Preparation of External Stimulus-Free Gelatin-Catechol Hydrogels with Injectability and Tunable Temperature Responsiveness.

Gelatin is one of the most versatile biopolymers in various biomedical applications. A gelatin derivative gelatin-catechol (Gel-C) was developed in this study to further optimize its chemical and physical properties such as thermal reversibility and injectability. We found that Gel-C remains in a solution state at room temperature, and the temperature-dependent gelation capability of gelatin is well preserved in Gel-C. Its gel-forming temperature decreased to about 10 °C (about 30 °C for gelatin), and a series of gelatin derivatives with different gel-forming temperatures (10-30 °C) were formed by mixing gelatin and Gel-C in different ratios. Additionally, irreversible Gel-C hydrogels could be made without the addition of external stimuli by combining the physical cross-linking of gelatin and the chemical cross-linking of catechol. At the same time, properties of Gel-C hydrogels such as thermal reversibility and injectability could be manipulated by controlling the temperature and pH of the precursor solution. By simulating the formation of an irreversible Gel-C hydrogel in vivo, an in situ gelling system was fabricated by lowering the local temperature of the hydrogel with cold shock, thus realizing targeted and localized molecular delivery with prolonged retention time. This simple system integrated with the temperature responsiveness of gelatin and chemical cross-linking of catechol groups thus provides a promising platform to fabricate an in situ gelling system for drug delivery.

Synthesis and experimental/computational characterization of sorghum procyanidins-gelatin nanoparticles.

In this research, sorghum procyanidins (PCs) and procyanidin B1 (PB1) were encapsulated in gelatin (Gel) to form nanoparticles as a strategy to maintain their stability and bioactivity and for possible applications as inhibitors of metalloproteinases (MMPs) of the gelatinase type. Encapsulation was carried out by adding either PCs or PB1 to an aqueous solution of A- or B-type Gel (GelA or GelB) at different concentrations and pH. Under this procedure, the nanoparticles PCs-GelA, PCs-GelB, PB1-GelA, and PB1-GelB were synthesized and subsequently characterized by experimental and computational methods. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed that all types of nanoparticles had sizes in the range of 22-138 nm and tended to adopt an approximately spherical morphology with a smooth surface, and they were immersed in a Gel matrix. Spectral analysis indicated that the nanoparticles were synthesized by establishing hydrogen bonds and hydrophobic interactions betweenGel and the PCs or PB1. Study of simulated gastrointestinal digestion suggested that PCs were not released from the Gel nanoparticles, and they maintained their morphology (SEM analysis) and antioxidant activity determined by Trolox-equivalent antioxidant capacity (TEAC) assay. Computational characterization carried out through molecular docking studies of PB1 with Gel or (pro-)metalloproteinase-2 [(pro-)MMP-2], as a model representative of the PCs, showed very favorable binding energies (around -5.0 kcal/mol) provided by hydrogen bonds, van der Waals interactions, and desolvation. Additionally, it was found that PB1 could act as a selective inhibitor of (pro-)MMP-2.

The composite microbeads of alginate, carrageenan, gelatin, and poly(lactic-co-glycolic acid): Synthesis, characterization and Density Functional Theory calculations.

Binary (AC, AG), ternary (ACG, ACP, AGP), quaternary (ACGP) composite beads of alginate (A), carrageenan (C), gelatin (G), and poly (lactic-co-glycolic acid) (P) were prepared. The dried beads had a 700 µm average diameter. The microspheres with and without P were characterized by FT-IR, TGA/DTA, SEM, and PZC analysis. The results proved that the features of the composites were completely different from their bare components. Density Functional Theory (DFT) calculations were performed at the B3LYP/6-311++G** level to enlighten the elementary physical and chemical properties of A, C, P, and G compounds. The vibrational modes obtained by calculations were compared with those observed in the FT-IR spectra. The Frontier Molecular Orbital (FMO) analyses showed that the component G was the softer and had smaller energy gap than the other components and vice versa for component P. NBO (Natural Bond Orbital) analyses implied that the n → П* (resonance) interactions for components A, G, and P contributed to the lowering of the molecular stabilization, whereas that the n → σ* (anomeric) interactions were responsible for decreasing of the stabilization of the component. From the obtained results, these kinds of components can be hoped the promising materials for usage in the many scientific fields, especially in medicine and in drug design.

Valorization of a By-Product from the Production of Mechanically Deboned Chicken Meat for Preparation of Gelatins.

In recent decades, food waste management has become a key priority of industrial and food companies, state authorities and consumers as well. The paper describes the biotechnological processing of mechanically deboned chicken meat (MDCM) by-product, rich in collagen, into gelatins. A factorial design at two levels was used to study three selected process conditions (enzyme conditioning time, gelatin extraction temperature and gelatin extraction time). The efficiency of the technological process of valorization of MDCM by-product into gelatins was evaluated by % conversion of the by-product into gelatins and some qualitative parameters of gelatins (gel strength, viscosity and ash content). Under optimal processing conditions (48-72 h of enzyme conditioning time, 73-78 °C gelatin extraction temperature and 100-150 min gelatin extraction time), MDCM by-product can be processed with 30-32% efficiency into gelatins with a gel strength of 140 Bloom, a viscosity of 2.5 mPa.s and an ash content of 5.0% (which can be reduced by deionization using ion-exchange resins). MDCM is a promising food by-product for valorization into gelatins, which have potential applications in food-, pharmaceutical- and cosmetic fields. The presented technology contributes not only to food sustainability but also to the model of a circular economy.

Expression, Purification and Characterization of CAR/NCOA-1 Tethered Protein in E. coli Using Maltose-Binding Protein Fusion Tag and Gelatinized Corn Starch.

The constitutive active/androstane receptor (CAR) is a nuclear receptor that functions as a xenobiotic sensor, which regulates the expression of enzymes involved in drug metabolism and of efflux transporters. Evaluation of the binding properties between CAR and a drug was assumed to facilitate the prediction of drug-drug interaction, thereby contributing to drug discovery. The purpose of this study is to construct a system for the rapid evaluation of interactions between CAR and drugs. We prepared recombinant CAR protein using the Escherichia coli expression system. Since isolated CAR protein is known to be unstable, we designed a fusion protein with the CAR binding sequence of the nuclear receptor coactivator 1 (NCOA1), which was expressed as a fusion protein with maltose binding protein (MBP), and purified it by several chromatography steps. The thus-obtained CAR/NCOA1 tethered protein (CAR-NCOA1) was used to evaluate the interactions of CAR with agonists and inverse agonists by a thermal denaturation experiment using differential scanning fluorometry (DSF) in the presence and absence of drugs. An increase in the melting temperature was observed with the addition of the drugs, confirming the direct interaction between them and CAR. DSF is easy to set up and compatible with multiwell plate devices (such as 96-well plates). The use of DSF and the CAR-NCOA1 fusion protein together allows for the rapid evaluation of the interaction between a drug and CAR, and is thereby considered to be useful in drug discovery.

Designing Gelatin Methacryloyl (GelMA)-Based Bioinks for Visible Light Stereolithographic 3D Biofabrication.

Bioinks play a key role in determining the capability of the biofabricatoin processes and the resolution of the printed constructs. Excellent biocompatibility, tunable physical properties, and ease of chemical or biological modifications of gelatin methacryloyl (GelMA) have made it an attractive choice as bioinks for biomanufacturing of various tissues or organs. However, the current preparation methods for GelMA-based bioinks lack the ability to tailor their physical properties for desired bioprinting methods. Inherently, GelMA prepolymer solution exhibits a fast sol-gel transition at room temperature, which is a hurdle for its use in stereolithography (SLA) bioprinting. Here, synthesis parameters are optimized such as solvents, pH, and reaction time to develop GelMA bioinks which have a slow sol-gel transition at room temperature and visible light crosslinkable functions. A total of eight GelMA combinations are identified as suitable for digital light processing (DLP)-based SLA (DLP-SLA) bioprinting through systematic characterizations of their physical and rheological properties. Out of various types of GelMA, those synthesized in reverse osmosis (RO) purified water (referred to as RO-GelMA) are regarded as most suitable to achieve high DLP-SLA printing resolution. RO-GelMA-based bioinks are also found to be biocompatible showing high survival rates of encapsulated cells in the photocrosslinked gels. Additionally, the astrocytes and fibroblasts are observed to grow and integrate well within the bioprinted constructs. The bioink's superior physical and photocrosslinking properties offer pathways of tuning the scaffold microenvironment and highlight the applicability of developed GelMA bioinks in various tissue engineering and regenerative medicine applications.

Feasibility Study of Gelatin Preparation from the Bioinspired Collagen Aggregates by a "Two-step" Facile Degradation Method.

Gelatin is the putative research hotspot of natural products, but gelatin prepared by traditional alkali methods has seriously affected its applications due to the worryingly low molecular weight and poor gel strength. Herein, we took the lead to extract the distinct gelatin from a kind of bioinspired collagen aggregate (CA) by a two-step controlled degradation method. Structural analysis suggested that the CA better preserves the natural aggregated structure of nature collagen (typical D-periodic cross-striated pattern). Compared with the gelatin gelatinized by the conventional alkali method (G-Al) and commercial gelatin (CG), the gelatin (G-CA) from CA had a wide molecular weight distribution range, high transparency, high viscosity, and strong gel strength as expected. Meanwhile, the G-CA film exhibited better mechanical performance and thermostability than CG and G-Al films, and water vapor permeability was also higher in the G-CA film, whereas water solubility was higher in the CG and G-Al films. Thus, the G-CA film is more conducive to the use of food packaging or edible films, exhibiting more potential market application prospects. Notably, G-CA based on CA from waste hide offal provides a way to reuse leather waste resources and further realize green and clean production in leather industry.

One-pot preparation of a multi-functional enzymatically generated gelatin hydrogel with controllable antibacterial and hemorheological properties.

The creation of multi-functional bio-hydrogels with tunable properties that meet in vivo demands is significant but remains challenging. Inspired by host-guest chemistry, a novel multi-functional gelatin-based bio-hydrogel with tunable antibacterial and hemorheological properties (TAH-GEL) is synthesized via an in situ one-pot strategy. TAH-GEL not only exhibits excellent mechanical properties but also shows promising self-healing and bio-compatibility features. For the first time, this biomaterial presents controllable antibacterial and hemorheological properties by controlling the TAH-GEL polypseudorotaxane motif. The resulting bio-hydrogel is easy to prepare and delivers superior performance, making it a powerful tool for bio-applications, such as hemostatic materials.

Evaluation of hyaluronic acid nanoparticle embedded chitosan-gelatin hydrogels for antibiotic release.

The development of chitosan-gelatin (CS-G) hydrogels embedded with ampicillin-loaded hyaluronic acid nanoparticles (HA-NPs) for wound dressing is proposed. It was aimed to provide controlled ampicillin delivery by incorporation of HA-NPs into biocompatible CS-G hydrogel structure. According to in vitro ampicillin release studies, 55% of ampicillin was released from CS-G/HA-NPs hydrogels after 5 days. Antibacterial performance of CS-G/HA-NPs hydrogels was proven with agar disc diffusion test. For cytotoxicity assay, fibroblast cell viability increased in CS-G/HA-NPs hydrogels compared with CS-G group after 24 hr incubation. Consequently, the potential ability of CS-G/HA-NPs hydrogels as a controlled drug delivery system has been verified.

Eco-friendly gelatin films with rosin-grafted cellulose nanocrystals for antimicrobial packaging.

We report on gelatin films incorporating rosin-grafted cellulose nanocrystals (r-CNCs), which fulfill the most relevant requirements for antimicrobial packaging applications. Transparent gelatin/r-CNCs bionanocomposite films (0.5-6 wt% r-CNCs) were obtained by solution casting and displayed high UV-barrier properties, which were superior to the most used plastic packaging films. The gelatin/r-CNCs films exhibited a moderate water vapor permeability (0.09 g mm/m2 h kPa), and high tensile strength (40 MPa) and Young's modulus (1.9 GPa). The r-CNCs were more efficient in improving the optical, water vapor barrier and tensile properties of gelatin films than conventional CNCs. Grafting of rosin on CNCs resulted in an antimicrobial nanocellulose that inhibited the growth of Staphylococcus aureus and Escherichia coli. The antibacterial properties of r-CNCs were sustained in the gelatin films, as demonstrated by agar diffusion tests and proof-of-principle experiments involving cheese storage. Overall, the incorporation of r-CNCs as active fillers in gelatin films is a suitable approach for producing novel eco-friendly, antimicrobial packaging materials.

Application of Nanoemulsions (W/O) of Extract of Opuntia oligacantha C.F. Först and Orange Oil in Gelatine Films.

Over the past decade, consumers have demanded natural, completely biodegradable active packaging serving as food containers. Bioactive plant compounds can be added to biopolymer-based films to improve their functionality, as they not only act as barriers against oxidation, microbiological, and physical damage, they also offer functionality to the food they contain. A water-in-oil (W/O) nanoemulsion was produced by applying ultrasound to xoconostle extract and orange oil, and was incorporated into gelatine films in different proportions 1:0 (control), 1:0.10, 1:0.25, 1:0.50, 1:0.75, and 1:1 (gelatine:nanoemulsion). The nanoemulsions had an average size of 118.80 ± 5.50 nm with a Z-potential of -69.9 ± 9.93 mV. The presence of bioactive compounds such as phenols, flavonoids, and betalains in the films was evaluated. The 1:1 treatment showed the highest presence of bioactive compounds, 41.31 ± 3.71 mg of gallic acid equivalent per 100 g (GAE)/100g for phenols, 28.03 ± 3.25 mg of quercetin equivalent per 100 g (EQ)/100g flavonoids and 0.014 mg/g betalains. Radical inhibition reached 72.13% for 2,20-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS), and 82.23% for 1,1-diphenyl-2-picrylhydrazyl (DPPH). The color of the films was influenced by the incorporation of nanoemulsions, showing that it was significantly different (p < 0.05) to the control. Mechanical properties, such as tensile strength, Young's modulus, and percentage elongation, were affected by the incorporation of nanoemulsified bioactive compounds into gelatine films. The obtained films presented changes in strength and flexibility. These characteristics could be favorable as packaging material.

Control the fate of human umbilical cord mesenchymal stem cells with dual-enzymatically cross-linked gelatin hydrogels for potential applications in nerve regeneration.

Stem-cell-based therapy is a promising strategy to treat challenging neurological diseases, while its application is hindered primarily by the low viability and uncontrolled differentiation of stem cell. Hydrogel can be properly engineered to share similar characteristics with the target tissue, thus promoting cell viability and directing cell differentiation. In this study, we proposed a new dual-enzymatically cross-linked and injectable gelatin hydrogel for regulating survival, proliferation, and differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) in a three-dimensional matrix. This injectable gelatin hydrogel was formed by oxidative coupling of gelatin-hydroxyphenyl acid conjugates catalyzed by hydrogen horseradish peroxidase (HRP) and choline oxidase (ChOx). Modulus and H2 O2 release can be well controlled by ChOx activity. Results from calcein-AM/PI staining and Ki67 immunofluorescence tests demonstrated that the survival and proliferation behavior of hUC-MSCs were highly enhanced in HRP1U ChOx0.25U hydrogel with lower modulus and less H2 O2 release compared with other groups. Attractively, the expression of neuron-specific markers ß-III tubulin, neurofilament light chain (NFL), and synapsin-1 was significantly increased in HRP1U ChOx0.25U hydrogel as well. Additionally, in vitro hemolysis test and in vivo HE staining data highlighted the good biocompatibility. Undoubtedly, this injectable gelatin hydrogel's ability to control hUC-MSCs' fate holds enormous potentials in nervous disorders' therapy and nerve regeneration.

3D Bioprinting of Carbohydrazide-Modified Gelatin into Microparticle-Suspended Oxidized Alginate for the Fabrication of Complex-Shaped Tissue Constructs.

Extrusion-based bioprinting of hydrogels in a granular secondary gel enables the fabrication of cell-laden three-dimensional (3D) constructs in an anatomically accurate manner, which is challenging using conventional extrusion-based bioprinting processes. In this study, carbohydrazide-modified gelatin (Gel-CDH) was synthesized and deposited into a new multifunctional support bath consisting of gelatin microparticles suspended in an oxidized alginate (OAlg) solution. During extrusion, Gel-CDH and OAlg were rapidly cross-linked because of the Schiff base formation between aldehyde groups of OAlg and amino groups of Gel-CDH, which has not been demonstrated in the domain of 3D bioprinting before. Rheological results indicated that hydrogels with lower OAlg to Gel-CDH ratios possessed superior mechanical rigidity. Different 3D geometrically intricate constructs were successfully created upon the determination of optimal bioprinting parameters. Human mesenchymal stem cells and human umbilical vein endothelial cells were also bioprinted at physiologically relevant cell densities. The presented study has offered a novel strategy for bioprinting of natural polymer-based hydrogels into 3D complex-shaped biomimetic constructs, which eliminated the need for cytotoxic supplements as external cross-linkers or additional cross-linking processes, therefore expanding the availability of bioinks.

Preparation of Electrospun Gelatin Mat with Incorporated Zinc Oxide/Graphene Oxide and Its Antibacterial Activity.

Current wound dressings have poor antimicrobial activities and are difficult to degrade. Therefore, biodegradable and antibacterial dressings are urgently needed. In this article, we used the hydrothermal method and side-by-side electrospinning technology to prepare a gelatin mat with incorporated zinc oxide/graphene oxide (ZnO/GO) nanocomposites. The resultant fibers were characterized by field emission environment scanning electron microscopy (FESEM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffractometry (XRD) and Fourier transform infrared spectroscopy (FTIR). Results indicated that the gelatin fibers had good morphology, and ZnO/GO nanocomposites were uniformly dispersed on the fibers. The loss of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) viability were observed to more than 90% with the incorporation of ZnO/GO. The degradation process showed that the composite fibers completely degraded within 7 days and had good controllable degradation characteristics. This study demonstrated the potential applicability of ZnO/GO-gelatin mats with excellent antibacterial properties as wound dressing material.

Injectable self-crosslinking hydrogels for meniscal repair: A study with oxidized alginate and gelatin.

Injectable in situ gelling hydrogels are viable treatment options for meniscal injuries occurring in athletes. The present study aims to develop an injectable hydrogel via borax complexation of oxidized alginate, followed by a self-crosslinking reaction with gelatin through a Schiff's base reaction. Gelation kinetics and degree of crosslinking could be controlled by changing the concentration of components and the formation of Schiff ;'s base formation was confirmed by Raman spectroscopy. The injectable alginate dialdehyde-gelatin (15ADA20G) hydrogel showed 423 ±â€¯20 % water uptake, had an average pore size of 48 µm and compressive strength 295 ±â€¯32 kPa. Phase contrast images, scanning electron micrographs and actin staining depicted adhesion, profuse proliferation, and distribution of fibrochondrocytes on the hydrogel demonstrating its cytocompatibility. Application of hydrogel at the pig meniscal tear ex vivo showed good integration with the host meniscal tissue. Further, the histology of 15ADA20G hydrogel filled meniscus showed retention of hydrogel in the close proximity of meniscal tear even after 3days in culture. The self-crosslinking injectable hydrogel offers a niche for the growth of fibrochondrocytes.

Designing an anti-inflammatory and tissue-adhesive colloidal dressing for wound treatment.

Wound dressing materials are widely used to protect wounds from the external environment and to promote wound healing. However, conventional wound dressings lack tissue adhesive properties and anti-inflammatory functions, which lead to fibrosis and stricture, in cases such as gastrointestinal wounds after endoscopic surgery. In the current study, we report tissue-adhesive and anti-inflammatory properties of a wound dressing composed of corticosteroid-modified gelatin particles. Hydrocortisone (HC), which is a class of anti-inflammatory corticosteroid, was used to modify Alaska-pollock gelatin (ApGltn) to synthesize HC-modified ApGltn (HC-ApGltn). Microparticles (MPs) of HC-ApGltn were fabricated by adding ethanol in HC-ApGltn aqueous solution and performing thermal crosslinking (TC) without the use of toxic surfactants and crosslinking reagents. Modification of ApGltn with hydrophobic HC containing cholesterol backbone structure improved its adhesion strength to gastric submucosal tissues under wet conditions owing to hydrophobic interactions. This retention of adhesive property under wet conditions allows for stable protection of wounds from the external environment. We found that HC-ApGltn MPs were taken up by macrophages and they effectively suppressed morphological changes of LPS-activated macrophages and the expression level of the inflammatory cytokine. Robust tissue adhesive and anti-inflammatory MPs may serve as an advanced wound dressing that can protect wounds and suppress inflammatory responses for promoting wound healing.

An efficient two-step preparation of photocrosslinked gelatin microspheres as cell carriers to support MC3T3-E1 cells osteogenic performance.

Gelatin microspheres have been commonly used in tissue engineering, but their application is often limited by the uncontrollability and potential cytotoxicity of traditional chemical cross-linking method. Methylacrylamide modification and photocrosslinking provide a controllable and cytocompatible cross-linking method for gelatin hydrogels, however, microspheres fabricated by this single photopolymerization process is uncontrollable. In this study, we show that increasing the gelling ability of gelatin methacrylamide (GMA) at low temperatures is vital to prepare photocrosslinked gelatin microspheres, which in turn improves the controllability and compatibility of conventional chemical cross-linking methods. We detailed characterized the rheological performance with varying temperature and demonstrated that the gelling capability of GMA could be improved by increasing GMA solution concentration and reducing methacrylate substitution. The physicochemical properties of the photocrosslinked microspheres can be modulated via methacrylamide modification, as evidenced by the positive correlation between the physicochemical optimization of the hydrogel bulk and the degree of methacrylate substitution. Next, we successfully fabricated GMA spheres by a two-step process of low-temperature gelation followed by photopolymerization crosslinking. Finally, we show that the microcarriers exhibited favorable supporting for MC3T3-E1 cell proliferation, spreading, and osteogenic differentiation. This study provided a controllable and cytocompatible photocrosslinking procedure for GMA microspheres with broad application prospects, of course, not limited to cell microcarriers.

In situ formation of silver nanoparticles-contained gelatin-PEG-dopamine hydrogels via enzymatic cross-linking reaction for improved antibacterial activities.

Hydrogels containing silver nanoparticles (AgNPs) were recently found to exhibit excellent antibacterial properties against both gram-negative/positive bacteria and fungi. In this study, we reported the synthesis of AgNPs-contained gelatin-polyethylene glycol-dopamine (AgNP-GPD) hydrogels via the in situ formation of AgNPs in GPD polypeptide solution, followed by an enzymatic cross-linking reaction to form hydrogels. The experimental results showed that the reducing reaction exerted by GPD polypeptides under physiological conditions can afford the formation of AgNPs in situ in the polypeptide solution without the need for additional reducing agents and/or processes such as UV or thermal treatments and then the hydrogelation of GPD polypeptide solution containing AgNPs were preceeded via enzymatic cross-linking reaction. It was found that the gelation time, hydrogel mechanical property, degree of swelling and degree of enzymatic degradation for both GPD and AgNP-GPD hydrogels can be tuned by varying enzyme/oxidative agent concentration, catechol content, and reducing reaction conditions such as reaction time and silver ion concentration. Importantly, AgNP-GPD hydrogels exhibit excellent antibacterial properties against gram-negative and gram-positive bacteria. This type of hydrogel is a promising biomaterial for biomedical applications including wound healing and tissue engineering.

Molecularly imprinted gelatin nanoparticles for DNA delivery and in-situ fluorescence imaging of telomerase activity.

DNA-loaded molecularly imprinted gelatin nanoparticles (GDMI-NPs) were prepared to deliver the Cy3- and Cy5-labelled DNA probe to a tumor region. This allows the activity of telomerase can be detected over 3-400 cells with a low detection limit (3 cells). Fluorescence images were acquired at an excitation wavelength of 535 nm and the emission from the green channel (550-580 nm; label Cy3) and the red channel (650-680 nm; label Cy5). HeLa cells and HepG2 cells were both used to test the performance of GDMI-NPs. Experimental results confirmed the GDMI-NPs has hardly retained in liver and spleen tissue, and its circulated time was longer than that of non-imprinted nanoparticles in blood. The ability of GDMI-NPs to resist immuno stress and anti-macrophage phagocytosis shows great potential for cancer diagnosis and as a drug carrier. Graphical abstract Highly DNA-loaded molecularly imprinted gelatin nanoparticles (GDMI-NPs) were prepared to deliver the Cy3-labelled DNA probe to a cancer region, and realization of telomerase in situ fluorescence imaging at the tumor site.

Multicellular Co-Culture in Three-Dimensional Gelatin Methacryloyl Hydrogels for Liver Tissue Engineering.

Three-dimensional (3D) tissue models replicating liver architectures and functions are increasingly being needed for regenerative medicine. However, traditional studies are focused on establishing 2D environments for hepatocytes culture since it is challenging to recreate biodegradable 3D tissue-like architecture at a micro scale by using hydrogels. In this paper, we utilized a gelatin methacryloyl (GelMA) hydrogel as a matrix to construct 3D lobule-like microtissues for co-culture of hepatocytes and fibroblasts. GelMA hydrogel with high cytocompatibility and high structural fidelity was determined to fabricate hepatocytes encapsulated micromodules with central radial-type hole by photo-crosslinking through a digital micromirror device (DMD)-based microfluidic channel. The cellular micromodules were assembled through non-contact pick-up strategy relying on local fluid-based micromanipulation. Then the assembled micromodules were coated with fibroblast-laden GelMA, subsequently irradiated by ultraviolet for integration of the 3D lobule-like microtissues encapsulating multiple cell types. With long-term co-culture, the 3D lobule-like microtissues encapsulating hepatocytes and fibroblasts maintained over 90% cell viability. The liver function of albumin secretion was enhanced for the co-cultured 3D microtissues compared to the 3D microtissues encapsulating only hepatocytes. Experimental results demonstrated that 3D lobule-like microtissues fabricated by GelMA hydrogels capable of multicellular co-culture with high cell viability and liver function, which have huge potential for liver tissue engineering and regenerative medicine applications.