Synthesis,characterization, and cytotoxicity of PCL–PEG–PCL diacrylate and agarose interpenetrating network hydrogels for cartilage tissue engineering

Hydrogels are suitable biomaterials for cartilage tissue engineering due to the excellent ability to retain water to provide suitable environment for the tissue, however, the insufficient mechanical properties often prevent their wider applications. The objective of this study was to fabricate biocompatible hydrogels with good mechanical performance, high-water content, and porous microstructure for cartilage regeneration. Photocrosslinked hydrogels are one of the most widely used systems in tissue engineering due to the superior mechanical properties. In this study, block copolymer, poly(ε -caprolactone)-poly(ethylene)-poly(ε -caprolactone) diacrylate (PCL–PEG–PCL; PEC), was prepared by ring-opening polymerization, and PEC hydrogels were made through free radical crosslinking mechanism. Agarose network is chosen as another component of the hydrogels, because of the high-swelling behavior and cartilage-like microstructure, which is helpful for chondrocytes growth. Interpenetrating networks (IPN) were fabricated by diffusing PEC into agarose network followed by photo-crosslinking process. It was noted that incorporating PEC into the agarose network increased the elastic modulus and the compressive failure properties of individual component networks. In addition, high-swelling ratio and uniform porosity microstructures were found in the IPN hydrogels. IPN and PEC showed low cytotoxicity and good biocompatibility in elution test method. The results suggest promising characteristics of IPN hydrogels as a potential biomaterial for cartilage tissue engineering.     

Surface treatment with amino acids of porous collagen based scaffolds to improve cell adhesion and proliferation

The purpose of this study was to improve the biocompatibility of glutaraldehyde (GA) cross‐linked chitosan coated collagen scaffold for cartilage tissue regeneration. In order to prevent the potential toxicity of GA, we treated the designed scaffold with either glutamic acid or glycine. Amino acid treated scaffolds were characterized by scanning electron microscopy (SEM) techniques. Afterward, chondrocyte interaction with the composite scaffold was investigated assessing cell adhesion and proliferation using Hoechst staining and MTT cell proliferation assay, respectively. The SEM analyses of the scaffolds’ surface and cross‐section confirmed the adhesion of amino acids on the surface of the scaffolds. We also observed that scaffolds’ porosity was reduced due to the coverage of the pores by chitosan and amino acids, leading to low porosity. The use of amino acid improved the chondrocyte adhesion and proliferation inside the scaffolds’ pores when cells were cultured onto the chitosan‐coated collagen scaffolds. Overall, our in vitro results suggest the use of amino acid to improve the biocompatibility of natural polymer composite scaffold being crosslinked with glutaraldehyde. Such scaffold has improved mechanical properties; biocompatibility thus may be useful for tissue regeneration such as cartilage.

     

Hydrogels for Engineering of Perfusable Vascular Networks

Hydrogels are commonly used biomaterials for tissue engineering. With their high-water content, good biocompatibility and biodegradability they resemble the natural extracellular environment and have been widely used as scaffolds for 3D cell culture and studies of cell biology. The possible size of such hydrogel constructs with embedded cells is limited by the cellular demand for oxygen and nutrients. For the fabrication of large and complex tissue constructs, vascular structures become necessary within the hydrogels to supply the encapsulated cells. In this review, we discuss the types of hydrogels that are currently used for the fabrication of constructs with embedded vascular networks, the key properties of hydrogels needed for this purpose and current techniques to engineer perfusable vascular structures into these hydrogels. We then discuss directions for future research aimed at engineering of vascularized tissue for implantation.     

Biodegradable-Glass-Fiber Reinforced Hydrogel Composite with Enhanced Mechanical Performance and Cell Proliferation for Potential Cartilage Repair

Polyvinyl alcohol (PVA) hydrogels are promising implants due to the similarity of their low-friction behavior to that of cartilage tissue, and also due to their non-cytotoxicity. However, their poor mechanical resistance and insufficient durability restricts their application in this area. With the development of biodegradable glass fibers (BGF), which show desirable mechanical performance and bioactivity for orthopedic engineering, we designed a novel PVA hydrogel composite reinforced with biodegradable glass fibers, intended for use in artificial cartilage repair with its excellent cytocompatibility and long-term mechanical stability. Using structure characterization and thermal properties analysis, we found hydrogen bonding occurred among PVA molecular networks as well as in the PVA–BGF interface, which explained the increase in crystallinity and glass transition temperature, and was the reason for the improved mechanical performance and better anti-fatigue behavior of the composites in comparison with PVA. The compressive strength and modulus for the PBGF-15 composite reached 3.05 and 3.97 MPa, respectively, equaling the mechanical properties of human articular cartilage. Moreover, the increase in BGF content was found to support the proliferation of chondrocytes in vitro, whilst the PVA hydrogel matrix was able to control the ion concentration by adjusting the ions released from the BGF. Therefore, this novel biodegradable-glass-fiber-reinforced hydrogel composite possesses excellent properties for cartilage repair with potential in medical application.     

Glycerol-modified poly(vinyl alcohol)/poly(ethylene glycol) self-healing hydrogel for artificial cartilage

Poly(vinyl alcohol) (PVA) hydrogels have shown potential applications in bionic articular cartilage due to their tissue-like viscoelasticity, good biocompatibility and low friction. However, their lack of adequate mechanical properties is a key obstacle for PVA hydrogels to replace natural cartilage. In this study, poly(ethylene glycol) (PEG) and glycerol were introduced into PVA, and a PVA/PEG–glycerol composite hydrogel was synthesized using a mixing physical crosslinking method. The mechanical properties, hydrophilicity and tribological behavior of the PVA/PEG–glycerol hydrogel were investigated by changing the concentration of glycerol in PEG. The results showed that the tensile strength of the hydrogel reached 26.6 MPa at 270% elongation at break with 20 wt% of glycerol plasticizer, which satisfied the demand of natural cartilage. In addition, the excellent hydrophilicity of glycerol provides good lubricating properties for the composite gel under dry friction. Meanwhile, self-healing and cellular immunity assays demonstrated that the composite gel could have good self-healing ability and excellent biocompatibility even in the absence of external stimuli. This study provides a new candidate material for the design of articular cartilage, which has the potential to facilitate advances in artificial joint cartilage repair. © 2022 Society of Industrial Chemistry.     

Enhanced function of chondrocytes in a chitosan-based hydrogel to regenerate cartilage tissues by accelerating degradability of the hydrogel via a hydrolysable crosslinker

Chitosan-based hydrogels as scaffolds for culturing chondrocytes were prepared using linkers with and without hydrolysable poly(dl -lactide) (PLA) segments. The evaluation of the cultured chondrocytes in them indicated that the accelerated degradation of the hydrogel via hydrolysis of the PLA slightly promoted production of the sulfated glycosaminoglycan and drastically improved that of collagen from the encapsulated chondrocytes, which are the chondrospecific extracellular matrix components. Furthermore, the accelerated degradability significantly upregulated the gene expression for Collagen II production and downregulated that for Collagen I production of the encapsulated chondrocytes. Because major component of the articular cartilage tissue is Collagen II-rich hyaline cartilage, these results suggest the degradation of the scaffolds is an important parameter in cartilage tissue regeneration and the accelerated degradability may have benefits on promotion of cartilage tissue regeneration especially from the viewpoint of hyaline cartilage-like collagen production. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 137, 48893.     

Thiol–norbornene photoclick hydrogels for tissue engineering applications

Thiol–norbornene (thiol–ene) photoclick hydrogels have emerged as a diverse material system for tissue engineering applications. These hydrogels are crosslinked through light‐mediated orthogonal reactions between multifunctional norbornene‐modified macromers [e.g., poly(ethylene glycol) (PEG), hyaluronic acid, gelatin] and sulfhydryl‐containing linkers (e.g., dithiothreitol, PEG–dithiol, biscysteine peptides) with a low concentration of photoinitiator. The gelation of thiol–norbornene hydrogels can be initiated by long‐wave UV light or visible light without an additional coinitiator or comonomer. The crosslinking and degradation behaviors of thiol–norbornene hydrogels are controlled through material selections, whereas the biophysical and biochemical properties of the gels are easily and independently tuned because of the orthogonal reactivity between norbornene and the thiol moieties. Uniquely, the crosslinking of step‐growth thiol–norbornene hydrogels is not oxygen‐inhibited; therefore, gelation is much faster and highly cytocompatible compared with chain‐growth polymerized hydrogels with similar gelation conditions. These hydrogels have been prepared as tunable substrates for two‐dimensional cell cultures as microgels and bulk gels for affinity‐based or protease‐sensitive drug delivery, and as scaffolds for three‐dimensional cell encapsulation. Reports from different laboratories have demonstrated the broad utility of thiol–norbornene hydrogels in tissue engineering and regenerative medicine applications, including valvular and vascular tissue engineering, liver and pancreas‐related tissue engineering, neural regeneration, musculoskeletal (bone and cartilage) tissue regeneration, stem cell culture and differentiation, and cancer cell biology. This article provides an up‐to‐date overview on thiol–norbornene hydrogel crosslinking and degradation mechanisms, tunable material properties, and the use of thiol–norbornene hydrogels in drug‐delivery and tissue engineering applications. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41563.     

Needle‐punched nonwoven matrix from regenerated collagen fiber for cartilage tissue engineering

In this study, a new application of needle‐punched three‐dimensional (3D) fiber matrix from regenerated collagen fiber in articular cartilage tissue engineering (TE) was developed. Scanning electron microscopy images showed that the arrangement of fibers well mimicked the transitional part of the zonal articular cartilage. The 3D matrices exhibited a high porosity (93.5 ± 2.3%) and a large pore size range from about 20 μm in the inner part to 200–300 μm on the surface. The interconnected pore structure and hydrophilicity of the fibers led to the rapid and desired water uptake capacity of the matrices. Although the tensile and compressive properties of the scaffold were slightly lower than those of the natural articular cartilage, the anisotropic and nonlinear tension–compression were highly similar. In vitro human bone marrow stromal cells proliferation and cell morphology revealed the well cytocompatibility of the matrix, indicating its great potential in articular cartilage TE. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40404.     

La3+ modified poly(γ-glutamic acid) hydrogels with high strength and anti-swelling property for cartilage regeneration

The applications of synthetic hydrogels in cartilage regeneration are usually limited by their weak mechanical properties, uncontrolled swelling/degradation, and insufficient osteogenic activity. Developing tough hydrogels have been attracting great attention in biomedical engineering. In this study, a high strength and tough poly(γ-glutamic acid) (γ-PGA) hydrogels with excellent anti-swelling property were developed by immersing as-prepared γ-PGA hydrogels in LaCl₃ aqueous solution. Results revealed that the concentration of LaCl₃ aqueous solution has great influence on the mechanical properties of γ-PGA hydrogels. The tensile strength of γ-PGA hydrogels improved from 0.12 ± 0.02 MPa to 14.65 ± 0.48 MPa when LaCl₃ concentration was 0.15 M. Moreover, the swelling ratio decreased from 1035.75 ± 33.16% to 18.21 ± 3.08%. The morphology and microstructure of La³⁺ reinforced γ-PGA hydrogels were characterized by SEM/EDS, FT-IR and XPS. Furthermore, in vitro cytocompatibility of La³⁺ reinforced γ-PGA hydrogels was evaluated via MC3T3-E1 cells. Finally, this study provides a facile and effective strategy for modifying the mechanical and swelling properties of γ-PGA-based hydrogels, which offers great potential applications in cartilage repair and regeneration.     

The synthesis,mechanisms, and additives for bio-compatible polyvinyl alcohol hydrogels: A review on current advances,trends, and future outlook

Vinyl polymers are widely used in biological, textile and industrial applications and are currently attracting research attention for specialized bio-based applications. Polyvinyl alcohol (PVA) hydrogels show great advantages as a material with high biocompatibility, permeability, hydrophilicity, and low-friction coefficient, allowing applications as smart materials, wound dressings, and flexible sensors. However, the poor mechanical properties of PVA hydrogels and biocompatibility less than natural polymers make them unsuitable in practical applications. Additives are often added to PVA hydrogels to enhance mechanical properties, endow more compatibility, functionality and expand their application range. Among them, bio-additives such as nanocellulose, natural polysaccharides and proteins are biodegradable, biocompatible, and inexpensive, broadening their applications in the biomedical and tissue engineering fields. This work reviews the synthesis of PVA hydrogels, methods to enhance their mechanical properties, types of bio-additives incorporated for biocompatibility, their mechanism of interaction with PVA and future prospects of PVA composite bio-hydrogels for application in various fields. Representative cases are carefully selected and discussed with regard to their composition and pros and cons are discussed. Finally, future requirements, as well as the opportunities and challenges of these bio-additives for improving the multifunctionality of PVA hydrogels are also presented.     

An Injectable Enzymatically Crosslinked and Mechanically Tunable Silk Fibroin/Chondroitin Sulfate Chondro-Inductive Hydrogel

An injectable hybrid hydrogel is synthesized, comprising silk fibroin (SF) and chondroitin sulfate (CS) through di-tyrosine formation bond of SF chains. CS and SF are reported with excellent biocompatibility as tissue engineering scaffolds. Nonetheless, the rapid degradation rate of pure CS scaffolds presents a challenge to effectively recreate articular cartilage. As CS is one of the cartilage extracellular matrix (ECM) components, it has the potential to enhance the biological activity of SF-based hydrogel in terms of cartilage repair. Therefore, altering the CS concentrations (i.e., 0 wt%, 0.25 wt%, 0.5 wt%, 1 wt%, and 2 wt%), which are interpenetrated between SF β-sheets and chains, can potentially adjust the physical, chemical, and mechanical features of these hybrid hydrogels. The formation of β-sheets by 30 days of immersion in de-ionized (DI) water can improve the compression strength of the SF/CS hybrid hydrogels in comparison with the same SF/CS hybrid hydrogels in the dried state. Biological investigation and observation depicts proper cell attachment, proliferation and cell viability for C28/I2 cells. Gene expression of sex-determining region YBox 9 (SOX9), Collagen II α1, and Aggrecan (AGG) exhibits positive C3H10T1/2 growth and expression of cartilage-specific genes in the 0.25 wt% and 0.5 wt% SF/CS hydrogels.     

Polymer-based hydrogel scaffolds for skin tissue engineering applications: a mini-review

Polymer hydrogels consist of a three-dimensional (3D) structure with cross-linked networks rich in a huge amount of water through hydrogen-bonding interactions, making them highly hydrophilic. Due to their impressive hydrophilic characteristics and cell non-cytotoxicity, polymer hydrogels are useful tissue engineering tools for the organization of cells and tissues and organ regeneration. Many biomedical engineers and researchers have recently begun to utilize polymer hydrogels as tissue or cell culture environments and as scaffolds for the stable growth of organs in tissue engineering and regeneration medicine. This paper focuses on skin regeneration in polymer hydrogels where skin is a means of protecting the body from infection or physical or chemical damage. Generally, skin tissue that has incurred minor damage or wounds can regenerate and heal in a relatively short time, while severe injuries may require transplantation or artificial skin. For those purposes, skin culturing in an in vitro environment is essential, and the environment produced using polymer hydrogel scaffolds needs to be both similar to the real environment and safe for skin cell growth. This paper reviews post-2000 skin regeneration research in the field of tissue engineering, focusing specifically on polymer hydrogels; it also discusses some of the central perspectives and key issues.     

Chondrogenic stimulation in mesenchymal stem cells using scaffold-based sustained release of platelet-rich plasma

Developing minimal invasive strategies via injectable hydrogels for effective repairing of cartilage defects is highly desired. Injectable hydrogels, which can simultaneously embed cell and growth factors (GFs), serve as in situ formed scaffolds and could support an accelerated tissue regeneration process. The purpose of this study is to fabricate a composite injectable hydrogel, based on alginate (Alg)/polyvinyl alcohol (PVA) incorporating platelet rich plasma (PRP)-encapsulated Alg sulfate (AlgS) microbeads, as a localized sustained release system of GFs, for the articular cartilage regeneration. The results show that synthesized AlgS microbeads support the sustained release of PRP GFs during 14 days, where preserve the bioactivity of them more than the free PRP. Rabbit adipose-derived mesenchymal stem cells in contact with PRP-loaded AlgS beads show more proliferation (2.7 folds) and have obviously higher deposition of collagen type ΙΙ and GAGs than free PRP treated ones. In addition, cells encapsulated into the hydrogel including PRP sustained release system show upregulated expression of collagen type ΙΙ (61 folds), Aggrecan (294 folds) and SOX9 (71.5 folds), as cartilage-critical genes, compared to the direct treatment by PRP. To summarize, the developed hybrid Alg/PVA hydrogel embedding with PRP-encapsulated AlgS microbeads is suggested as a potential in situ formed scaffold for cartilage regeneration.     

Chemical synthesis of biomimetic hydrogels for tissue engineering

Owing to their high water content, porous structure, biocompatibility and tissue‐like viscoelasticity, hydrogels have become attractive and promising biomaterials for use in drug delivery, three‐dimensional cell culture and tissue engineering applications. Various chemical approaches have been developed for hydrogel synthesis using monomers or polymers carrying reactive functional groups. For in vivo tissue repair and in vitro cell culture purposes, it is desirable that the crosslinking reactions occur under mild conditions, do not interfere with biological processes and proceed at high yield with exceptional selectivity. Additionally, the crosslinking reaction should allow straightforward incorporation of bioactive motifs or signaling molecules, at the same time providing tunability of the hydrogel microstructure, mechanical properties and degradation rates. In this review, we discuss various chemical approaches applied to the synthesis of complex hydrogel networks, highlighting recent developments from our group. The discovery of new chemistries and novel materials fabrication methods will lead to the development of the next generation of biomimetic hydrogels with complex structures and diverse functionalities. These materials will likely facilitate the construction of engineered tissue models that may bridge the gap between two‐dimensional experiments and animal studies, providing preliminary insight prior to in vivo assessments. © 2017 Society of Chemical Industry     

Self-healable polyacrylic acid-polyacrylamide-ferric ion dual-crosslinked hydrogel with good biotribological performance as a load-bearing surface

Although having been widely investigated, polymer hydrogels still have many defects like poor tribological properties and insufficient durability, hindering their further applications in biomedical fields. In this study, we present a simple method to synthesize polyacrylic acid-polyacrylamide-ferric ion (PAA-PAAm-Fe³⁺) dual-crosslinked hydrogels with self-healing abilities and “soft-hard” hydrogel-polyetheretherketone (PEEK) combined load-bearing surfaces with low friction coefficients. After analytical characterizations, the results demonstrated that the hydrogels could repair themselves without any external stimuli. Because of the excellent biphasic and aqueous lubrication provided by the hydrogel layer and the load-bearing capacity provided by the PEEK substrate, the friction coefficient of a load-bearing surface was as low as 0.048 in water, much lower than a pristine PEEK block or a hydrogel block sample. This work fabricated self-healable PAA-PAAm-Fe³⁺ hydrogels and low friction bearing surfaces, successfully improving the tribology properties of hydrogels, hopefully promoting their applications as biomedical materials such as articular cartilage. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48499.     

功能性明胶基水凝胶的分类及研究进展

明胶基水凝胶作为一类具有三维网络结构的天然高分子软物质材料,因具有良好的生物相容性、生物可降解性以及生物安全性,且含水量高、结构和性能与细胞外基质相似而受到研究者的广泛关注.该文概述了明胶基水凝胶的结构与性质,并按功能性将明胶基水凝胶进行分类,重点阐述了自修复型、抗菌型、刺激响应型、导电型以及抗冻型明胶基水凝胶的特点、...     

Alginate Based Scaffolds for Cartilage Tissue Engineering: A Review

Abstract

Many people, especially old and middle-aged, suffer from pain and disabilities caused by cartilage degradation. There are many surgical methods for cartilage treatment, however, none of them have shown acceptable results in long-term. Tissue engineering would be an acceptable approach for cartilage treatment. This includes cells, a carrier such as a matrix scaffold and signaling molecule. An ideal scaffold for cartilage tissue engineering should meet some requirements includes biocompatibility, biodegradability, and sufficient mechanical characteristic. While there are many suitable scaffolds made by natural and synthesis polymers, alginate- a natural polymer- has received good attention. Alginate offers many advantages for cartilage treatment; it has great potential in having tunable mechanical properties and easy manufacturing process. In the present paper, focusing on alginate as main scaffold constructive component, different studies on alginate based scaffolds in the form of physically, chemically and biologically crosslinked hydrogel, sponge, fiber, micro/nano particles and 3?D printed for articular cartilage tissue engineering are discussed and reviewed.     

Biocomposites: Their multifunctionality

During the last decade, tissue engineering has shown a considerable promise in providing more viable alternatives to surgical procedures for harvested tissues, implants and prostheses. Due to the fast development on nano- and biomaterial technologies, it is now possible for doctors to use patients' cells to repair orthopaedic defects such as focal articular cartilage lesions. In order to support the three-dimensional tissue formation, scaffolds made by biocompatible and bioresorbable polymers and composite materials, for providing temporary support of damaged body and cell structures, have been developed recently. Although ceramic and metallic materials have been widely accepted for the development of implants, their non-resorbability and necessity of second surgical operation (like for bone repair), which induce extra pain for the patients, limit their wide applications. The development of different types of biocomposites for biomedical engineering applications is described. These biocomposites include (i) basic biomaterials; (ii) natural fiber-reinforced biocomposites and (iii) nanoparticle-reinforced biocomposites. Their multifunctionality is discussed in terms of the control of mechanical properties, biodegradability and bioresorbability.     

Enzymatic and ionic crosslinked gelatin/K‐carrageenan IPN hydrogels as potential biomaterials

Hydrogels of a natural origin have attracted considerable attention in the field of tissue engineering due to their resemblance to ECM, defined degradability and compatibility with biological systems. In this study, we introduced carrageenan into a gelatin network, creating IPN hydrogels through biological methods of enzymatic and ionic crosslinking. Their gelation processes were monitored and confirmed by rheology analysis. The combination of biochemical and physical crosslinking processes enables the formation of biohydrogels with tunable mechanical properties, swelling ratios and degradation behaviors while maintaining the biocompatibilities of natural materials. The mechanical strength increased with an increase in carrageenan content while swelling ratio and degradability decreased correspondingly. In addition, the IPN hydrogels were shown to support adhesion and proliferation of L929 cell line. All the results highlighted the use of biological crosslinked gelatin‐carrageenan IPN hydrogels in the context of tissue engineering. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 10.1002/app.40975.     

An assessment of biopolymer‐ and synthetic polymer‐based scaffolds for bone and vascular tissue engineering

The promise of tissue engineering is the combination of a scaffold with cells to initiate the regeneration of tissues or organs. Engineering of scaffolds is critical for success and tailoring of polymer properties is essential for their good performance. Many different materials of natural and synthetic origins have been investigated, but the challenge is to find those that have the right mix of mechanical performance, biodegradability and biocompatibility for biological applications. This article reviews key polymeric properties for bone and vascular scaffold eligibility with focus on biopolymers, synthetic polymers and their blends. The limitations of these polymeric systems and ways and means to improve scaffold performance specifically for bone and vascular tissue engineering are discussed. © 2013 Society of Chemical Industry