Scaffolds For Cartilage Tissue Engineering Research Articles

Silk Fibroin Films with Microscopic Surface Pattern as Promising Cartilage Tissue Engineering Scaffold in Supporting MSCs Amplification and Chondrogenic Differentiation

ABSTRACT Mesenchymal stem cells (MSCs) with multipotential differentiation ability are main cell source for cartilage tissue engineering, but its application is restricted by the difficulty in regulating committed chondrogenic differentiation. Study revealed that certain substrate topographic structure can enhance the chondrogenic differentiation of MSCs. This study prepared silk fibroin (SF) films with gradient microscopic surface patterns based on P280-P7000 type specification sandpapers, and investigated the adhesion, proliferation and chondrogenic differentiation of human adipose-derived stem cells (hADSCs) on the SF films. The results showed that the SF films surface had convex dots with diameter of 3–50 μm, and hADSCs could adhere and proliferate well on the film. Transcriptomics analysis indicated that hADSCs tend to differentiate differently on the SF films, among which hADSCs on P1000 SF films had higher expression of chondrogenic markers Sox-9 and Col-II, with activated yes associated protein (YAP) and restricted F-actin. In vitro co-culture of human OA knee cartilage with hADSCs seeded SF films found satisfactory cartilage protection effect in P1000 group. Overall, this study preliminarily revealed the microscopic surface pattern of SF films may induce chondrogenic differentiation of hADSCs through extracellular matrix mediated Hippo/Yap pathway, and provided a new strategy for cartilage repair.

Nanoenabled Immunomodulatory Scaffolds for Cartilage Tissue Engineering

AbstractArticular cartilage regeneration is a challenge in tissue engineering. Although diverse materials have been developed for this purpose, cartilage regeneration remains suboptimal. The integration of nanomaterials into 3D network materials holds great potential in the improvement of key mechanical properties, particularly important for osteochondral replacement scaffolds and even to function as carriers for disease‐modifying drugs or other regulatory signals. In this study, a simple yet effective cell‐free nanoenabled Col‐PLA scaffold specially designed to enhance cartilage regeneration and modulate inflammatory response is proposed, by incorporating poly(lactic‐co‐glycolic acid) (PLGA) ibuprofen nanoparticles (NPs) into a collagen/polylactide (Col‐PLA) matrix. The developed nanoenabled scaffold successfully decreases IL‐1β release and leads to primary human chondrocytes survival, ultimately restoring extracellular matrix (ECM) production under inflammatory conditions. The nanoenabled Col‐PLA scaffolds secretome effectively decreases macrophage invasion in vitro, as well as neutrophil infiltration and inflammatory mediators’, namely the complement component C5/C5a, C‐reactive protein, IL‐1β, MMP9, CCL20, and CXCL1/KC production in vivo in a rodent air‐pouch model. Overall, the established nanoenabled scaffold has the potential to support chondrogenesis as well as modulate inflammatory response, overcoming the limitations of traditional tissue engineering strategies.

The potential of an electrospun poly (3-hydroxybutyrate)/poly (ethylene glycol) blend fibrous scaffold for cartilage tissue engineering

Poly (3-hydroxybutyrate) and poly (ethylene glycol) are frequently used as materials for designing scaffolds in tissue engineering. This work aimed the production of P3HB-PEG scaffolds in different weight ratio of PEG using electrospinning and characterized. Samples also were examined for rabbit chondrocyte cell assay. 5 wt% of PEG decreased the nanofiber diameter with more uniformity and enhanced hydrophilicity. The biodegradation rate increased while the mechanical features tended to decrease. In vitro tests showed good chondrocyte cell viability and adhesion. These results manifested that electrospun the P3HB-PEG scaffolds could enhance cartilage regeneration, showing their marvelous prospect as scaffolds for cartilage tissue engineering.

Construction of Integral Decellularized Cartilage Using a Novel Hydrostatic Pressure Bioreactor.

The decellularized extracellular matrix (ECM) of cartilage is a widely used natural bioscaffold for constructing tissue-engineered cartilage due to its good biocompatibility and regeneration properties. However, current decellularization methods for accessing decellularized cartilaginous tissues require multiple steps and a relatively long duration to produce decellularized cartilage. In addition, most decellularization strategies lead to damage of the microstructure and loss of functional components of the cartilaginous matrix. In this study, a novel decellularization strategy based on a hydrostatic pressure (HP) bioreactor was introduced, which aimed to improve the efficiency of producing integral decellularized cartilage pieces by combining physical and chemical decellularization methods in a perfusing manner. Two types of cartilaginous tissues, auricular cartilage (AC) and nucleus pulposus (NP) fibrocartilage, were selected for comparison of the effects of ordinary, positive, and negative HP-based decellularization according to the cell clearance ratio, microstructural changes, ECM components, and mechanical properties. The results indicated that applying positive HP improved the efficiency of producing decellularized AC, but no significant differences in decellularization efficiency were found between the ordinary and negative HP-treated groups. However, compared with the ordinary HP treatment, the application of the positive or negative HP did not affect the efficiency of decellularized NP productions. Moreover, neither positive nor negative HP influenced the preservation of the microstructure and components of the AC matrix. However, applying negative HP disarranged the fibril distribution of the NP matrix and reduced glycosaminoglycans and collagen type II contents, two essential ECM components. In addition, the positive HP was beneficial for maintaining the mechanical properties of decellularized cartilage. The recellularization experiments also verified the good biocompatibility of the decellularized cartilage produced by the present bioreactor-based decellularization method under positive HP. Overall, applying positive HP-based decellularization resulted in a superior effect on the production of close-to-natural scaffolds for cartilage tissue engineering. Impact statement In this study, we successfully constructed a novel hydrostatic pressure (HP) bioreactor and used this equipment to produce decellularized cartilage by combining physical and chemical decellularization methods in a perfusing manner. We found that positive HP-based decellularization could improve the production efficiency of integral decellularized cartilage pieces and promote the maintenance of matrix components and mechanical properties. This new decellularization strategy exhibited a superior effect in the production of close-to-natural scaffolds and positively impacts cartilage tissue engineering.

Biomimetic Injectable Hydrogel Based on Methacrylate-Modified Silk Fibroin Embedded with Kartogenin for Superficial Cartilage Regeneration.

The weak regeneration ability of chondrocytes is one of the main reasons that limit the therapeutic effect of clinical cartilage injury. Injectable hydrogels are potential scaffolds for cartilage tissue engineering with advantages such as minimally invasive surgery, porous structure, and drug sustained-release ability. At present, many biomaterials have been developed for the repair of deep cartilage defects. However, cartilage injury often begins on the surface, which requires us to propose a treatment strategy suitable for superficial cartilage injury repair. In this study, we fabricated a biomimetic injectable hydrogel based on methacrylate-modified silk fibroin (SilMA) embedded with kartogenin (KGN). The SilMA/KGN hydrogels have good biohistocompatibility and the ability to promote cartilage differentiation. In addition, SEM results show that it has a porous structure conducive to cell adhesion and proliferation. Most importantly, it has demonstrated remarkable superficial cartilage repair ability in vivo, showing potential in cartilage tissue engineering.

In Vitro Degradation of 3D-Printed Poly(L-lactide-Co-Glycolic Acid) Scaffolds for Tissue Engineering Applications.

The creation of scaffolds for cartilage tissue engineering has faced significant challenges in developing constructs that can provide sufficient biomechanical support and offer suitable degradation characteristics. Ideally, such tissue-engineering techniques necessitate the fabrication of scaffolds that mirror the mechanical characteristics of the articular cartilage while degrading safely without damaging the regenerating tissues. The aim of this study was to create porous, biomechanically comparable 3D-printed scaffolds made from Poly(L-lactide-co-glycolide) 85:15 and to assess their degradation at physiological conditions 37 °C in pH 7.4 phosphate-buffered saline (PBS) for up to 56 days. Furthermore, the effect of scaffold degradation on the cell viability and proliferation of human bone marrow mesenchymal stem cells (HBMSC) was evaluated in vitro. To assess the long-term degradation of the scaffolds, accelerated degradation tests were performed at an elevated temperature of 47 °C for 28 days. The results show that the fabricated scaffolds were porous with an interconnected architecture and had comparable biomechanical properties to native cartilage. The degradative changes indicated stable degradation at physiological conditions with no significant effect on the properties of the scaffold and biocompatibility of the scaffold to HBMSC. Furthermore, the accelerated degradation tests showed consistent degradation of the scaffolds even in the long term without the notable release of acidic byproducts. It is hoped that the fabrication and degradation characteristics of this scaffold will, in the future, translate into a potential medical device for cartilage tissue regeneration.

3D-printed fish gelatin scaffolds for cartilage tissue engineering.

Knee osteoarthritis is a chronic disease caused by the deterioration of the knee joint due to various factors such as aging, trauma, and obesity, and the nonrenewable nature of the injured cartilage makes the treatment of osteoarthritis challenging. Here, we present a three-dimensional (3D) printed porous multilayer scaffold based on cold-water fish skin gelatin for osteoarticular cartilage regeneration. To make the scaffold, cold-water fish skin gelatin was combined with sodium alginate to increase viscosity, printability, and mechanical strength, and the hybrid hydrogel was printed according to a pre-designed specific structure using 3D printing technology. Then, the printed scaffolds underwent a double-crosslinking process to enhance their mechanical strength even further. These scaffolds mimic the structure of the original cartilage network in a way that allows chondrocytes to adhere, proliferate, and communicate with each other, transport nutrients, and prevent further damage to the joint. More importantly, we found that cold-water fish gelatin scaffolds were nonimmunogenic, nontoxic, and biodegradable. We also implanted the scaffold into defective rat cartilage for 12 weeks and achieved satisfactory repair results in this animal model. Thus, cold-water fish skin gelatin scaffolds may have broad application potential in regenerative medicine.

Evaluation of the effects of chitosan nanoparticles on polyhydroxy butyrate electrospun scaffolds for cartilage tissue engineering applications

In this study, we synthesized and incorporated chitosan nanoparticles (Cs) into polyhydroxy butyrate (PHB) electrospun scaffolds for cartilage tissue engineering. The Cs nanoparticles were synthesized via an ionic gel interaction between Cs powder and tripolyphosphate (TPP). The mechanical properties, hydrophilicity, and fiber diameter of the PHB scaffolds with varying concentrations of Cs nanoparticles (1–5 wt%) were evaluated. The results of these evaluations showed that the scaffold containing 1 wt% Cs nanoparticles (P1Cs) was the optimum scaffold, with increased ultimate strength from 2.6 to 5.2 MPa and elongation at break from 5.31 % to 12.6 %. Crystallinity, degradation, and cell compatibility were also evaluated. The addition of Cs nanoparticles decreased crystallinity and accelerated hydrolytic degradation. MTT assay results showed that the proliferation of chondrocytes on the scaffold containing 1 wt% Cs nanoparticles were significantly higher than that on pure PHB after 7 days of cultivation. These findings suggest that the electrospun P1Cs scaffold has promising potential as a substrate for cartilage tissue engineering applications. This combination offers a promising approach for the fabrication of biomimetic scaffolds with enhanced mechanical properties, hydrophilicity, and cell compatibility for tissue engineering applications.

Dynamically adaptive scaffolds for cartilage tissue engineering

AbstractAlthough cartilage regeneration is a continuously changing, complex process, most studies on cartilage tissue regeneration have focused on specific stages, thereby promoting regeneration in only a limited period rather than over the entire repair process. The ideal adaptive scaffolds for cartilage tissue engineering should exhibit dynamic and spatiotemporally controlled evolution according to the physiological microenvironments during each phase of cartilage repair. Moreover, scaffolds should have a specific feedback regulation capability, including long‐term feedback for the controlled delivery of multiple factors, which would better meet the requirements of cartilage regeneration. Adaptive scaffolds matching the whole evolving stages of cartilage repair are still under exploration. This review highlights the development of dynamically adaptive scaffolds to provide the appropriate microenvironments for cartilage regeneration, including scaffolds that manipulate the immune microenvironments from the inflammation stage to the regeneration stage, scaffolds that release cytokines on demand across several stages, and scaffolds that adapt according to the mechanical properties of extracellular matrices over time, and also discusses the future challenges and development directions of adaptive scaffolds in cartilage tissue regeneration. This understanding is expected to inspire the application of adaptive scaffolds for cartilage repair.

Poly-ε-caprolactone (PCL)/poly-l-lactic acid (PLLA) nanofibers loaded by nanoparticles-containing TGF-β1 with linearly arranged transforming structure as a scaffold in cartilage tissue engineering.

This study aimed to present a novel three-dimensional nanocomposite scaffold using poly-ε-caprolactone (PCL), containing transforming growth factor-beta 1 (TGF-β1)-loaded chitosan-dextran nanoparticles and poly-l-lactic acid (PLLA), to make use of nanofibers and nanoparticles simultaneously. The electrospinning method fabricated a bead-free semi-aligned nanofiber composed of PLLA, PCL, and chitosan-dextran nanoparticles containing TGF-β1. A biomimetic scaffold was constructed with the desired mechanical properties, high hydrophilicity, and high porosity. Transmission electron microscopy findings showed a linear arrangement of nanoparticles along the core of fibers. Based on the results, burst release was not observed. The maximum release was achieved within 4 days, and sustained release was up to 21 days. The qRT-PCR results indicated an increase in the expression of aggrecan and collagen type Ι genes compared to the tissue culture polystyrene group. The results indicated the importance of topography and the sustained release of TGF-β1 from bifunctional scaffolds in directing the stem cell fate in cartilage tissue engineering.

Chondrogenic Differentiation of Adipose-Derived Stromal Cells Induced by Decellularized Cartilage Matrix/Silk Fibroin Secondary Crosslinking Hydrogel Scaffolds with a Three-Dimensional Microstructure.

Finding an ideal scaffold is always an important issue in the field of cartilage tissue engineering. Both decellularized extracellular matrix and silk fibroin have been used as natural biomaterials for tissue regeneration. In this study, a secondary crosslinking method of γ irradiation and ethanol induction was used to prepare decellularized cartilage extracellular matrix and silk fibroin (dECM-SF) hydrogels with biological activity. Furthermore, the dECM-SF hydrogels were cast in custom-designed molds to produce a three-dimensional multi-channeled structure to improve internal connectivity. The adipose-derived stromal cells (ADSC) were seeded on the scaffolds, cultured in vitro for 2 weeks, and implanted in vivo for another 4 and 12 weeks. The double crosslinked dECM-SF hydrogels exhibited an excellent pore structure after lyophilization. The multi-channeled hydrogel scaffold presents higher water absorption ability, surface wettability, and no cytotoxicity. The addition of dECM and a channeled structure could promote chondrogenic differentiation of ADSC and engineered cartilage formation, confirmed by H&E, safranin O staining, type II collagen immunostaining, and qPCR assay. In conclusion, the hydrogel scaffold fabricated by the secondary crosslinking method has good plasticity and can be used as a scaffold for cartilage tissue engineering. The multi-channeled dECM-SF hydrogel scaffolds possess a chondrogenic induction activity that promotes engineered cartilage regeneration of ADSC in vivo.

STUDY OF SILK STRAIN AND ITS PREPARATION FOR MENISCUS TISSUE ENGINEERING SCAFFOLD

Silk fibroin (SF) has been used as a scaffold for cartilage tissue engineering. Different silkworms strain produced different protein. Also, molecular weight of SF depends on extraction method. We hypothesised that strain of silkworm and method of SF extraction would effect biological properties of SF scaffold. Therefore, cell viability and chondrogenic gene expression of human chondrogenic progenitor cells (HCPCs) treated with SF from 10 silkworm strains and two common SF extraction methods were investigate in this study.Twenty g of 10 strains silk cocoons were separately degummed in 0.02M Na2CO3 solution and dissolved in 100๐C for 30 minutes. Half of them were then dissolved in CaCl2/Ethanol/H2O [1:2:8 molar ratio] at 70±5๐C (method 1) and other half was dissolved in 46% w/v CaCl2 at 105±5๐C (method 2) for 4 hours. HCPCs were cultured in SF added cultured medial according to strain and extraction method. Cell viability at day 1, 3, and 7, were determined. Expression of collagen I, collagen II, and aggrecan at day 7 and 14, was studied. All experiment were done in triplicated samples.Generally, method 1 SF extraction showed higher cell viability in all strains. Cell viability from Nanglai Saraburi, Laung Saraburi and Nangtui strains were higher than those without SF in every time point while Wanasawan and J108 had higher viability at day 1 and decreased by time. Expression in collagen 1, collagen 2 and aggrecan in method 1 are higher at day 7 and day 14. Collagen 1 expression was highest in Nangnoi Srisaket, followed by Laung Saraburi and Nanglai Saraburi in day 7. Nangnoi Srisaket also had highest expression at day 14, followed by Nanglai Saraburi and Laung Saraburi respectively. Nangseaw had highest collagen 2 expression, follow by Laung Saraburi and Nangnoi Srisaket respectively. Higher aggrecan gene expression of Tubtimsiam, Wanasawan, UB 1 and Nangnoi Srisaket was observed at day 7 and increased expression of all strains at day 14.SF extraction using CaCl2/Ethanol/H2O offered better cell viability and chondrogenic expression. Nangseaw, Laung Saraburi and Nangnoi Srisaket strains expressed more chondrogenic phenotype.

Scaffolds for Cartilage Tissue Engineering from a Blend of Polyethersulfone and Polyurethane Polymers

In recent years, one of the main goals of cartilage tissue engineering has been to find appropriate scaffolds for hyaline cartilage regeneration, which could serve as a matrix for chondrocytes or stem cell cultures. The study presents three types of scaffolds obtained from a blend of polyethersulfone (PES) and polyurethane (PUR) by a combination of wet-phase inversion and salt-leaching methods. The nonwovens made of gelatin and sodium chloride (NaCl) were used as precursors of macropores. Thus, obtained membranes were characterized by a suitable structure. The top layers were perforated, with pores over 20 µm, which allows cells to enter the membrane. The use of a nonwoven made it possible to develop a three-dimensional network of interconnected macropores that is required for cell activity and mobility. Examination of wettability (contact angle, swelling ratio) showed a hydrophilic nature of scaffolds. The mechanical test showed that the scaffolds were suitable for knee joint applications (stress above 10 MPa). Next, the scaffolds underwent a degradation study in simulated body fluid (SBF). Weight loss after four weeks and changes in structure were assessed using scanning electron microscopy (SEM) and MeMoExplorer Software, a program that estimates the size of pores. The porosity measurements after degradation confirmed an increase in pore size, as expected. Hydrolysis was confirmed by Fourier-transform infrared spectroscopy (FT-IR) analysis, where the disappearance of ester bonds at about 1730 cm-1 wavelength is noticeable after degradation. The obtained results showed that the scaffolds meet the requirements for cartilage tissue engineering membranes and should undergo further testing on an animal model.

Double network microcrystalline cellulose hydrogels with high mechanical strength and biocompatibility for cartilage tissue engineering scaffold

Cartilage tissue regeneration is tremendously tough, it has become a major clinical challenge for the orthopedic medical community. Because of their bionic structure, high water content, biocompatibility, and biodegradability, hydrogels derived from natural polysaccharide are excellent candidates for cartilage tissue engineering. However, these materials often face problems such as poor mechanical strength and excessive swelling, which limit their clinical application. This study used a chemical-physical multi-step cross-linking strategy to create double-network (DN) microcrystalline cellulose (MCC) hydrogels. The hydrogels' intrinsic biomimetic macroporous shape and high water content made them ideal for chondrocyte adhesion and proliferation. The performance requirements for cartilage tissue engineering scaffolds are met by DN hydrogels, which have a sufficiently high compressive strength (4.53 MPa), superior compression recovery, and fatigue resistance, compared to single-network (SN) hydrogels. According to in vitro findings, DN hydrogels could boost cell adhesion and proliferation due to their safe and non-toxic nature. Hydrogels were demonstrated to be stable over the long-term performance, to degrade slowly, and to have strong histocompatibility by in vivo implantation. To construct cartilage tissue engineering scaffold and conduct three-dimensional cell culture, DN hydrogels have significant potential.

From materials to clinical use: advances in 3D-printed scaffolds for cartilage tissue engineering.

Osteoarthritis caused by articular cartilage defects is a particularly common orthopedic disease that can involve the entire joint, causing great pain to its sufferers. A global patient population of approximately 250 million people has an increasing demand for new therapies with excellent results, and tissue engineering scaffolds have been proposed as a potential strategy for the repair and reconstruction of cartilage defects. The precise control and high flexibility of 3D printing provide a platform for subversive innovation. In this perspective, cartilage tissue engineering (CTE) scaffolds manufactured using different biomaterials are summarized from the perspective of 3D printing strategies, the bionic structure strategies and special functional designs are classified and discussed, and the advantages and limitations of these CTE scaffold preparation strategies are analyzed in detail. Finally, the application prospect and challenges of 3D printed CTE scaffolds are discussed, providing enlightening insights for their current research.

Effects of decellularized extracellular matrix on Polyhydroxybutyrate electrospun scaffolds for cartilage tissue engineering

ABSTRACT The current study was carried out to evaluate the effects of cartilage-decellularized extracellular matrix (ECM) on Poly (3-hydroxybutyrate) (PHB) electrospun scaffold for cartilage tissue engineering. In this study, PHB scaffolds with three different percentages of ECM (1%, 2%, and 3%) were fabricated. Mechanical, chemical, and physical properties of scaffolds along with their interactions with cells were assessed. Results showed that by adding of ECM to the scaffolds, the Physiomechanical properties of combined scaffolds were modified, compared to the pure PHB scaffold. In the other hand, the biological behavior was increased by increasing the ECM ratio to the PHB.

Immunomodulatory PEG-CRGD Hydrogels Promote Chondrogenic Differentiation of PBMSCs.

Cartilage damage is a common injury. Currently, tissue engineering scaffolds with composite seed cells have emerged as a promising approach for cartilage repair. Polyethylene glycol (PEG) hydrogels are attractive tissue engineering scaffold materials as they have high water absorption capacity as well as nontoxic and nutrient transport properties. However, PEG is fundamentally bio-inert and lacks intrinsic cell adhesion capability, which is critical for the maintenance of cell function. Cell adhesion peptides are usually added to improve the cell adhesion capability of PEG-based hydrogels. The suitable cell adhesion peptide can not only improve cell adhesion capability, but also promote chondrogenesis and regulate the immune microenvironment. To improve the interactions between cells and PEG hydrogels, we designed cysteine-arginine-glycine-aspartic acid (CRGD), a cell adhesion peptide covalently cross-linked with PEG hydrogels by a Michael addition reaction, and explored the tissue-engineering hydrogels with immunomodulatory effects and promoted chondrogenic differentiation of mesenchymal stem cells (MSCs). The results indicated that CRGD improved the interaction between peripheral blood mesenchymal stem cells (PBMSCs) and PEG hydrogels. PEG hydrogels modified with 1 mM CRGD had the optimal capacity to promote chondrogenic differentiation, and CRGD could induce macrophage polarization towards the M2 phenotype to promote tissue regeneration and repair. PEG-CRGD hydrogels combined with PBMSCs have the potential to be suitable scaffolds for cartilage tissue engineering.

Effect of Glutaraldehyde Concentration Variation on Diclofenac Sodium Scaffolds as Cross-Linking Agent

Chitosan and gelatin were used as polymer scaffolds for cartilage tissue engineering. The scaffold was used as a biodegradable drug delivery system for diclofenac sodium to treat cartilage defects on osteoarthritis (OA). The materials were composed of diclofenac sodium, chitosan, gelatin, and cross-linking agent-glutaraldehyde (GTA) were form as scaffold. The purpose of this study to investigate the effect of GTA concentration variations (0.00%; 0.25%; 0.50%; 1.00%; 2.50%) on characteristics and the release of diclofenac sodium from chitosan-gelatin scaffold. The scaffolds were made by using the pre-freezing method with a temperature of -56 ± 5°C for 24 hours and characterized by porosity, pore size, swelling, degradation, toxicity test, and diclofenac sodium released from chitosan-gelatin scaffolds at pH and temperature body. The results showed, the addition of GTA increased the swelling ratio from 195.79 ± 7.04% to 793.49 ± 6.92% and minimized weight loss up to 50.98 ± 0.82%, percentage of living cells >60%, optimal porosity at 106.94 ± 9.38 % with pore size 135.48 ± 89.70 µm, diclofenac sodium as sustained release drug completed in 542 hours and the release was following zero-order kinetic. Chitosan-gelatin scaffold is a potential candidate for cartilage tissue engineering and drug delivery system for diclofenac sodium.

Enhancing cartilage repair with optimized supramolecular hydrogel-based scaffold and pulsed electromagnetic field.

Functional tissue engineering strategies provide innovative approach for the repair and regeneration of damaged cartilage. Hydrogel is widely used because it could provide rapid defect filling and proper structure support, and is biocompatible for cell aggregation and matrix deposition. Efforts have been made to seek suitable scaffolds for cartilage tissue engineering. Here Alg-DA/Ac-β-CD/gelatin hydrogel was designed with the features of physical and chemical multiple crosslinking and self-healing properties. Gelation time, swelling ratio, biodegradability and biocompatibility of the hydrogels were systematically characterized, and the injectable self-healing adhesive hydrogel were demonstrated to exhibit ideal properties for cartilage repair. Furthermore, the new hydrogel design introduces a pre-gel state before photo-crosslinking, where increased viscosity and decreased fluidity allow the gel to remain in a semi-solid condition. This granted multiple administration routes to the hydrogels, which brings hydrogels the ability to adapt to complex clinical situations. Pulsed electromagnetic fields (PEMF) have been recognized as a promising solution to various health problems owing to their noninvasive properties and therapeutic potentials. PEMF treatment offers a better clinical outcome with fewer, if any, side effects, and wildly used in musculoskeletal tissue repair. Thereby we propose PEMF as an effective biophysical stimulation to be 4th key element in cartilage tissue engineering. In this study, the as-prepared Alg-DA/Ac-β-CD/gelatin hydrogels were utilized in the rat osteochondral defect model, and the potential application of PEMF in cartilage tissue engineering were investigated. PEMF treatment were proven to enhance the quality of engineered chondrogenic constructs in vitro, and facilitate chondrogenesis and cartilage repair in vivo. All of the results suggested that with the injectable self-healing adhesive hydrogel and PEMF treatment, this newly proposed tissue engineering strategy revealed superior clinical potential for cartilage defect treatment.

Preparation of chitosan-sodium alginate/bioactive glass composite cartilage scaffolds with high cell activity and bioactivity

Chitosan-sodium alginate/bioactive glass (CSB) composite cartilage scaffold with outstanding in vitro mineralization property and cytocompatibility is synthesized by freeze drying method. The effect of bioactive glass (BG) addition on the microstructure, porosity, swelling/degradation ratio, in vitro mineralization property and cytocompatibility of CSB scaffold is investigated by the characterization techniques of SEM, XRD, FTIR and BET. Results showed that CSB composite cartilage scaffold had a three-dimensional (3D) porous structure, and both porosity and average pore size met the requirements of cartilage tissue repair. Among, the typical CSB-1.0 had the largest overall pore size and lowest compressive modulus (1.083 ± 0.002 MPa). As the amount of BG increased, pore volume and porosity of CSB scaffolds gradually decreased, and the swelling and degradation ratios gradually reduced. After immersing in SBF for 3 d, cauliflower like hydroxyapatite (HA) was formed on CSB surface, indicating that the scaffold had good in vitro mineralization property. Moreover, the introduction of BG into the composite scaffold can improve the relative cell viability of MC3T3-E1 cells, and CSB-1.0 has the strongest ability to promote the proliferation of cells. Therefore, the as-obtained CSB scaffold can be used as a strong candidate for cartilage tissue engineering scaffold to meet clinical needs.