Hyaluronic acid hydrogels for biomedical applications

Hyaluronic acid (HA), an immunoneutral polysaccharide that is ubiquitous in the human body, is crucial for many cellular and tissue functions and has been in clinical use for over thirty years. When chemically modified, HA can be transformed into many physical forms-viscoelastic solutions, soft or stiff hydrogels, electrospun fibers, non-woven meshes, macroporous and fibrillar sponges, flexible sheets, and nanoparticulate fluids-for use in a range of preclinical and clinical settings. Many of these forms are derived from the chemical crosslinking of pendant reactive groups by addition/condensation chemistry or by radical polymerization. Clinical products for cell therapy and regenerative medicine require crosslinking chemistry that is compatible with the encapsulation of cells and injection into tissues. Moreover, an injectable clinical biomaterial must meet marketing, regulatory, and financial constraints to provide affordable products that can be approved, deployed to the clinic, and used by physicians. Many HA-derived hydrogels meet these criteria, and can deliver cells and therapeutic agents for tissue repair and regeneration. This progress report covers both basic concepts and recent advances in the development of HA-based hydrogels for biomedical applications.     

Hyaluronic acid and silver sulfadiazine-impregnated polyurethane foams for wound dressing application

Five different kinds of PU foam wound dressings were prepared to investigate their wound healing capability. They include (i) PU+silver sulfadiazine (AgSD), (ii) PU+alginate (Al), (iii) PU+Al+AgSD, (iv) PU+hyaluronic acid (HA), and (v) PU+HA+AgSD. Physical properties and in vitro behaviors of AgSD release and fibroblast adhesion on those dressings were evaluated. From the drug release and fibroblast adhesion studies, it was observed that PU foam impregnated with both HA and AgSD shows good drug release behavior and low adhesion of the cells. Furthermore, the HA and AgSD-containing PU foam showed excellent wound healing effect without any inflammation or yellow cluster. The wound size decreased around 77% after 1 week application of that foam dressing onto a rat skin defect.     

In vitro release dynamics of thiram fungicide from starch and poly(methacrylic acid)-based hydrogels

In order to make the judicious use of pesticide/fungicide and to maintain the environment and ecosystem we have developed the starch and poly(methacrylic acid)-based agrochemical delivery system for their controlled and sustained release. The delivery device was prepared by using N,N'-methylenebisacrylamide (N,N'-MBAAm) as crosslinker and was characterized with FTIR, TGA and with swelling studies as a function of time and crosslinker concentration. This article discusses the swelling kinetics of polymer matrix and release dynamics of thiram (fungicide) from hydrogels for the evaluation of the diffusion mechanism and diffusion coefficients. The values of the diffusion exponent 'n' for both cases, that is the swelling of hydrogels and for the release of thiram from the hydrogels have been observed between 0.7 and 0.9 when the concentration of the crosslinker in the polymers were varied from 6.49x10(-3) to 32.43x10(-3) moles/L. It is inferred from the values of the 'n' that Non-Fickian diffusion mechanism has occurred in both the cases.     

Hyaluronic acid hydrogels incorporating platelet lysate enhance human pulp cell proliferation and differentiation

The restoration of dentine-pulp complex remains a challenge for dentists; nonetheless, it has been poorly addressed. An ideal system should modulate the host response, as well as enable the recruitment, proliferation and differentiation of relevant progenitor cells. Herein was proposed a photocrosslinkable hydrogel system based on hyaluronic acid (HA) and platelet lysate (PL). PL is a cocktail of growth factors (GFs) and cytokines involved in wound healing orchestration, obtained by the cryogenic processing of platelet concentrates, and was expected to provide the HA hydrogels specific biochemical cues to enhance pulp cells’ recruitment, proliferation and differentiation. Stable HA hydrogels incorporating PL (HAPL) were prepared after photocrosslinking of methacrylated HA (Met-HA) previously dissolved in PL, triggered by the Ultra Violet activated photoinitiator Irgacure 2959. Both the HAPL and plain HA hydrogels were shown to be able to recruit cells from a cell monolayer of human dental pulp stem cells (hDPSCs) isolated from permanent teeth. The hDPCs were also seeded directly over the hydrogels (5?×?10⁴ cells/hydrogel) and cultured in osteogenic conditions. Cell metabolism and DNA quantification were higher, in all time-points, for PL supplemented hydrogels (p?<?0,05). Alkaline phosphatase (ALPL) activity and calcium quantification peaks were observed for the HAPL group at 21 days (p?<?0,05). The gene expression for ALPL and COLIA1 was up-regulated at 21 days to HAPL, compared with HA group (p?<?0,05). Within the same time point, the gene expression for RUNX2 did not differ between the groups. Overall, data demonstrated that the HA hydrogels incorporating PL increased the cellular metabolism and stimulate the mineralized matrix deposition by hDPSCs, providing clear evidence of the potential of the proposed system for the repair of damaged pulp/dentin tissue and endodontics regeneration.     

Linear poly(ethylene oxide)-based polyurethane hydrogels: polyurethane-ureas and polyurethane-amides

Over the last 30 years, water-swellable and water-insoluble hydrogels have been extensively investigated and developed, leading to a large family of materials which have found uses in a wide range of biomedical applications. While hydrogels usually present a crosslinked structure, linear polyurethane-ureas (PUUs) based on poly(ethylene oxide) have been shown to be able to absorb and swell with aqueous media without dissolving. This behavior is due to the phase separated domain morphology, where hydrogen bonded urethane/urea hard segment domains are dispersed in a PEO soft segment domain. This work investigates the possibility of obtaining linear poly(ethylene oxide)-based polyurethane-amide (PUA) hydrogels using two amide diols as chain extenders, a mono amide diol (AD) and a diamide diol (DD), and a dicarboxylic acid (maleic acid, MA). Poly(ethylene oxide) based PUAs were obtained using a “one-shot” bulk polymerization technique. The chemicophysical characterization and water-solubility tests showed that these materials, while having molecular weights similar to the PUUs, do not possess sufficient phase separation, hydrogen bonding and hydrophobicity of the hard segment domains to exhibit hydrogel behavior. Crosslinked PUAs using maleic acid as chain extender show interesting hydrogel properties. © 1999 Kluwer Academic Publishers     

Poly(HEMA) hydrogels with controlled pore architecture for tissue regeneration applications

The technique for fabrication of soft porous hydrogels, in which both the size and the orientation of inner pores can be controlled, was developed. Three-dimensional hydrophilic gels based on poly[2-hydroxyethyl methacrylate] are designed as scaffolds for regeneration of soft tissues, e.g., nerve tissue. Anisotropic macropores of the size ranging from 10 to 50 μm were formed (1) by using a porogen-leaching method with a solid organic porogen, (2) by phase-separation during gelation in solvent-nonsolvent mixture, or (3) by combination of solid porogen elimination and phase-separation. As a porogen, poly(l-lactide) fibers were applied and consequently washed away under mild conditions to obtain desired spatial orientation of pores. Highly water-swollen polymer gels were characterized with high pressure (low vacuum) scanning electron microscopy (AquaSEM). The morphology of voids remaining after removing the solid PLLA porogen (the macropores) was clearly shown.     

Biological evaluation of human hair keratin scaffolds for skin wound repair and regeneration

The cytocompatibility, in vivo biodegradation and wound healing of keratin biomaterials were investigated. For the purposes, three groups of keratin scaffolds were fabricated by freeze-drying reduced solutions at 2 wt.%, 4 wt.% and 8 wt.% keratins extracted from human hairs. These scaffolds exhibited evenly distributed high porous structures with pore size of 120–220 μm and the porosity > 90%. NIH3T3 cells proliferated well on these scaffolds in culture lasting up to 22 days. Confocal micrographs stained with AO visually revealed cell attachment and infiltration as well as scaffold architectural stability. In vivo animal experiments were conducted with 4 wt.% keratin scaffolds. Early degradation of subcutaneously implanted scaffolds occurred at 3 weeks in the outermost surface, in concomitant with inflammatory response. At 5 weeks, the overall porous structure of scaffolds severely deteriorated while the early inflammatory response in the outermost surface obviously subsided. A faster keratin biodegradation was observed in repairing full-thickness skin defects. Compared with the blank control, keratin scaffolds gave rise to more blood vessels at 2 weeks and better complete wound repair at 3 weeks with a thicker epidermis, less contraction and newly formed hair follicles. These preliminary results suggest that human hair keratin scaffolds are promising dermal substitutes for skin regeneration.     

Assessment of reinforced poly(ethylene glycol) chitosan hydrogels as dressings in a mouse skin wound defect model

Wound dressings of chitosan are biocompatible, biodegradable, antibacterial and hemostatic biomaterials. However, applications for chitosan are limited due to its poor mechanical properties. Here, we conducted an in vivo mouse angiogenesis study on reinforced poly(ethylene glycol) (PEG)-chitosan (RPC) hydrogels. RPC hydrogels were formed by cross-linking chitosan with PEGs of different molecular weights at various PEG to chitosan ratios in our previous paper. These dressings can keep the wound moist, had good gas exchange capacity, and was capable of absorbing or removing the wound exudate. We examined the ability of these RPC hydrogels and neat chitosan to heal small cuts and full-thickness skin defects on the backs of male Balb/c mice. Histological examination revealed that chitosan suppressed the infiltration of inflammatory cells and accelerated fibroblast proliferation, while PEG enhanced epithelial migration. The RPC hydrogels promoted wound healing in the small cuts and full layer wounds. The optimal RPC hydrogel had a swelling ratio of 100% and a water vapor transmission rate (WVTR) of about 2000 g/m²/day. In addition, they possess good mechanical property and appropriate degradation rates. Thus, the optimal RPC hydrogel formulation functioned effectively as a wound dressing and promoted wound healing.     

羟基磷灰石(HA)表面改性对HA/聚乳酸复合材料力学性能的影响

采用三种高分子有机物聚乙二醇(PEG)、聚乙烯醇(PVA)和聚丙烯酸(PAA)对羟基磷灰石(HA)颗粒表面进行改性处理,以便改善两相在复合时的界面结合强度,从而获得力学性能优良的复合材料.首先,将三种高分子聚合物分别溶于含HA颗粒的水溶液中,均匀分散后经喷雾干燥获得改性的粉体;然后,利用流延法获得HA/PLLA复合材料薄膜.研究了3种高分子表面改性HA颗粒后复合材料的力学性能,以及自然断面的界面结合情况.结果表明HA表面经PEG改性后,HA与PLLA间的界面结合状态优良,HA/PEG/PLLA的断裂强度较未经表面改性处理的HA颗粒与PLLA的复合材料的断裂强度提高了31%.     

Characteristics of curcumin-loaded poly (lactic acid) nanofibers for wound healing

Curcumin (Cur) is a well-known extract of the root of Curcuma longa L. that has multi biological functions such as anti-oxidation, anti-inflammatory, anti-cancer, and wound healing properties. In the present study, poly (lactic acid) (PLA) nanofibers were used as a carrier for Cur because PLA nanofibers are biocompatible and have a high-specific surface area and high porosity, which can enhance the functional properties of Cur. The chemical and biological characteristics of Cur/PLA blended nanofibers containing varied amounts of Cur were examined. An increase from 0.125 to 6.250 wt% Cur in PLA caused a decrease in the diameters of the nanofibers from 971 ± 274 to 562 ± 177 nm. At Cur concentrations of <1.250 wt%, PLA and Cur showed good miscibility in the blended nanofibers, as shown by FTIR analysis and tensile tests. The inclusion of Cur in the blended nanofibers at concentration as low as 0.125 wt% promotes the attachment and proliferation of cells. The in vivo wound healing capability of Cur-loaded PLA nanofibers was assessed in a mouse model; treatment with Cur-loaded PLA nanofibers significantly increased the rate of wound closure (87 %) by day 7 compared with that of PLA nanofibers (58 %). The results of this study suggest that Cur-loaded nanofibers with appropriate Cur concentration are nontoxic and have potential as component of wound-healing patches.     

The synthesis and degradation of collagenase-degradable poly(2-hydroxyethyl methacrylate)-based hydrogels and sponges for potential applications as scaffolds in tissue engineering

A collagenase-cleavable peptide-based crosslinking agent was synthesized and was incorporated into PHEMA sponges, and P[HEMA-co-MeO-PEGMA] gels and sponges [HEMA 2-hydroxyethyl methacrylate, PHEMA = poly(2-hydroxyethyl methacrylate), MeO-PEGMA = poly(ethylene glycol) monomethyl ether methacrylate]. PHEMA and P[HEMA-co-MeO-PEGMA] sponges had polymer droplet morphologies where the dimensions of the morphological features were three to five times larger compared to sponges that were crosslinked with tetraethylene glycol dimethacrylate (TEGDMA), while the P[HEMA-co-MeO-PEGMA] gels had similar morphologies regardless of the crosslinking agent. The differences in the dimensions of the morphologies of the sponges were attributed to differences in hydrophilicities of the crosslinking agent. When incubated in a collagenase solution, PHEMA sponges did not degrade, but P[HEMA-co-MeO-PEGMA] gels took 28 days to degrade and the P[HEMA-co-MeO-PEGMA] sponges took 101 days to degrade to 8% dry weight remaining. A cytotoxicity assay showed that the hydrogels do not elicit any cytotoxic response in vitro.     

Physico-chemical and mechanical characterization of hydrogels of poly (vinyl alcohol) and hyaluronic acid

Hydrogels are three-dimensional polymeric networks very similar to biological tissues and potentially useful as soft tissue substitutes and drug delivery systems. Many synthetic polymers can be used to make hydrogels: poly (vinyl alcohol) is widely employed to make hydrogels for biomedical applications. Improvements in the biocompatibility characteristics of synthetic materials could be achieved by the addition of biological macromolecules. The resulting materials named bioartificial polymeric materials could possess the good mechanical properties of the synthetic component and adequate biocompatibility due to the biological component. We have used poly (vinyl alcohol) to make hydrogels containing various amounts of hyaluronic acid. These bioartificial materials were studied to investigate the effect of the presence of the hyaluronic acid on the structural properties of the hydrogels. Thermal, mechanical, morphological and X-ray analyses were performed. A close correspondence between the network consistency and the degree of crystallinity developed in the matrix suggested that the hyaluronic acid, when its content is about 20%, could provide heterogeneous crystallization nuclei for poly (vinyl alcohol) thus increasing the crystallization degree, and consequently, the storage modulus.     

Mineralization of poly(l-lactic acid)-based foams induced by cellulose nanofibrils

Cellulose nanofibrils (CNFs) were blended with poly(l-lactic acid) (PLLA) to produce CNFs/PLLA composite solid foams. The dispersed CNFs’ phase was partially embedded in the PLLA matrix. The CNFs not only reduced the water contact angle of the composite, but also induced the formation of hydroxyapatite (HA) on the walls of its inner pores. After incubation for 7 days in 3× simulated body fluid, a large number of HA particles were formed throughout the CNFs/PLLA composite foams. HA particles have diameters ranging from 200 nm to 2 μm and a Ca/P ratio of 1.42. The spatial distribution of calcium and phosphorus elements was uniform. A porosity of approximately 92 % was achieved after mineralization of the CNFs/PLLA composite foams. The mass of HA grown over CNFs/PLLA foams increased faster than in the case of PLLA foams. The ternary polymeric foams have potential applications in tissue engineering.     

Swelling and aspirin release study: cross-linked pH-sensitive vinyl acetate-co-acrylic acid (VAC-co-AA) hydrogels

The objective of this work was to develop new pH-sensitive hydrogels to deliver gastric mucosal irritating drugs to the lower part of the gastrointestinal tract. For this purpose, cross-linked vinyl acetate-co-acrylic acid (VAC-co-AA) hydrogels were synthesized by using N, N, methylene bisacrylamide (MBAAm) as a cross-linking agent. Different ratios of 90:10, 70:30, 50:50, 30:70, and 10:90 of VAC-co-AA were synthesized. All of the compositions were cross-linked using 0.15, 0.30, 0.45, and 0.60 mol percent MBAAm. Swelling and aspirin release were studied for 8 hour period. The drug release data were fitted into various kinetic models like the zero-order, first-order, Higuchi, and Peppas. Hydrogels were characterized by Fourier transform infrared spectroscopy and scanning electron microscopy. In addition to the above, these hydrogels were loaded with 2%, 8% and 14% w/v aspirin solutions, keeping the monomeric composition and degree of cross-linking constant. In conclusion, it can be said that aspirin can be successfully incorporated into cross-linked VAC/AA hydrogels and its swelling and drug release can be modulated by changing the mole fraction of the acid component in the gels.     

Preparation and characterization of thermoresponsive poly(N-isopropylacrylamide-co-acrylic acid) hydrogels: studies with electroactive probes

A new method of preparing thermoresponsive hydrogels consisting of N-isopropylacrylamide (NIPA) and acrylic acid (AA), with a well-defined concentration of an electroactive probe, 1,1'-ferrocenedimethanol (Fc(MeOH)2), is described, and a comparison of the physical and electrochemical properties of NIPA-AA gels with those of aqueous solutions is presented. The NIPA-AA gels undergo a discontinuous volume phase transition at 45 degrees C; this transition results in a release of approximately 40% of the solution mass from the gel phase. Characterization of hydrogels with electroactive probes is performed using electroanalytical techniques and FTIR and UV/vis spectroscopies. Steady-state voltammetry and chronoamperometry at platinum disk microelectrodes are used to measure the diffusion coefficient of Fc(MeOH)2 in gels under a wide range of experimental conditions. Similar diffusion coefficients for Fc(MeOH)2 in NIPA-AA gels are obtained by either electroanalytical technique at temperatures lower than 20 degrees C. The uncertainty in the Fc(MeOH)2 concentration in the gels, resulting from the discontinuous volume change transition, necessitated the use of concentration-independent chronoamperometric data (i.e. the chronoamperometric response divided by the steady-state current obtained at sufficiently long times) to obtain reliable diffusion coefficient values for Fc(MeOH)2. For temperatures above the volume phase transition, changes of concentration of Fc(MeOH)2 are detected in a copolymeric collapsed phase.     

PEDOT:PSS/聚(丙烯酰胺-甲基丙烯酸)导电水凝胶的制备与性能

通过原位自由基溶液聚合法制备了聚(3,4-乙撑二氧噻吩):聚苯乙烯磺酸盐(PEDOT:PSS)/聚(丙烯酰胺-甲基丙烯酸)(P(AAM-MAA))导电水凝胶.采用FTIR、Raman、TGA和SEM分别表征了PEDOT:PSS/P(AAM-MAA)的组成与形貌,测定了PEDOT:PSS/P(AAM-MAA)的透光率、电...     

Composite nanofiber of bioactive glass nanofiller incorporated poly(lactic acid) for bone regeneration

A novel composite nanofiber of poly(lactic acid) (PLA) incorporated with the nanocomponent of bioactive glass was exploited using an electrospinning method. Small concentrations of the bioactive glass phase added up to 10% facilitated the generation of a nanofibrous matrix with hundreds of nanometers in diameter without a formation of beads. The addition of the bioactive glass phase greatly enhanced the in vitro apatite formation on the nanofiber surface under a body simulating medium. Osteoblastic cells were demonstrated to adhere well on the composite nanofiber and grow actively with culturing time, suggesting its usefulness as a supporting matrix for the hard tissue regeneration.     

Thermo- and pH-sensitive behavior of hydrogels based on oligo (ethylene glycol) methacrylates and acrylic acid

Novel dual stimuli-responsive hydrogels were prepared by free-radical polymerization of 2-(2-methoxyethoxy) ethyl methacrylate (MEO₂MA) and oligo (ethylene glycol) methacrylate (OEGMA), as thermosensitive monomers, and acrylic acid (AAc), as a pH-sensitive monomer. Due to the thermosensitive monomers introduced in the macromolecular network, the synthesized materials possessed tunable thermal behavior. In addition, as well as by introducing in the polymerization feed pH-sensitive monomer AAc, the equilibrium swelling properties of the hydrogels can be tuned by three comonomers. Moreover, the de-swelling kinetics was studied by changing temperature and/or pH, and they could be well described with a first-order kinetics equation. Especially, the faster shrinking rates of hydrogels were observed when the simultaneous temperature and pH stimuli changed from pH 8/18 to pH 2/55 °C because of the cooperative thermo-/pH responses. The prepared dual temperature-/pH-sensitive PMOA hydrogels as a new material candidate may provide significant valuable information for various potential applications.     

Injectable melatonin-loaded carboxymethyl chitosan (CMCS)-based hydrogel accelerates wound healing by reducing inflammation and promoting angiogenesis and collagen deposition

Carboxymethyl chitosan(CMCS)-based hydrogels have antibacterial activity,and have shown the abilities of preventing wound infection,promoting cell proliferation,accelerating collagen deposition,and stimulating hyaluronic acid formation during wound healing.As a hormone produced by the pineal gland in humans and animals,melatonin promotes skin wound healing by regulating the release of inflammatory mediators and accelerating the proliferation and migration of cells,angiogenesis,and collagen deposition.However,the combined effects of CMCS and melatonin on wound healing remain unclear.Injectable CMCS-based hydrogels containing melatonin were prepared,and their healing effects were evaluated using a full-thickness cutaneous wound model in rats.Compared with the control and the hydrogel with no melatonin groups,the melatonin-loaded hydrogel significantly increased the percentage of wound closure,promoted the proliferation of granulation tissue and re-epithelialization,and accelerated collagen deposition.Additionally,the melatonin-loaded hydrogel promoted angiogenesis and vascular endothelial growth factor receptor protein expression and increased the expression of cyclooxygenase-2 and inducible nitric oxide synthase.The melatonin-loaded hydrogel also markedly increased the expression of collagen III,α-smooth muscle actin,and transforming growth factor-β1 proteins and reduced collagen I expression.These results suggest that the melatonin-loaded hydrogel promoted granulation tissue formation and accelerated wound healing by reducing inflammation and promoting angiogenesis and collagen deposition.