Gelatin Methacryloyl Hydrogel Research Articles

Synthesis and Modification of Gelatin Methacryloyl (GelMA) with Antibacterial Quaternary Groups and Its Potential for Periodontal Applications.

Gelatin methacryloyl (GelMA) hydrogels have been widely used for different biomedical applications due to their tunable physical characteristics and appropriate biological properties. In addition, GelMA could be modified with the addition of functional groups providing inherent antibacterial capabilities. Here, GelMA-based hydrogels were developed through the combination of a GelMA unmodified and modified polymer with quaternary ammonium groups (GelMAQ). The GelMAQ was synthesized from GelMA with a low degree of substitution of methacrylamide groups (DSMA) and grafted with glycidyltrimethylammonium chloride in the free amine groups of the lysine moieties present in the original gelatin. GelMAs with high DSMA and GelMAQ were combined 50/50% or 25/75% (w/w), respectively, and compared to controls GelMA and GelMA with added chlorhexidine (CHX) at 0.2%. The different hydrogels were characterized using 1H-NMR spectroscopy and swelling behavior and tested in (1) Porphyromonas gingivalis to evaluate their antibacterial properties and (2) human gingival fibroblast to evaluate their cell biocompatibility and regenerative properties. GelMA/GelMAQ 25/75% showed good antibacterial properties but also excellent biocompatibility and regenerative properties toward human fibroblasts in the wound healing assay. Taken together, these results suggest that the modification of GelMA with quaternary groups could facilitate periodontal tissue regeneration, with good biocompatibility and added antibacterial properties.

Epidermal growth factor-loaded microspheres/hydrogel composite for instant hemostasis and liver regeneration

Rapid hemostasis and effective healing for the non-compressible liver wounds which are not able to be sewn, especially for those large-area wounds, remain great clinical challenges. In this study, we fabricated epidermal growth factor (EGF)-loaded chitosan microspheres (CM) and then incorporated them into a photo-crosslinking gelatin methacryloyl (GelMA) hydrogel. The results showed that the EGF-loaded CM/GelMA precursor solution could transform into a hydrogel and cease bleeding at laceration sites without external stress. Subsequently, the sustained release of EGF accelerated wound closure and promoted liver regeneration. The in vitro experiments demonstrated that the microsphere/hydrogel composite could promote the proliferation and migration of L02 ​cells. Moreover, the histological and immunohistological analyses indicated that EGF-CM/GelMA composite could alleviate inflammation in the mouse liver and promote liver remodeling. Overall, this multi-functional microsphere/hydrogel composite will inspire the development of clinical applications for noncompressible hemostasis and successive wound closure.

Bioprinting of 3D Adipose Tissue Models Using a GelMA-Bioink with Human Mature Adipocytes or Human Adipose-Derived Stem Cells.

Adipose tissue is related to the development and manifestation of multiple diseases, demonstrating the importance of suitable in vitro models for research purposes. In this study, adipose tissue lobuli were explanted, cultured, and used as an adipose tissue control to evaluate in vitro generated adipose tissue models. During culture, lobule exhibited a stable weight, lactate dehydrogenase, and glycerol release over 15 days. For building up in vitro adipose tissue models, we adapted the biomaterial gelatin methacryloyl (GelMA) composition and handling to homogeneously mix and bioprint human primary mature adipocytes (MA) and adipose-derived stem cells (ASCs), respectively. Accelerated cooling of the bioink turned out to be essential for the homogeneous distribution of lipid-filled MAs in the hydrogel. Last, we compared manual and bioprinted GelMA hydrogels with MA or ASCs and the explanted lobules to evaluate the impact of the printing process and rate the models concerning the physiological reference. The viability analyses demonstrated no significant difference between the groups due to additive manufacturing. The staining of intracellular lipids and perilipin A suggest that GelMA is well suited for ASCs and MA. Therefore, we successfully constructed physiological in vitro models by bioprinting MA-containing GelMA bioinks.

Superwettable and injectable GelMA-MSC microspheres promote cartilage repair in temporomandibular joints.

Temporomandibular disorders (TMD) can be treated by promoting cartilage regeneration with biomaterials. However, there are deficiencies in the infiltration function of bone filler biological materials. In this study, stems cells were loaded onto gelatin methacryloyl (GelMA) hydrogel microspheres endowed with superwettable properties and TGF-β sustained-release function, which can quickly infiltrate the irregular surface of the temporomandibular joint (TMJ) bone defect area and accelerate cartilage healing. First, to improve cell adhesion and spreading function, the BMSCs-coated GelMA microspheres were endowed with superwetting property. At the same time, the swelling adsorption characteristics of gelatin microspheres could be used to load recombinant TGF-β within the microspheres, which could in turn promote the chondrogenic differentiation of multi-potent bone marrow mesenchymal stem cells. The SEM imaging demonstrated that BMSCs-coated GelMA microsphere has superwettable and superhydrophilic property, which enabled rapid adaptation to the bone defect surface morphology, which is conducive to tissue repair. Furthermore, the cartilage defect model showed that rBMSCs-coated GelMA microspheres promote temporomandibular joint arthritis repair. In conclusion, our study established that BMSC-coated GelMA microspheres endowed with superwetting properties, can colonize the bone defect repair site better with sustained release of growth factors, thus providing an innovative strategy for promoting cartilage regeneration.

Gelatin-methacryloyl hydrogels containing turnip mosaic virus for fabrication of nanostructured materials for tissue engineering.

Current tissue engineering techniques frequently rely on hydrogels to support cell growth, as these materials strongly mimic the extracellular matrix. However, hydrogels often need ad hoc customization to generate specific tissue constructs. One popular strategy for hydrogel functionalization is to add nanoparticles to them. Here, we present a plant viral nanoparticle the turnip mosaic virus (TuMV), as a promising additive for gelatin methacryloyl (GelMA) hydrogels for the engineering of mammalian tissues. TuMV is a flexuous, elongated, tubular protein nanoparticle (700–750 nm long and 12–15 nm wide) and is incapable of infecting mammalian cells. These flexuous nanoparticles spontaneously form entangled nanomeshes in aqueous environments, and we hypothesized that this nanomesh structure could serve as a nanoscaffold for cells. Human fibroblasts loaded into GelMA-TuMV hydrogels exhibited similar metabolic activity to that of cells loaded in pristine GelMA hydrogels. However, cells cultured in GelMA-TuMV formed clusters and assumed an elongated morphology in contrast to the homogeneous and confluent cultures seen on GelMA surfaces, suggesting that the nanoscaffold material per se did not favor cell adhesion. We also covalently conjugated TuMV particles with epidermal growth factor (EGF) using a straightforward reaction scheme based on a Staudinger reaction. BJ cells cultured on the functionalized scaffolds increased their confluency by approximately 30% compared to growth with unconjugated EGF. We also provide examples of the use of GelMA-TuMV hydrogels in different biofabrication scenarios, include casting, flow-based-manufacture of filaments, and bioprinting. We envision TuMV as a versatile nanobiomaterial that can be useful for tissue engineering.

Ellipsoidal porous patch with anisotropic cell inducing ability for inhibiting skin scar formation

• An ellipsoidal porous patch is fabricated for inhibiting skin scar formation. • The anisotropic surface microstructure of patch showed dramatic capability in guiding cell growth and arrangement by reconstructing cell morphology. • The proliferation of fibroblasts growing on the stretched inverse opal patch was significantly suppressed without cell cytotoxicity. • The patch effectively reduce the scar formation by inhibiting excessive fibroblast proliferation of and type-I collagen deposition. Scar formation has always been a difficult point to overcome in the field of clinical wound care. Here, we present an ellipsoidal porous patch with cell inducing ability for inhibiting scar formation. The patch was prepared by stretching a poly (lactic-co-glycolic acid) (PLGA) inverse opal film at the glass transition temperature to form a neatly arranged three-dimensional ellipsoidal porous structure. Such anisotropic structure showed dramatic capability in directing cell growth and arrangement by reconstructing cell morphology. Besides, the proliferation of cells growing on the stretched patch was significantly suppressed without cell cytotoxicity. In addition, benefitting from the abundant and connected nanopores, the patch could be imparted with a potent ability to promote cell migration by encapsulating fibroblast growth factor 2 (FGF2) via the second filling of functional gelatin methacryloyl (GelMA) hydrogel into its scaffold. In a typical scar model, we have demonstrated that the resultant patch performed well in inhibiting scar formation characterized by inhibiting the excessive proliferation of fibroblasts, decreasing the deposition of type I collagen, reducing the scar index and achieved complete tissue reconstruction. These results indicate the anisotropic inverse opal patch has an excellent application prospect in inhibiting scar formation during wound repair.

Investigating Autoregressive and Machine Learning-based Time Series Modeling with Dielectric Spectroscopy for Predicting Quality of Biofabricated Constructs

Advances in biofabrication processes need to be complemented with appropriate nondestructive quality engineering techniques that can be integrated into scalable engineered tissue manufacturing systems. Previous studies have demonstrated the feasibility of dielectric spectroscopy (DS) as a inline, real time biological quality monitoring alternative. Time series modeling can help improve the efficiency and accuracy of quality prediction by analyzing trends in DS data as the biofabricated constructs mature over time. These models can help forecast potential future deviations in quality attributes and provide opportunities to take preemptive, corrective actions, leading to better yields and higher quality of final products. In this study, we investigated time series modeling of DS data to characterize the effects of two critical biofabrication parameters on constructs of gelatin methacryloyl (GelMA) hydrogel containing human adipose-derived stem cells (hASC) over 11 days of in vitro culture. The performance of standard autoregressive time series models (Exponential Smoothing, ARMA, ARIMA, SARIMA) and conventional sequence-based machine learning (ML) models (SVM, ANN, CNN and LSTM) were analyzed to forecast trends in Δɛ, a key DS metric that directly correlates to the volume of viable cells in constructs. The ML-based time series models, in general, showed superior performance in predicting future trends in Δɛ, with LSTM providing the lowest least mean square errors (MSE) in Δɛ forecasts. The outcomes of this study highlight the benefits of using DS and time series modeling synergistically for efficient quality monitoring in biofabrication.

Galunisertib-Loaded Gelatin Methacryloyl Hydrogel Microneedle Patch for Cardiac Repair after Myocardial Infarction.

Uncontrolled and excessive fibrosis after myocardial infarction (MI) in the peri-infarct zone leads to left ventricular remodeling and deterioration of cardiac function. Inhibiting fibroblast activation during the mature phase of cardiac repair improves cardiac remodeling and function after MI. Here, we engineered a biocompatible microneedle (MN) patch using gelatin methacryloyl and loaded it with galunisertib, a transforming growth factor-beta (TGF-β)-specific inhibitor, to treat excessive cardiac fibrosis after MI. The MN patch could sustainably release galunisertib for more than 2 weeks and provide mechanical support for the fragile ventricular wall. After being applied to a rat model of MI, the galunisertib-loaded MN patch improved long-term cardiac function and reduced cardiac fibrosis by effectively inhibiting TGF-β depending on fibroblast activation. This strategy shows the potential of the MN patch as an advanced platform to locally deliver direct antifibrotic drugs to prevent myocardial fibrosis for the treatment of MI and the promotion of cardiac repair.

Study on properties of 3D-printed GelMA hydrogel scaffolds with different nHA contents

Biological 3D printing is a reliable technology for 3D printing bone repair scaffolds with simple operation, high efficiency, and relatively low cost. Gelatin methacryloyl (GelMA) hydrogels have attracted much attention due to their good biocompatibility, but the poor mechanical properties limit their application in bone reconstruction engineering. In this study, nano-hydroxyapatite (nHA) particles were added to GelMA hydrogels, and the performances of composite hydrogel scaffolds with different nHA contents were investigated in terms of rheological properties, light transmission properties, surface morphology, mechanical properties, and biocompatibility. The experimental results showed that the incorporation of nHA particles could effectively improve the printability and mechanical properties of the scaffolds, the scaffold fibers had better resistance to deformation, improved degradation rate, and biological experiments confirmed that nHA particles had no significant cytotoxicity. However, the addition of HA particles also reduced the light transmission properties of the slurry, and when its content exceeds a certain value, the hydrogel scaffolds show incomplete curing and eventually affect their test performance. The results can offer guidance and reference for the selection of ink and function for 3D printing bone repair scaffold.

Mechanosensation mediates volume adaptation of cardiac cells and spheroids in 3D.

With the adoption of 3-dimensional (3D) cell culture for in vitro modelling of cardiac function and regenerative medicine applications, there is an increased need to understand cardiomyocyte mechanosensation in 3D. With existing studies of cardiomyocyte mechanosensation primarily focussed on the behaviour of individual cells in a 2-Dimensional context, it is unclear whether mechanosensation is the same in a 3D, multicellular context. In this study, H9C2 cardiac-derived myoblasts were encapsulated as individual cells and as cell spheroids within stiffness gradient gelatin methacryloyl (GelMA) hydrogels to investigate individual and collective cardiac cell mechanosensation in 3D. Over a 3.68–17.52 ​kPa stiffness range, it was found that H9C2 cells have a limited capacity to adapt their volume to increasing substrate stiffness, demonstrated by the lack of changes in cell volume and shape across the stiffness gradient. Morphological trends were reflected by the expression of the mechanomarkers YAP, MRTF-A and Lamin-A, which were better correlated with cell and nuclear volume than with substrate stiffness. The localisation of YAP and MRTF-A were dependent on the relative volumes of the cytoplasm and nucleus while Lamin-A expression was elevated with increasing cytoplasmic and nuclear volumes. When cultured as spheroids rather than as individual cells, H9C2 cells adopted a distinct morphology with comparably smaller nuclei than individually cultured cells, while retaining the same overall cell volume. As spheroids, H9C2 cells were sensitive to stiffness cues, shown by decreasing YAP and MRTF-A nuclear localisation, increasing Lamin-A expression, and increasing vinculin expression with increasing substrate stiffness. Like the individually cultured H9C2 cells, mechanomarker expression was correlated to volume adaptation. With increasing cytoplasmic volume, YAP and MRTF-A became less nuclear localised, vinculin expression was increased, and with increasing nuclear volume, the Lamin-A expression fincreased. Together, these data suggest that cardiac cell volume adaptation may be enhanced by cell-cell interactions.

Curcumin loaded hydrogel with anti-inflammatory activity to promote cartilage regeneration in immunocompetent animals

Stable cartilage regeneration in immunocompetent animals remains a huge challenge, mainly ascribing to the in vivo implantation of tissue-engineered cartilage inevitably arousing inflammatory reactions, resulting in cartilage-specific extracellular matrix erosion, chondrogenic niche destruction, and chondrocyte deterioration. Herein, we developed an anti-inflammatory platform, namely, Cur/GelMA hydrogel, by loading a potent anti-inflammatory drug of curcumin (Cur) into gelatin methacryloyl (GelMA) hydrogel. The Cur/GelMA hydrogel exhibited satisfactory Cur release kinetics in vitro and exerted favorable anti-inflammatory effects when cocultured with lipopolysaccharide-induced RAW264.7 macrophages in vitro. Furthermore, the Cur/GelMA hydrogel showed gratifying biocompatibility and supported cartilage regeneration in vitro when colonized with rabbit- and goat-derived chondrocytes. In addition, the in vitro engineered cartilages in the Cur/GelMA hydrogel were able to maintain a cartilaginous phenotype and achieved stable cartilage regeneration when subcutaneously implanted in autologous rabbits and goats for 2 and 4 weeks compared to the GelMA hydrogel. Furthermore, our data revealed that the in vivo-generated cartilage in the Cur/GelMA group apparently alleviated the inflammatory reaction compared to its GelMA counterpart, suggesting that the locally released Cur endowed the Cur/GelMA hydrogel with potent anti-inflammatory capacity. This study provides a reliable anti-inflammatory platform for stable cartilage regeneration in immunocompetent animals, significantly advancing the clinical application of tissue-engineered cartilage.

Matrix Metalloproteases from Adipose Tissue-Derived Stromal Cells Are Spatiotemporally Regulated by Hydrogel Mechanics in a 3D Microenvironment.

Adipose tissue-derived stromal cells (ASCs) are of interest in tissue engineering and regenerative medicine (TERM) due to their easy acquisition, multipotency, and secretion of a host of factors that promote regeneration. Retention of ASCs in or around lesions is poor following direct administration. Therefore, for TERM applications, ASCs can be ‘immobilized’ via their incorporation into hydrogels such as gelatine methacryloyl (GelMA). Tweaking GelMA concentration is a common approach to approximate the mechanical properties found in organs or tissues that need repair. Distinct hydrogel mechanics influence the ability of a cell to spread, migrate, proliferate, and secrete trophic factors. Mesenchymal cells such as ASCs are potent remodellers of the extracellular matrix (ECM). Not only do ASCs deposit components, they also secrete matrix metalloproteases (MMPs) which degrade ECM. In this work, we investigated if GelMA polymer concentration influenced the expression of active MMPs by ASCs. In addition, MMPs’ presence was interrogated with regard to ASCs morphology and changes in hydrogel ultrastructure. For this, immortalised ASCs were embedded in 5%, 10%, and 15% (w/v) GelMA hydrogels, photopolymerised and cultured for 14 d. Zymography in situ indicated that MMPs had a variable, hydrogel concentration-dependent influence on ASCs-secreted MMPs. In 5% GelMA, ASCs showed a high and sustained expression of MMPs, while, in 10% and 15% GelMA, such expression was almost null. ASCs morphology based on F-actin staining showed that increasing GelMA concentrations inhibit their spreading. Scanning electron microscopy (SEM) showed that hydrogel ultrastructure in terms of pore density, pore size, and percentage porosity were not consistently influenced by cells. Interestingly, changes in ultrastructural parameters were detected also in cell-free materials, albeit without a clear trend. We conclude that hydrogel concentration and its underlying mechanics influenced MMP expression by ASCs. The exact MMPs that respond to these mechanical cues should be defined in follow-up experiments.

Osteoimmunity-Regulating Biomimetically Hierarchical Scaffold for Augmented Bone Regeneration.

Engineering a proper immune response following biomaterial implantation is essential to bone tissue regeneration. Herein, a biomimetically hierarchical scaffold composed of deferoxamine@poly(ε-caprolactone) nanoparticles (DFO@PCL NPs), manganese carbonyl (MnCO) nanosheets, gelatin methacryloyl hydrogel, and a polylactide/hydroxyapatite (HA) matrix is fabricated to augment bone repair by facilitating the balance of the immune system and bone metabolism. First, a 3D printed stiff scaffold with a well-organized gradient structure mimics the cortical and cancellous bone tissues; meanwhile, an inside infusion of a soft hydrogel further endows the scaffold with characteristics of the extracellular matrix. A Fenton-like reaction between MnCO and endogenous hydrogen peroxide generated at the implant-tissue site triggers continuous release of carbon monoxide and Mn2+ , thus significantly lessening inflammatory response by upregulating the M2 phenotype of macrophages, which also secretes vascular endothelial growth factor to induce vascular formation. Through activating the hypoxia-inducible factor-1α pathway, Mn2+ and DFO@PCL NP further promote angiogenesis. Moreover, DFO inhibits osteoclast differentiation and synergistically collaborates with the osteoinductive activity of HA. Based on amounts of data in vitro and in vivo, strong immunomodulatory, intensive angiogenic, weak osteoclastogenic, and superior osteogenic abilities of such an osteoimmunity-regulating scaffold present a profound effect on improving bone regeneration, which puts forward a worthy base and positive enlightenment for large-scale bone defect repair.

Gelatin Methacryloyl Hydrogels for Musculoskeletal Tissue Regeneration.

Musculoskeletal disorders are a significant burden on the global economy and public health. Hydrogels have significant potential for enhancing the repair of damaged and injured musculoskeletal tissues as cell or drug delivery systems. Hydrogels have unique physicochemical properties which make them promising platforms for controlling cell functions. Gelatin methacryloyl (GelMA) hydrogel in particular has been extensively investigated as a promising biomaterial due to its tuneable and beneficial properties and has been widely used in different biomedical applications. In this review, a detailed overview of GelMA synthesis, hydrogel design and applications in regenerative medicine is provided. After summarising recent progress in hydrogels more broadly, we highlight recent advances of GelMA hydrogels in the emerging fields of musculoskeletal drug delivery, involving therapeutic drugs (e.g., growth factors, antimicrobial molecules, immunomodulatory drugs and cells), delivery approaches (e.g., single-, dual-release system), and material design (e.g., addition of organic or inorganic materials, 3D printing). The review concludes with future perspectives and associated challenges for developing local drug delivery for musculoskeletal applications.

In vitro and in vivo evaluation of 3D constructs engineered with human iPSC-derived chondrocytes in gelatin methacryloyl hydrogel.

Articular cartilage defects have limited healing potential and, when left untreated, can lead to osteoarthritis. Tissue engineering focuses on regenerating the damaged joint surface, preferably in an early stage. Here, we investigate the regenerative potential of three-dimensional (3D) constructs consisting of human induced pluripotent stem cell (iPSC)-derived chondrocytes in gelatin methacryloyl (GelMA) hydrogel for stable hyaline cartilage production. iPSC-derived chondrocytes are encapsulated in GelMA hydrogel at low (1 × 107 ml-1 ) and high (2 × 107 ml-1 ) density. In a conventional medium, GelMA hydrogel supports the chondrocyte phenotype, as opposed to cells cultured in 3D in absence of hydrogel. Moreover, encapsulated iPSC-derived chondrocytes preserve their in vivo matrix formation capacity after 21 days in vitro. In differentiation medium, hyaline cartilage-like tissue forms after 21 days, demonstrated by highly sulfated glycosaminoglycans and collagen type II. Matrix deposition is delayed at low encapsulation density, corroborating with lower transcript levels of COL2A1. An ectopic assay in nude mice demonstrates further maturation of the matrix deposited in vitro. Direct ectopic implantation of iPSC-derived chondrocyte-laden GelMA, without in vitro priming, also generates hyaline cartilage-like tissue, albeit less mature. Since it is unclear what maturity upon implantation is desired for joint surface regeneration, this is an attractive technology to generate immature and more mature hyaline cartilage-like tissue.

Visible-Light Stiffness Patterning of GelMA Hydrogels Towards In Vitro Scar Tissue Models.

Variations in mechanical properties of the extracellular matrix occurs in various processes, such as tissue fibrosis. The impact of changes in tissue stiffness on cell behaviour are studied in vitro using various types of biomaterials and methods. Stiffness patterning of hydrogel scaffolds, through the use of stiffness gradients for instance, allows the modelling and studying of cellular responses to fibrotic mechanisms. Gelatine methacryloyl (GelMA) has been used extensively in tissue engineering for its inherent biocompatibility and the ability to precisely tune its mechanical properties. Visible light is now increasingly employed for crosslinking GelMA hydrogels as it enables improved cell survival when performing cell encapsulation. We report here, the photopatterning of mechanical properties of GelMA hydrogels with visible light and eosin Y as the photoinitiator using physical photomasks and projection with a digital micromirror device. Using both methods, binary hydrogels with areas of different stiffnesses and hydrogels with stiffness gradients were fabricated. Their mechanical properties were characterised using force indentation with atomic force microscopy, which showed the efficiency of both methods to spatially pattern the elastic modulus of GelMA according to the photomask or the projected pattern. Crosslinking through projection was also used to build constructs with complex shapes. Overall, this work shows the feasibility of patterning the stiffness of GelMA scaffolds, in the range from healthy to pathological stiffness, with visible light. Consequently, this method could be used to build in vitro models of healthy and fibrotic tissue and study the cellular behaviours involved at the interface between the two.

Free radical-scavenging composite gelatin methacryloyl hydrogels for cell encapsulation

Gelatin methacryloyl (GelMA) hydrogels have been widely used for cell encapsulation in tissue engineering due to their cell adhesiveness and biocompatibility. However, free radicals generated during gelation decrease the viability of the encapsulated cells by increasing intracellular oxidative stress, so appropriate strategies for scavenging free radicals need to be developed. To meet that need, we developed composite GelMA hydrogels incorporating nanofiber particles (EF) coated with epigallocatechin-gallate (EGCG). The GelMA composite hydrogels were successfully fabricated and had a storage modulus of about 5 kPa, which is similar to that of pristine GelMA hydrogel, and the drastic free radical scavenging activity of EGCG was highly preserved after gelation. In addition, human adipose-derived stem cells encapsulated within our composite hydrogels had better viability (about 1.5 times) and decreased intracellular oxidative stress (about 0.3 times) compared with cells within the pristine GelMA hydrogel. We obtained similar results with human dermal fibroblasts and human umbilical vein endothelial cells, indicating that our composite hydrogels are suitable for various cell types. Furthermore, we found that the ability of the encapsulated cells to spread and migrate increased by 5 times within the composite hydrogels. Collectively, our results demonstrate that incorporating EF into GelMA hydrogels is a promising way to enhance cell viability by reducing free-radical-derived cellular damage when fabricating 3D tissue ex vivo. Statement of significanceGelatin methacryloyl (GelMA) hydrogels have been widely applied to various tissue engineering applications because of their biocompatibility and cell interactivity. However, free radicals generated during the GelMA hydrogel fabrication decrease the viability of encapsulated cells by elevating intracellular oxidative stress. Here, we demonstrate radical scavenging GelMA hydrogels incorporating epigallocatechin-gallate (EGCG)-coated nanofiber particles (EF). The composite GelMA hydrogels are successfully fabricated, maintaining their mechanical properties, and the viability of encapsulated human adipose-derived stem cells is greatly improved after the gelation, indicating that our composite GelMA hydrogel alleviates damages from free radicals. Collectively, the incorporation of EF within GelMA hydrogels may be a promising way to enhance the viability of encapsulated cells, which could be applied to 3D tissue fabrication.

Substrate-Independent, Mechanically Tunable, and Scalable Gelatin Methacryloyl Hydrogel Coating with Drag-Reducing and Anti-Freezing Properties

Substrate-Independent, Mechanically Tunable, and Scalable Gelatin Methacryloyl Hydrogel Coating with Drag-Reducing and Anti-Freezing Properties

3D Printing of Diatomite Incorporated Composite Scaffolds for Skin Repair of Deep Burn Wounds.

Deep burn injury always causes severe damage of vascular network and collagen matrix followed by delayed wound healing process. In this study, natural diatomite (DE) microparticles with porous nanostructure were separated based on the particles size through a dry sieving method and combined with gelatin methacryloyl (GelMA) hydrogel to form a bioactive composite ink. The DE-containing inorganic/organic composite scaffolds, which were successfully prepared through three-dimensional (3D) printing technology, were used as functional burn wound dressings. The scaffolds incorporated with DE are of great benefit to several cellular activities, including cell spreading, proliferation, and angiogenesis-related gene expression in vitro, which can mainly be attributed to the positive effect of bioactive silicon (Si) ions released from the embedded DE. Moreover, due to establishment of bioactive ionic environment, the deep burn wounds treated with 3D-printed DE incorporated scaffolds exhibited rapid wound healing rate, enhanced collagen deposition, and dense blood vessel formation in vivo. Therefore, the present study demonstrates that the cost-effective DE can be used as biocompatible Si source to significantly promote the bioactivities of wound dressings for effective tissue regeneration.

Standardised quantitative ultrasound imaging approach for the contact-less three-dimensional analysis of neocartilage formation in hydrogel-based bioscaffolds

In this work we present a standardised quantitative ultrasound imaging (SQUI) approach for the non-destructive three-dimensional imaging and quantification of cartilage formation in hydrogel based bioscaffolds. The standardised concept involves the processing of ultrasound backscatter data with respect to an acellular phantom in combination with the compensation of sound speed mismatch diffraction effects between the bioscaffold and the phantom. As a proof-of-concept, the SQUI approach was tested on a variety of bioscaffolds with varying degree of neocartilage formation. These were composed of Gelatine Methacryloyl (GelMA) hydrogels laden with human adipose-derived stem cells (hADSCs). These were cultured under chondrogenic stimulation following a previously established protocol, where the degree of the neocartilage formation was modulated using different GelMA network densities (6, 8, 10 % w/v) and culture time (0, 14, 28 days). Using the SQUI approach we were able to detect marked acoustic and morphological changes occurring in the bioscaffolds a result of their different chondrogenic outcome. We defined an acoustic neocartilage indicator, the sonomarker, for the selective imaging and quantification of neocartilage formation. The sonomarker, of backscatter intensity logIBC -2.4, was found to correlate with data obtained via standard destructive bioassays. The ultrasonic evaluation of human specimens confirmed the sonomarker as a relevant intensity, although it was found to shift to higher intensity values in proportion to the cartilage condition as inferred from sound speed measurements. This study demonstrates the potential of the SQUI approach for the realization of non-destructive analysis of cartilage regeneration over-time. Statement of significanceAs tissue engineering strategies for neocartilage regeneration evolve towards clinical implementation, alternative characterisation approaches that allow the non-destructive monitoring of extracellular matrix formation in implantable hydrogel based bioscaffolds are needed. In this work we present an innovative standardized quantitative ultrasound imaging (SQUI) approach that allows the non-destructive, volumetric, and quantitative evaluation of neocartilage formation in hydrogel based bioscaffolds. The standardised concept aims to provide a robust approach that accounts for the dynamic changes occurring during the conversion from a cellular bioscaffold towards the formation of a neocartilage construct. We believe that the SQUI approach will be of great benefit for the evaluation of constructs developing neocartilage, not only for in-vitro applications but also potentially applicable to in-vivo applications.