Concentrations Of Hydrogel Research Articles

Hydrogel composition and mechanical stiffness of 3D bioprinted cell-loaded scaffolds promote cartilage regeneration

ObjectiveTo investigate the impact of different component ratios and mechanical stiffness of the gelatin-sodium alginate composite hydrogel scaffold, fabricated through 3D bioprinting, on the viability and functionality of chondrocytes.MethodsThree different concentrations of hydrogel, designated as low, medium, and high, were prepared. The rheological properties of the hydrogel were characterized to optimize printing parameters. Subsequently, the printability and shape fidelity of the cell-loaded hydrogel scaffolds were statistically evaluated, and the chondrocyte viability was observed. Dynamic mechanical analysis was conducted to measure the modulus, thereby assessing the scaffold’s stiffness. Following a 21-day culture period, RT-PCR, histological staining, and immunostaining were employed to assess chondrocyte activity, chondrosphere aggregates formation, and cartilage matrix production.ResultsBased on rheological analysis, optimal printing temperatures for each group were determined as 27.8°C, 28.5°C, and 30°C. The optimized printing parameters could ensure the molding effect of the scaffolds on the day of printing, with the actual grid area of the scaffolds was close to the theoretical grid area. And the scaffolds exhibited good cell viability (93.24% ± 0.99%, 92.04% ± 1.49%, and 88.46% ± 1.53%). After 7 days of culture, the medium and high concentration groups showed no significant change in grid area compared to the day of printing (p > 0.05), indicating good morphological fidelity. As the hydrogel’s bicomponent ratio increased, both the storage modulus and loss modulus increased, while the loss factor remained relatively constant. The highest number of chondrocytes-formed chondrosphere aggregates in the medium concentration group was observed by light microscopy. RT-PCR results indicated significantly higher expression levels of chondrogenic genes SOX9, Agg, and Col-II in the low and medium concentration groups compared to the high concentration group (p < 0.05). Histological staining results showed that the middle concentration group formed the highest number of typical cartilage lacunae.ConclusionThe aforementioned results indicate that in 3D bioprinted cell-loaded GA-SA composite hydrogel scaffolds, the scaffolds with the composition ratio (10:3) and mechanical stiffness (∼155 kPa) exhibit sustained morphological fidelity, effectively preserve the hyaline phenotype of chondrocytes, and are more conducive to cartilage regeneration.

Engineering Cyborg Pathogens through Intracellular Hydrogelation.

Synthetic biology primarily focuses on two kinds of cell chassis: living cells and nonliving systems. Living cells are autoreplicating systems that have active metabolism. Nonliving systems, including artificial cells and nanoparticles, are nonreplicating systems typically lacking active metabolism. In recent work, Cyborg bacteria that are nonreplicating-but-metabolically active have been engineered through intracellular hydrogelation. Intracellular hydrogelation is conducted by infusing gel monomers and photoactivators into cells, followed by the activation of polymerization of the gel monomers inside the cells. However, the previous work investigated only Escherichia coli cells. Extending the Cyborg-Cell method to pathogenic bacteria could enable the exploitation of their pathogenic properties in biomedical applications. Here, we focus on different strains of Pseudomonas aeruginosa, Staphylococcus aureus, and Klebsiella pneumoniae. To synthesize the Cyborg pathogens, we first reveal the impact of different hydrogel concentrations on the metabolism, replication, and intracellular gelation of Cyborg pathogens. Next, we demonstrate that the Cyborg pathogens are taken up by macrophages in a similar magnitude as wild-type pathogens through confocal microscopy and real-time PCR. Finally, we show that the macrophage that takes up the Cyborg pathogen exhibits a similar phenotypic response to the wild-type pathogen. Our work generalizes the intracellular hydrogelation approach from lab strains of E. coli to bacterial pathogens. The new Cyborg pathogens could be applied in biomedical applications ranging from drug delivery to immunotherapy.

Natural loofah sponge inspired 3D printed bionic scaffolds promote personalized bone defect regeneration

Critical-sized bone defects pose serious health concerns for patients. Clinically, the use of functionalized bone implants has emerged as an effective solution. However, the rapid advancement in drug and biomaterials has led to an increasing design cost, triggering discussions in the field about how to efficiently create customized functional bone implants. Inspired by the unique structure of natural loofah sponges that effectively deliver nutrients to seeds, we designed a functionalized bone implant emulating this structure. Drug-release gradients were achieved through the application of different concentrations of hydrogels within the composite scaffold. This approach allowed active substances to be released outwardly during the early stage of bone repair, sustaining a local drug micro-environment within the implant scaffold that promotes angiogenesis and osteogenic differentiation in damaged areas. In vivo experiments showed that our loofah sponge bionic scaffold outperformed traditional hydroxyapatite scaffolds by promoting both bone and vascular regeneration. We expect the design of loofah sponge bionic scaffold could potentially deliver an effective strategy in the development of functionalized bone implants.

Hydrogel Composition Effects on Performance as Single-Walled Carbon Nanotube Purification Media.

Hydrogel microsphere media allows for postsynthetic purification of single-walled carbon nanotubes (SWNTs), affording characterization and application of their unique (n,m) chirality-dependent properties. This work reports the characterization of five hydrogel resins, Sephacryl S-100, S-200, S-300, S-400, and S-500, and the implementation of each as a SWNT purification medium. The physiochemical properties of each resin were explored spectroscopically through elemental analyses and with both light and electron microscopy. Both surface porosity and hydrogel swelling ratio were found to increase as the concentration of component allyl dextran (aDEX) decreased, each with an increasing Sephacryl S-number. Conversely, invariant properties included a hydrogel microsphere size distribution and concentrations of components methylenebisacrylamide and ammonium persulfate. When employed within gel-based SWNT purification schemes in overloading conditions, Sephacryl formulations of larger S-number adsorbed fewer SWNTs, but the chirality dependence of SWNT adsorption and elution was approximately consistent across all resins. In underloading conditions, approximately one-third of introduced SWNTs passed through each resin unabsorbed, while the resins showed varying chirality-dependent adsorption efficiencies. These observations collectively identify aDEX-rich gel regions as being responsible for SWNT purification, along with a SWNT-exclusive parameter other than chirality (speculated as length) that convolutes the effectiveness of gel-based single-chirality purification.

Enhanced hepatotoxicity assessment through encapsulated HepG2 spheroids in gelatin hydrogel matrices: Bridging the gap from 2D to 3D culture

Conventional 2D drug screening often fails to accurately predict clinical outcomes. We present an innovative approach to improve hepatotoxicity assessment by encapsulating HepG2 spheroids in gelatin hydrogel matrices with different mechanical properties. Encapsulated spheroids exhibit sustained liver-specific functionality, enhanced expression of drug-metabolizing enzymes, and increased drug sensitivity compared to 2D cultures. The platform detects critical variations in drug response, with significant differences in IC50 values between 2D and spheroid cultures ranging from 1.3-fold to > 13-fold, particularly for acetaminophen. Furthermore, drug-metabolizing enzyme expression varies across hydrogel concentrations, suggesting a role for matrix mechanical properties in modulating hepatocyte function. This novel spheroid-hydrogel platform offers a transformative approach to hepatotoxicity assessment, providing increased sensitivity, improved prediction, and a more physiologically relevant environment. The use of such advanced in vitro models can accelerate drug development, reduce animal testing, and contribute to improved patient safety and clinical outcomes.

Comparative Analysis of the Cytotoxic Effects of Modified Triple Antibiotic Hydrogel: Insights From Experimental Models.

The effectiveness of intracanal medicaments (ICMs) in root canal therapy is critical for successful dental treatments, yet their cytotoxic effects pose significant challenges. This research uses zebrafish embryos and dental pulp stem cells (DPSCs) to identify the optimal concentration that balances antibacterial efficacy with minimal toxicity. This study aims to address the need for effective ICMs in dentistry by formulating and assessing the embryotoxicity and cytocompatibility of a novel carrageenan-based modified triple antibiotic paste (MTAP) hydrogel at different concentrations (1, 5, and 10 mg/mL) using a zebrafish model and cell culture assay. The hydrogel was formulated by combining antibiotic solutions (ciprofloxacin, metronidazole, and amoxicillin) with carrageenan and xanthan gum. Zebrafish embryos were exposed to varying concentrations of MTAP hydrogel, chlorhexidine (CHX), calcium hydroxide (CaOH2), and plain carrageenan to assess developmental toxicity, survival rate, heart rate, hatching rate, and macrophage migration. The cytotoxicity against DPSCswas examined within a timeframe of 6, 24, and 72 hours with the use of the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay. The analysis revealed developmental toxicity with malformations observed at higher concentrations of MTAP hydrogel, CaOH2, and CHXmedicaments, indicating potential toxicity. Significant impacts on survival, heart rate, and hatching rate were noted in the CaOH2 and CHX groups, as well as at higher MTAP hydrogel concentrations, emphasizing the importance of dosage considerations. The neutral red assay confirmed toxicity, with macrophage migration observed in CaOH2, CHX, and higher MTAP hydrogel concentrations. Lower concentrations, particularly at 1 mg/mL, showed no adverse effects on zebrafish embryos and larvae. These findings align with cell viability investigations, which demonstrated that higher antibiotic concentrations resulted in decreased cell proliferation and viability over time. Conversely, at a lower concentration of 1 mg/mL, cell proliferation notably increased after 72 hours.Plain MTAP and CHX exhibited the highest toxicity levels in the MTTassay. The study concludes that while higher concentrations of MTAP hydrogel exhibit toxic effects, the hydrogel at 1 mg/mL demonstrates no adverse impact on zebrafish embryos, larvae, and DPSCs. These findings underscore the necessity of optimizing ICM concentrations to balance antibacterial efficacy and minimal cytotoxicity.

Extrusion-based bioprinting: considerations toward gelatin-alginate bioink

PurposeThis study aims to develop a simplified bioink preparation method that can be applied to most hydrogel bioinks used in extrusion-based techniques.Design/methodology/approachThe parameters of the bioprinting process significantly affect the printability of the bioink and the viability of cells. In turn, the bioink formulation and its physicochemical properties may influence the appropriate range of printing parameters. In extrusion-based bioprinting, the rheology of the bioink affects the printing pressure, cell survival and structural integrity. Three concentrations of alginate-gelatin hydrogel were prepared and printed at three different flow rates and nozzle gauges to investigate the print parameters. Other characterizations were performed to evaluate the hydrogel structure, printability, gelation time, swelling and degradation rates of the bioink and cell viability. An experimental design was used to determine optimal parameters. The analyses included live/dead assays, rheological measurements, swelling and degradation.FindingsThe experimental design results showed that the hydrogel flow rate substantially influenced printing accuracy and pressure. The best hydrogel flow rate in this study was 10 ml/h with a nozzle gauge of 18% and 4% alginate. Three different concentrations of alginate-gelatin hydrogels were found to exhibit shear-thinning behavior during printing. After seven days, 46% of the structure in the 4% alginate-5% gelatin sample remained intact. After printing, the viability of skin fibroblast cells for the optimized sample was 91%.Originality/valueThis methodology offers a straightforward bioink preparation method applicable to the majority of hydrogels used in extrusion-based procedures. This can also be considered a prerequisite for cell printing.

Effect of super absorbent hydrogel on hydro-physical properties of soil under deficit irrigation

Due to water scarcity challenges, efficient management of irrigation water is becoming crucial. Water use efficiency (WUE) involves increasing crop productivity without increasing water consumption. This study was carried out to study the effect of hydrogel, deficit irrigation and soil type on WUE, soil hydro-physical properties and lettuce productivity. For this purpose, four irrigation treatments (100%, 85%, 70% and 60% of full irrigation requirements), four hydrogel concentrations (0, 0.1, 0.2 and 0.3% w/w) and three soil textural classes (clay, loamy sand, and sandy-clay soil) were conducted in pot experiment at open field during two consecutive seasons. The results revealed that crop growth parameters and soil hydro-physical properties were significantly affected by hydrogel application rates. Hydrogel addition significantly enhanced head fresh and dry weights, chlorophyll content, number of leaves and WUE. Application of hydrogel at 0.3% and 85% of irrigation requirements achieved the highest WUE without significant yield reductions. Changes in the studied hydro-physical properties of soil were more dependent on soil texture and hydrogel application rate than on the amount of irrigation water. The significant decrease in soil saturated hydraulic conductivity and bulk density confirms that super absorbent hydrogels could be recommended to improve soil water retention and enhance water use efficiency under deficit irrigation conditions.

Pinecone failure, hydrogel, fertilizer and volume of containers in the production of araucaria seedlings

The objective of this work was the evaluation of initial growth and quality of araucaria seedlings under the influence of alterations to different proprieties of the substrate. Four different experiments were installed, comparing volume of tubes, concentrations of controlled release fertilizer (FLC), proportion of crushed pinecone failures (substrate characteristics were also evaluated) and the effect of hydrogel concentrations in seedlings derived from different mother trees. The evaluations were carried out 10 months after sowing, varying according to the experiment, and were comprised of height, diameter, height/diameter ratio (h/d), survival rate and germination speed index. The seedlings developed inside the larger tubes (280 cm3) averaged higher in height, diameter and h/d ratio. The 6 g L-1 dose of fertilizer generated higher averages for all analyzed variables. The addition of pinecone failure to the substrate improved some physical attributes such as density, total porosity and aeration space, however it worsened the attributes related to water availability in the substrate. As for the growth of seedlings, the addition of pinecone failure did not change the height, diameter and h/d ratio. The use of hydrogel (3 g L-1 and 6 g L-1) generated better results for height. It is recommended the use of 180 cm³ tubes and 6 g L-1 of the contloed release fertilizer. Pinecone can be used in the substrate, as well as hydrogel as a additional resource.

Analysis of a Poloxamer Hydrogel Composed of Human Mesenchymal Stromal Cells for Reepithelization of Skin Injuries

The body's natural response to tissue damage is wound healing. Hydrogels have compelling benefits in the healing of wounds, not only because of their permeability, biodegradability, and biocompatibility, but also because they offer an optimal environment for cell migration and proliferation. The primary goal of this work was to create and characterize a hydrogel that contained human mesenchymal stromal cells (hMSCs) in order to facilitate the healing of superficial skin injuries. hMSCs were embedded using a biocompatible biomaterial called Poloxamer 407®. The hydrogel that was created, with 20 percent (w/w) of polymer, had the best formulation in terms of its mechanical, morphological, biological, and physical characteristics.. The hydrogel's remarkable swelling capacity spoke to its ability to absorb wound exudate. The LIVE/DEAD® assay verifies that when hMSCs were injected into the hydrogels, they stayed viable for at least 48 hours. The keratinocytes' ability to proliferate and mend was unaffected by the addition of progressively higher concentrations of hMSC-loaded hydrogel to the epithelium; the entire wound healed in less than a day. Our research provides new avenues for the application of poloxamer hydrogels as skin superficial wound carriers.

Nanocomposite Hydrogels Based on Poly(vinyl alcohol) and Carbon Nanotubes for NIR-Light Triggered Drug Delivery.

Photothermal nanocomposite hydrogels are promising materials for remotely triggering drug delivery by near-infrared (NIR) radiation stimuli. In this work, a novel hydrogel based on poly(vinyl alcohol), poly(vinyl methyl ether-alt-maleic acid), poly(vinyl methyl ether), and functionalized multiwalled carbon nanotubes (MWCNT-f) was prepared by the freeze/thaw method. A comparative characterization of materials (with and without MWCNT-f) was carried out by infrared spectroscopy, differential scanning calorimetry, scanning electron microscopy, mechanical assays, swelling kinetics measurements, and photothermal analysis under NIR irradiation. Hydrophilic chemotherapeutic 5-fluorouracil (5-FU) and hydrophobic ibuprofen drugs were independently loaded into hydrogels, and the drug release profiles were obtained under passive and NIR-irradiation conditions. The concentration-dependent cytotoxicity of materials was studied in vitro using noncancerous cells and cancer cells. Notable changes in the microstructure and physicochemical properties of hydrogels were observed by adding a low content (0.2 wt %) of MWCNT-f. The cumulative release amounts of 5-FU and ibuprofen from the hydrogel containing MWCNT-f were significantly increased by 21 and 39%, respectively, through the application of short-term NIR irradiation pulses. Appropriate concentrations of the nanocomposite hydrogel loaded with 5-FU produced cytotoxicity in cancer cells without affecting noncancerous cells. The overall properties of the MWCNT-f-containing hydrogel and its photothermal behavior make it an attractive material to promote the release of hydrophilic and hydrophobic drugs, depending on the treatment requirements.

Comparison of Antibacterial Efficacy of Triple Antibiotic-Loaded Hydrogel Versus Modified Triple Antibiotic-Loaded Hydrogel as Intracanal Medicament Against Enterococcus faecalis: An In vitro Study.

Triple antibiotic paste (TAP) is known to have an essential role in the success of endodontic treatment by eliminating pathogens from the root canal system. Unfortunately, it causes discolouration and cytotoxicity at high concentrations. The objective of this research was to assess and compare the antimicrobial effectiveness of various concentrations (1 mg, 5 mg, 10 mg) of TAP, TAP hydrogel (TAPH), M-TAP, and M-TAP hydrogel (MTAPH) against Enterococcus faecalis. The agar well diffusion method was used to assess the antibiotic sensitivity of the following intracanal medicaments: TAP (ciprofloxacin, metronidazole, and minocycline) mixed in a ratio of 1: 1: 1; TAPH, M-TAP (ciprofloxacin, metronidazole, and amoxicillin), M-TAPH and plain hydrogel. Each tested medicament was individually evaluated for its antimicrobial activity against Enterococcus faecalis. Structural and topographical characterisation were analysed using a Scanning Electron Microscope (SEM) and interpreted using ImageJ software. A microdilution broth test was performed to examine the minimum inhibitory concentration and minimum bactericidal concentration (MBC) of M-TAP and TAP. Except for the plain hydrogel, M-TAP and hydrogel and TAP and hydrogel showed significantly varied inhibitory zones at different concentrations. M-TAPH showed the highest mean zone of inhibition of 21.6, 33.33 and 38.0 mm at a concentration of 1, 5, and 10 mg/mL when compared to TAPH, which showed a mean zone of inhibition of 3.3 mm,12.3 mm, 21.3 mm at the respective concentrations. The MIC study shows that more than 75% of Enterococcus faecalis growth was inhibited by M-TAP at a concentration of 5 μg/mL, whereas TAP showed inhibition at a concentration of 35 μg/mL. MBC results indicate that almost 99.9% of the bacterial population was killed at a concentration of 100 μg/mL (10-1) for TAP and 10 μg/mL (10-2) for M-TAP. The antibacterial efficacy of M-TAP was significantly higher than TAP. Application of M-TAP at lower doses is advised to overcome the disadvantages seen with TAP.

Hidrogel e sistema arduino no transplantio de Schinus terebinthifolia para arborização urbana

Abstract Urban forest provide medium and large vegetation cover in urban areas. Planting native trees on sidewalks is a viable approach to reduce damages caused by extensive urbanization. The use of hydrogels seems to increase the success of seedling transplantation in urban environments. Thus, this study aims to evaluate the efficacy of Schinus terebinthifolia in urban afforestation, focusing on its adaptation and post-transplant survival using hydrogel and being monitored by an arduino system. The concentrations of the commercial hydrogel used were 0.75, 1.5, 3.0, and 6.0 g L-1, and two controls were also established: a control treatment without irrigation and without hydrogel, and a control with daily water irrigation. The evaluation was continuous for 14 days, and the parameters analyzed were substrate temperature and moisture, relative water content (RWC), electrolyte extravasation (EEE), chlorophyll content, and biochemical compound. A completely randomized design was adopted, consisting of 6 treatments with 9 replicates each. The results indicate that S. terebinthifolia is a highly resilient species suitable for urban afforestation, showing remarkable tolerance to transplantation and water restriction. It was observed that the use of hydrogels significantly contributes to maintaining substrate moisture, resulting in greater stability of the transplanted seedlings. The Arduino system allowed for continuous and precise evaluation of substrate conditions, optimizing the management of urban afforestation and validating the efficiency of the applied treatments. Positive responses were observed when using hydrogels in terms of relative water content, membrane stability, and antioxidant activity, even under water restriction. The viability of Schinus terebinthifolia for urban afforestation stands out, through the application of hydrogels and the use of the Arduino system to monitor parameters such as temperature and humidity.

Poly (ε-caprolactone)-Based Scaffolds with Multizonal Architecture: Synthesis, Characterization, and In Vitro Tests.

Tissue engineering is vital in treating injuries and restoring damaged tissues, aiming to accelerate regeneration and optimize the complex healing process. In this study, multizonal scaffolds, designed to mimic tissues with bilayer architecture, were prepared using the rotary jet spinning technique (RJS scaffolds). Polycaprolactone and different concentrations of alginate hydrogel (2, 4, and 6% m/v) were used. The materials were swollen in pracaxi vegetable oil (PO) (Pentaclethra macroloba) and evaluated in terms of surface morphology, wettability, functional groups, thermal behavior, crystallinity, and cytotoxicity. X-ray diffraction (XRD) showed the disappearance of the diffraction peak 2θ = 31.5° for samples from the polycaprolactone/pracaxi/alginate (PCLOA) group, suggesting a reduction of crystallinity according to the presence of PO and semi-crystalline structure. Wettability gradients (0 to 80.91°) were observed according to the deposition layer and hydrogel content. Pore diameters varied between 9.27 μm and 37.57 μm. Molecular interactions with the constituents of the formulation were observed via infrared spectra with Fourier transform (FTIR), and their influence was detected in the reduction of the maximum degradation temperature within the groups of scaffolds (polycaprolactone/alginate (PCLA) and PCLOA) about the control. In vitro tests indicated reduced cell viability in the presence of alginate hydrogel and PO, respectively.

Microfiber-reinforced hydrogels prolong the release of human induced pluripotent stem cell-derived extracellular vesicles to promote endothelial migration

Extracellular vesicle (EV)-based approaches for promoting angiogenesis have shown promising results. Yet, further development is needed in vehicles that prolong EV exposure to target organs. Here, we hypothesized that microfiber-reinforced gelatin methacryloyl (GelMA) hydrogels could serve as sustained delivery platforms for human induced pluripotent stem cell (hiPSC)-derived EV. EV with 50–200 nm size and typical morphology were isolated from hiPSC-conditioned culture media and tested negative for common co-isolated contaminants. hiPSC-EV were then incorporated into GelMA hydrogels with or without a melt electrowritten reinforcing mesh. EV release was found to increase with GelMA concentration, as 12 % (w/v) GelMA hydrogels provided higher release rate and total release over 14 days in vitro, compared to lower hydrogel concentrations. Release profile modelling identified diffusion as a predominant release mechanism based on a Peppas-Sahlin model. To study the effect of reinforcement-dependent hydrogel mechanics on EV release, stress relaxation was assessed. Reinforcement with highly porous microfiber meshes delayed EV release by prolonging hydrogel stress relaxation and reducing the swelling ratio, thus decreasing the initial burst and overall extent of release. After release from photocrosslinked reinforced hydrogels, EV remained internalizable by human umbilical vein endothelial cells (HUVEC) over 14 days, and increased migration was observed in the first 4 h. EV and RNA cargo stability was investigated at physiological temperature in vitro, showing a sharp decrease in total RNA levels, but a stable level of endothelial migration-associated small noncoding RNAs over 14 days. Our data show that hydrogel formulation and microfiber reinforcement are superimposable approaches to modulate EV release from hydrogels, thus depicting fiber-reinforced GelMA hydrogels as tunable hiPSC-EV vehicles for controlled release systems that promote endothelial cell migration.

Protein Folding Stability and Kinetics in Alginate Hydrogels.

Proteins are commonly encapsulated in alginate gels for drug delivery and tissue-engineering applications. However, there is limited knowledge of how encapsulation impacts intrinsic protein properties such as folding stability or unfolding kinetics. Here, we use fast relaxation imaging (FReI) to image protein unfolding in situ in alginate hydrogels after applying a temperature jump. Based on changes in the Förster resonance energy transfer (FRET) response of FRET-labeled phosphoglycerate kinase (PGK), we report the quantitative impact of multiple alginate hydrogel concentrations on protein stability and folding dynamics. The gels stabilize PGK by increasing its melting temperature up to 18.4 °C, and the stabilization follows a nonmonotonic dependence on the alginate density. In situ kinetic measurements also reveal that PGK deviates more from two-state folding behavior in denser gels and that the gel decreases the unfolding rate and accelerates the folding rate of PGK, compared to buffer. Phi-value analysis suggests that the folding transition state of an encapsulated protein is structurally similar to that of folded protein. This work reveals both beneficial and negative impacts of gel encapsulation on protein folding, as well as potential mechanisms contributing to altered stability.

Visible light-mediated cross-linking of injectable gellan gum hydrogels embedding human chondrocytes

Gellan gum, a polysaccharide with structural similarities to glycosaminoglycans, is gaining attention in cartilage tissue engineering. It can be chemically modified with methacrylic moieties to produce a photo-crosslinkable formulation, called methacrylated gellan gum (GGMA), envisaging the injection and the in situ cross-linking through light on the cartilage surface.This study aimed to investigate the visible light cross-linking of GGMA, underlying the impact of steam sterilization on its chemical and rheological properties and cross-linking efficiency. After an initial assessment of the sterilization effects, we evaluated a combination of hydrogel concentrations (1 and 2 % w/v), ruthenium/sodium persulfate concentrations (0.1/1, 0.2/2 and 0.5/5 mM ratios), and light exposure times (30, 60 and, 120 s). The formulations with the lowest sol fraction were characterized for mechanical and showed self-healing properties. Biological characterization on encapsulated chondrocytes in hydrogels showed high cell viability with increased metabolic activity over two weeks. The expression of genes encoding collagen type II and cartilage oligomeric matrix protein was upregulated in the softer formulation (E = 0.95 ± 0.48 kPa) composed of 2 % w/v GGMA, 0.1/1 mM Ru/SPS, and photo-crosslinked for 60 s. These findings represent an interesting overview of the potential application of visible-light crosslinked GGMA in the clinical scenario to treat cartilage defects.

Development and characterization of a poloxamer hydrogel composed of human mesenchymal stromal cells (hMSCs) for reepithelization of skin injuries

Wound healing is a natural physiological reaction to tissue injury. Hydrogels show attractive advantages in wound healing not only due to their biodegradability, biocompatibility and permeability but also because provide an excellent environment for cell migration and proliferation. The main objective of the present study was the design and characterization of a hydrogel loaded with human mesenchymal stromal cells (hMSCs) for use in would healing of superficial skin injures. Poloxamer 407® was used as biocompatible biomaterial to embed hMSCs. The developed hydrogel containing 20 % (w/w) of polymer resulted in the best formulation with respect to physical, mechanical, morphological and biological properties. Its high swelling capacity confirmed the hydrogel’s capacity to absorb wounds’ exudate. LIVE/DEAD® assay confirm that hMSCs remained viable for at least 48 h when loaded into the hydrogels. Adding increasing concentrations of hMSCs-loaded hydrogel to the epithelium did not affect keratinocytes’ viability and healing capacity and all wound area was closed in less than one day. Our study opens opportunities to exploit poloxamer hydrogels as cell carriers for the treatment of skin superficial wound.

The molecular mechanisms of extracellular matrix-derived hydrogel therapy in idiopathic pulmonary fibrosis models

Idiopathic Pulmonary Fibrosis (IPF) is a progressively debilitating lung condition characterized by oxidative stress, cell phenotype shifts, and excessive extracellular matrix (ECM) deposition. Recent studies have shown promising results using decellularized ECM-derived hydrogels produced through pepsin digestion in various lung injury models and even a human clinical trial for myocardial infarction. This study aimed to characterize the composition of ECM-derived hydrogels, assess their potential to prevent fibrosis in bleomycin-induced IPF models, and unravel their underlying molecular mechanisms of action. Porcine lungs were decellularized and pepsin-digested for 48 h. The hydrogel production process, including visualization of protein molecular weight distribution and hydrogel gelation, was characterized. Peptidomics analysis of ECM-derived hydrogel contained peptides from 224 proteins. Probable bioactive and cell-penetrating peptides, including collagen IV, laminin beta 2, and actin alpha 1, were identified. ECM-derived hydrogel treatment was administered as an early intervention to prevent fibrosis advancement in rat models of bleomycin-induced pulmonary fibrosis. ECM-derived hydrogel concentrations of 1 mg/mL and 2 mg/mL showed subtle but noticeable effects on reducing lung inflammation, oxidative damage, and protein markers related to fibrosis (e.g., alpha-smooth muscle actin, collagen I). Moreover, distinct changes were observed in macroscopic appearance, alveolar structure, collagen deposition, and protein expression between lungs that received ECM-derived hydrogel and control fibrotic lungs. Proteomic analyses revealed significant protein and gene expression changes related to cellular processes, pathways, and components involved in tissue remodeling, inflammation, and cytoskeleton regulation. RNA sequencing highlighted differentially expressed genes associated with various cellular processes, such as tissue remodeling, hormone secretion, cell chemotaxis, and cytoskeleton engagement. This study suggests that ECM-derived hydrogel treatment influence pathways associated with tissue repair, inflammation regulation, cytoskeleton reorganization, and cellular response to injury, potentially offering therapeutic benefits in preventing or mitigating lung fibrosis.

A mist-based crosslinking technique for coaxial bioprinting of hollow hydrogel fibers

In this paper, a mist-based method for coaxial three-dimensional bioprinting of ionically crosslinking hydrogel hollow fibers is presented. Unlike current techniques of coaxial bioprinting that utilize the crosslinker in liquid or sacrificial form, the developed method introduces the core crosslinking agent in mist form. The use of mist as a core flow provides adequate pressure and sufficient crosslinking to maintain the tubular shape of a hollow fiber. Through controlled exposure of crosslinker, the developed system prevents poor resolution and layer adhesion caused by the accumulation of liquid crosslinker on the printbed. Furthermore, it eliminates additional processing steps, such as partial crosslinking of the hydrogel prior- or removal of sacrificial material post-printing. The printability and mechanical properties of hollow fiber scaffolds printed using various mist and hydrogel concentrations are studied. It is shown that mist concentration influences the gelation rate of the hollow fiber, impacting the shape fidelity, layer adhesion, and mechanical properties of the printed structures. Moreover, the effects of printing parameters, including the mist core pressure and hydrogel flowrate, on the diameter and wall thickness of the hollow fiber are investigated. Finally, scaffolds printed and crosslinked using mist exhibit over 90% cell viability. The developed mist-based coaxial system enables direct printing of continuous hollow fibers.