model, is developed for simulation of responsive behavior of the electric-sensitive hydrogels when they are immersed into a bathing solution subject to an externally applied electric field. In the developed model, with chemo-electro-mechanical coupling effects, the convection-diffusion equations for concentration distribution of diffusive ions incorporate the influence of electric potential. The electroneutrality condition is replaced by the Poisson equation for distribution of electric potential. The steady and transient analyses of hydrogel deformation are easily carried out by the continuity and momentum equations of the mixture phase. Further, the computational domain of the present model covers both the hydrogel and the surrounding solution. In order to solve the present mathematical model consisting of multi-field coupled nonlinear partial differential governing equations, a hierarchical iteration technique is proposed and a meshless Hermite-Cloud method (HCM) is employed. The steady-state simulation of the electric-stimulus responsive hydrogel is numerically conducted when it is subjected to an externally applied electric field. The hydrogel deformation and the ionic concentrations as well as electric potentials of both the hydrogel and external solution are investigated. The parameter influences on the swelling behaviors of the hydrogel are also discussed in detail. The simulating results are in good agreement with the experimental data and they validate the presently developed model.

Biosensing Techniques↗

Biodegradable dextran hydrogels crosslinked by stereocomplex formation for the controlled release of pharmaceutical proteins.

Hydrogels are based on hydrophilic polymers, which are crosslinked to prevent dissolution in water. Because hydrogels can contain large amounts of water, they are interesting devices for the delivery of proteins. In this contribution a biodegradable dextran hydrogel is described which is based on physical interactions and is particularly suitable for the controlled delivery of pharmaceutically active proteins. The unique feature of our system is that the preparation of the hydrogels takes place in an all-aqueous solution, by which the use of organic solvents is avoided. Furthermore, chemical crosslinking agents are not needed to create the hydrogels, since crosslinking is established physically by stereocomplex formation between enantiomeric oligomeric lactic acid chains. The hydrogel is simply obtained after mixing aqueous solutions of dextran(l)-lactate and dextran(d)-lactate. In this contribution, the formation of the hydrogels as well as their protein release properties and degradation behavior are discussed.

Biocompatible Materials↗

The effects of electrically assisted methods on transdermal delivery of nalbuphine benzoate and sebacoyl dinalbuphine ester from solutions and hydrogels.

The aim of this study was to assess the effects of iontophoresis and electroporation on transdermal delivery of nalbuphine (NA) and its two novel prodrugs: nalbuphine benzoate (NAB) and sebacoyl dinalbuphine ester (SDN) from solutions as well as from hydrogels. Hydroxypropyl cellulose (HPC) and carboxymethyl cellulose (CMC) were used in hydrogel formulations to evaluate their feasibility for delivery of NA and its prodrugs. Application of iontophoresis or electroporation significantly enhanced the in vitro permeation of NA and its prodrugs. The enhancement effect was more pronounced after applying iontophoresis. The combination of two electrically assisted methods enhanced the delivery of NA; however, no such enhancement was observed for the permeation of NAB and SDN. Hydrogels containing low concentration HPC did not affect the passive as well as electrically assisted permeation of NA and its prodrugs. The increase of hydrogel concentration as well as molecular weight significantly decreased the electrically assisted permeation of NA, whereas the permeation of NAB and SDN remained unchanged. For the electrically assisted permeation from CMC-based hydrogels, the reduced permeation from higher percentage of CMC hydrogels may be attributed the viscosity effect as well as the ion competition effect. The above results demonstrated that lipophilicity and molecular size, as well as hydrogel compositions had significant effects on skin permeation of NA, NAB and SDN via passive diffusion or under the electric field.

Administration, Cutaneous↗

Release of theophylline from polymer blend hydrogels.

Polymer blending is a simple yet attractive method to obtain combined physical and mechanical properties of polymers. In this paper, three types of blend hydrogels were prepared, each by physically blending two different natural polymers, and a model drug, theophylline (TPH), was immobilized into these hydrogels for the studies of drug release. The release profiles of TPH from various types of hydrogels were determined by UV-vis absorption measurement at 272 nm. The experimental results show that the releases of TPH from these hydrogels are dependent upon the composition of the hydrogel, the type of component, the possible interactions between two component polymers, as well as external temperature. All the release profiles clearly demonstrate a temperature effect. Among the three blend hydrogels, the slowest release was observed from the blend hydrogel of gelatin and agar with a weight ratio of 1:1. The drug release patterns and release mechanisms have been discussed by considering the possible molecular interactions and gel network structures.

Carrageenan↗

A thermo- and pH-sensitive hydrogel composed of quaternized chitosan/glycerophosphate.

The quaternized chitosan was synthesized by the reaction of chitosan and glycidyltrimethylammonium chloride (GTMAC) and named as N-[(2-hydroxy-3-trimethylammonium) propyl] chitosan chloride (HTCC). A novel hydrogel system composed of HTCC/glycerophosphate (HTCC/GP) with thermo- and pH-sensitivity was synthesized and used as an intelligent drug carrier. The formulation was solution below or at room temperature, which allowed it injectable and to incorporate living cells, proteins, enzymes or other therapeutic drugs easily. Once the surrounding temperature was up to 37 degrees C, the system was transformed to a non-flowing hydrogel, and the formed hydrogel can release the trapped drug as a function of pH values. The swelling behavior of the system and the release profiles of doxorubicin hydrochloride (DX) as a model drug at different pH values were investigated. At acidic condition the hydrogel dissolved and released drug quickly, while it absorbed water and released drug slowly at neutral or basic conditions. Hydrogel composed of chitosan hydrochloride and glycerophosphate (CS/GP) was also prepared to compare with HTCC/GP hydrogel. The HTCC/GP hydrogel in this study was transparent which made it suitable for some specific uses such as ocular drug formulation.

Chitosan↗

Synergistic effect of TGFbeta-3 on chondrogenic differentiation of rabbit chondrocytes in thermo-reversible hydrogel constructs blended with hyaluronic acid by in vivo test.

In this study, a hydrogel composite, based on the thermo-reversible hydrogel of p(NiPAAm-co-AAc) and hyaluronic acid (HA) was used as an injectable cell and growth factor carrier for cartilage tissue engineering applications. Rabbit chondrocytes were embedded in blended hydrogel composites co-encapsulated with the transforming growth factor beta-3 (TGFbeta-3). The blended hydrogel with the embedded chondrocytes and HA co-encapsulating unloaded growth factors and those with the thermo-reversible hydrogel were used as the controls to examine the effects of TGFbeta-3 on neocartilage formation. The blended hydrogel loaded with TGFbeta-3 embedded with chondrocytes were injected subcutaneously into the nude mice. The mice were monitored for 8 weeks after the injection. Both the differentiation and level of cartilage-specific ECM production were significantly higher in the presence of HA and growth factor than in the control without the growth factor. The level of cartilage associated ECM proteins was examined by immunohistochemical staining (collagen types II and X) as well as by Safranin-O and Alcian blue (GAG) staining. The results showed the potential application of blended hydrogel mixed with the growth factor to neocartilage formation.

Acrylamides↗

In vitro and in vivo evaluation of recombinant silk-elastinlike hydrogels for cancer gene therapy.

The objectives of this study were to evaluate: (i). the influences of hydrogel geometry, DNA molecular weight, and DNA conformation on DNA release from a silk-elastinlike protein polymer (SELP) hydrogel, (ii). the bioactivity and transfection efficiency of encapsulated DNA over time in vitro, (iii). the delivery and transfection of a reporter gene in a murine model of human breast cancer in vivo, and (iv). the in vitro release and bioactivity of adenovirus containing the green fluorescent protein (gfp) gene as a marker of gene transfer. Plasmid DNA was released from SELP hydrogels in a size-dependent manner, with the average effective diffusivity ranging from 1.70+/-0.52 x 10(-12) cm(2)/s for a larger plasmid (11 kbp) to 2.55+/-0.51 x 10(-10) cm(2)/s for a smaller plasmid (2.6 kbp). Plasmid conformation also influenced the rate of release, with the rank order linear>supercoiled>open-circular. DNA retained bioactivity in vitro, after encapsulation in a SELP hydrogel for up to 28 days. Delivery of pRL-CMV from a SELP hydrogel resulted in increased transfection in a murine model of human breast cancer by 1-3 orders of magnitude, as compared to naked DNA. The release of a bioactive adenoviral vector was related to the concentration of the polymer in the hydrogel. These studies indicate that genetically engineered SELP hydrogels have potential as matrices for controlled nonviral and viral gene delivery.

Amino Acid Sequence↗

PEG-grafted chitosan as an injectable thermosensitive hydrogel for sustained protein release.

Thermosensitive polymer hydrogels that undergo a sol-to-gel transition in response to temperature changes are of great interest in therapeutic delivery and tissue engineering as injectable depot systems. A chitosan-based, injectable thermogel was prepared by grafting an appropriate amount of PEG onto the chitosan backbone and studied for drug release in vitro using bovine serum albumin (BSA) as a model protein. When more than approximately 40 wt.% of PEG was grafted to chitosan chains via covalent bonding, the aqueous solution of the resultant copolymer was an injectable liquid at low temperature and transformed to a semisolid hydrogel at body temperature. After an initial burst release in the first 5 h, a steady linear release of protein from the hydrogel was achieved for a period of approximately 70 h. Prolonged quasi-linear release of protein up to 40 days was achieved by crosslinking the hydrogel with genipin in situ, in a fashion suitable for protein encapsulation while maintaining the injectability of the hydrogel. The crosslinkage transformed the copolymer from a physical gel to an insoluble chemical gel and substantially reduced the initial burst release of protein. Both high performance liquid chromatography (HPLC) and gel electrophoresis indicated that the primary structure of BSA released from the hydrogels with or without genipin-crosslinking was generally conserved. The hydrogel can be prepared in solutions with a physiological pH, allowing the safe incorporation of bioactive molecules for a broad range of medical applications, particularly for sustained in vivo drug release and tissue engineering.

Chitosan↗

The repair of brain lesion by implantation of hyaluronic acid hydrogels modified with laminin.

The hyaluronic acid (HA) hydrogels modified with laminin were used for implantation in rat brain in present study, in order to investigate its effects in reparation of injury in the CNS. Cross-linked HA hydrogels were synthesized and their characteristics were analyzed. Laminin, an extracellular matrix protein, which participates in neuronal development and survival, was immobilized on the backbone of the hydrogels. Hydrogels unmodified and modified with laminin were implanted into cortical defects mechanically created in rats and their ability to improve tissue reconstruction was then evaluated. After 6 and 12 weeks of implantation, sections of brains were processed with Nissl and Glees staining for revealing neural cell bodies and fibers, with DAB histochemistry for detecting the blood vessels, as well as with immunocytochemistry for recognizing GFAP. The sections were also taken to SEM and TEM for ultrastructral examination. The results showed that the HA hydrogels synthesized had mechanical properties and rheological behavior similar to the brain tissue. After being implanted into the lesion of the cortex, the porous hydrogels created a scaffold, which could support cell infiltration and angiogenesis, and simultaneously inhibit the formation of glial scar. In addition, HA hydrogels modified with laminin could promote neurite extension. It seems possible that the tissue engineering technique may pave the way to repair injury in the CNS as suggested by the results in present study.

Animals↗