Gelatin-based Hydrogels Research Articles

Growth factor free, peptide-functionalized gelatin hydrogel promotes arteriogenesis and attenuates tissue damage in a murine model of critical limb ischemia

Critical limb ischemia (CLI) occurs when blood flow is restricted through the arteries, resulting in ulcers, necrosis, and chronic wounds in the downstream extremities. The development of collateral arterioles (i.e. arteriogenesis), either by remodeling of pre-existing vascular networks or de novo growth of new vessels, can prevent or reverse ischemic damage, but it remains challenging to stimulate collateral arteriole development in a therapeutic context. Here, we show that a gelatin-based hydrogel, devoid of growth factors or encapsulated cells, promotes arteriogenesis and attenuates tissue damage in a murine CLI model. The gelatin hydrogel is functionalized with a peptide derived from the extracellular epitope of Type 1 cadherins. Mechanistically, these “GelCad” hydrogels promote arteriogenesis by recruiting smooth muscle cells to vessel structures in both ex vivo and in vivo assays. In a murine femoral artery ligation model of CLI, delivery of in situ crosslinking GelCad hydrogels was sufficient to restore limb perfusion and maintain tissue health for 14 days, whereas mice treated with gelatin hydrogels had extensive necrosis and autoamputated within 7 days. A small cohort of mice receiving the GelCad hydrogels were aged out to 5 months and exhibited no decline in tissue quality, indicating durability of the collateral arteriole networks. Overall, given the simplicity and off-the-shelf format of the GelCad hydrogel platform, we suggest it could have utility for CLI treatment and potentially other indications that would benefit from arteriole development.

Adhesivity-tuned bioactive gelatin/gellan hybrid gels drive efficient wound healing

Gelatin-based hydrogels have been widely used for wound healing applications. However, increase in ligand density and reduction in pore size with increasing gelatin concentration may delay wound healing by limiting cell infiltration. In this study, we address this shortcoming by combining gelatin with gellan—which is super hydrophilic and non-adhesive to cells. We show that UV crosslinked hybrid gels composed of methacrylated gelatin (GelMA) and methacrylated gellan gum (mGG), possess considerably larger pores and improved mechanical properties compared to GelMA gels. Reduced spreading and reduced formation of focal adhesions on hybrid gels combined with lower contractility and faster detachment upon trypsin-induced de-adhesion suggests that hybrid gels are less adhesive than GelMA gels. Gradual release of fibroblast growth factor (FGF) and silver nanoparticles (AgNPs) incorporated in hybrid gels not only boosts cell migration, but also confers anti-bacterial activity against gram-positive and gram-negative bacteria at concentrations nontoxic to cells. Full thickness wound healing in Wistar rats revealed increased granulation tissue formation in hybrid gels, fastest epithelialization and highest collagen deposition in rats treated with FGF entrapped hybrid gels. Together, our results demonstrate how adhesive tuning and incorporation of bioactive factors can be synergistically combined for achieving complete wound healing.

A Gelatin-Based Hydrogel Electrode With High Moisturizing Ability for Wearable EEG Recording

A Gelatin-Based Hydrogel Electrode With High Moisturizing Ability for Wearable EEG Recording

A systematic investigation of the effects of TGF-β3 and mechanical stimulation on tenogenic differentiation of mesenchymal stromal cells in a poly(ethylene glycol)/gelatin-based hydrogel

BackgroundHigh post-surgical failure rates following tendon injury generate high medical costs and poor patient recovery. Cell-based tendon tissue engineering has the potential to produce fully functional replacement tissue and provide new strategies to restore tendon function and healing. In this endeavour, the application of mesenchymal stromal cells (MSCs) encapsulated in biomaterial scaffolds has shown great promise. However, a consensus on optimal promotion of tenogenic differentiation of MSCs has yet to be reached, although growth factors and mechanical cues are generally acknowledged as important factors. MethodsIn this study, we prepared a hydrogel cell culture system consisting of methacrylated poly(d,l-lactic acid-ethylene glycol-d,l-lactic acid) (P(LA-EG-LA)) and gelatin methacrylate (GelMA) to encapsulate human bone marrow-derived MSCs (hBMSCs). We further systematically investigated the influence of static and intermittent cyclic uniaxial strain mechanical stimulation, in combination with transforming growth factor-β3 (TGF-β3) supplementation, on tenogenic differentiation of hBMSCs. ResultsIncreased TGF-β3 concentration upregulated the tenogenic genes Scleraxis (SCX) and collagen type I (COL1A1) but showed no effects on tenascin-c (TNC) and collagen type III (COL3A1) expression. Mechanical stimulation had no observable effect on gene expression, but intermittent cyclic uniaxial strain stimulation improved matrix deposition. Together, these data provide new insights into how TGF-β3 and mechanical stimulation regulate MSC tenogenesis, with TGF-β3 promoting the expression of key tenogenic genes whilst mechanical stimulation aided matrix deposition in the engineered tissue. Furthermore, intermittent cyclic uniaxial strain at 3% elongation and 0.33 ​Hz for 1 ​h/day showed improved matrix effects compared to static strain. ConclusionTogether, the most promising result for tenogenic differentiation of hBMSCs was identified as treatment with 5 ​ng/ml TGF-β3 under intermittent cyclic uniaxial strain (3% strain; 0.33 ​Hz; 1 ​h/day). This knowledge is of importance for the development of an improved protocol for tenogenic differentiation of MSCs and thereby for tendon tissue engineering. The translational potential of this articleTissue-engineered strategies for tendon repair require a consensus on the differentiation of mesenchymal stromal cells to tenocytes, which is currently lacking. This article provides a systematic investigation of two main tenogenic differentiation conditions to further development of a tenogenic differentiation protocol.

Engineering Stem Cell Fate Controlling Biomaterials to Develop Muscle Connective Tissue Layered Myofibers.

Skeletal muscle connective tissue (MCT) surrounds myofiber bundles to provide structural support, produce force transduction from tendons, and regulate satellite cell differentiation during muscle regeneration. Engineered muscle tissue composed of myofibers layered within MCT has not yet been developed. Herein, a bioengineering strategy to create MCT-layered myofibers through the development of stem cell fate-controlling biomaterials that achieve both myogenesis and fibroblast differentiation in a locally controlled manner at the single construct is introduced. The reciprocal role of transforming growth factor-beta 1 (TGF-β1) and its inhibitor as well as 3D matrix stiffness to achieve co-differentiation of MCT fibroblasts and myofibers from a human-induced pluripotent stem cell (hiPSC)-derived paraxial mesoderm is studied. To avoid myogenic inhibition, TGF-β1 is conjugated on the gelatin-based hydrogel to control the fibroblasts' populations locally; the TGF-β1 degrades after 2 weeks, resulting in increased MCT-specific extracellular matrix (ECM) production. The locations of myofibers and fibroblasts are precisely controlled by using photolithography and co-axial wet spinning techniques, which results in the formation of MCT-layered functional myofibers in 3D constructs. This advanced engineering strategy is envisioned as a possible method for obtaining biomimetic human muscle grafts for various biomedical applications.

Increasing hydrogel complexity from 2D towards 3D towards intestinal tissue engineering

There is a high demand for in vitro intestinal models towards studying intestinal disfunctions, such as gastrointestinal diseases (Crohn's disease, irritable bowel syndrome, colorectal cancer, etc.) which are commonly diagnosed in modern society. In parallel, in vitro models are utilized for the evaluation of novel food supplements and pharmaceuticals in the intestinal micro-environment. Gelatin-based hydrogels can offer appropriate mechanical cues (down to G’ < 10 kPa) and cell interactivity to obtain physiologically relevant soft tissue models. Therefore, the current paper focused on the development of a novel, gelatin-methacryloyl-aminoethyl-methacrylate (gel-MA-AEMA)-based in vitro intestinal model, which also provides sufficient permeability towards nutrients and drugs. Moreover, the effect of the hydrogel morphology on the cell response was assessed by comparing the formation of a functional intestinal cell monolayer on flat hydrogel films versus 3D hydrogel scaffolds maintaining a close morphological resemblance with the intestinal architecture.A gel-MA-AEMA-based biomaterial ink was formulated and processed towards flat hydrogel films, while digital light processing was exploited to replicate the intestinal microarchitecture. The mechanical assessment of the 2D films confirmed physiologically relevant scaffold stiffness (G’ = 3.30±1.07 kPa). To evaluate the permeability of the hydrogels towards a medium size marker molecule (FITC-dextran 4 kg·mol−1), a static diffusion setup was exploited with custom-made adjustable inserts, which evidenced that both the 2D films and 3D constructs exhibited sufficient permeability. Finally, the combination of the hydrogels with a Caco-2/HT29-MTX co-culture evidenced hydrogel biocompatibility, enabling the formation of a functional cell monolayer after 21 days on the 2D hydrogel films, proven by transepithelial electrical resistance measurements and immunohistochemistry, while the 3D hydrogel constructs did not achieve confluency within 35 days.

Design and researching of biologically active polymeric hydrogel transdermal materials modified by humic acid

Biologically active polymer hydrogel transdermal materials based on gelatin, sodium alginate, modified by humic acids, were designed and researched. Literature review was carried out and it was proved that humic acids using is perspective for the functional effect on the biologically active polymer hydrogel transdermal properties. It has been found that effective processes for receiving biologically active polymer hydrogel transdermal materials based on gelatin, hydroxypropyl cellulose and sodium alginate can be carried out in different humic acids concentration while achieving an effective increase in hydrogel polymers structuring processes. It can be seen efficiently increasing biologically active polymeric hydrogel transdermal materials based on gelatin and sodium alginate conditional viscosity and the specific electrical conductivity with an increase of humic acids content: from 114 to 135 sec and from 2350 to 2850 mkS/cm, respectively. Humic acids modification in biologically active polymeric hydrogel transdermal materials based on gelatin and sodium alginate caused structure formation with high density, and resistance and with larger agglomerates in hydrogels. It was found that the gelatin-based biopolymer hydrogels modification by humic acids makes it possible to receive biologically active polymeric hydrogel transdermal materials with higher swelling degree. It is shown that the application of new biologically active polymeric hydrogel transdermal materials based on gelatin and sodium alginate modified by humic acids allows improving the skin moisture-lipid balance. From the initial values of 34-36% moisture and 8-10 skin fat, they increase to 58-66% and 52-60%. So, designed polymer hydrogel based on gelatin, sodium alginate, modified by humic acids, are transdermal materials with good properties.

Design and fabrication of gelatin-based hydrogel loaded with modified amniotic extracellular matrix for enhanced wound healing

Trauma can damage the structural integrity of skin leading to its function being affected. There is an urgent clinical need for innovative therapeutic wound dressings. However, several challenges persist despite the current demands. The development and application of functional dressings offer a novel approach to address skin and subcutaneous soft tissue defects. Amniotic membrane as an ideal biological multifunctional material covering wound surface has been reported in clinic. However, current clinical applications of amniotic membrane still have limitations, such as thinness and mechanically weak. In this paper, we employed decellularized human amniotic membrane (dHAM) as a bioactive extracellular matrix (ECM) and modified it through methacrylate (MA) grafting for engineering purposes, resulting in the photosensitive dECMMA. Subsequently, we utilized a photosensitizer to achieve photopolymerization of dECMMA with GelMA hydrogel, successfully creating a novel composite hydrogel termed dECMMA/GelMA. This composite hydrogel not only inherits the favorable physicochemical properties of hydrogels but also maintains comparable levels of bioactivity to dHAM itself, supporting cell proliferation, migration, angiogenesis, and retaining significant anti-inflammatory capacity.Additionally, we evaluated the reparative effect of the designed dECMMA/GelMA composite hydrogel on rabbit wound defects. We demonstrated that the dECMMA/GelMA promoted wound healing and re-epithelization. These findings highlight the substantial benefits and therapeutic potential of the dECMMA/GelMA composite hydrogel as a practical solution for clinical applications in the treatment of soft tissue damage. Furthermore, this research provides a new strategy for designing and manufacturing bioactive dressings with exceptional clinical efficacy in the future.

Gelatin-based hydrogel with antiseptic properties: synthesis and properties

The work presents the results of research on the formation of a hydrogel polymer matrix during the gelatin crosslinking with a dioxirane derivative of poly(ethylene glycol) and experiments on studying their properties. The crosslinking process of type A gelatin using poly(ethylene glycol) diglycidyl ether (PEGDE 500) was optimized by changing the gelatin: PEGDE ratio without using any other factors. The properties of the obtained hydrogels were studied, in particular the gel fraction amount, the degree of swelling in different media, mechanical properties, pore size, cytotoxicity, the ability to absorb and release antiseptic drugs, and bactericidal properties of hydrogel samples filled with an antiseptic agent (chlorhexidine). The crosslinking reaction has been found to be the most efficient at gelatin: PEGDE 500 ratio ranging from 1:3 to 1:5. In this case, the formation of highly elastic hydrogels with maximum fraction amount and high mechanical properties was observed (the samples were not destroyed under compression). Moreover, only within this ratio range, the obtained hydrogels retained their shape when heated to 40 °C. The pore sizes of the swollen hydrogel clearly depend on the crosslinker content. The high values of the degree of swelling in the exudate-simulating fluid in combination with mechanical strength allow the development of wound dressings based on hydrogels of the developed composition. Crosslinking of gelatin with PEGDE 500 allows to obtain gelatin-based hydrogels in a controlled manner which does not require subsequent purification steps. The resulting material does not exhibit cytotoxicity during in vitro studies on living cells. The hydrogel filled with an antiseptic drug is capable of releasing it by 50% within the first day into the medium of saline and exudate-simulating fluid. The antimicrobial properties of the hydrogel containing chlorhexidine are confirmed by the effective inhibition of the growth of Escherichia coli and Staphylococcus aureus bacteria.

Skin-interfaced self-powered pressure and strain sensors based on fish gelatin-based hydrogel for wireless wound strain and human motion detection

Biomass-based hydrogels, due to their excellent biocompatibility, can behave as wearable monitoring platforms for healthcare applications and triboelectric sensing devices. However, low tissue adhesiveness, poor stretchability, and bacterial susceptibility cause them to fail to adapt to specific joints with complex movements. Herein, we report a novel biomass-based hydrogel by integrating fish gelatin into polymer networks with in-situ formation of silver nanoparticles (denoted as FG-Ag hydrogel), which achieves great stretchability (2600%), excellent sensitivity (gauge factor = 4), and strong self-adhesion simultaneously. The FG-Ag hydrogel also features a robust antibacterial activity for Escherichia coli and Staphylococcus aureus. Benefitting from these advantages, we have developed a FG-Ag hydrogel-based wearable strain sensor for diverse human-motion detections such as elbow, knee, finger, and wrist bending. Moreover, we have demonstrated an FG-Ag hydrogel-based prototype with an integrated wireless system for real-time strain monitoring on the wound sites. The FG-Ag hydrogel could significantly reduce bacterial infection in vivo and effectively promote wound healing. Additionally, the self-powered pressure sensor and the biomechanical energy harvester also have been demonstrated by the FG-Ag hydrogel based triboelectric nanogenerator (TENG). Accordingly, such FG assistance of hydrogel-based skin-interfaced electronics could provide adequately delicate biomechanical information related to the general health, which furnishes essential technical support for cost-effective, all-green, and highly precise personalized health assessment.

Corneal stromal filler injection of gelatin-based photocurable hydrogels for maintaining the corneal thickness and reconstruction of corneal stroma

Refractive errors manifest as thinning of the central cornea, which can seriously impair vision. The corneal stromal filler is an innovative concept for treating refractive problems. However, corneal stromal regeneration poses a challenge for the tissue-engineered cornea. Hence, materials for corneal reconstruction with high transparency and biocompatibility are urged to be developed. This study aimed to establish a novel therapeutic approach by injecting gelatin-based photocurable hydrogel fillers into the corneal stroma to maintain corneal thickness and promote corneal stromal regeneration for refractive error. The hydrogel possesses high transparency and suitable mechanical qualities, which degrade slowly in the cornea. Furthermore, the hydrogel presents high biocompatibility for cellular adhesion and proliferation. The hydrogel can also provide space for cells to develop and facilitate cell migration when cells are infused into it. In vivo experiments showed that the hydrogel could effectively fortify corneal thickness and induce corneal stromal regeneration. The photocurable hydrogel showed potential in terms of corneal reconstruction.

Rate-dependent fracture behavior of gelatin-based hydrogels

Rate-dependent fracture behavior of gelatin-based hydrogels

The Mechanical and Biological Performance of Photopolymerized Gelatin-Based Hydrogels as a Function of the Reaction Media.

From the first experiments with biomaterials to mimic tissue properties, the mechanical and biochemical characterization have evolved extensively. A number of properties can be described, however, what should be essential is to conduct a proper and physiologically relevant characterization. Herein, we describe the influence of the reaction media and buffer media -phosphate buffer saline (PBS) and Dulbecco's modified Eagle's medium (DMEM) with two different glucose concentrations- in GelMA hydrogel mechanics and in the biological behavior of two tumoral cell lines (Caco-2 and HCT-116). All scaffolds were photocrosslinked using UV light under identical conditions and evaluated for mass swelling ratio and stiffness. Our results indicate that stiffness is highly susceptible to the reaction media, but not to the swelling media. In addition, PBS-prepared hydrogels exhibited a higher photopolymerization degree as confirmed by HR-MAS NMR. These findings correlate with the biological response of Caco-2 and HCT-116 cells seeded on the substrates, which demonstrated flatter morphologies on stiffer hydrogels. Overall, cell viability and proliferation were excellent for both cell lines, and Caco-2 cells displayed a characteristic apical-basal polarization as observed in F-actin/Nuclei fluorescence images. These characterization experiments highlight the importance of conducting mechanical testing of biomaterials in the same medium as cell culture. This article is protected by copyright. All rights reserved.

Gelatin-based hydrogels and ferrogels as smart drug delivery systems: synthesis, characterization and drug release kinetics

Gelatin-based hydrogels and ferrogels as smart drug delivery systems: synthesis, characterization and drug release kinetics

Double network cross-linked hydrogel coating membrane with photocatalytic self-cleaning performance for efficient oil-water separation

Oil-water separation is one of the most attractive areas in the field of energy and the environment. However, the synthesis of oil-water separation membranes with high separation efficiency and anti-fouling via simple methods is still a challenge. In this work, a double network with tannic acid and g-C3N4 cross-linked gelatin hydrogel coating membrane was prepared via ultrasound and coating. The prepared hydrogel had excellent mechanical properties and swelling resistance and also endowed the membrane with underwater superoleophobicity. For stratified oil-water mixtures, the separation flux achieved 40,355 ± 1957 L·m−2·h−1 and separation efficiency higher than 99 %. Moreover, after 40 cycles, there was no decrease significantly in separation performance. For oil-in-water emulsions, the separation efficiency is >99.90 % under gravity. The adsorption of MB by the GE-CNTA-PP membrane conformed to a pseudo-first-order kinetic process (R2 = 0.99906), which was consistent with the Langmuir isothermal adsorption model (R2 = 0.99932). The contaminated membrane could self-clean via photocatalytic reaction, which mechanism was explained through theoretical investigation of the band structure of g-C3N4 by DFT. Due to these obvious advantages, the practical application of the gelatin-based hydrogel membrane in the purification of oily wastewater is promising.

Gelatin-Based Hydrogels Containing Microcrystalline and Nanocrystalline Cellulose as Moisture Absorbers for Food Packaging Applications

Sustainable hydrogels are an innovative biodegradable alternative to traditional packaging materials. They offer exceptional water absorption capacity and high biocompatibility, making them ideal food absorbents to reduce plastic waste, extend shelf life and ensure the safety and quality of packaged foods. In this study, hydrogels based on gelatin, microcrystalline cellulose (MCC), and nanocrystalline cellulose (NCC) were developed, characterized, and applied in the packaging of chicken breasts. For this, MCC was isolated from the banana pseudostem and commercial NCC was incorporated into a gelatin solution to produce the hydrogel materials by film casting. The resulting hydrogels were analyzed in terms of morphology, structural properties, water absorption capacity, mechanical strength, and color properties. The results showed that the incorporation of MCC and NCC significantly improved the mechanical integrity of the hydrogels, which prevented premature deformation of the hydrogels when they absorbed moisture. In addition, changes in the color properties of chicken breast samples in contact with the hydrogels were observed, indicating their ability to preserve food quality. Subsequently, the effectiveness of the hydrogels for chicken breast storage at 4 °C for 4 days was validated. The results demonstrated that the hydrogels developed in this study are a sustainable and environmentally friendly alternative to traditional packaging materials that can extend the shelf life of food products while maintaining their physical and microbiological integrity.

Photocuring and Gelatin-Based Antibacterial Hydrogel for Skin Care.

The development of moisturizing, antibacterial, and biocompatible multifunctional hydrogels is essential to protect skin and promote skin defects recovery. Gelatin has admired potential to be applied for skin care as a hydrogel in virtue of its hydrophilic biocompatible and biodegradable properties. In this study, triclosan-grafted gelatin and photo-cross-linkable methacrylated gelatin were synthesized and then combined to construct the semi-interpenetrating network and antibacterial hydrogels with the aid of a visible blue light. The antimicrobial test demonstrated that the resulting hydrogel obtained excellent inactivation capacity against E. coli, S. aureus, T. rubrum, and C. albicans with sterilizing rates of 99.998%, 99.998%, 99.19%, and 99.64%, respectively. In addition, the cytotoxicity, hemolysis, skin irritation, and rat skin wound healing experiments proved the good biocompatibility of the hydrogel. Therefore, this investigation sheds light on the development of multifunctional hydrogels in skin care.

The Efficacy of Transplanting Human Umbilical Cord Mesenchymal Stem Cell Sheets in the Treatment of Myocardial Infarction in Mice.

The transplantation of mesenchymal stem cell (MSC) sheets derived from human umbilical cords (hUCs) was investigated in this study as a potential application in treating myocardial infarction (MI). Two groups of hUC-MSC sheets were formed by populating LunaGelTM, which are 3D scaffolds of photo-crosslinkable gelatin-based hydrogel with two different cell densities. An MI model was created by ligating the left anterior descending coronary artery of healthy BALB/c mice. After two weeks, the cell sheets were applied directly to the MI area and the efficacy of the treatment was evaluated over the next two weeks by monitoring the mice's weight, evaluating the left ventricle ejection fraction, and assessing the histology of the heart tissue at the end of the experiment. Higher cell density showed significantly greater efficiency in MI mice treatment in terms of weight gain and the recovery of ejection fraction. The heart tissue of the groups receiving cell sheets showed human-CD44-positive staining and reduced fibrosis and apoptosis. In conclusion, the hUC-MSC sheets ameliorated heart MI injury in mice and the efficacy of the cell sheets improved as the number of cells increased.

UV-polymerizable methacrylated gelatin (GelMA)-based hydrogel containing tannic acids for wound healing

Gelatin-based photopolymerizable methacrylate hydrogel (GelMA) is a promising biomaterial for in situ drug delivery, while aqueous extract of Punica granatum (AEPG) peel fruit rich in gallic acid and ellagic acid is used to improve wound healing. The aim of this study was to develop and analyze the healing properties of GelMA containing AEPG, gallic acid, or ellagic acid in a rodent model. GelMA hydrogels containing 5% AEPG (GelMA-PG), 1.6% gallic acid (GelMA-GA), or 2.1% ellagic acid (GelMA-EA) were produced and their mechanical properties, enzymatic degradation, and thermogravimetric profile determined. Wound closure rates, healing histological grading, and immunohistochemical counts of myofibroblasts were assessed over time. The swelling of hydrogels varied between 50 and 90%, and GelMA exhibited a higher swelling than the other groups. The GPG samples showed higher compression and Young’s moduli than GelMA, GGA, and GAE. All samples degraded around 95% in 48 h. GPG and GGA significantly accelerated wound closure, improved collagenization, increased histological grading, and hastened myofibroblast differentiation in comparison to the control, GelMA, and GEA. GelMA containing AEPG (GPG) improved wound healing, and although gallic acid is the major responsible for such biological activity, a potential synergic effect played by other polyphenols present in the extract is evident.Graphical

Multilayer 3D bioprinting and complex mechanical properties of alginate-gelatin mesostructures

In the biomedical field, extrusion-based 3D bioprinting has emerged as a promising technique to fabricate tissue replacements. However, a main challenge is to find suitable bioinks and reproducible procedures that ensure good printability and generate final printed constructs with high shape fidelity, similarity to the designed model, and controllable mechanical properties. In this study, our main goal is to 3D print multilayered structures from alginate-gelatin (AG) hydrogels and to quantify their complex mechanical properties with particular focus on the effects of the extrusion process and geometrical parameters, i.e. different mesostructures and macroporosities. We first introduce a procedure including a pre-cooling step and optimized printing parameters to control and improve the printability of AG hydrogels based on rheological tests and printability studies. Through this procedure, we significantly improve the printability and flow stability of AG hydrogels and successfully fabricate well-defined constructs similar to our design models. Our subsequent complex mechanical analyses highlight that the extrusion process and the mesostructure, characterized by pore size, layer height and filament diameter, significantly change the complex mechanical response of printed constructs. The presented approach and the corresponding results have important implications for future 3D bioprinting applications when aiming to produce replacements with good structural integrity and defined mechanical properties similar to the native tissue, especially in soft tissue engineering. The approach is also applicable to the printing of gelatin-based hydrogels with different accompanying materials, concentrations, or cells.