Biocompatible Hydrogel Material Research Articles

Physicochemical and in vitro biological evaluation of an injectable self-healing quaternized chitosan/oxidized pectin hydrogel for potential use as a wound dressing material

Injectable self-healing hydrogels are attractive materials for use as wound dressings. To prepare such hydrogels, the current study used quaternized chitosan (QCS) to improve the solubility and antibacterial activity and oxidized pectin (OPEC) to introduce aldehyde groups for Schiff's base reaction with the amine groups from QCS. Self-healing hydrogels were made by co-injection of polymer solutions at specific polymer concentrations and reagent ratios that optimized both Schiff's base reactions and ionic interactions. The optimal hydrogel displayed self-healing 30 min after cutting and continuous self-healing during continuous step strain analysis, rapid gelation (< 1 min), a storage modulus of 394 Pa, and hardness of 700 mN, and compressibility of 162 mN s. The adhesiveness of this hydrogel (133 Pa) was within a suitable range for application as a wound dressing. The extraction media from the hydrogel displayed no cytotoxicity to NCTC clone 929 cells and higher cell migration than the control. While the extraction media from the hydrogel was found not to have antibacterial properties, QCS was verified as having MIC50 of 0.04 mg/mL against both E. coli and S. aureus. Therefore, this injectable self-healing QCS/OPEC hydrogel has the potential use as a biocompatible hydrogel material for wound management.

Preparation of a Cellulosic Photosensitive Hydrogel for Tubular Tissue Engineering.

Since the concept of tissue engineering was proposed, biocompatible hydrogel materials have attracted the attention of researchers. With the help of three-dimensional (3D) printing technology, precise shaping of hydrogels can be realized. In this paper, we synthesized a cellulosic photosensitive acrylamide (AM)/N,N-methylenebisacrylamide (MBA) hydrogel. With the high-efficiency water-soluble photoinitiator TPO@Tw developed by our research group, the efficient photocuring cross-linking process of the hydrogel can be realized under 405 nm visible light. In consideration of the viscosity, curing mass, curing depth, and break distance of the hydrogel, we screened out hydroxypropyl cellulose (HPC) as the preferred tackifier of the material. The addition of HPC greatly improved the mechanical properties of the hydrogel. The compressive modulus of the optimal sample AM-HPC-5 increased by 709.2% and the tensile strength increased by 76.7% compared with the blank control group. By adding a PEGDA shell to the surface of the material, the water retention capacity of the hydrogel was effectively improved. The water loss rate was greatly reduced. The 3D wooden-pile structure model was printed by a DIW 3D printer. Further, through coaxial extrusion, the microtubule structure that may be applied in tissue engineering was obtained. Cell experiment results showed high biocompatibility of the hydrogel. NIH 3T3 cells could adhere and grow on the surface of microtubules.

Development and Characterization of Highly Stable Silver NanoParticles as Novel Potential Antimicrobial Agents for Wound Healing Hydrogels.

Recurrent microbial infections are a major cause of surgical failure and morbidity. Wound healing strategies based on hydrogels have been proposed to provide at once a barrier against pathogen microbial colonization, as well as a favorable environment for tissue repair. Nevertheless, most biocompatible hydrogel materials are more bacteriostatic than antimicrobial materials, and lack specific action against pathogens. Silver-loaded polymeric nanocomposites have efficient and selective activity against pathogenic organisms exploitable for wound healing. However, the loading of metallic nanostructures into hydrogels represents a major challenge due to the low stability of metal colloids in aqueous environments. In this context, the aim of the present study was the development of highly stable silver nanoparticles (AgNPs) as novel potential antimicrobial agents for hyaluronic acids hydrogels. Two candidate stabilizing agents obtained from natural and renewable sources, namely cellulose nanocrystals and ulvan polysaccharide, were exploited to ensure high stability of the silver colloid. Both stabilizing agents possess inherent bioactivity and biocompatibility, as well as the ability to stabilize metal nanostructures thanks to their supramolecular structures. Silver nitrate reduction through sodium borohydride in presence of the selected stabilizing agents was adopted as a model strategy to achieve AgNPs with narrow size distribution. Optimized AgNPs stabilized with the two investigated polysaccharides demonstrated high stability in phosphate buffer saline solution and strong antimicrobial activity. Loading of the developed AgNPs into photocrosslinked methacrylated hyaluronic acid hydrogels was also investigated for the first time as an effective strategy to develop novel antimicrobial wound dressing materials.

Injectable, self-healable and adhesive hydrogels using oxidized Succinoglycan/chitosan for pH-responsive drug delivery

We prepared chitosan (CS) based multifunctional hydrogels using oxidized succinoglycan (OSG) with a semi-dissolving acidified sol-gel transition method. OSG cross-linked CS hydrogels (OSG/CS) was prepared by aldehyde-amine Schiff-base reaction. OSG/CS increased not only thermal stability but also improved mechanical strength by 5.75 times. Through the tensile and strain sweep test, OSG/CS showed excellent self-healing properties by 98.82% and 99.89%, respectively. It showed the high compressive stress of 173 kPa at 60% strain, the adhesive strength of 2763 kPa, and the antibacterial effect of 90%. Furthermore, OSG/CS showed a pH-controlled drug release pattern, where a change of pH from 7.4 to 2.0 accelerated for 5-fluorouracil release from 60% to 90%. WST-8 assay demonstrated that OSG/CS maintained 97.30% cell viability and 98.84% cell proliferation after 7 days, indicating the potential as biocompatible hydrogel materials such as wound healing, tissue engineering and drug release systems.

Glycerol-modified γ-PGA and gellan composite hydrogel materials with tunable physicochemical and thermal properties for soft tissue engineering application

Biocompatible hydrogel materials are considered to be a crucial and indispensable character that transitorily supplants the natural skeleton and acts as a preferable microenvironment. In this study, two modifiers – poly(γ-glutamic acid) (γ-PGA) and glycerol (Gly) – were incorporated to form a rigid and coarse structure, which significantly ameliorates the mechanical and biological properties as compared with original gellan gum (GG). The surface characterization and thermal and mechanical properties of the modified compositions were scrutinized using SEM, FTIR, TGA, DSC, and tensile testing. The cytocompatibility and cell viability were reported by regulating fibroblast cells (L929). As a result, it is found that adequate γ-PGA modification considerably enhances the thermal and mechanical properties. The glycerol additives advance the elongation by decreasing the elastic modulus, and the modification did not cause a pernicious effect on chemical structure and non-toxicity according to a cellular response. The adhered fibroblast cells on suitable γ-PGA (0.5–0.7 w/v%) and glycerol (0–2 w/v%) modified structures present better proliferation and higher adhesion than those of pure GG and other study groups. These useful combinations inspired the prospective development of biodegradable hydrogel materials, which were enhanced with excellent material stability and biological activity, for utilization in soft tissue engineering.

Engineered cell-laden alginate microparticles for 3D culture.

Advanced microfabrication technologies and biocompatible hydrogel materials facilitate the modeling of 3D tissue microenvironment. Encapsulation of cells in hydrogel microparticles offers an excellent high-throughput platform for investigating multicellular interaction with their surrounding microenvironment. Compartmentalized microparticles support formation of various unique cellular structures. Alginate has emerged as one of the most dominant hydrogel materials for cell encapsulation owing to its cytocompatibility, ease of gelation, and biocompatibility. Alginate hydrogel provides a permeable physical boundary to the encapsulated cells and develops an easily manageable 3D cellular structure. The interior structure of alginate hydrogel can further regulate the spatiotemporal distribution of the embedded cells. This review provides a specific overview of the representative engineering approaches to generate various structures of cell-laden alginate microparticles in a uniform and reproducible manner. Capillary nozzle systems, microfluidic droplet systems, and non-chip based high-throughput microfluidic systems are highlighted for developing well-regulated cellular structure in alginate microparticles to realize potential drug screening platform and cell-based therapy. We conclude with the discussion of current limitations and future directions for realizing the translation of this technology to the clinic.

Synthesis and characterization of site selective photo-crosslinkable glycidyl methacrylate functionalized gelatin-based 3D hydrogel scaffold for liver tissue engineering

The presented work outlined the development of a new biocompatible hydrogel material that has potential applications in soft tissue engineering. As a proof of concept, human hepatocytes were used to demonstrate the suitability of this material in providing conducive environment for cellular growth and functional development. Herein, a detailed synthesis of novel gelatin derivatives - photo-crosslinkable glycidyl methacrylate (GMA) functionalized gelatins (Gelatin-GMA), and preparation of three-dimensional (3D) hydrogel scaffolds for the encapsulated Huh-7.5 cells is reported. The Gelatin-GMA biopolymers were synthesized at two different pH values of 3.5 (acidic) and 10.5 (basic) where two different photo-crosslinkable polymers were formed utilizing –COOH & –OH groups in acidic pH, and –NH2 & –OH groups in basic pH. The hydrogels were prepared using an initiator (Irgacure I2959) in the presence of UV light. The Gelatin-GMA biopolymers were characterized using spectroscopic studies which confirmed the successful preparation of the polymer derivatives. Rheological measurement was carried out to characterize the mechanical properties and derive the mesh sizes of the 3D hydrogels. Subsequently, detailed in vitro hepatocyte compatibility and functionality studies were performed in the 3D cell seeded hydrogel platform. The 3D hydrogel design with larger mesh sizes utilizes the advantage of the excellent diffusion properties of porous platform, and enhanced cell-growth was observed, which in turn elicited favorable Huh-7.5 response. The hydrogels led to improved cellular functions such as differentiation, viability and proliferation. Overall, it showed that the Gelatin-GMA based hydrogels presented better results compared to control sample (GelMA) because of the higher mesh sizes in Gelatin-GMA based hydrogels. Additionally, the functional group studies of the two Gelatin-GMA samples revealed that the cell functionalities are almost unaffected even after the tripeptide - Arg-Gly-Asp (RGD) in Gelatin-GMA synthesized at pH 3.5 is no longer completely available.

Refractive Index Measurement of Corneal-Shaped Hydrogel Material and Elasmobranch Corneas by Shack–Hartmann Wavefront Sensor System

The biocompatible hydrogel materials are used for the fabrication of artificial cornea. The refractive index of the hydrogel material can affect the optical quality of the artificial cornea. It is challenging to measure the refractive index of the hydrogel material because it is between solid and liquid status during the fabrication processes. In this work, we propose using the Shack-Hartmann wavefront sensor system to measure the wavefront images and to calculate the corresponding aberrations of optical material. The wavefront aberrations can be used to estimate the refractive index of the optical material. To prove the concept, the standard N-BK7 bi-convex lenses with focal lengths of 100 and 150 mm at wavelengths 780, 532, and 450 nm have been measured, and the results show that the relative errors of the average measured refractive indices are about 0.1%-0.17%. We used this proposed method to measure the refractive index of the P407-DA hydrogel material. We found that the average measured refractive indices of the P407-DA hydrogel material are 1.3175, 1.3516, and 1.3645 at wavelengths 780, 532, and 450 nm, respectively. Compared with the refractive index of human cornea, the P407-DA hydrogel can be used as the artificial cornea material. The Cauchy dispersion formula can be used to calculate the Abbe number of the P407-DA hydrogel material, which is about 11, lower than the human cornea's Abbe number. It means that the P407-DA hydrogel material has higher chromatic dispersion than the human cornea material. Moreover, the elasmobranch (shark) corneas, which are known as the suitable material for xenotransplantation, are measured by the Shack-Hartmann wavefront sensor system. The ABBE refractometer (ATAGO DR-M4) is used to measure the flat P407-DA hydrogel sample and it is found that the refractive index is slightly different when compared with the measured results by the Shack-Hartmann wavefront sensor system. The refractive indices of other biocompatible hydrogels poly(acrylic acid) (PAA) and poly(2-hydroxyethyl methacrylate) (PHEMA) are also measured by our proposed method, and it is found that the refractive index of P407-DA is more suitable for designing corneal samples than other hydrogels. It is found that the refractive indices of the elasmobranch corneas are similar to the P407-DA material. Our proposed method to measure the refractive index of hydrogel material using the Shack-Hartmann wavefront sensor system is useful. It is a non-contact, minimally invasive, and time-efficient method for shaped structures. The results can be used as a guide for developing better biocompatible material for artificial cornea.

Reactive Ion Plasma Modification of Poly(Vinyl-Alcohol) Increases Primary Endothelial Cell Affinity and Reduces Thrombogenicity.

Bulk material properties and luminal surface interaction with blood determine the clinical viability of vascular grafts, and reducing intimal hyperplasia is necessary to improve their long-term patency. Here, the authors report that the surface of a biocompatible hydrogel material, poly(vinyl alcohol) (PVA) can be altered by exposing it to reactive ion plasma (RIP) in order to increase primary endothelial cell attachment. The power and the carrier gas of the RIP treatment are varied and the resultant surface nitrogen, water contact angle, as well as the ability of the RIP-treated surfaces to support primary endothelial colony forming cells is characterized. Additionally, in a clinically relevant shunt model, the amounts of platelet and fibrin attachment to the surface were quantified during exposure to non-anticoagulated blood. Treatments with all carrier gases resulted in an increase in the surface nitrogen. Treating PVA with O2 , N2 , and Ar RIP increased affinity to primary endothelial colony forming cells. The RIP treatments did not increase the thrombogenicity compared to untreated PVA and had significantly less platelet and fibrin attachment compared to the current clinical standard of expanded polytetrafluoroethylene (ePTFE). These findings indicate that RIP-treatment of PVA could lead to increased patency in synthetic vascular grafts.

Fabrication of alginate/humic acid/Fe-aminoclay hydrogel composed of a grafted-network for the efficient removal of strontium ions from aqueous solution

Abstract A polymer hydrogel composed of alginate/humic acid/Fe-aminoclay was fabricated and utilized as a potential adsorbent for the removal of strontium ions (Sr 2 + ) from aqueous solution. The effects of adsorbate concentration, reaction pH, and time on the removal efficiency of Sr 2 + were investigated. The results revealed that the adsorption capacity increases as the Sr 2 + concentration increased from 1 to 10 ppm, which might be due to the more available adsorption sites. An additional increase in the Sr 2 + concentration had no effect. The optimal pH and reaction time for the removal of Sr 2 + were identified as pH 7 and 24 h, respectively, with an adsorbent dosage of 20 mg. The adsorption isotherm was fitted to the Freundlich model with a maximum Sr 2 + adsorption capacity of 45.65 mg g−1. The kinetic study showed that the adsorption behavior followed a pseudo-second-order kinetics. The hydrogel showed good selectivity toward Sr 2 + even in the presence of competitive cations having a higher concentration than Sr 2 + . From the above results, it is expected that this biocompatible hydrogel material can be used as an adsorbent for the removal of radioactive strontium.

A hydrogel-based intravascular microgripper manipulated using magnetic fields

This study presents a magnetic hydrogel-based microgripper that can be wirelessly manipulated using magnetic fields. The proposed device can move freely in liquids when driven by direct current (dc) magnetic fields, and perform a gripping motion by using alternating current (ac) magnetic fields. The device is fabricated from a biocompatible hydrogel material that can be employed for intravascular applications. The actuation mechanism for gripping motions is realized by controlling the exposure dose on the hydrogel composite during the lithography process. The preliminary characterization of the device is also presented. The measurement results show that the gripping motion reached a full stroke at approximately 38°C. By dispersing multiwall carbon nanotubes (MWCNT) into the material, the overall response time of the gripping motion decreases by approximately 2-fold. Device manipulations such as the gripping motion, translational motion, and rotational motion are also successfully demonstrated on a polyvinyl chloride (PVC) tube and in a polydimethylsiloxane (PDMS) microfluidic channel.

Three-dimensional printed trileaflet valve conduits using biological hydrogels and human valve interstitial cells

Tissue engineering has great potential to provide a functional de novo living valve replacement, capable of integration with host tissue and growth. Among various valve conduit fabrication techniques, three-dimensional (3-D) bioprinting enables deposition of cells and hydrogels into 3-D constructs with anatomical geometry and heterogeneous mechanical properties. Successful translation of this approach, however, is constrained by the dearth of printable and biocompatible hydrogel materials. Furthermore, it is not known how human valve cells respond to these printed environments. In this study, 3-D printable formulations of hybrid hydrogels are developed, based on methacrylated hyaluronic acid (Me-HA) and methacrylated gelatin (Me-Gel), and used to bioprint heart valve conduits containing encapsulated human aortic valvular interstitial cells (HAVIC). Increasing Me-Gel concentration resulted in lower stiffness and higher viscosity, facilitated cell spreading, and better maintained HAVIC fibroblastic phenotype. Bioprinting accuracy was dependent upon the relative concentrations of Me-Gel and Me-HA, but when optimized enabled the fabrication of a trileaflet valve shape accurate to the original design. HAVIC encapsulated within bioprinted heart valves maintained high viability, and remodeled the initial matrix by depositing collagen and glyosaminoglycans. These findings represent the first rational design of bioprinted trileaflet valve hydrogels that regulate encapsulated human VIC behavior. The use of anatomically accurate living valve scaffolds through bioprinting may accelerate understanding of physiological valve cell interactions and progress towards de novo living valve replacements.

Assembly of complex cell microenvironments using geometrically docked hydrogel shapes

Cellular communities in living tissues act in concert to establish intricate microenvironments, with complexity difficult to recapitulate in vitro. We report a method for docking numerous cellularized hydrogel shapes (100-1,000 µm in size) into hydrogel templates to construct 3D cellular microenvironments. Each shape can be uniquely designed to contain customizable concentrations of cells and molecular species, and can be placed into any spatial configuration, providing extensive compositional and geometric tunability of shape-coded patterns using a highly biocompatible hydrogel material. Using precisely arranged hydrogel shapes, we investigated migratory patterns of human mesenchymal stem cells and endothelial cells. We then developed a finite element gradient model predicting chemotactic directions of cell migration in micropatterned cocultures that were validated by tracking ∼2,500 individual cell trajectories. This simple yet robust hydrogel platform provides a comprehensive approach to the assembly of 3D cell environments.

Micron-scale spatially patterned, covalently immobilized vascular endothelial growth factor on hydrogels accelerates endothelial tubulogenesis and increases cellular angiogenic responses.

Spontaneous formation of endothelial tubules was restricted to patterned micron-scale regions presenting cell adhesion ligands and angiogenic signaling protein on poly(ethylene glycol) hydrogels. Arginine-glycine-aspartic acid-serine (RGDS), an integrin ligand, and vascular endothelial growth factor (VEGF), a rate-limiting signaling protein involved in angiogenesis, were covalently bound through photopolymerization via laser scanning lithography to the surface of poly(ethylene glycol) hydrogels in patterned micron-scale regions. Endothelial cells cultured in this restricted environment underwent accelerated tubulogenesis, forming endothelial tubes within 2 days, whereas cells cultured on larger patterned areas remained spread and did not form tubules by day 2. Tubules formed in 2 days on RGDS and VEGF patterns were observed to possess lumens; however, tubule-like structures formed in 2 days on RGDS-only control patterns did not have observable lumens. Additionally, tubules that formed on restricted areas of RGDS and VEGF expressed more VEGF receptor 1, VEGF receptor 2, and ephA7 surface markers, in addition to higher expression of laminin, than cells remaining spread on wide patterned lines. This work reports spatial control and acceleration of endothelial tubule formation using biocompatible hydrogel materials to allow the formation of highly organized vascularized tissues.

Boundary Lubricant - Functionalized PVA Gels for Biotribological Applications

AbstractA novel material design was developed by functionalizing a biocompatible hydrogel material with organic boundary lubricants. Polyvinyl alcohol was functionalized with varying molar ratios of 0.2, 0.5, and 1.0 moles of lauroyl chloride. Tribological and mechanical characterization was performed by means of nanofriction testing and nanoindentation to determine the influence of the hydrocarbon chains on the friction coefficient and elastic modulus of the hydrogels. It was found that fusing of the lubricant to the polymer material has a positive effect on the surface friction properties, yet an unfavorable effect on stiffness properties of the gel due to the processing method.

Direct Cell Printing With Microfabricated Quill-Pen Cantilevers

AbstractA novel direct cell printing technique has been developed to control and manipulate the position of cells on solid surfaces. The method utilizes microfabricated polymeric “quill-pen” cantilevers to transfer living cells onto a wide variety of surfaces. In contrast with existing cell deposition methods, such as ink jet or laser ablation methods, the quill-pen approach imparts minimal thermal and shear stress to cells, preserving cell viability and biological functionality. Deposition of both bacterial and mammalian cells into defined patterns has been demonstrated using this method. The size of printed, cell-containing droplets could be controlled by varying the geometry of the quill-pen stylus and by varying printing conditions such as contact time, relative humidity, and surface hydrophobicity. Initial experiments using 10 μm diameter polymer beads demonstrated that the number of beads per droplet could be controlled by varying spot size and particle concentration in the printing solution. Spots could be printed ranging from 20 μm and 100 μm in diameter with approximate volumes ranging from 1-250 pL. We demonstrated deposition of both cells and beads onto a variety of solid surfaces including agarose gel, polystyrene, polyethylene, and glass. Printed cells have also been immobilized on glass and polymer surfaces using biocompatible hydrogel materials (both alginic acid and hyaluronic acid-based matrices) as well as poly-L-lysine. Similar to polymer beads, the number of cells in printed droplets was shown to be dependent upon the size of the droplet, and could be varied by adjusting the concentration of cells present in the printing fluid. As few as one cell per spot could be achieved by adjusting these parameters. The viability and proliferation of printed cells has been evaluated using live optical imaging to observe cell growth and division. Both bacterial cells (Escherichia coli) and mammalian cells were able to divide and proliferate for at least 96 hr post-printing (experiments were discontinued after 96 hr). Live/dead staining was also used to confirm the viability of printed cells. Rat mammary adenocarcinoma MTLn3 cells and mouse embryonic stem cells were also shown to survive the printing process for at least 24 - 96 hr post-printing. These results demonstrate the feasibility of the printing method and its compatibility with a wide range of cell types. It is especially noteworthy that embryonic stem cells could survive the printing process (and proliferate on the printing substrate). This novel printing method has applications for tissue engineering, cell-to-cell signaling studies, and for directly interfacing cells with nanodevices and biosensors.

Gatekeeper Therapy: An Endoscopic Treatment for GERD: Randomized, Sham-Controlled Multi-Center Trial Overview

Background: The Gatekeeper ™ Reflux Repair System (Medtronic, Shoreview, MN) is a promising endoscopic therapy that is designed to augment the functioning of the lower esophageal sphincter through the implantation of prostheses into the submucosal layer of the LES in an outpatient procedure. These prostheses, made of a biocompatible hydrogel material, expand upon contact with moisture, creating a mechanical barrier between the stomach and the esophagus, restricting the flow of gastric contents into the esophagus. Preliminary data from 68 patients in a European Multi-center study showed implantation of approximately 4 prostheses at the LES improved GERD symptoms in a majority of PPI dependent GERD patients. Aim: To evaluate the effects of the Gatekeeper Therapy in symptomatic GERD patients in a large, prospective, multi-center, randomized, sham-controlled study. Method: Recruitment is ongoing for a Multicenter U.S./European study. Inclusion criteria include: Symptomatic GERD with symptom control on PPI therapy. Baseline 24 hr with pH<=4.0 =>4% of time. Baseline GERD-HRQL >=20 off PPI. Exclusion criteria: Barrett's > 2cm, strictures, varices, grade 3 or greater esophagitis, hiatal hernia > 3cm, prior anti-reflux surgery. Procedure: Implant 4 prostheses circumferentially in LES. 144 patients will be randomized by a sealed envelope at ratio of 2:1 implant vs. sham. Blind will be broken at 6 months, with sham patients offered Gatekeeper therapy. Outcome parameters being evaluated include HRQL, esophagitits, medication use, 24 hr pH, and implant persistence. Results: Enrollment started 09/24/03 with 210 subjects enrolled and 52 subjects randomized and implanted to date (12/01/04). Summary: The study is ongoing with enrollment completion anticipated by April, 2005.