Scaffolds For Cartilage Regeneration Research Articles

Multicomponent 3D-printed Collagen-based Scaffolds for Cartilage Regeneration: Recent Progress, Developments, and Emerging Technologies

Abstract: Cartilage tissue presents challenges in terms of repair and regeneration due to its inherent limitations in self-healing and the scarcity of available donors. Cartilage damage can result in the development of joint problems characterized by symptoms, such as pain, swelling, and osteoarthritis. Collagen scaffolds are extensively used as biomimetic substances for cartilage engineering due to their ability to offer structural, biochemical, and mechanical signals for chondrocytes. Nevertheless, traditional techniques for producing collagen scaffolds frequently yield inadequate pore architecture, diminished mechanical robustness, and restricted form accuracy. Hence, 3D printing is a developing method that can surpass these restrictions by allowing accurate manipulation of the shape, porousness, and makeup of the scaffold. 3D printing has the capability to include various materials and cells in the scaffolds, resulting in the production of intricate and personalized tissue structures. This research examines the latest progress in utilizing 3D printing to create collagen scaffolds for the purpose of regenerating cartilage. This text discusses the different sources of collagen, methods of cross-linking, techniques for printing, and strategies for post-processing that are employed to improve the performance of scaffolds. Furthermore, it discusses the difficulties and potential future paths of utilizing 3D printing to create collagen scaffolds for the purpose of regenerating cartilage.

Multipatterned Chondrocytes' Scaffolds by FL-MOPL with a BSA-GMA Hydrogel to Regulate Chondrocytes' Morphology.

Repairing articular cartilage damage is challenging due to its low regenerative capacity. In vitro, cartilage regeneration is a potential strategy for the functional reconstruction of cartilage defects. A hydrogel is an advanced material for mimicking the extracellular matrix (ECM) due to its hydrophilicity and biocompatibility, which is known as an ideal scaffold for cartilage regeneration. However, chondrocyte culture in vitro tends to dedifferentiate, leading to fibrosis and reduced mechanical properties of the newly formed cartilage tissue. Therefore, it is necessary to understand the mechanism of modulating the chondrocytes' morphology. In this study, we synthesize photo-cross-linkable bovine serum albumin-glycidyl methacrylate (BSA-GMA) with 65% methacrylation. The scaffolds are found to be suitable for chondrocyte growth, which are fabricated by homemade femtosecond laser maskless optical projection lithography (FL-MOPL). The large-area chondrocyte scaffolds have holes with interior angles of triangle (T), quadrilateral (Q), pentagon (P), hexagonal (H), and round (R). The FL-MOPL polymerization mechanism, swelling, degradation, and biocompatibility of the BSA-GMA hydrogel have been investigated. Furthermore, cytoskeleton and nucleus staining reveals that the R-scaffold with larger interior angle is more effective in maintaining chondrocyte morphology and preventing dedifferentiation. The scaffold's ability to maintain the chondrocytes' morphology improves as its shape matches that of the chondrocytes. These results suggest that the BSA-GMA scaffold is a suitable candidate for preventing chondrocyte differentiation and supporting cartilage tissue repair and regeneration. The proposed method for chondrocyte in vitro culture by developing biocompatible materials and flexible fabrication techniques would broaden the potential application of chondrocyte transplants as a viable treatment for cartilage-related diseases.

Recent Advances on Scaffolds: A Comprehensive Review of Materials, Fabrication Techniques, and Applications

Scaffolds offer a three-dimensional framework supporting cell growth, proliferation, and differentiation of cells which are used to repair and regenerate tissues. Recent advancements in scaffold technology have significantly exploited the field of tissue engineering and regenerative medicine. This comprehensive review provides in-depth exploration of scaffold materials, fabrication techniques, and their recent progress in applications. Composite scaffolds have promising applications in bone and dental tissue regeneration due to their greater mechanical properties and ability to promote cell growth. The inherent crosslinking present in hydrogels allows them to maintain their integrity and three-dimensional structure without dissolving. However, there is a growing interest in smart hydrogels which can respond to changes in their external surroundings like pH, ionic strength, temperature, or specific molecules. dECM scaffold is an alternative potential technique for reconstructing the functional organs/tissues by excluding the cell-associated antigens while maintaining the native ECM compositions like growth factors, basement membrane structural proteins, and GAG’s. The degree of porosity in scaffolds can be increased by various fabrication techniques such as TIPS, SCPL, gas foaming, and freeze drying. GelMA hydrogels have shown promising potential in cell proliferation and tissue regeneration. In addition, graphene and its derivatives have been instrumental in the fabrication of bioactive scaffolds for cartilage regeneration. The introduction of additive manufacturing technologies, specifically 3D bioprinting, has significantly improved the precision and control of scaffold fabrication.

Controlled degradation and kinetics response in calcium silicate doped with sodium alginate/functionalized multi-walled carbon nanotube composite 3D scaffolds for cartilage regeneration

Development of bio-mimetic scaffolds with controlled degradation is a key research area in cartilage regeneration. Therefore, novel scaffold composites and favorable techniques to fabricate essential 3D scaffolds are crucial. In this investigation, we achieved controlled biodegradation of composite scaffolds by incorporating calcium silicate (CS) into sodium alginate (SA)/functionalized with multi-walled carbon nanotubes (f-MWCNT). The SA/f-MWCNT and scaffolds with various amounts of CS (5, 10, and 15%) were shown to enhance scaffold properties. We conducted analyses of the composites using X-ray diffraction and scanning electron microscopy. The findings indicated that CS was successfully loaded onto SA/f-MWCNT, and after inclusion the CS crystal structure remained unaltered. We also evaluated the composites using differential scanning calorimetry, and we determined the mechanical strength, swelling, porosity, degradation, and pH properties. The 15% CS-SA/f-MWCNT scaffolds demonstrated improved mechanical properties, controlled degradation and swelling ratio, increased material density, and strengthened bonds that resulted in a more compact structure and better control of porosity. Furthermore, kinetic investigations of the composite scaffolds to potentially absorb water could be accurately modeled using the pseudo-first-order, pseudo-second-order, Elovich, and diffusion kinetic models. The gradual water absorption rate observed in 15% CS-SA/f-MWCNT could be attributed to controlled intraparticle and micropore diffusion. Further biocompatibility studies showed that the scaffold encouraged chondrocyte attachment, spreading, and division. Additionally, collagen deposition and biomineralization of the 15% CS-SA/f-MWCNT scaffolds were slightly higher compared to the SA/f-MWCNT, 5% CS-SA/f-MWCNT, and 10% CS-SA/f-MWCNT scaffolds. The results indicated that the 15% CS-SA/f-MWCNT fabricated scaffolds are the most suitable for cartilage regeneration.

3D Bioprinted Xanthan Hydrogels with Dual Antioxidant and Chondrogenic Functions for Post-traumatic Cartilage Regeneration.

Intra-articular trauma typically initiates the overgeneration of reactive oxidative species (ROS), leading to post-traumatic osteoarthritis and cartilage degeneration. Xanthan gum (XG), a branched polysaccharide, has shown its potential in many biomedical fields, but some of its inherent properties, including undesirable viscosity and poor mechanical stability, limit its application in 3D printed scaffolds for cartilage regeneration. In this project, we developed 3D bioprinted XG hydrogels by modifying XG with methacrylic (MA) groups for post-traumatic cartilage therapy. Our results demonstrated that the chemical modification optimized the viscoelasticity of the bioink, improved printability, and enhanced the mechanical properties of the resulting scaffolds. The XG hydrogels also exhibit decent ROS scavenging capacities to protect stem cells from oxidative stress. Furthermore, XGMA(H) (5% MA substitution) exhibited superior chondrogenic potential in vitro and promoted cartilage regeneration in vivo. These dual-functional XGMA hydrogels may provide a new opportunity for cartilage tissue engineering.

A 3D printed biomimetic scaffold for cartilage regeneration with lubrication, load-bearing, and adhesive fixation properties

Reconstructing the tri-layer structure of articular cartilage (AC) and replicating its functions pose significant challenges. In response, we propose a novel approach utilizing 3D printing technology to replicate and reconstruct the complex structure of AC. This is achieved by introducing three distinct functional hydrogels, each providing lubrication, load-bearing, and adhesive fixation functions. We have systematically characterized the structure, chemical composition, and function of each layer. The results demonstrate the effectiveness of the tri-layer scaffold, which exhibits low friction, mechanical properties similar to natural AC, excellent adhesion capacity, and good biocompatibility. This study paves the way for innovation in the traditional methods of preparing biomimetic AC and has broad applications in osteochondral (OC) defect repair.

Magnetothermal spider silk-based scaffolds for cartilage regeneration

Developing biocompatible, magnetically controlled polymers is a multifunctional solution to many surgical complications. By combining nanoparticle technology with the latest advancements in polymer materials science, we created a multicomponent hybrid system comprised of a robust native spider silk-based matrix; a Mn0.9Zn0.1Fe2O4 nanoparticles coating to provide a controlled thermal trigger for drug release; and liposomes, which act as drug carriers. Fluorescent microscope images show that the dye loaded into the liposomes is released when the system is exposed to an alternating magnetic field due to heating of ferromagnetic nanoparticles, which had a low Curie temperature (40–46°С). The silk matrix also demonstrated outstanding biocompatibility, creating a favorable environment for human postnatal fibroblast cell adhesion, and paving the way for their directed growth. This paper describes a complex approach to cartilage regeneration by developing a spider silk-based scaffold with anatomical mechanical properties for controlled drug delivery in a multifunctional autologous matrix-induced chondrogenesis.

3D Printed Chondrogenic Functionalized PGS Bioactive Scaffold for Cartilage Regeneration.

Tissue engineering is emerging as a promising approach for cartilage regeneration and repair. Endowing scaffolds with cartilaginous bioactivity to obtain bionic microenvironment and regulating the matching of scaffold degradation and regeneration play a crucial role in cartilage regeneration. Poly(glycerol sebacate) (PGS) is a representative thermosetting bioelastomer known for its elasticity, biodegradability, and biocompatibility and is widely used in tissue engineering. However, the modification and drug loading of the PGS scaffold is still a key challenge due to its high temperature curing conditions and limited reactive groups, which seriously hinders its further functional application. Here, a simple versatile new strategy of super swelling-absorption and cross-linked networks locking is presented to successfully create the 3Dprinted PGS-CS/Gel scaffold for the first time based on FDA-approved PGS, gelatin (Gel) and chondroitin sulfate (CS). The PGS-CS/Gel scaffold exhibits the desirable synergistic properties of well-organized hierarchical structures, excellent elasticity, improved hydrophilicity, and cartilaginous bioactivity, which can promote the adhesion, proliferation, and migration of chondrocytes. Importantly, the rate of cartilage regeneration can be well-matched with degradation of PGS-CS/Gel scaffold, and achieve uniform and mature cartilage tissue without scaffold residual. The bioactive scaffold can successfully repair cartilage in a rabbit trochlear groove defect model indicating a promising prospect of clinical transformation.

Magnetic Microcarriers with Accurate Localization and Proliferation of Mesenchymal Stem Cell for Cartilage Defects Repairing.

Magnetic biomaterials are widely used in the field of tissue engineering because of their functions such as drug delivery and targeted therapy. In this study, a magnetically responsive composite microcarrier was prepared through in situ polymerization of dopamine with Fe3O4 (MS) to form a complex. The magnetic composite microcarriers are paramagnetic and have certain magnetic responsiveness, suitable pore size porosity for cell growth, and good blood compatibility and biocompatibility. The bone marrow mesenchyml stem cells (BMSCs) were cultured on magnetic composite microcarriers, and a static magnetic field (SMF) was applied. The results showed that BMSCs adhered to the microcarriers proliferated under the action of horizontal and vertical forces. Magnetic composite microcarriers loaded with BMSCs were implanted into the SD rat model of cartilage defect, and a magnet was added to the operative side. After 12 weeks, cartilage regeneration was observed. The results of gross observation and histological immunostaining 1 month, 2 months, and 3 mounths after operation showed that the magnetic composite microcarriers of loaded cells promoted the early maturation of cartilage and collagen secretion, and the effect of cartilage repair was significantly better than that of the control group. Gait analysis showed that implanting magnetic composite microcarriers loaded with stem cells can reduce postoperative pain and promote limb recovery in SD rats. In conclusion, this study suggests that magnetic composite microcarriers are promising tissue-engineered scaffolds for cartilage regeneration and repair.

Graphene Hydrogel as a Porous Scaffold for Cartilage Regeneration.

Porous scaffolds have widely been exploited in cartilage tissue regeneration. However, it is often difficult to understand how the delicate hierarchical structure of the scaffold material affects the regeneration process. Graphene materials are versatile building blocks for robust and biocompatible porous structures, enabling investigation of structural cues on tissue regeneration otherwise challenging to ascertain. Here, we utilize a graphene hydrogel with stable and tunable structure as a model scaffold to examine the effect of porous structure on matrix remodeling associated with ingrowth of chondrocytes on scaffolds. We observe much-accelerated yet balanced cartilage remodeling correlating the ingrowth of chondrocytes into the graphene scaffold with an open pore structure on the surface. Importantly, such an enhanced remodeling selectively promotes the expression of collagen type II fibrils over proteoglycan aggrecan, hence clearly illustrating that chondrocytes maintain a stable phenotype when they migrate into the scaffold while offering new insights into scaffold design for cartilage repair.

Hydrogel nanosheets confined 2D rhombic ice: a new platform enhancing chondrogenesis

Nanoconfinement within flexible interfaces is a key step towards exploiting confinement effects in several biological and technological systems wherein flexible 2D materials are frequently utilized but are arduous to prepare. Hitherto unreported, the synthesis of 2D hydrogel nanosheets (HNSs) using a template- and catalyst-free process is developed representing a fertile ground for fundamental structure-property investigations. In due course of time, nucleating folds propagating along the edges trigger co-operative deformations of HNS generating regions of nanoconfinement within trapped water islands. These severely constricting surfaces force water molecules to pack within the nanoscale regime of HNS almost parallel to the surface bringing about phase transition into puckered rhombic ice with AA and AB Bernal stacking pattern, which was mostly restricted to molecular dynamics studies so far. Interestingly, under high lateral pressure and spatial inhomogeneity within nanoscale confinement, bilayer rhombic ice structures were formed with an in-plane lattice spacing of 0.31 nm. In this work, a systematic exploration of rhombic ice formation within HNS has been delineated using high-resolution transmission electron microscopy, and its ultrathin morphology was examined using atomic force microscopy. Scanning electron microscopy images revealed high porosity while mechanical testing presented young’s modulus of 155 kPa with ∼84% deformation, whereas contact angle suggested high hydrophilicity. The combinations of nanosheets, porosity, nanoconfinement, hydrophilicity, and mechanical strength, motivated us to explore their application as a scaffold for cartilage regeneration, by inducing chondrogenesis of human Wharton Jelly derived mesenchymal stem cells. HNS promoted the formation of cell aggregates giving higher number of spheroid formation and a marked expression of chondrogenic markers (ColI, ColII, ColX, ACAN and S-100), thereby providing some cues for guiding chondrogenic differentiation.

A Photoannealed Granular Hydrogel Facilitating Hyaline Cartilage Regeneration via Improving Chondrogenic Phenotype.

Hydrogel-based chondrocyte implantation presents a promising tissue engineering strategy for cartilage repair. However, the widely used elastic hydrogels usually restrict cell volume expansion and induce the dedifferentiation of encapsulated chondrocytes. To address this limitation, a photoannealed granular hydrogel (GH) composed of hyaluronic acid, polyethylene glycol, and gelatin was formulated for cartilage regeneration in this study. The unannealed GH prepared by Diels-Alder cross-linked microgels could be mixed with chondrocytes and delivered to cartilage defects by injection, after which light was introduced to anneal the scaffold, leading to the formation of a stable and microporous chondrocyte deploying scaffold. The in vitro studies showed that GH could promote the volume expansion and morphology recovery of chondrocytes and significantly improve their chondrogenic phenotype compared to the nongranular hydrogel (nGH) with similar compositions. Further in vivo studies of subcutaneous culture and the rat full-thickness cartilage defect model proved that chondrocyte loaded GH could significantly stimulate hyaline cartilage matrix deposition and connection, therefore facilitating hyaline-like cartilage regeneration. Finally, the mechanistic study revealed that GH might improve chondrogenic phenotype via activating the AMP-activated protein kinase/glycolysis axis. This study proves the great feasibility of GHs as in situ chondrocyte deploying scaffolds for cartilage regeneration and brings new insights in designing hydrogel scaffold for cartilage tissue engineering.

Direct 3D printing of decellularized matrix embedded composite polycaprolactone scaffolds for cartilage regeneration.

Treatment options for large osteochondral injuries (OCIs) are limited by donor tissue scarcity, morbidity, and anatomic mismatch. 3D printing technology can produce patient-specific scaffolds to address these large defects. Thermoplastics like polycaprolactone (PCL) offer necessary mechanical properties, but lack bioactivity. We fabricated 3D printed PCL scaffolds embedded with polylactic acid microspheres containing decellularized cartilage matrix (DM). DM incorporation within polylactic acid microspheres prevented its thermal degradation during the 3D printing process. The scaffolds replicated the mechanical properties of native cartilage and demonstrated controlled release of DM proteins. Human mesenchymal stem cells (hMSCs) seeded on the composite scaffolds with DM and cultured in basal media self-assembled into aggregates mimicking mesenchymal condensates during embryonic development. The DM composite scaffolds also induced higher expression of biochemical markers of cartilage development than controls, providing evidence for their translational application in the treatment of OCIs. The present study demonstrates the potential of direct incorporation of DM with thermoplastics for 3D printing of patient-specific scaffolds for osteochondral regeneration.

Double – network hydrogel based on exopolysaccharides as a biomimetic extracellular matrix to augment articular cartilage regeneration

Cartilage regeneration remains a current challenge with no satisfactory strategy in surgery. Hydrogels with structurally and biochemically biomimicking characteristics have been regarded as a promising approach for the success of cartilage regeneration. Naturally sourced hydrogels from exopolysaccharides are ideal candidates for the construction of biomimetic extracellular matrix (ECM) because of their biomimetic networks, high water content, cytocompatibility, and biodegradability. Here, an approach that integrates covalent and ionic bonds in a hydrogel system is shown to form a natural polymeric hydrogel double network (DN) for promoting the adhesion and proliferation of chondrocytes and supporting the formation of matured cartilage tissue. DN hydrogels comprised of chemically crosslinked hyaluronan (HA) and physically crosslinked gellan gum (GG) were developed for potential scaffold fabrication. Compared with HA single network (SN) hydrogel and GG SN hydrogel, the obtained HA/GG DN hydrogel with Young's modulus of 28.6 kPa exhibited adequate compressive strength (208.9 kPa) and high toughness (dissipated energy 2837 J/m3) and thus can be used as a biomimetic extracellular matrix for minimal invasively repairing cartilage. In vitro studies showed that HA/GG DN hydrogel-based ECM promoted the proliferation of chondrocytes. The HA/GG DN hydrogel significantly supported the deposition of cartilage ECM-specific sulfated glycosaminoglycan and type II collagen and facilitated the formation of cartilage tissues. In a rabbit osteochondral defect model, HA/GG DN hydrogel significantly improved cartilage regeneration. The HA/GG DN hydrogel as a biomimetic ECM is a promising candidate as a biomaterial scaffold for cartilage regeneration and repair. Statement of significanceThe fabrication of a biomaterial scaffold as an artificial extracellular matrix (ECM) for cartilage regeneration remains a big challenge. In this work, we fabricated a double-network (DN) hydrogel based on hyaluronan and gellan gum (HA/GG) through a sequential chemical and physical cross-linking process. The HA/GG DN hydrogel exhibited high compressive strength, high toughness, stiffness, and good self-recovery property. The HA/GG DN hydrogel can support chondrocyte proliferation and new ECM deposition correlated with the enhanced mechanical properties, good cytocompatibility, and biodegradability. In vivo animal experiments demonstrated that this HA/GG DN hydrogel facilitates hyaline-like cartilage regeneration. These findings imply that the developed HA/GG DN hydrogel as a biomimetic ECM offers a hopeful new platform for cartilage tissue engineering.

Cartilage regeneration using improved surface electrospun bilayer polycaprolactone scaffolds loaded with transforming growth factor-beta 3 and rabbit muscle-derived stem cells.

Polycaprolactone (PCL) has recently received significant attention due to its mechanical strength, low immunogenicity, elasticity, and biodegradability. Therefore, it is perfectly suitable for cartilage tissue engineering. PCL is relatively hydrophobic in nature, so its hydrophilicity needs to be enhanced before its use in scaffolding. In our study, first, we aimed to improve the hydrophilicity properties after the network of the bilayer scaffold was formed by electrospinning. Electrospun bilayer PCL scaffolds were treated with ozone and further loaded with transforming growth factor-beta 3 (TGFβ3). In vitro studies were performed to determine the rabbit muscle-derived stem cells’ (rMDSCs) potential to differentiate into chondrocytes after the cells were seeded onto the scaffolds. Statistically significant results indicated that ozonated (O) scaffolds create a better environment for rMDSCs because collagen-II (Coll2) concentrations at day 21 were higher than non-ozonated (NO) scaffolds. In in vivo studies, we aimed to determine the cartilage regeneration outcomes by macroscopical and microscopical/histological evaluations at 3- and 6-month time-points. The Oswestry Arthroscopy Score (OAS) was the highest at both mentioned time-points using the scaffold loaded with TGFβ3 and rMDSCs. Evaluation of cartilage electromechanical quantitative parameters (QPs) showed significantly better results in cell-treated scaffolds at both 3 and 6 months. Safranin O staining indicated similar results as in macroscopical evaluations—cell-treated scaffolds revealed greater staining with safranin, although an empty defect also showed better results than non-cell-treated scaffolds. The scaffold with chondrocytes represented the best score when the scaffolds were evaluated with the Mankin histological grading scale. However, as in previous in vivo evaluations, cell-treated scaffolds showed better results than non-cell-treated scaffolds. In conclusion, we have investigated that an ozone-treated scaffold containing TGFβ3 with rMDSC is a proper combination and could be a promising scaffold for cartilage regeneration.

Subchondral bone-inspired hydrogel scaffold for cartilage regeneration

Promoting the in situ regeneration of cartilage without additional cells or cytokines remains challenging. Here, inspired by the unique microstructures of subchondral bone, a cell and cytokine free hydrogel scaffold for cartilage regeneration was developed via a strategy of directional lyophilization and postcrosslinking. This strategy achieved intersecting microchannels in an orderly arrangement and an aligned ladder-like texture in a semi-interpenetrating hydrogel network. The resulting hydrogel had similar mechanical properties to the native cartilage extracellular matrix. Incorporating chitosan into the rigid network also endowed the hydrogel with excellent hemostatic properties. By delicately tuning the components and postcrosslinking conditions, the hydrogel was further endowed with suitable swelling and degradation properties for cartilage regeneration. In vitro tests showed that the highly biocompatible hydrogel scaffold could facilitate the migration and chondrogenic differentiation of bone marrow mesenchymal stem cells. In vivo results further verified that the hydrogel could promote the in situ regeneration of cartilage in a rat model of osteochondral defects. In summary, the subchondral bone-like hydrogel revealed promising prospects in cartilage regeneration and a variety of bioremediation applications.

3D Contour Printing of Anatomically Mimetic Cartilage Grafts with Microfiber-Reinforced Double-Network Bioink.

Bioprinting is an emerging technology for fabricating cell-laden scaffolds with custom shapes that resemble the complex architecture of human tissues, however, construction of mechanically competent tissue grafts which mimic irregular cartilage defect is still a big challenge. Here, 3D printing of short fiber-reinforced double-network bioink to generate anatomically accurate and mechanical tunable scaffold for cartilage regeneration is reported. Poly(lactic acid) (PLLA) short fibers are first prepared by electrospinning and then fragmented through aminolysis reaction. Composite inks are constructed with an incorporation of fragmented microfibers with varied amounts and lengths into oxidized alginate bioink. The results show that incorporation of PLLA short fibers not only improves the printing fidelity but also facilitates in generating mechanically strong constructs. By incorporating gelatin methacryloyl (GelMA) and optimizing the bioink composition, the fabricated constructs with a compressive stress of ≈150 kPa even after 100 cyclical compression loading (up to 40% of strain) are achieved. In addition, this mechanically reinforced alginate/GelMA double-network bioink displays good biocompatibility and supports bone marrow-derived stromal cell chondrogenesis in vitro. Collectively, these findings demonstrate this approach is capable of printing engineered grafts which resemble the irregular size and mechanical properties of cartilage and thus hold potential for functional tissue regeneration.

The effect of decellularized cartilage matrix scaffolds combined with endometrial stem cell-derived osteocytes on osteochondral tissue engineering in rats.

Since decellularized tissues may offer the instructive niche for cell differentiation and function, their use as cell culture scaffolds is a promising approach for regenerative medicine. To repair osteochondral tissues, developing a scaffold with biomimetic structural, compositional, and functional characteristics is vital. As a result of their heterogeneous structure, decellularized articular cartilage matrix from allogeneic and xenogeneic sources are considered appropriate scaffolds for cartilage regeneration. We developed a scaffold for osteochondral tissue engineering by decellularizing sheep knee cartilage using a chemical technique. DNA content measurements and histological examinations revealed that this protocol completely removed cells from decellularized cartilage. Furthermore, SEM, MTS assay, and H&E staining revealed that human endometrial stem cells could readily adhere to the decellularized cartilage, and the scaffold was biocompatible for their proliferation. Besides, we discovered that decellularized scaffolds could promote EnSC osteogenic differentiation by increasing bone-specific gene expression. Further, it was found that decellularized scaffolds were inductive for chondrogenic differentiation of stem cells, evidenced by an up-regulation in the expression of the cartilage-specific gene. Also, in vivo study showed the high affinity of acellularized scaffolds for cell adhesion and proliferation led to an improved regeneration of articular lesions in rats after 4weeks. Finally, a perfect scaffold with high fidelity is provided by the developed decellularized cartilage scaffold for the functional reconstruction of osteochondral tissues; these types of scaffolds are helpful in studying how the tissue microenvironment supports osteocytes and chondrocytes differentiation, growth, and function to have a good osteochondral repair effect.

Acellular cartilage matrix biomimetic scaffold with immediate enrichment of autologous bone marrow mononuclear cells to repair articular cartilage defects.

Functional repair of articular cartilage defects is always a great challenge in joint surgery clinically. Tissue engineering strategies that combine autologous cell implantation with three-dimensional scaffolds have proven effective for repairing articular cartilage tissue. However, it faces the problem of cell sources and scaffold materials. Autologous chondrocytes and bone marrow are difficult to popularize clinically due to limited donor sources and low mononuclear cell (MNC) concentrations, respectively. The density gradient centrifugation method can increase the concentration of MNCs in fresh bone marrow by nearly a hundredfold and achieve immediate enrichment. In addition, acellular cartilage matrix (ACM), with good biocompatibility and a cartilage-specific microenvironment, is considered to be an ideal candidate scaffold for cartilage regeneration. In this study, hybrid pigs were used to establish articular cartilage defect models of different sizes to determine the feasibility and maximum scope of application of ACM-based biomimetic scaffolds combined with MNCs for inducing articular cartilage regeneration. Importantly, ACM-based biomimetic scaffolds instantly enriched MNCs could improve the repair effect of articular cartilage defects in situ, which established a new model of articular cartilage regeneration that could be applied immediately and suited for large-scale clinical promotion. The current study significantly improves the repair effect of articular cartilage defects, which provides scientific evidence and detailed insights for future clinical applications of ACM-based biomimetic scaffolds combined with MNCs.

BC enhanced photocurable hydrogel based on 3D bioprinting for nasal cartilage repair

The repair of nasal cartilage lesions and defects is still a difficult problem in nasal surgery, and nasal cartilage tissue engineering will be an effective way to solve this problem. Hydrogel has excellent application potential in tissue engineering. In order to produce a 3D printable scaffold for cartilage regeneration, we prepared gelatin methacryloyl (GelMA)/hyaluronic acid methacryloyl (HAMA)/bacterial cellulose (BC) composite hydrogel. The composite hydrogel was characterized by swelling, mechanical properties, and printing performance test. Compared with GelMA/HAMA hydrogel, the addition of BC not only significantly enhanced the mechanical properties of the hydrogels, but also improved the printing fidelity. At the same time, the mouse chondrocyte cell line (ATDC5) was used to evaluate the biocompatibility of the composite hydrogel. After 7 d of culture, the cells encapsulated within the composite hydrogels proliferated significantly and maintain the expression of specific proteins. In addition, the cell viability of the encapsulated ATDC5 cells in the 3D bioprinted hybrid constructs was evaluated. In summary, these results indicate that BC-enhanced GelMA/HAMA hydrogel is a promising candidate material for nasal cartilage repair.