Hydrogels For Tissue Engineering Applications Research Articles

Publisher Correction: An Injectable In Situ Forming Collagen/Alginate/CaSO4 Composite Hydrogel for Tissue Engineering Applications: Optimization, Characterization and In Vitro Assessments

Publisher Correction: An Injectable In Situ Forming Collagen/Alginate/CaSO4 Composite Hydrogel for Tissue Engineering Applications: Optimization, Characterization and In Vitro Assessments

Microfluidic Platforms to Screen Granular Hydrogel Microenvironments for Tissue Regeneration

AbstractGranular hydrogels have emerged as a promising class of biomaterials in medical research, enabling independent control of matrix stiffness within a porous biomaterial. Such microgel packings comprise interconnected pores and high surface‐to‐volume ratios. These features facilitate cell viability and nutrient diffusion, which are critical in enabling tissue regeneration. Despite the current interest in granular hydrogels for tissue engineering applications, only a few in vitro platforms are used to investigate cell interactions, limiting their design, and translation. In this study, microfluidic platforms able to reproducibly confine and immobilize microgels without the need for secondary cross‐linking are developed. Protocols are established for the generation of human bone marrow‐derived mesenchymal stem/stromal cells (hBM‐MSC)‐infiltrated microporous substrates and early‐time responses of cells to their environment are studied. Further, a tissue invasion assay is established, where cells infiltrate granular materials at different rates depending on growth factor presence or material properties. This platform is compatible with a range of different granular materials, and it is envisioned to have significant utility as a pre‐clinical tool for the rational design of materials for tissue healing applications.

An Injectable In Situ Forming Collagen/Alginate/CaSO4 Composite Hydrogel for Tissue Engineering Applications: Optimization, Characterization and In Vitro Assessments

An Injectable In Situ Forming Collagen/Alginate/CaSO4 Composite Hydrogel for Tissue Engineering Applications: Optimization, Characterization and In Vitro Assessments

Superparamagnetic freeze-thawed PVA hydrogel for applications in tissue engineering, drug delivery and bioimaging

The prerequisite of super-paramagnetic materials in biomedical applications necessitate the optimum magnetic saturation and negligible cytotoxic characteristics. Especially, the development of super-paramagnetic hydrogels in tissue engineering has been rather a scarce documentation. In this context, the present study aims to integrate Fe2+/3+ in PVA hydrogels through repetitive freeze-thawing technique followed by lyophilisation. A detailed physiochemical characterization of the developed hydrogels has been evaluated using X-ray diffraction, TGA, Raman and XPS techniques. The binding of iron with PVA has been confirmed through FTIR analysis and magnetic measurements at room temperature ensured the super-paramagnetic nature of obtained hydrogels. The release profiles of gentamycin drug loaded PVA hydrogel showcased an 8% release in 12 h and this profile followed the non- Fickian diffusion process. The maximum tensile and compression strength of hydrogels are respectively determined as 3.2 and 4.6 MPa. The presence of Fe in the hydrogel deliberated T2 MRI imaging characteristics. In vitro test results affirmed the cell viability of more than 60% for all the hydrogels. The overall findings from the study signified the applications of FePVA hydrogels in tissue engineering applications.

Pristine gelatin incorporation as a strategy to enhance the biofunctionality of poly(vinyl alcohol)-based hydrogels for tissue engineering applications.

Synthetic polymers, such as poly(vinyl alcohol) (PVA), are popular biomaterials for the fabrication of hydrogels for tissue engineering and regenerative medicine (TERM) applications, as they provide excellent control over the physico-chemical properties of the hydrogel. However, their bioinert nature is known to limit cell-biomaterial interactions by hindering cell infiltration, blood vessel recruitment and potentially limiting their integration with the host tissue. Efforts in the field have therefore focused on increasing the biofunctionality of synthetic hydrogels, without limiting the advantages associated with their tailorability and controlled release capacity. The aim of this study was to investigate the suitability of pristine gelatin to enhance the biofunctionality of tyraminated PVA (PVA-Tyr) hydrogels, by promoting cell infiltration and host blood vessel recruitment for TERM applications. Pure PVA-Tyr hydrogels and PVA-Tyr hydrogels incorporated with vascular endothelial growth factor (VEGF), a well-known pro-angiogenic stimulus, were used for comparison. Incorporating increasing concentrations of VEGF (0.01-10 μg mL-1) or gelatin (0.01-5 wt%) did not influence the physical properties of PVA-Tyr hydrogels. However, their presence within the polymer network (>0.1 μg mL-1 VEGF and >0.1 wt% gelatin) promoted endothelial cell interactions with the hydrogels. The covalent binding of unmodified gelatin or VEGF to the PVA-Tyr network did not hamper their inherent bioactivity, as they both promoted angiogenesis in a chick chorioallantoic membrane (CAM) assay, performing comparably with the unbound VEGF control. When the PVA-Tyr hydrogels were implanted subcutaneously in mice, it was observed that cell infiltration into the hydrogels was possible in the absence of gelatin or VEGF at 1- or 3-weeks post-implantation, highlighting a clear difference between in vitro an in vivo cell-biomaterial interaction. Nevertheless, the presence of gelatin or VEGF was necessary to enhance blood vessel recruitment and infiltration, although no significant difference was observed between these two biological molecules. Overall, this study highlights the potential of gelatin as a standalone pro-angiogenic cue to enhance biofunctionality of synthetic hydrogels and provides promise for their use in a variety of TERM applications.

Glycosaminoglycan-mimetic infernan grafted with poly(N-isopropylacrylamide): Toward a thermosensitive polysaccharide

Glycosaminoglycans (GAGs) are essential constituents of the cell surface and extracellular matrix, where they are involved in several cellular processes through their interactions with various proteins. For successful tissue regeneration, developing an appropriate matrix supporting biological activities of cells in a similar manner than GAGs remains still challenging. In this context, this study aims to design a thermosensitive polysaccharide that could further be used as hydrogel for tissue engineering applications. For this purpose, infernan, a marine bacterial exopolysaccharide (EPS) endowed with GAG-mimetic properties was grafted with a thermosensitive polymer, poly(N-isopropylacrylamide) (pNIPAM). Eight grafted polysaccharides were obtained by varying EPS/pNIPAM molar ratio and the molecular weight of pNIPAM. Their physicochemical characteristics and their thermosensitive properties were determined using a multi-technique, experimental approach. In parallel, molecular dynamics and Monte Carlo simulations were applied at two different scales to elucidate, respectively, the molecular conformation of grafted infernan chain and their ability to form an infinite network undergoing a sol-gel transition near the percolation, a necessary condition in hydrogel formation. It comes out from this study that thermosensitive infernan was successfully developed and its potential use in tissue regeneration as a hydrogel scaffold will further be assessed.

In situ forming aldehyde-modified xanthan/gelatin hydrogel for tissue engineering applications: synthesis, characterization, and optimization

In situ forming aldehyde-modified xanthan/gelatin hydrogel for tissue engineering applications: synthesis, characterization, and optimization

Additive Manufacturing and Physicomechanical Characteristics of PEGDA Hydrogels: Recent Advances and Perspective for Tissue Engineering.

In this brief review, we discuss the recent advancements in using poly(ethylene glycol) diacrylate (PEGDA) hydrogels for tissue engineering applications. PEGDA hydrogels are highly attractive in biomedical and biotechnology fields due to their soft and hydrated properties that can replicate living tissues. These hydrogels can be manipulated using light, heat, and cross-linkers to achieve desirable functionalities. Unlike previous reviews that focused solely on material design and fabrication of bioactive hydrogels and their cell viability and interactions with the extracellular matrix (ECM), we compare the traditional bulk photo-crosslinking method with the latest three-dimensional (3D) printing of PEGDA hydrogels. We present detailed evidence combining the physical, chemical, bulk, and localized mechanical characteristics, including their composition, fabrication methods, experimental conditions, and reported mechanical properties of bulk and 3D printed PEGDA hydrogels. Furthermore, we highlight the current state of biomedical applications of 3D PEGDA hydrogels in tissue engineering and organ-on-chip devices over the last 20 years. Finally, we delve into the current obstacles and future possibilities in the field of engineering 3D layer-by-layer (LbL) PEGDA hydrogels for tissue engineering and organ-on-chip devices.

Engineering synthetic poly(ethylene) glycol-based hydrogels compatible with injection molding biofabrication.

Hydrogel injection molding is a biofabrication method that is useful for the rapid generation of complex cell-laden hydrogel geometries, with potential utility in biomanufacturing products for tissue engineering applications. Hydrogel injection molding requires that hydrogel polymers have sufficiently delayed crosslinking times to enable injection and molding prior to gelation. In this work, we explore the feasibility of injection molding synthetic poly(ethylene) glycol (PEG)-based hydrogels functionalized with strain promoted azide-alkyne cycloaddition click chemistry functional groups. We evaluate the mechanical properties of a PEG-based hydrogel library, including time to gelation and successful generation of complex geometries via injection molding. We evaluate the binding and retention of adhesive ligand RGD within the library matrices and characterize the viability and function of encapsulated cells. This work demonstrates the feasibility of injection molding synthetic PEG-based hydrogels for tissue engineering applications, with potential utility in the clinic and biomanufacturing.

Gelatin modified with alkoxysilanes (GelmSi) forms hybrid hydrogels for bioengineering applications

Biopolymers are ideal candidates for the development of hydrogels for tissue engineering applications. However, chemical modifications are required to further improve their mechanical properties, in particular to cross-link them for long-lasting applications or biofabrication. Herein, we developed a novel gelatin-based hydrogel precursor, “GelmSi” which consist on modified gelatin with triethoxysilyl groups. Gelatin was chosen as starting material because of its biocompatibility and bioactivity, favouring cell adhesion and migration. Alkoxysilane moieties were introduced in a controlled manner on the lysine side chains of gelatin to obtain a hybrid precursor which reacts in physiological conditions, forming covalent siloxane bonds and allowing the formation of a three-dimensional chemical network. On the contrary to unmodified gelatin, siloxane covalent network dramatically increases the stiffness and the thermal stability of the resulting gelatin-based hydrogel, making it suitable for cell encapsulation and cell culture. The biorthogonality and versatility of the GelmSi hybrid hydrogel unlock a broad range of gelatin-based bioengineering applications.

Recent Developments in Biopolymer-Based Hydrogels for Tissue Engineering Applications.

Hydrogels are being investigated for their application in inducing the regeneration of various tissues, and suitable conditions for each tissue are becoming more apparent. Conditions such as the mechanical properties, degradation period, degradation mechanism, and cell affinity can be tailored by changing the molecular structure, especially in the case of polymers. Furthermore, many high-functional hydrogels with drug delivery systems (DDSs), in which drugs or bioactive substances are contained in controlled hydrogels, have been reported. This review focuses on the molecular design and function of biopolymer-based hydrogels and introduces recent developments in functional hydrogels for clinical applications.

Synthesis and evaluation of alginate, gelatin, and hyaluronic acid hybrid hydrogels for tissue engineering applications

Tissue engineering (TE) has been proposed extensively as a potential solution to the worldwide shortages of donor organs needed for transplantation. Over the years, numerous hydrogel formulations have been studied for various TE endeavours, including bone, cardiac or neural TE treatment strategies. Amongst the materials used, organic and biocompatible materials which aim to mimic the natural extracellular matrix of the native tissue have been investigated to create biomimicry regenerative environments. As such, the comparison between studies using the same materials is often difficult to accomplish due to varying material concentrations, preparation strategies, and laboratory settings, and as such these variables have a huge impact on the physio-chemical properties of the hydrogel systems. The purpose of the current study is to investigate popular biomaterials such as alginate, hyaluronic acid and gelatin in a variety of concentrations and hydrogel formulations. This aims to provide a clear and comprehensive understanding of their behaviours and provide a rational approach as to the appropriate selection of natural polysaccharides in specific targeted TE strategies.

Species-Based Differences in Mechanical Properties, Cytocompatibility, and Printability of Methacrylated Collagen Hydrogels.

Collagen methacrylation is a promising approach to generate photo-cross-linkable cell-laden hydrogels with improved mechanical properties. However, the impact of species-based variations in amino acid composition and collagen isolation method on methacrylation degree (MD) and its subsequent effects on the physical properties of methacrylated collagen (CMA) hydrogels and cell response are unknown. Herein, we compared the effects of three collagen species (bovine, human, and rat), two collagen extraction methods (pepsin digestion and acid extraction), and two photoinitiators (lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) and Irgacure-2959 (I-2959)) on the physical properties of CMA hydrogels, printability and mesenchymal stem cell (MSC) response. Human collagen showed the highest MD. LAP was more cytocompatible than I-2959. The compressive modulus and cell viability of rat CMA were significantly higher (p < 0.05) than bovine CMA. Human CMA yielded constructs with superior print fidelity. Together, these results suggest that careful selection of collagen source and cross-linking conditions is essential for biomimetic design of CMA hydrogels for tissue engineering applications.

Knowledge domain and hotspots concerning photosensitive hydrogels for tissue engineering applications: A bibliometric and visualized analysis (1996-2022).

Objective: The aim of tissue engineering (TE) is to replace the damaged tissues or failed organs, or restore their missing functions. The important means to achieve this aim is to integrate biomaterials and life elements. Hydrogels are very attractive biomaterials in the field of TE. In particular, engineering extracellular matrices (ECMs) formed by photosensitive hydrogels have captivated much attention, because photopolymerization has many advantages over traditional polymerization approaches, such as rapidity of reaction, spatiotemporal controllability of polymerization process, and operability at physiological temperature, especially it can realize the fabrications of engineering ECMs in the presence of living cells. There have been many excellent reviews on the applications of photosensitive hydrogels in TE in recent years, however, it is inevitable that researchers may have left out many important facts due to exploring the literature from one or a few aspects. It is also a great challenge for researchers to explore the internal relationships among countries, institutions, authors, and references from a large number of literatures in related fields. Therefore, bibliometrics may be a powerful tool to solve the above problems. A bibliometric and visualized analysis of publications concerning the photosensitive hydrogels for TE applications was performed, and the knowledge domain, research hotspots and frontiers in this topic were identified according to the analysis results. Methods: We identified and retrieved the publications regarding the photosensitive hydrogels for TE applications between 1996 and 2022 from Web of Science Core Collection (WoSCC). Bibliometric and visualized analysis employing CiteSpace software and R-language package Bibliometrix were performed in this study. Results: 778 publications meeting the eligibility criteria were identified and retrieved from WoSCC. Among those, 2844 authors worldwide participated in the studies in this field, accompanied by an average annual article growth rate of 15.35%. The articles were co-authored by 800 institutions from 46 countries/regions, and the United States published the most, followed by China and South Korea. As the two countries that published the most papers, the United States and China could further strengthen cooperation in this field. Univ Colorado published the most articles (n = 150), accounting for 19.28% of the total. The articles were distributed in 112 journals, among which Biomaterials (n = 66) published the most articles, followed by Acta Biomaterialia (n = 54) and Journal of Biomedical Materials Research Part A (n = 42). The top 10 journals published 47.8% of the 778 articles. The most prolific author was Anseth K (n = 33), followed by Khademhosseini A (n = 29) and Bryant S (n = 22). A total of 1443 keywords were extracted from the 778 articles and the keyword with the highest centrality was "extracellular matrix" (centrality: 0.12). The keywords appeared recently with strong citation bursts were "gelatin", "3d printing" and "3d bioprinting", representing the current research hotspots in this field. "Gelma", "3d printing" and "thiol-ene" were the research frontiers in recent years. Conclusion: This bibliometric and visualized study offered a comprehensive understanding of publications regarding the photosensitive hydrogels for TE applications from 1996 to 2022, including the knowledge domain, research hotspots and frontiers in this filed. The outcome of this study would provide insights for scholars in the related research filed.

3D-Printed Hybrid Collagen/GelMA Hydrogels for Tissue Engineering Applications.

Bioprinting is an emerging technology involved in the fabrication of three-dimensional tissue constructs for the repair and regeneration of various tissues and organs. Collagen, a natural protein found abundantly in the extracellular matrix of several tissues, can be extracted from collagen-rich tissues of animals such as sheep, cows, rats, pigs, horses, birds, and marine animals. However, due to the poor printability of collagen bioinks, biocompatible collagen scaffolds that mimic the extracellular matrix (ECM) are difficult to fabricate using bioprinting techniques. Gelatin methacrylate (GelMA), a semi-synthetic polymer with tunable physical and chemical properties, has been found to be a promising biomaterial in various bioprinting applications. The printability of collagen can be improved by combining it with semi-synthetic polymers such as GelMA to develop hybrid hydrogels. Such hybrid hydrogels printed have also been identified to have enhanced mechanical properties. Hybrid GelMA meshes have not previously been prepared with collagen from ovine sources. This study provides a novel comparison between the properties of hybrid meshes with ovine skin and bovine hide collagen. GelMA (8% w/v) was integrated with three different concentrations (0.5%, 1%, and 2%) of bovine and ovine collagen forming hybrid hydrogels inks that were printed into meshes with enhanced properties. The maximum percentage of collagen suitable for integration with GelMA, forming hybrid hydrogels with a stable degradation rate was 1%. The water-soluble nature of ovine collagen promoted faster degradation of the hybrid meshes, although the structural crosslinking was identified to be higher than bovine hybrid meshes. The 1% bovine collagen hybrid meshes stood out in terms of their stable degradation rates.

Recent Advances in Cellulose-Based Hydrogels for Tissue Engineering Applications

In recent years, cellulose has attracted much attention because of its excellent properties, such as its hydrophilicity, mechanical properties, biodegradability, biocompatibility, low cost and low toxicity. In addition, cellulose and its derivatives contain abundant hydrophilic functional groups (such as hydroxyl, carboxyl and aldehyde groups), which are good raw materials for synthesizing biocompatible hydrogels. In this paper, the application prospects of cellulose and its derivatives-based hydrogels in biomedical tissue engineering are summarized and discussed through the analysis of recent research. Firstly, we discuss the structure and properties of cellulose, nano celluloses (NC) from different sources (including cellulose nanocrystals (CNC), cellulose nanofibrils (CNF) and bacterial nano celluloses (BNC)) and cellulose derivatives (including cellulose ethers and cellulose esters) obtained by different modification methods. Then, the properties and preparation methods of physical and chemical cellulose hydrogels are described, respectively. The application of cellulose-based hydrogels as a tissue engineering scaffold (skin, bone and cartilage) in the biomedical field is introduced. Finally, the challenges and prospects of cellulose-based hydrogels in tissue engineering are summarized.

Three-dimensional tailor-made collagen-like proteins hydrogel for tissue engineering applications.

Despite the potential tunable properties of blank slate collagen-like proteins (CLP), an alternative to animal-originated collagen, assembling them into a stable 3D hydrogel to mimic extracellular matrix is a challenge. To address this constraint, the CLP (without hydroxyproline, CLPpro) and its variants encoding functional unnatural amino acids such as hydroxyproline (CLPhyp) and 3,4-dihydroxyphenylalanine (CLPdopa) were generated through genetic code engineering for 3D hydrogel development. The CLPhyp and CLPdopa were chosen to enhance the intermolecular hydrogen bond interaction through additional hydroxyl moiety and thereby facilitate the self-assembly into a fibrillar network of the hydrogel. Hydrogelation was induced through genipin as a cross-linker, enabling intermolecular cross-linking to form a hydrogel. Spectroscopic and rheological analyses confirmed that CLPpro and its variants maintained native triple-helical structure, which is necessary for its function, and viscoelastic nature of the hydrogels, respectively. Unlike CLPpro, the varients (CLPhyp and CLPdopa) increased pore size formation in the hydrogel scaffold, facilitating 3T3 fibroblast cell interactions. DSC analysis indicated that the stability of the hydrogels got increased upon the genetic incorporation of hydroxyproline (CLPhyp) and dopa (CLPdopa) in CLPpro. In addition, CLPdopa hydrogel was found to be relatively stable against collagenase enzyme compared to CLPpro and CLPhyp. It is the first report on 3D biocompatible hydrogel preparation by tailoring CLP sequence with non-natural amino acids. These next-generation tunable CLP hydrogels open a new venue to design synthetic protein-based biocompatible 3D biomaterials for tissue engineering applications.

Preparation and Characterization of Plasma-Derived Fibrin Hydrogels Modified by Alginate di-Aldehyde.

Fibrin hydrogels are one of the most popular scaffolds used in tissue engineering due to their excellent biological properties. Special attention should be paid to the use of human plasma-derived fibrin hydrogels as a 3D scaffold in the production of autologous skin grafts, skeletal muscle regeneration and bone tissue repair. However, mechanical weakness and rapid degradation, which causes plasma-derived fibrin matrices to shrink significantly, prompted us to improve their stability. In our study, plasma-derived fibrin was chemically bonded to oxidized alginate (alginate di-aldehyde, ADA) at 10%, 20%, 50% and 80% oxidation, by Schiff base formation, to produce natural hydrogels for tissue engineering applications. First, gelling time studies showed that the degree of ADA oxidation inhibits fibrin polymerization, which we associate with fiber increment and decreased fiber density; moreover, the storage modulus increased when increasing the final volume of CaCl2 (1% w/v) from 80 µL to 200 µL per milliliter of hydrogel. The contraction was similar in matrices with and without human primary fibroblasts (hFBs). In addition, proliferation studies with encapsulated hFBs showed an increment in cell viability in hydrogels with ADA at 10% oxidation at days 1 and 3 with 80 µL of CaCl2; by increasing this compound (CaCl2), the proliferation does not significantly increase until day 7. In the presence of 10% alginate oxidation, the proliferation results are similar to the control, in contrast to the sample with 20% oxidation whose proliferation decreases. Finally, the viability studies showed that the hFB morphology was maintained regardless of the degree of oxidation used; however, the quantity of CaCl2 influences the spread of the hFBs.

The Role of Aldehyde-Functionalized Crosslinkers on the Property of Chitosan Hydrogels.

Chitosan has been utilized as a popular biopolymer to fabricate hydrogels for biomedical applications. However, chitosan hydrogels are generally too brittle to mimic the deformability of the extracellular matrix in many tissues and organs. In particular, the role of the varied crosslinkers in determining the elasticity of chitosan hydrogels is lack of discussion. Here, three aldehyde-functionalized crosslinkers (i.e., aldehyde-modified poly(xylitol sebacate)-co-poly(ethylene glycol) (APP), glutaraldehyde (GA), and polydextran aldehyde (PDA)) are used to react with quaternized chitosan (QCS) through imine bonds to form hydrogels. The microstructures, mechanical performances, and cytocompatibility of the three hydrogels are systematically investigated. The APP/QCS hydrogels presented the best compressive and stretch properties among the three hydrogels. The mechanical property and antibacterial activity of APP/QCS hydrogels can be further modulated using varied QCS amounts, where more QCS contributed higher stiffness and stretchability as well as better bacterial inhibition to the APP/QCS hydrogels. Taken together, it is demonstrated that the inherent elastomeric characteristic of APP crosslinker provides the desirable elasticity and stretchability to QCS hydrogels compared to the other aldehyde-functionalized crosslinkers of GA and PDA. This strategy of using multivalent elastomeric crosslinkers to fabricate deformable chitosan hydrogels can expand the use of chitosan hydrogels in tissue engineering applications.

Gelatin Methacrylate Hydrogel for Tissue Engineering Applications-A Review on Material Modifications.

To recreate or substitute tissue in vivo is a complicated endeavor that requires biomaterials that can mimic the natural tissue environment. Gelatin methacrylate (GelMA) is created through covalent bonding of naturally derived polymer gelatin and methacrylic groups. Due to its biocompatibility, GelMA receives a lot of attention in the tissue engineering research field. Additionally, GelMA has versatile physical properties that allow a broad range of modifications to enhance the interaction between the material and the cells. In this review, we look at recent modifications of GelMA with naturally derived polymers, nanomaterials, and growth factors, focusing on recent developments for vascular tissue engineering and wound healing applications. Compared to polymers and nanoparticles, the modifications that embed growth factors show better mechanical properties and better cell migration, stimulating vascular development and a structure comparable to the natural-extracellular matrix.