Mechanical Properties Of Hydrogels Research Articles

Rational design and engineering of carbon nano-onions reinforced natural protein nanocomposite hydrogels for biomedical applications

In the current study, poly 4-hydroxyphenyl methacrylate-carbon nano-onions (PHPMA-CNOs = f-CNOs) are synthesized and reinforced with natural protein gelatin (GL) to engineer GL/f-CNOs composite hydrogels under the sonochemical method. The influence of f-CNOs content on the mechanical properties of hydrogels is examined. Cytotoxicity of hydrogels is measured with the human osteoblast cells. The results revealed good cell viability, cell growth, and attachment on the surface of the hydrogels, and results are f-CNOs dose-dependent. Specifically, the GL/f-CNOs (2 mg/mL) hydrogel showed the highest cell viability, enhanced tensile strength, elastic modulus, and yield strength as compared to pristine GL and GL/f-CNOs (1 mg/mL) hydrogels. It reveals the extent of physisorption and degree of colloidal stability of f-CNOs within the gel matrix. Furthermore, GL/f-CNOs hydrogels efficiently load the 5-fluorouracil (5-FU) and show a pH-responsive sustained drug release over 15 days. Nevertheless, these CNOs based composite hydrogels offer a potential prospect to use them in diverse biomedical applications.

Measurements of Elastic Properties of Biological Hydrogels using Atomic Force Microscopy

Biological hydrogels are widely used as extracellular environment for encapsulating and growing cells. Mechanical properties of hydrogels can influence cell function, mechanotransduction and cellular behavior such as growth, migration, adhesion, differentiation and morphology. Microenvironmental modulus of hydrogels dictates cell behavior and growth. The main purpose of the current study was to determine elastic modulus of two well-known hydrogels (Agarose and Gelatin Methacryloyl) using Atomic Force Microscope (AFM). The elastic modulus was calculated from force-deflection curve obtained by indenting an indenter in the direction normal to the plane of the gel surface. The elastic moduli of the prescribed gels were found to be strongly influenced by the level of concentration in the gel. Elastic modulus of 1% agarose and 2% agarose was found comparable with 10% GelMA and 20% GelMA, respectively. Results obtained from indentation experiments and those published in the literature revealed that AFM can be successfully and confidently used to determine elastic response of hydrogels in the solid state.

Supramolecular Peptide Gels: Influencing Properties by Metal Ion Coordination and Their Wide-Ranging Applications.

Supramolecular gels have been an area of interest in many research fields. They provide a means to understand assembly of nanostructures, and through the use of amino acid- and peptide-based gelators they can give insights into the similar assembly pathways of their more complex structural counterparts. Bio-inspired metal coordination, such as histidine–copper coordination, in the supramolecular assembly of these gelators is one method for furthering our understanding and development of these materials. Metal–gelator coordination mimics biologically relevant metal–peptide coordination, thus influencing hydrogel self-assembly and mechanical properties, including biodegradability, biocompatibility, tunability, and recyclablity, while the metal coordination can functionalize the gels to allow for widespread applications in biomedical industries (e.g., drug delivery), waste management, and catalysis. This review aims to discuss recent insights into the supramolecular assembly of gels involving metal ions, as well as a few key areas of application using metal interactions and incorporation.

Human Atrial Fibroblast Adaptation to Heterogeneities in Substrate Stiffness.

Fibrosis is associated with aging and many cardiac pathologies. It is characterized both by myofibroblast differentiation and by excessive accumulation of extracellular matrix proteins. Fibrosis-related tissue remodeling results in significant changes in tissue structure and function, including passive mechanical properties. This research area has gained significant momentum with the recent development of new tools and approaches to better characterize and understand the ability of cells to sense and respond to their biophysical environment. We use a novel hydrogel, termed CyPhyGel, to provide an advanced in vitro model of remodeling-related changes in tissue stiffness. Based on light-controlled dimerization of a Cyanobacterial Phytochrome, it enables contactless and reversible tuning of hydrogel mechanical properties with high spatial and temporal resolution. Human primary atrial fibroblasts were cultured on CyPhyGels. After 4 days of culturing on stiff (~4.6 kPa) or soft (~2.7 kPa) CyPhyGels, we analyzed fibroblast cell area and stiffness. Cells grown on the softer substrate were smaller and softer, compared to cells grown on the stiffer substrate. This difference was absent when both soft and stiff growth substrates were combined in a single CyPhyGel, with the resulting cell areas being similar to those on homogeneously stiff gels and cell stiffnesses being similar to those on homogeneously soft substrates. Using CyPhyGels to mimic tissue stiffness heterogeneities in vitro, our results confirm the ability of cardiac fibroblasts to adapt to their mechanical environment, and suggest the presence of a paracrine mechanism that tunes fibroblast structural and functional properties associated with mechanically induced phenotype conversion toward myofibroblasts. In the context of regionally increased tissue stiffness, such as upon scarring or in diffuse fibrosis, such a mechanism could help to prevent abrupt changes in cell properties at the border zone between normal and diseased tissue. The light-tunable mechanical properties of CyPhyGels and their suitability for studying human primary cardiac cells make them an attractive model system for cardiac mechanobiology research. Further investigations will explore the interactions between biophysical and soluble factors in the response of cardiac fibroblasts to spatially and temporally heterogeneous mechanical cues.

Probing mechanical properties and failure mechanisms of fibrils of self-assembling peptides.

Self-assembling peptides (SAPs) are a promising class of biomaterials amenable to easy molecular design and functionalization. Despite their increasing usage in regenerative medicine, a detailed analysis of their biomechanics at the nanoscale level is still missing. In this work, we propose and validate, in all-atom dynamics, a coarse-grained model to elucidate strain distribution, failure mechanisms and biomechanical effects of functionalization of two SAPs when subjected to both axial stretching and bending forces. We highlight different failure mechanisms for fibril seeds and fibrils, as well as the negligible contribution of the chosen functional motif to the overall system rupture. This approach could lay the basis for the development of “more” coarse-grained models in the long pathway connecting SAP sequences and hydrogel mechanical properties.

Hierarchically structured hydrogels utilizing multifunctional assembling peptides for 3D cell culture.

Approaches for the creation of soft materials, particularly hydrogels, with hierarchical structure are of interest in a variety of applications owing to their unique properties. In the context of tissue mimics, hydrogels with multiscale structures more accurately capture the complexities of tissues within the body (e.g., fibrous collagen-rich microenvironments). However, cytocompatible fabrication of such materials with hierarchical structures and independent control of mechanical and biochemical properties remains challenging and is needed for probing and directing cell-microenvironment interactions for three-dimensional (3D) cell encapsulation and culture applications. To address this, we have designed innovative multifunctional assembling peptides: these unique peptides contain a core block that mimics the structure of collagen for achieving relevant melting temperatures; 'sticky' ends to promote assembly of long fibrils; and a biocompatible reactive handle that is orthogonal to assembly to allow the formation of desired multiscale structures and their subsequent rapid, light-triggered integration within covalently crosslinked synthetic hydrogels. Nano- to micro-fibrils were observed to form in physiologically-relevant aqueous solutions, where both underlying peptide chemical structure and assembly conditions were observed to impact the resulting fibril sizes. These assembled structures were 'locked' into place and integrated as linkers within cell-degradable, bioactive hydrogels formed with photoinitiated thiol-ene 'click' chemistry. Hydrogel compositions were identified for achieving robust mechanical properties like those of soft tissues while also retaining higher ordered structures after photopolymerization. The utility of these innovative materials for 3D cell culture was demonstrated with human mesenchymal stem cells, where cell morphologies reminiscent of responses to assembled native collagen were observed now with a fully synthetic material. Using a bottom-up approach, a new materials platform has been established that combines the advantageous properties of covalent and assembling chemistries for the creation of synthetic hydrogels with controllable nanostructure, mechanical properties, and biochemical content.

Chromophore amphiphile–polyelectrolyte hybrid hydrogels for photocatalytic hydrogen production

Incorporating supramolecular assemblies of chromophore amphiphiles into covalent hydrogels creates a reusable, robust host for photocatalysis.

EAK Hydrogels Cross-Linked by Disulfide Bonds: Cys Number and Position Are Matched to Performances.

Hydrogels produced by self-assembling peptides are intrinsically biocompatible and thus appropriate for many biomedical purposes. Their application field may be even made wider by reducing the softness and improving the hydrogel mechanical properties through cross-linking treatments. To this aim, modifications of EAK16-II sequence by including Cys residues in its sequence were here investigated in order to obtain hydrogels cross-linkable through a disulfide bridge. Two sequences, namely, C-EAK and C-EAK-C, that contain Cys residues at the N-terminus or at both ends were characterized. Fiber-forming abilities and biological and dynamic mechanical properties were explored before and after the oxidative treatment. In particular, the oxidized version of C-EAK presents a good cell viability and sustains osteoblast proliferation. Furthermore, molecular dynamics (MD) simulations on monomeric and assembled forms of the peptides were performed. MD simulations explained how a specific Cys functionalization was better than the other one. In particular, the results suggested that EAK16-II functionalization with a single Cys residue, instead of two, together with biocompatible cross-linking may be considered an intriguing strategy to obtain a support with better dynamic mechanical properties and biological performances.

Mechanically improved porous hydrogels with polysaccharides via polyelectrolyte complexation for bone tissue engineering

Bone tissue engineering aims to design mechanically improved macroporous hydrogels with fibrous topologies using polysaccharides that can provide an appropriate microenvironment in bone defects in order to enhance bone regeneration similar to the native bone extracellular matrix. Herein, we developed hydrogels by intercalation of chitosan (CS) and sodium alginate (SA)-based polyelectrolyte complexes (PECs) (in situ formation using glucuronic acid delta-galactone as an acidifying agent (GDL)) within the poly(acrylamide) (PAM)-crosslinked network (PEC-PAM) during free radical polymerization. The structure and interactions of PEC-PAM were confirmed by FTIR and XRD experiments. The PEC greatly influenced the porosity, pore size, and mechanical properties of hydrogels. Importantly, the PEC within the hydrogels possessed a macroporous structure with a ladder-like fibrous topology, which may provide better cell growth and adhesion. Moreover, the hydrogels showed good bio-mineralization capacity in simulated body fluid solutions as confirmed by FTIR, XRD, FE-SEM, and SEM-EDX. The in vitro performance of the PEC-PAM hydrogels was assessed towards human bone osteoblasts cells in terms of cell proliferation, biocompatibility, and cell adhesion. All of the results suggest that PEC-PAM hydrogels have good potential in bone tissue engineering.

Preparation and properties of a highly elastic galactomannan- poly(acrylamide- N, N-bis (acryloyl) cysteamine) hydrogel with reductive stimuli-responsive degradable properties

An oxidation-reduction responsive degradable highly elastic galactomannan hydrogel was synthesized from galactomannan (GA), N,N-bis (acryloyl) cysteamine (BAC) and acrylamide by grafting polymerization in aqueous solution. The microstructure, degradability and mechanical properties of the hydrogels were emphatically investigated using Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM), ultraviolet spectroscopy and differential scanning calorimetry (DSC). The mechanical properties of hydrogels can be improved by adjusting the content of GA. Continuous cyclic compression tests showed that the hydrogel did not rupture under 60 %,70 %,80 % strain and quickly recovered to its original shape. The degradation rate and drug release rate of hydrogel can be adjusted by the concentration of the reductant and the reduction time. These hydrogels broaden the scope of application of GA and can be tuned with a broad range of mechanical, degradation and release properties and therefore hold potential applications in drug carriers, tissue engineering scaffolds, extracellular matrix and other fields.

Stereocomplex Poly(Lactic Acid) Amphiphilic Conetwork Gel with Temperature and pH Dual Sensitivity.

A novel stereocomplex poly(lactic acid) amphiphilic conetwork gel with temperature and pH dual sensitivity was synthesized by ring-opening polymerization (ROP) and free radical copolymerization. The chemical structure and composition of hydrogel were characterized by Fourier transform infrared spectroscopy (FT-IR), proton nuclear magnetic resonance (1H NMR) and X-ray diffraction (XRD). The temperature and pH sensitivity and good amphiphilicity of hydrogel were studied using digital photos, the swelling ratios and a scanning electron microscope (SEM). The thermal stability and mechanical properties of hydrogel were studied by differential scanning calorimeter (DSC) and dynamic viscoelastic spectrometer. The results indicated that the hydrogel has amphiphilicity, temperature and pH sensitivity, good thermal stability and mechanical strength.

Alcohol‐triggered silk fibroin hydrogels having random coil and β‐turn structures enhanced for cytocompatible cell response

ABSTRACTAlcohol additive is one of the stimulants that induces the fast gelation of silk fibroin solution. Based on our previous report, different alcohol types influence the gelation kinetic and the properties of resulting silk fibroin hydrogels. Here, the effects of alcohol concentrations on the silk fibroin gelation and cell response were reported. All fibroin hydrogels prepared with various alcohol additives showed cell biocompatibility, especially the fibroin hydrogel prepared with 10 wt % n‐butanol. Results on the mechanical properties of hydrogels, n‐butanol additive enhanced a higher compressive modulus up to ~ 22 times in comparison to non‐alcoholic fibroin hydrogel. Fourier transform infrared analysis and peak deconvolution showed a possible formation of more β‐turn linkage and random coil structure of fibroin segments in alcoholic fibroin hydrogel. So, the micro‐segmental structure of fibroin hydrogel caused the higher compressive modulus, prolonged deformation of the hydrogels, and efficient cell growth on the fibroin hydrogel. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020, 137, 48731.

Preparation and evaluation of microfluidic magnetic alginate microparticles for magnetically templated hydrogels

Our aim is to develop a hydrogel-based scaffold containing porous microchannels that mimic complex tissue microarchitecture and provide physical cues to guide cell growth for scalable, cost-effective tissue repair. These hydrogels are patterned through the novel process of magnetic templating where magnetic alginate microparticles (MAMs) are dispersed in a hydrogel precursor and aligned in a magnetic field before hydrogel crosslinking and subsequent MAM degradation, leaving behind an aligned, porous architecture. Here, a protocol for fabricating uniform MAMs using microfluidics was developed for improved reproducibility and tunability of templated microarchitecture. Through iron quantification, we find that this approach allows control over magnetic iron oxide loading of the MAMs. Using Brownian dynamics simulations and nano-computed tomography of templated hydrogels to examine MAM chain length and alignment, we find agreement between simulated and measured areal densities of MAM chains. Oscillatory rheology and stress relaxation experiments demonstrate that magnetically templated microchannels alter bulk hydrogel mechanical properties. Finally, in vitro studies where rat Schwann cells were cultured on templated hydrogels to model peripheral nerve injury repair demonstrate their propensity for providing cell guidance along the length of the channels. Our results show promise for a micro-structured biomaterial that could aid in tissue repair applications.

In situ miRNA delivery from a hydrogel promotes osteogenesis of encapsulated mesenchymal stromal cells

Hydrogels are attractive candidates for use in tissue-engineering and the encapsulation and subsequent differentiation of mesenchymal stem/stromal cells (MSCs) is a strategy that holds great promise for the repair and regeneration of bone and cartilage. However, MSCs are well-known for their sensitivity to mechanical cues, particularly substrate stiffness, and so the inherent softness of hydrogels is poorly matched to the mechanical cues that drive efficient osteogenesis. One approach to overcome this limitation is to harness mechanotransductive signalling pathways and override the signals cells receive from their environment. Previous reports demonstrate that mechanosensitive miRNAs, miR-100–5p and miR-143–3p can enhance MSC osteogenesis, using a complex multi-step procedure to transfect, encapsulate and differentiate the cells.In this study, we develop and characterise a facile system for in situ transfection of MSCs encapsulated within a light-crosslinkable gelatin-PEG hydrogel. Comparing the influence of different transfection agents and hydrogel compositions, we show that particle size, charge, and hydrogel mechanical properties all influence the diffusion of embedded transfection agent complexes. By incorporating both MSCs and transfection agents into the hydrogels we demonstrate successful in situ transfection of encapsulated MSCs. Comparing the efficacy of pre- and in situ transfection of miR-100–5p/miR-143–3p on the osteogenic capacity of hydrogel-encapsulated MSCs, our data demonstrates superior mineralisation and osteogenic gene expression following in situ transfections. Overall, we demonstrate a simple, one-pot system for in situ transfection of miRNAs to enhance MSC osteogenic potential and thus demonstrates significant promise to improve the efficiency of MSC differentiation in hydrogels for bone tissue-engineering applications. Statement of significanceMesenchymal stromal cells (MSCs) are sensitive to cues from their surrounding microenvironment. Osteogenesis is enhanced in MSCs grown on stiffer substrates, but this is limited when using hydrogels for bone tissue-engineering. Modulating pro-osteogenic genes with mechanosensitive microRNAs (miRNAs) represents a potential tool to overcome this challenge. Here we report a hydrogel platform to deliver miRNAs to encapsulated MSCs. We characterise effects of hydrogel composition and transfection agent type on their mobility and transfection efficiency, demonstrating successful in situ transfection of MSCs and showing that miRNAs can significantly enhance osteogenic mineral deposition and marker gene expression. This system was simpler and more effective than conventional 2D transfection prior to encapsulation and therefore holds promise to improve MSC differentiation in bone tissue-engineering.

Injectable supramolecular polymer-nanoparticle hydrogels enhance human mesenchymal stem cell delivery.

Stem cell therapies have emerged as promising treatments for injuries and diseases in regenerative medicine. Yet, delivering stem cells therapeutically can be complicated by invasive administration techniques, heterogeneity in the injection media, and/or poor cell retention at the injection site. Despite these issues, traditional administration protocols using bolus injections in a saline solution or surgical implants of cell‐laden hydrogels have highlighted the promise of cell administration as a treatment strategy. To address these limitations, we have designed an injectable polymer–nanoparticle (PNP) hydrogel platform exploiting multivalent, noncovalent interactions between modified biopolymers and biodegradable nanoparticles for encapsulation and delivery of human mesenchymal stem cells (hMSCs). hMSC‐based therapies have shown promise due to their broad differentiation capacities and production of therapeutic paracrine signaling molecules. In this work, the fundamental hydrogel mechanical properties that enhance hMSC delivery processes are elucidated using basic in vitro models. Further, in vivo studies in immunocompetent mice reveal that PNP hydrogels enhance hMSC retention at the injection site and retain administered hMSCs locally for upwards of 2 weeks. Through both in vitro and in vivo experiments, we demonstrate a novel scalable, synthetic, and biodegradable hydrogel system that overcomes current limitations and enables effective cell delivery.

Plastic-like Hydrogels with Reversible Conversion of Elasticity and Plasticity and Tunable Mechanical Properties.

The development of hydrogels with excellent mechanical properties is highly desirable in both fundamental studies and practical applications. But it is difficult to construct hydrogels that are both tough and stiff at the same time as these properties often contradict each other. Here, we report a facile and efficient method for producing ultrastiff and tough poly(N-isopropylacrylamide) (PNIPAM)/clay plastic-like hydrogels (PHs) by immersing PNIPAM/clay hydrogel into NaCl aqueous solution. The optimized PH-2-6 presented superior strength, modulus, and toughness (4.1 ± 0.2 MPa, 41.6 ± 8 MPa, and 15.85 ± 0.8 MJ m-3, respectively). The unique mechanical properties are attributed to the synergistic effect of the osmotic pressure and the strong affinity between Na+ ion and the PNIPAM chain, which lead to a high degree of PNIPAM chain entanglement and fixing. Note that the PHs were molded into any required shape under an applied force, and retained permanently their shapes even if the load was removed, thus displaying typical plasticity. However, the deformed PHs could return to their original size and softness of hydrogel when immersed in pure water, which is a kind of shape-memory effect. The reversible conversion of elasticity and plasticity and shape memory arise from a kind of dynamic physical across-linking of Na+ and PNIPAM molecular chains, which could exist in the salt aqueous and disintegrate in water reversibly. Moreover, the mechanical properties of hydrogel can be tuned by adjusting the salt concentration and immersion time. The facile strategy may provide further avenue in developing hydrogels with such versatile dynamic behaviors to expand their applications.

Soluble Signals and Remodeling in a Synthetic Gelatin-Based Hematopoietic Stem Cell Niche.

Hematopoietic stem cells (HSCs) reside in the bone marrow within niches that provide microenvironmental signals in the form of biophysical cues, bound and diffusible biomolecules, and heterotypic cell-cell interactions that influence HSC fate decisions. This study seeks to inform the development of a synthetic culture platform that promotes ex vivo HSC expansion without exhaustion. A library of methacrylamide-functionalized gelatin (GelMA) hydrogels is used to explore remodeling and crosstalk from mesenchymal stromal cells (MSCs) on the expansion and quiescence of murine HSCs. The use of a degradable GelMA hydrogel enables MSC-mediated remodeling, yielding dynamic shifts in the matrix environment over time. An initially low-diffusivity hydrogel for co-culture of hematopoietic stem and progenitor cells to MSCs facilitates maintenance of an early progenitor cell population over 7 days. Excitingly, this platform promotes retention of a quiescent HSC population compared to HSC monocultures. These studies reveal MSC-density-dependent upregulation of MMP-9 and changes in hydrogel mechanical properties (ΔE = 2.61 ± 0.72) suggesting MSC-mediated matrix remodeling may contribute to a dynamic culture environment. Herein, a 3D hydrogel is reported for ex vivo HSC culture, in which HSC expansion and quiescence is sensitive to hydrogel properties, MSC co-culture, and MSC-mediated hydrogel remodeling.

Peptide-/Drug-Directed Self-Assembly of Hybrid Polyurethane Hydrogels for Wound Healing.

Drug-loading hydrogels are promising candidates in the bioengineering research field; nevertheless, hydrophobic drug loading into a hydrophilic carrier system remains unsolved and is full of challenges. In this work, following the potential dual interactions between peptides and aromatic drugs, we developed a potent hybrid hydrogel formation method, namely, "peptide-/drug-directed self-assembly". The hybrid hydrogels were synthesized using polyethylene glycol (PEG)-based Fmoc-FF peptide hybrid polyurethane, in which curcumin could be encapsulated through self-assembly with Fmoc-FF peptide via π-π stacking. On the basis of this, curcumin loading capacity could be improved to as high as 3.3 wt % with sustained release. In addition, the curcumin loading enhanced the hydrogel mechanical properties from 4 kPa to over 10 kPa, similar to that of natural soft tissues. Furthermore, the hydrogels were injectable with self-healing properties since the Fmoc-FF peptide/curcumin coassembly was noncovalent and reversible. Spectroscopy results confirmed the existence of the coassembly of Fmoc-FF peptide/curcumin. Further in vivo experiments effectively demonstrated that the hydrogels could improve the cutaneous wound healing in a full-thickness skin defected model. This peptide-/drug-directed self-assembly of hybrid polyurethane hydrogel could be used as a promising platform for tissue-engineering scaffold and biomedical application.

Textural and tensile properties of thermo-responsive poly(2-(2-methoxyethoxy)ethyl methacrylate) hydrogel

ABSTRACTHydrogel dressings should be adhesive and elastic, while providing excellent functional and aesthetic features. Owing to weak intermolecular forces, various measures are undertaken to improve the hydrogel mechanical properties. Thermo-responsive poly(2-(2-methoxyethoxy)ethyl methacrylate) (poly(MEO2MA)) hydrogel fabricated with different amounts of di(ethylene glycol) dimethacrylate (DEGDMA) cross-linker was characterised using texture profile analysis. More cross-linking sites incorporated within the hydrogel limit the swelling ability where 1.0 and 10 mol-% of DEGDMA swell by 8.03 and 1.92 times, at 5°C. Hydrogel textural and tensile property correlated with the cross-linker amount. Meanwhile, network structure disentanglement within swollen hydrogel reduced the force required for deformation. Thus, the swelling behaviour, textural and tensile properties were investigated to discern the possible biomedical applications suitable for this hydrogel.

Gelatin-Methacryloyl (GelMA) Formulated with Human Platelet Lysate Supports Mesenchymal Stem Cell Proliferation and Differentiation and Enhances the Hydrogel's Mechanical Properties.

Three-dimensional (3D) cell culture is a major focus of current research, since cultivation under physiological conditions provides more reliable information about in vivo cell behavior. 3D cell cultures are used in basic research to better understand intercellular and cell-matrix interactions. Moreover, 3D cell culture plays an increasingly important role in the in vitro testing of bioactive substances and tissue engineering. Gelatin-methacryloyl (GelMA) hydrogels of different degrees of functionalization (DoFs) are a versatile tool for 3D cell culture and related applications such as bioprinting. Human platelet lysate (hPL) has already demonstrated positive effects on 2D cell cultures of different cell types and has proven a valuable alternative to fetal calf serum (FCS). Traditionally, all hydrogels are formulated using buffers. In this study, we supplemented GelMA hydrogels of different DoF with hPL during adipose tissue-derived mesenchymal stem cell (AD-MSCs) encapsulation. We studied the effect of hPL supplementation on the spreading, proliferation, and osteogenic differentiation of AD-MSCs. In addition, the influence of hPL on hydrogel properties was also investigated. We demonstrate that the addition of hPL enhanced AD-MSC spreading, proliferation, and osteogenic differentiation in a concentration-dependent manner. Moreover, the addition of hPL also increased GelMA viscosity and stiffness.