Control of cell cultivation and cell sheet detachment on the surface of polymer/clay nanocomposite hydrogels

Cell cultivation on the surface of a novel poly(N-isopropylacrylamide) (PNIPA) hydrogel (N-NC gel), consisting of a specific type of organic (PNIPA)/inorganic (clay) network, was studied using three cell types; HepG2 human hepatoma cells, human dermal fibroblasts, and human umbilical vein endothelial cells. For the first time, it was found that cells could be cultured to be confluent on the surfaces of PNIPA hydrogels using N-NC gels, regardless of gel thickness. Cell adhesion and proliferation on N-NC gels exhibit strong dependencies on clay concentration (C(clay)), and the numbers of cultured cells are maximum at about C(clay) = 6 x 10(-2) mol (45.72 g)/1 L of H(2)O. On the contrary, it was almost impossible to culture cells on conventional, chemically crosslinked PNIPA hydrogels, regardless of their cross-linker concentration. The reasons why cells cultured only on the surfaces of N-NC gels with their specific network structure and composition were discussed in terms of water content, protein adsorption, surface flatness, hydrophobicity of dehydrated PNIPA chains, and the anionic charge on exfoliated clay. Finally, it was found that cells cultured on the surfaces of N-NC gels could be detached in the forms of sheets of cells without trypsin treatment, but by just decreasing the temperature to 20 degrees C.     

Synthesis of biocompatible polymeric hydrogels with tunable adhesion to both hydrophobic and hydrophilic surfaces

We report the synthesis of biocompatible polymeric hydrogels based on poly(vinyl acetate) (PVAc) and poly(methyl vinyl ether-co-maleic anhydride) (PMVE-MA). These polymeric hydrogels show strong and tunable adhesion to both hydrophobic and hydrophilic surfaces and should be ideal candidates as bioadhesives for applications such as denture adhesion. PVAc was partially hydrolyzed and then mixed with PMVE-MA. Crosslinking between these two polymers through reactions between hydroxyl groups in partially hydrolyzed PVAc and maleic anhydride groups in PMVE-MA increased their compatibility and prevented phase separation so transparent hydrogels were formed. The adhesion of these polymeric hydrogels to hydrophobic and hydrophilic surfaces was tailored by regulating the degree of hydrolysis of PVAc and the molecular weights of the polymers. In the vicinity of critical gel point, where the elastic modulus G' and the viscous modulus G' scale as G' approximately G' approximately omega (0.3), polymeric hydrogels show optimal adhesion. Transparent gels are formed in mixed solvents of water and ethanol. The content of ethanol in the mixed solvent can be partially replaced by propylene glycol, glycerol, or poly(ethenyl glycol)-400, and the composition of appropriate mixed solvents can be determined by the calculation of solubility parameters.     

Photopolymerized thermosensitive hydrogels: synthesis, degradation, and cytocompatibility

In situ forming hydrogels based on thermosensitive polymers have attractive properties for tissue engineering. However, the physical interactions in these hydrogels are not strong enough to yield gels with sufficient stability for many of the proposed applications. In this study, additional covalent cross-links were introduced by photopolymerization to improve the mechanical properties and the stability of thermosensitive hydrogels. Methacrylate groups were coupled to the side chains of triblock copolymers (ABA) with thermosensitive poly( N-(2-hydroxypropyl) methacrylamide lactate) A blocks and a hydrophilic poly(ethylene glycol) B block. These polymers exhibit lower critical solution temperature (LCST) behavior in aqueous solution and the cloud point decreased with increasing amounts of methacrylate groups. These methacrylate groups were photopolymerized above the LCST to render covalent cross-links within the hydrophobic domains. The mechanical properties of photopolymerized hydrogels were substantially improved and their stability was prolonged significantly compared to nonphotopolymerized hydrogels. Whereas non-UV-cured gels disintegrated within 2 days at physiological pH and temperature, the photopolymerized gels degraded in 10 to 25 days depending on the degree of cross-linking. To assess biocompatibility, goat mesenchymal stem cells were seeded on the hydrogel surface or encapsulated within the gel and they remained viable as demonstrated by a LIVE/DEAD cell viability/cytotoxicity assay. Expression of alkaline phosphatase and production of collagen I demonstrated the functionality of the mesenchymal stem cells and their ability to differentiate upon encapsulation. Due to the improved mechanical properties, stability, and adequate cytocompatibility, the photopolymerized thermosensitive hydrogels can be regarded as highly potential materials for applications in tissue engineering.     

Synthesis, swelling behavior, and biocompatibility of novel physically cross-linked polyurethane-block-poly(glycerol methacrylate) hydrogels

Physically cross-linked novel block copolymer hydrogels with tunable hydrophilic properties for biomedical applications were synthesized by controlled radical polymerization of polyurethane macroiniferter and (2,2-dimethyl-1,3-dioxolane) methyl methacrylate. The block copolymers were converted to hydrogels by the selective hydrolysis of poly[(2,2-dimethyl-1,3-dioxolane) methyl methacrylate] block to poly(glycerol methacrylate). The block copolymerization has been monitored by monomer conversion and molecular weight increase as a function of time. It was observed that the polymerization proceeded with a characteristic "living" behavior where both monomer conversion and molecular weight increased linearly, with increasing reaction time. The resulting hydrogels were investigated for their equilibrium water content (EWC), dynamic water contact angles, swelling kinetics, thermodynamic interaction parameters, plasma protein adsorption, and platelet adhesion. Similar to our previous mechanically responsive hydrogels (Mequanint, K.; Sheardown, H. J. Biomater. Sci. Polym. Ed. 2005, 10, 1303-1318), the present results indicated that block copolymer hydrogels have excellent hydrophilicity and swelling behavior with improved modulus of elasticity. The equilibrium swelling was affected by the hydrolysis time, block length of poly(glycerol methacrylate), temperature, and the presence of soluble salts. Fibrinogen adsorption and platelet adhesion were significantly lower for the hydrogels than for the control polyurethane, whereas albumin adsorption increased for the hydrogels in proportion to the contents of poly(glycerol methacrylate). These hydrogels have potential in a number of biomedical applications such as drug delivery and scaffolds for tissue engineering.     

Antifouling properties of tough gels against barnacles in a long-term marine environment experiment

In the marine environment, the antifouling (AF) properties of various kinds of hydrogels against sessile marine organisms (algae, sea squirts, barnacles) were tested in a long-term experiment. The results demonstrate that most hydrogels can endure at least 2 months in the marine environment. In particular, mechanically tough PAMPS/PAAm DN and PVA gels exhibited AF activity against marine sessile organisms, especially barnacles, for as long as 330 days. The AF ability of hydrogels toward barnacles is explained in terms of an ‘easy-release’ mechanism in which the high water content and the elastic modulus of the gel are two important parameters.     

Biodegradable thermoresponsive hydrogels for aqueous encapsulation and controlled release of hydrophilic model drugs

A series of hydrogels with both thermoresponsive and completely biodegradable properties was developed for aqueous encapsulation and controlled release of hydrophilic drugs in response to temperature change. The hydrogels were prepared in phosphate-buffered saline (pH 7.4) through free radical polymerization of N-isopropylacrylamide (NIPAAm) monomer and a dextran macromer containing multiple hydrolytically degradable oligolactate-2-hydroxyethyl methacrylate units (Dex-lactateHEMA). Swelling measurement results demonstrated that four gels with feeding weight ratios of NIPAAm:Dex-lactateHEMA = 7:2, 6:3, 5:4, and 4:5 (w/w) were thermoresponsive by showing a lower critical solution temperature at approximately 32 degrees C. The swelling and degradation of the hydrogels strongly depended on temperature and hydrogel composition. An empirical mathematical model was established to describe the fast water absorption at the early stage and deswelling at the late stage of the hydrogels at 37 degrees C. Two hydrophilic model drugs, methylene blue and bovine serum albumin, were loaded into the hydrogels during the synthesis process. The molecular size of the drugs, the hydrophilicity and degradation of the hydrogels, and temperature played important roles in controlling the drug release.     

The use of immobilized Saccharomyces cerevisiae on radiation crosslinked acrylamide–maleic acid hydrogel carriers for production of ethyl alcohol

Radiation crosslinked acrylamide/maleic acid (AAm/MA) copolymers were prepared by γ-irradiation. They were used in experiments on swelling, diffusion, and immobilization of yeast cells (Saccharomyces cerevisiae) for the production of ethyl alcohol. AAm/MA hydrogels containing different amount of MA, irradiated at different doses, were used for swelling and diffusion studies. The parameters of swelling, diffusional exponents, network constants, diffusion coefficients and percent porosity of the hydrogel/penetrant systems were calculated and evaluated. Yeast cells were immobilized on to the hydrogels by adsorption during multiplication, and ethyl alcohol production by the hydrogels was investigated. Swelling of AAm/MA increased with increase in MA content. Ethyl alcohol production also increased with increasing MA in the hydrogels but decreased with an increase of irradiation dose.     

Injectable and moldable hydrogels for use in sensitive and wide range strain sensing applications

Recently, the use of hybrid double network (DN) hydrogels has become prominent due to their enhanced mechanical properties, which has opened the door for new applications of these soft materials. Only a few of these gels have demonstrated both injectable and moldable capabilities. In this work, we report the mechanical properties, gauge factor (GF) values and demonstrate both the injectability and moldability of a gelatin/polyacrylamide DN hydrogel. We optimized several parameters, such as, gelatin to polyacrylamide ratio, reactant concentrations and metal ion concentration, to produce a gelatin/polyacrylamide hydrogel with superior mechanical properties. The highest water content gel was capable of withstanding strains of 5000% before failure. These gels were facilely injected into molds where they effectively changed shape and maintained similar properties prior to remolding. When 20 mM calcium was doped into a similar gel, a tensile strength of 1.71 MPa was achieved. Aside from improving the mechanical properties of the gels, both Ca²⁺ and Mg²⁺ also improved their conductivity, so they were tested for use as strain sensors. The sensitivity of the hydrogel strain sensors were measured using the GF. For the 20 mM Ca²⁺ hydrogel, these GF values ranged from 1.63 to 6.85 for strains of 100% to 2100% respectively. Additionally, the sensors showed good stability over continuous cyclic stretching, demonstrating their long term reliability for strain sensing.     

Modulate the phase transition temperature of hydrogels with both thermosensitivity and biodegradability

The synthesis and characterization of thermoresponsive hydrogels on the basis of N-isoproplyarylamide (NIPAAm) and acrylamide (AAm) copolymers crosslinked with a novel biodegradable crosslinker (PEG-co-PLA) were carried out in this study. Swelling measurement results demonstrated that four gels of PNAM5, PNAM10, PNAM12 and PNAM15 are thermoresponsive. The equilibrium swelling ratio and degradation of the hydrogels strongly depend on hydrogels composition. The morphology of the hydrogels was observed by scanning electron microscopy (SEM), and their thermal property was characterized by differential scanning calorimetry (DSC). The results show that the proportion of AAm in the copolymer has notable effect on the low critical solution temperature (LCST) of the hydrogel. When the molar ratio of AAm to NIPAAm was increased from 1:10 to 3:10 the LCST of the copolymer increased from 39.7 to 64.2 °C. The compression modulus of PNAM15 is of the highest among other hydrogels, because PNAM15 hydrogel has a more compact structure.     

Swelling and de-swelling kinetics of gelatin hydrogels in ethanol-water marginal solvent

Controlled osmotic swelling and de-swelling measurements have been performed on gelatin, a polyampholyte, hydrogels suspended in water-ethanol marginal solvent at room temperature (20 degrees C) where the alcohol concentration was changed from 0 to 100% (v/v). The change in gel mass was monitored as function of time until osmotic equilibrium was established with the surrounding solvent. It was observed that osmotic pressure of polymer-solvent mixing, pi(m)相似文献   

Antibacterial Peptide-Based Gel for Prevention of Medical Implanted-Device Infection

Implanted medical devices are prone to infection. Designing new strategies to reduce infection and implant rejection are an important challenge for modern medicine. To this end, in the last few years many hydrogels have been designed as matrices for antimicrobial molecules destined to fight frequent infection found in moist environments like the oral cavity. In this study, two types of original hydrogels containing the antimicrobial peptide Cateslytin have been designed. The first hydrogel is based on alginate modified with catechol moieties (AC gel). The choice of these catechol functional groups which derive from mussel’s catechol originates from their strong adhesion properties on various surfaces. The second type of gel we tested is a mixture of alginate catechol and thiol-terminated Pluronic (AC/PlubisSH), a polymer derived from Pluronic, a well-known biocompatible polymer. This PlubisSH polymer has been chosen for its capacity to enhance the cohesion of the composition. These two gels offer new clinical uses, as they can be injected and jellify in a few minutes. Moreover, we show these gels strongly adhere to implant surfaces and gingiva. Once gelled, they demonstrate a high level of rheological properties and stability. In particular, the dissipative energy of the (AC/PlubisSH) gel detachment reaches a high value on gingiva (10 J.m^-2) and on titanium alloys (4 J.m^-2), conferring a strong mechanical barrier. Moreover, the Cateslytin peptide in hydrogels exhibited potent antimicrobial activities against P. gingivalis, where a strong inhibition of bacterial metabolic activity and viability was observed, indicating reduced virulence. Gel biocompatibility tests indicate no signs of toxicity. In conclusion, these new hydrogels could be ideal candidates in the prevention and/or management of periimplant diseases.     

The role of bacterial cell wall hydrophobicity in adhesion.

In this study, the adhesion of bacteria differing in surface hydrophobicity was investigated. Cell wall hydrophobicity was measured as the contact angle of water on a bacterial layer collected on a microfilter. The contact angles ranged from 15 to 70 degrees. This method was compared with procedures based upon adhesion to hexadecane and with the partition of cells in a polyethylene glycol-dextran two-phase system. The results obtained with these three methods agreed reasonably well. The adhesion of 16 bacterial strains was measured on sulfated polystyrene as the solid phase. These experiments showed that hydrophobic cells adhered to a greater extent than hydrophilic cells. The extent of adhesion correlated well with the measured contact angles (linear regression coefficient, 0.8).     

The role of bacterial cell wall hydrophobicity in adhesion

In this study, the adhesion of bacteria differing in surface hydrophobicity was investigated. Cell wall hydrophobicity was measured as the contact angle of water on a bacterial layer collected on a microfilter. The contact angles ranged from 15 to 70 degrees. This method was compared with procedures based upon adhesion to hexadecane and with the partition of cells in a polyethylene glycol-dextran two-phase system. The results obtained with these three methods agreed reasonably well. The adhesion of 16 bacterial strains was measured on sulfated polystyrene as the solid phase. These experiments showed that hydrophobic cells adhered to a greater extent than hydrophilic cells. The extent of adhesion correlated well with the measured contact angles (linear regression coefficient, 0.8).     

Beyond bread and beer: whole cell protein extracts from baker’s yeast as a bulk source for 3D cell culture matrices

Here, we present a novel approach to form hydrogels from yeast whole cell protein. Countless hydrogels are available for sophisticated research, but their fabrication is often difficult to reproduce, with the gels being complicated to handle or simply too expensive. The yeast hydrogels presented here are polymerized using a four-armed, amine reactive crosslinker and show a high chemical and thermal resistance. The free water content was determined by measuring swelling ratios for different protein concentrations, and in a freeze-drying approach, pore sizes of up to 100 μm in the gel could be created without destabilizing the 3D network. Elasticity was proofed to be adjustable with the help of atomic force microscopy by merely changing the amount of used protein. Furthermore, the material was tested for possible cell culture applications; diffusion rates in the network are high enough for sufficient supply of human breast cancer cells and adenocarcinomic human alveolar basal epithelial cells with nutrition, and cells showed high viabilities when tested for compatibility with the material. Furthermore, hydrogels could be functionalized with RGD peptide and the optimal concentration for sufficient cell adhesion was determined to be 150 μM. Given that yeast protein is one of the cheapest and easiest available protein sources and that hydrogels are extremely easy to handle, the developed material has highly promising potential for both sophisticated cell culture techniques as well as for larger scale industrial applications.

     

A new strategy for the preparation of supramolecular neutral hydrogels

This paper demonstrates that miscible blends from water-insoluble polymers, such as poly(2,4,4-trimethylhexamethylene terephthalamide) (1), methylamine imidized poly(methyl methacrylate) (2), and aromatic poly(ether sulfone) (3) and water-soluble polymers, such as poly(2-ethyl-2-oxazoline) (4) and poly(N-vinyl pyrrolidone) (5), respectively, represent a new class of supramolecular hydrogels. When the degree of polymerization (DP) of the water-soluble polymer is larger than that of water-insoluble polymer, the resulting hydrogels adsorb extremely high amounts of water (i.e., 229 wt % in the case of the hydrogel 1/4) and remain mechanically tough. The high water uptake capability of these blends is explained by a supramolecular network structure generated by H-bonding and/or other noncovalent interactions between the water-insoluble hydrophobic polymer and water-soluble hydrophilic segments as reversible cross-linking points interconnected by hydrophilic water soluble segments. The glass transition temperatures of these hydrogels are tailored via the ratio between the weight percent of the two polymers and by the glass transition temperature of the parent polymers. These supramolecular hydrogels can be processed from melt or solution and maintain excellent mechanical properties both in dry and in the water swollen state. This class of hydrogels is of interest for areas such as membranes, contact lenses, tissue engineering, and other biomedical applications.     

Bio-based hydrogels prepared by cross-linking of microbial poly(gamma-glutamic acid) with various saccharides

Novel bio-based hydrogels were prepared by cross-linking of microbial poly(gamma-glutamic acid) (PGA) with saccharides such as glucose, maltotriose, and cyclodextrin (CD) in the presence of water-soluble carbodiimide in dimethyl sulfoxide (DMSO) by one-pot synthesis at 25 degrees C for 24 h. The degradation of the gels in alkaline solution (pH 9) at 37 degrees C was also investigated. The PGA gels cross-linked with various neutral saccharides were obtained in relatively high recovery yields by use of a base like 4,4-(dimethylamino)pyridine. The PGA gel cross-linked by glucose showed the highest water absorption of 3000 g/g. The PGA gels cross-linked by CDs showed higher water absorption than those cross-linked by the corresponding linear saccharides. It was revealed that the water absorption of the PGA gel was affected by the cross-linker content and also the structure of cross-linkers as they had an effect on the cross-linking density of the PGA gel. The PGA gels were hydrolyzed under alkaline condition (pH 9) at 37 degrees C. The degradation rate was higher when the cross-linker content of the gel was lower.     

Dextran and hyaluronan methacrylate based hydrogels as matrices for soft tissue reconstruction

Polysaccharide hydrogels have become increasingly studied as matrices in soft tissue engineering because of their known cytocompatibility. In this work cross-linkable dextran methacrylates and hyaluronan methacrylate were synthesized and their transformation into stable hydrogels was studied. The in vitro degradation behaviour of the formed hydrogels could be controlled by the polysaccharide structure and the cross-linking density. Under in vitro conditions, the formed gels had no cytotoxic effects against fibroblasts, but cells could adhere only inefficiently in long term experiments. The use of composite gels improved the adherence of cells. Different scaffold architectures were studied including porous structures and perforated gel layers. Selected hydrogels were examined in an in vivo pilot study using a rabbit model to evaluate their biocompatibility, stability and degradation. No signs of inflammation were seen and with prolonged duration the material was degraded and lacunas were formed by immigrating or ingrowing cells. Optimizing their mechanical properties, the formed hydrogels represent promising candidates as matrices for soft tissue reconstruction.     

Rheology and nanostructure of hydrophobically modified alginate (HMA) gels and solutions

The effect of hydrophobic modification on the mechanical and structural characteristics of hydrophobically modified alginate (HMA) solutions and hydrogels were evaluated. The HMA systems consisted of alkyl chains, C₈, grafted onto alginate backbones. With an increase in degree of substitution of hydrophobic tails, the association became stronger in solution, but same was not true for gels. The contribution of ionic crosslinking was found to be the dominant factor in determining the mechanical strength of hydrogels. Rheological measurements of 2 wt% HMA gels reveal formation of a strongly crosslinked network with an elastic modulus close to 100 kPa. Small-angle X-ray scattering (SAXS) experiments indicate that HMA assembles into a disordered structure with regions rich in the hydrophobic domain surrounded by a crosslinked hydrophilic network.     

Molecular dynamics study of the influence of solvents on the structure and mechanical properties of poly(vinyl alcohol) gels

Molecular dynamics (MD) simulations were employed to study the influence of solvents on the structure and mechanical properties of physically crosslinked poly(vinyl alcohol) (PVA) gels. Firstly, three kinds of PVA precursor gels were made by adding water, dimethyl sulfoxide (DMSO) and a mixture of DMSO and water (4:1 by weight), respectively. The solvents in the precursor gels were then exchanged with water to obtain three kinds of PVA hydrogels. Solvent in the precursor gel with a mixture of DMSO and water was also exchanged with ethanol and DMSO, respectively. It was found that the tensile strength and failure strain of the PVA hydrogel prepared from precursor gel with a mixture of DMSO and water was the highest, and the polymer network was more homogeneous than the other two PVA hydrogels. The polymer network of PVA gel with ethanol or with DMSO was more heterogenous than with water, and the tensile strength and failure strain were much lower. The torsional activity of polymer chains of PVA gel with ethanol was much stronger than PVA gel with water and DMSO.