There is a clear need for novel in vitro models, especially for neuronal applications. Development of in vitro models is a multiparameter task consisting of cell‐, biomaterial‐, and environment‐related parameters. Here, three different human origin neuronal cell sources are studied and cultured in various hydrogel 3D scaffolds. For the efficient evaluation of complex results, an indexing method for data is developed and used in principal component analysis (PCA). It is found that no single hydrogel is superior to other hydrogels, and collagen I (Col1) and hyaluronan–poly(vinyl alcohol) (HA1‐PVA) gels are combined into an interpenetrating network (IPN) hydrogel. The IPN gel combines cell supportiveness of the collagen gel and stability of the HA1‐PVA gel. Moreover, cell adhesion is studied in particular and it is found that adhesion of neurons differs from that observed for fibroblasts. In conclusion, the HA1‐PVA‐col1 hydrogel is a suitable scaffold for neuronal cells and supports adhesion formation in 3D. 相似文献
The vascular system represents the key supply chain for nutrients and oxygen inside the human body. Engineered solutions to produce sophisticated alternatives for autologous or artificial vascular implants to sustainably replace diseased vascular tissue still remain a key challenge in tissue engineering. In this paper, cell‐laden 3D bioplotted hydrogel vessel‐like constructs made from alginate di‐aldehyde (ADA) and gelatin (GEL) are presented. The aim is to increase the mechanical stability of fibroblast‐laden ADA‐GEL vessels, tailoring them for maturation under dynamic cell culture conditions. BaCl2 is investigated as a crosslinker for the oxidized alginate‐gelatin system. Normal human dermal fibroblast (NHDF)‐laden vessel constructs are optimized successfully in terms of higher stiffness by increasing ADA concentration and using BaCl2, with no toxic effects observed on NHDF. Contrarily, BaCl2 crosslinking of ADA‐GEL accelerates cell attachment, viability, and growth from 7d to 24h compared to CaCl2. Moreover, alignment of cells in the longitudinal direction of the hydrogel vessels when extruding the cell‐laden hydrogel crosslinked with Ba2+ is observed. It is possible to tune the stiffness of ADA‐GEL by utilizing Ba2+ as crosslinker. In addition, a customized, low‐cost 3D printed polycarbonate (PC) perfusion chamber for perfusion of vessel‐like constructs is introduced. 相似文献
3D porous scaffolds fabricated from binary and ternary blends of silk fibroin (SF), gelatin (G), and hyaluronan (HA) and crosslinked by the carbodiimide coupling reaction were developed. Water-stable scaffolds can be obtained after crosslinking, and the SFG and SFGHA samples were stable in cell culture medium up to 10 days. The presence of HA in the scaffolds with appropriate crosslinking conditions greatly enhanced the swellability. The microarchitecture of the freeze-dried scaffolds showed high porosity and interconnectivity. In particular, the pore size was significantly larger with an addition of HA. Biological activities of NIH/3T3 fibroblasts seeded on SFG and SFGHA scaffolds revealed that both scaffolds were able to support cell adhesion and proliferation of a 7-day culture. Furthermore, cell penetration into the scaffolds can be observed due to the interconnected porous structure of the scaffolds and the presence of bioactive materials which could attract the cells and support cell functions. The higher cell number was noticed in the SFGHA samples, possibly due to the HA component and the larger pore size which could improve the microenvironment for fibroblast adhesion, proliferation, and motility. The developed scaffolds from ternary blends showed potential in their application as 3D cell culture substrates in fibroblast-based tissue engineering. 相似文献
Hydrogels are interesting as wound dressing for burn wounds to maintain a moist environment. Especially gelatin and alginate based wound dressings show strong potential. Both polymers are modified by introducing photocrosslinkable functionalities and combined to hydrogel films (gel‐MA/alg‐MA). In one protocol gel‐MA films are incubated in alg‐MA solutions and crosslinked afterward into double networks. Another protocol involves blending both and subsequent photocrosslinking. The introduction of alginate into the gelatin matrix results in phase separation with polysaccharide microdomains in a protein matrix. Addition of alg(‐MA) to gel‐MA leads to an increased swelling compared to 100% gelatin and similar to the commercial Aquacel Ag dressing. In vitro tests show better cell adhesion for films which have a lower alginate content and also have superior mechanical properties. The hydrogel dressings exhibit good biocompatibility with adaptable cell attachment properties. An adequate gelatin‐alginate ratio should allow application of the materials as wound dressings for several days without tissue ingrowth. 相似文献
Hydrogels are encouraging for different clinical purposes because of their high water absorption and mechanical relation to native tissues. Injectable hydrogels can modify the invasiveness of utilization, which decreases recovery and surgical costs. Principal designs applied to create injectable hydrogels incorporate in situ formation owing to chemical or/and physical crosslinking. Here, we report nontoxic, thermosensitive, injectable hydrogels composed of gelatin (GEL) and oxidized alginate (OA) reinforced by silicon carbide nanoparticles (SiC NPs) and crosslinked with N-hydroxysuccinimide (NHS) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC). The mechanical characteristics of the hydrogels were examined via rheological analysis. The outcomes reveal that extending the SiC NPs contents enhances the mechanical properties around five times. The cross-sectional microstructure of the scaffolds comprising 0.25, 1.0, and 1.5% SiC NPs was scrutinized by FESEM, verifying porous structure with interconnected pores. Because of the smaller pore sizes in the hydrogels, the swelling rate has reduced at the higher content of SiC, which diminishes the water uptake. Additionally, the biodegradation study unveils that the hydrogels with SiC are more long-lasting than the hydrogel without SiC. By adding SiC NPs, a decrease is observed in the biodegradation and swelling ratio. The scaffold with a higher SiC NPs content (1.5%) manifested better cell attachment and was less cytotoxic than hydrogel without SiC. OA/GEL composites embedded SiC NPs have manifested excellent physical properties for tissue engineering in comparison with hydrogel without nanoparticles. 相似文献
Hyaluronic acid (HA) is a high‐molecular‐weight glycosaminoglycan and extracellular matrix component that promotes cell proliferation. This study aimed to evaluate the effects of HA on alkali‐injured human corneal epithelial cells in vitro, and to elucidate the mechanisms by which HA mediates corneal cell protection. A human corneal epithelial cell line (HCE‐2) was treated with sodium hydroxide before incubation with low‐molecular‐weight HA (LMW‐HA, 127 kDa) or high‐molecular‐weight HA (HMW‐HA, 1525 kDa). A global proteomic analysis was then performed. Our data indicated that HA treatment protects corneal epithelial cells from alkali injury, and that the molecular weight of HA is a crucial factor in determining its effects. Only HMW‐HA reduced NaOH‐induced cytotoxic effects in corneal cells significantly and increased their migratory and wound healing ability. Results from 2D‐DIGE and MALDI‐TOF/TOF MS analyses indicated that HMW‐HA modulates biosynthetic pathways, cell migration, cell outgrowth, and protein degradation to stimulate wound healing and prevent cell death. To our knowledge, our study is the first to report the possible mechanisms by which HMW‐HA promotes repair in alkali‐injured human corneal epithelial cells. 相似文献
Bacterial cellulose (BC)/GEL composites were prepared in situ by adding gelatin into BC-producing culture medium. The addition of gelatin interfered with the formation of the BC pellicle structure and thus made the BC yield and growth rate quite different from that of pure BC. Scanning electron microscope images showed that the width of cellulose ribbons became narrower than that of pure BC and the gelatin filled in the pores of BC to form a dense structure. The addition level of gelatin significantly influences the yield of BC/GEL composites. An optimum value of 0.5 wt/v% gelatin was attained, with which the highest yield of 0.0541 g/100 mL was achieved. Under this condition, the weight percentage of gelatin in BC/GEL composite was 65 wt%. BC/GEL composites were treated with glutaraldehyde to crosslink BC fibrils and gelatin. The crosslinking degree, determined by the concentration of glutaraldehyde and crosslinking time, could affect the swelling behavior, thermal stability and mechanical properties of composites. With increasing of the crosslinking degree, the crystallinity index and swelling behavior of the composites decreased. The increase in the crosslinking degree also descreased the composite’s strain at break in elongation but increased the compressive and tensile strength. Covalent bonding between BC and gelatin provides good strength retention to the glutaraldehyde-treated composites with a high crosslinking degree. Considering the cytocompatibility and properties of composites, the most appropriate concentration of glutaraldehyde and crosslinking time were 1.0 wt/v% and 24 h, respectively. 相似文献
Papidly curable hydrogels were prepared through chemical crosslinking of gelatin with poly(carboxylic acid)s including poly (L-glutamic acid) (PLGA), hyaluronic acid (HA), and poly (acrylic acid) (PAA) by use of water-soluble carbodiimide (WSC). The effects of the nature of added poly (carboxylic acid)s on gelation of mixed gelatinpoly (carboxylic acid) aqueous solutions and adhesion of the resulting hydrogels to the mouse skin were evaluated. The addition of poly (carboxylic acid) s reduced the gelation time of gelatin aqueous solutions except for HA. Mixed gelatin-PLGA solutions were cured more rapidly than other mixed solutions and the gelation time was shortened with the increasing PLGA molecular weight. The resulting gelatin-PLGA hydrogels exhibited stronger adhesion to the mouse skin than gelatin-HA and gelatin-PAA hydrogels. The bonding strength increased with the increase in PLGA molecular weight up to 83,000 and thereafter decreased. The longer gelation time and lower adhesion of the gelatin-PAA hydrogels than the gelatin-PLGA hydrogels seem to be due to poorer compatibility of gelatin with PAA than with PLGA. The mixed gelatin-PLGA solution underwent phase separation when the concentration and molecular weight of PLGA became higher than a threshold. The insignificant or suppressive effect of HA addition might be ascribed to the HA-WSC reaction which was the least effective in hydrogel formation. 相似文献
A new methodology is developed to conjugate hyaluronic acid (HA) hydrogel with novel nano‐fibrous architectures via non‐covalent assembly that specifically allows for targeted adipose‐derived stem cells (ASCs) differentiation and soft tissue engineering. The assembly of non‐covalently associated hydrogel network produced via the interaction of a low molecular weight heparin (LMWH) modified HA derivative and heparin interacting protein (HIP). The multifunctional star poly(ethylene glycol) (PEG) and HIP copolymer has the capability to mediate the non‐covalent assembly of nano‐fibrous HA hydrogel networks via affinity interactions with LMWH. The effect of the HIP mediation on in vitro gelation, rheological characteristics, degradation, equilibrium swelling, adipose‐derived stem cells (ASCs) proliferation and differentiation of nano‐fibrous hydrogel is examined. The results suggest the potential utility of this unique design of the bioactive nano‐fibrous HA hydrogel in directing the differentiation of ASCs and adipogenesis in ECM‐mimetic scaffolds in vitro. These studies demonstrate that this nano‐fibrous HA hydrogel can render the formulation of a therapeutically effective platform for in vitro adipogenesis applications.
Hyaluronan is a polysaccharide that is increasingly investigated for its role in cellular adhesion and for the preparation of biomimetic matrices for tissue engineering. Hyaluronan gels are prepared for application as space fillers, whereas hyaluronan films are usually obtained by adsorbing or grafting a single hyaluronan layer onto a biomaterial surface. Here, we examine the possibility to employ the layer-by-layer technique to deposit thin films of cationic-modified hyaluronan (HA+) and hyaluronan (HA) of controlled thicknesses. The buildup conditions are investigated, and growth is compared to that of other polyelectrolyte multilayer films containing either HA as the polyanion or HA+ as the polycation. The films could be formed in a low ionic strength medium but are required to be cross-linked prior to contact with a physiological medium. NIH3T3 fibroblasts were perfectly viable on self-assembled hyaluronan films with, however, a preference for hyaluronan ending films. These findings point out the possibility to tune the thickness of thin hyaluronan films at the nanometer scale. Such architectures could be employed for investigating cell/substrate interactions or for functionalizing biomaterial surfaces. 相似文献
Hydrogels are attractive biomaterials for three-dimensional cell culture and tissue engineering applications. The preparation of hydrogels using alginate and gelatin provides cross-linked hydrophilic polymers that can swell but do not dissolve in water. In this work, we first reinforced pure alginate by using polyoxyethylene as a supporting material. In an alginate/PEO sample that contains 20 % polyoxyethylene, we obtained a stable hydrogel for cell culture experiments. We also prepared a stable alginate/gelatin hydrogel by cross-linking a periodate-oxidized alginate with another functional component such as gelatin. The hydrogels were found to have a high fluid uptake. In this work, preparation, characterization, swelling, and surface properties of these scaffold materials were described. Lyophilized scaffolds obtained from hydrogels were used for cell viability experiments, and the results were presented in detail. 相似文献
Cell therapies for age-related macular degeneration (AMD) treatment have been developed by integrating hydrogel-based biomaterials. Until now, cell activity has been observed only in terms of the modulus of the hydrogel. In addition, cell behavior has only been observed in the 2D environment of the hydrogel and the 3D matrix. As time-dependent stress relaxation is considered a significant mechanical cue for the control of cellular activities, it is important to optimize hydrogels for retinal tissue engineering (TE) by applying this viewpoint. Herein, a gellan Gum (GG)/Hyaluronic acid (HA) hydrogel was fabricated using a facile physical crosslinking method. The physicochemical and mechanical properties were controlled by forming a different composition of GG and HA. The characterization was performed by conducting a mass swelling study, a sol fraction study, a weight loss test, a viscosity test, an injection force study, a compression test, and a stress relaxation analysis. The biological activity of the cells encapsulated in 3D constructs was evaluated by conducting a morphological study, a proliferation test, a live/dead analysis, histology, immunofluorescence staining, and a gene expression study to determine the most appropriate material for retinal TE biomaterial. Hydrogels with moderate amounts of HA showed improved physicochemical and mechanical properties suitable for injection into the retina. Moreover, the time-dependent stress relaxation property of the GG/HA hydrogel was enhanced when the appropriate amount of HA was loaded. In addition, the cellular compatibility of the GG/HA hydrogel in in vitro experiments was significantly improved in the fast-relaxing hydrogel. Overall, these results demonstrate the remarkable potential of GG/HA hydrogel as an injectable hydrogel for retinal TE and the importance of the stress relaxation property when designing retinal TE hydrogels. Therefore, we believe that GG/HA hydrogel is a prospective candidate for retinal TE biomaterial. 相似文献
Innovative biomaterial‐based concepts are required to improve wound healing of damaged vascularized tissues especially in elderly multimorbid patients. To develop functional hydrogels as 3D cellular microenvironments and as carrier or scavenging systems, e.g., for mediator proteins or proinflammatory factors, collagen fibrils are embedded into a network of photo‐crosslinked acrylated hyaluronan (HA), chondroitin sulfate (CS), or sulfated HA (sHA). After lyophilization, the gels show a porous structure and an improved stability against degradation via hyaluronidase. Gels with CS and sHA bind significantly more lysozyme than HA/collagen gels and retard its release. The proliferation and metabolic activity of endothelial cells are significantly increased on sHA gels compared to CS‐ or only HA‐containing hydrogels. These findings highlight the potential of HA/collagen hydrogels with sulfated glycosaminoglycans to tune the protein binding and release behavior and to directly modulate cellular response. This can be easily translated into biomimetic biomaterials with defined properties to stimulate wound healing. 相似文献
This letter focuses on the first result the preparation and its swelling behavior of a novel hybrid gelatin hydrogel with carbon nanotubes. A novel hybrid gelatin hydrogel with carbon nanotubes was synthesized by physical mixing method. The structure of the novel hydorgel obtained was characterized by SEM. Besides, the swelling behavior of the hydrogel synthesized was measured at different temperature. The results indicated that carbon nanotubes added could maintain the stability of the hybrid hydrogel at 37 °C. This suggests that the hybrid gelatin hydrogel with carbon nanotubes could be used in biomedical field. Besides, its application in protein concentrating has been discussed. 相似文献
A well‐defined poly(ethylene glycol) based hyperbranched thermoresponsive copolymer with high content of acrylate vinyl groups was synthesized via a “one‐pot and one‐step” deactivation enhanced atom transfer radical polymerization approach, which provided an injectable and in situ crosslinkable system via Michael‐type thiol‐ene reaction with a thiol‐modified hyaluronan biopolymer. The hyperbranched structure, molecular weight, and percentage of vinyl content of the copolymer were characterized by gel permeation chromatography and 1H NMR. The lower critical solution temperature of this copolymer is close to body temperature, which can result in a rapid thermal gelation at 37 °C. The scanning electron microscopy analysis of crosslinked hydrogel showed the network formation with porous structure, and 3D cell culture study demonstrated the good cell viability after the cells were embedded inside the hydrogel. This injectable and in situ crosslinking hybrid hydrogel system offers great promise as a new class of hybrid biomaterials for tissue engineering. 相似文献
Hydrogels have been employed in regenerative treatments for decades because of their biocompatibility and structural similarity to the native extracellular matrix. Injectable hydrogels with interconnected porosity and specific internal structures are momentous for tissue engineering. Here, we develop a group of injectable hydrogels comprised of oxidized alginate (OA)/gelatin (GEL) strengthened by modifying the amount of Zn2SiO4 nanoparticles. The physicochemical characteristics of OA/GEL/Zn2SiO4 hydrogels were studied by mechanical strength, swelling ratio, and morphology. The outcomes revealed that the mechanical characteristics of hydrogels containing a higher amount of Zn2SiO4 (0.12 wt%) improved more than five times than the hydrogels fabricated without Zn2SiO4. The in vitro degradation outcomes manifested the degradation of the hydrogel comprising 0.12 wt% Zn2SiO4 NPs was slower than one without NPs, and remaining masses of hydrogels depend on different contents of Zn2SiO4 NPs. The hydrogel containing Zn2SiO4 NPs exhibited less cytotoxicity and good cell attachment than the hydrogels prepared without the nanoparticles. The cell viability and attachment show that the nanocomposite hydrogels are biocompatible (>96%) with great cell adhesion to osteosarcoma cell line MG63 depending on the presence of Zn2SiO4. The superior physical, chemical as well as mechanical characteristics of the hydrogels containing Zn2SiO4 NPs along with their cytocompatibility suggest that they can introduce as good candidates as scaffolds in tissue engineering. 相似文献
Synthesis of hydrogel at mild conditions is considered one most important challenge, especially if the hydrogel will be used for hosting bioactive materials or drugs. The procedure of hydrogel preparation should have no effect on the properties of the hosted materials. Hyaluronic acid (HA) was modified by adding dialdehyde groups to its structure to facilitate formation of hydrogel at very mild conditions. Dialdehyde HA (DHA) was prepared through oxidation of HA using sodium metaperiodate as oxidizing agent. The prepared DHA was characterized by Fourier‐transform infrared (FTIR) spectroscopy and X‐ray diffraction (XRD) and aldehyde content. A hydrogel was prepared using different chitosan/DHA molar ratio and fixed amount of glutaraldehyde at 25°C. The prepared hydrogel has tunable properties and pores size depending on the chitosan/DHA molar ratio. Sodium diclofenac was loaded on the hydrogel as a model drug. The hydrogel was characterized by FTIR spectroscopy, swelling rate, gel fraction, drug release profile, and cytotoxicity. The results obtained indicated that the properties of the prepared hydrogel, including gelling time, gel fraction, swelling, pores size, and drug release profile are highly tuned depending on the chitosan/DHA molar ratio. The drug loading efficiency was in the range of 70% to 85%. The cytotoxicity results reveal that the prepared hydrogel has a very low toxicity in presence and absence of sodium diclofenac. 相似文献
Nowadays, despite remarkable progress in developing bone tissue engineering products, the fabrication of an ideal scaffold that could meet the main criteria, such as providing mechanical properties and suitable biostability as well as mimicking the bone extracellular matrix, still seems challenging. In this regard, utilizing combinatorial approaches seems more beneficial. Here, we aim to reinforce the mechanical characteristics of gelatin hydrogel via a combination of Genipin‐based chemical cross‐linking and incorporation of the poly l ‐lactic acid (PLLA) nanocylinders for application as bone scaffolds. Amine‐functionalized nanocylinders are prepared via the aminolysis procedure and incorporated in gelatin hydrogel. The nanocylinder content (0, 1, 2, 3, and 4 wt%) and cross‐linking density (0.1, 0.5, and 1 wt/vol%) are optimized to achieve suitable morphology, swelling ratio, degradation rate, and mechanical behaviors. The results indicate that hydrogel scaffold cross‐linking by 0.5 wt% of Genipin shows optimized morphological feathers with a pore size of around 300 to 500 μm as well as an average degradation rate (40.09% ± 3.08%) during 32 days. Besides, the incorporation of 3 wt% PLLA nanocylinders into the cross‐linked gelatin scaffold provides an optimized mechanical reinforcement as compressive modulus, and compressive strength show a 4‐ and 2.6‐fold increase, respectively. 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay indicates that the scaffold does not have any cytotoxicity effect. In conclusion, gelatin composite reinforced with 3 wt% PLLA nanocylinders cross‐linked via 0.5 wt/vol% Genipin is suggested as a potential scaffold for bone tissue engineering applications. 相似文献