Cartilage Tissue Engineering Applications Research Articles

Advanced Hydrogels for Cartilage Tissue Engineering: Recent Progress and Future Directions.

Cartilage is a tension- and load-bearing tissue and has a limited capacity for intrinsic self-healing. While microfracture and arthroplasty are the conventional methods for cartilage repair, these methods are unable to completely heal the damaged tissue. The need to overcome the restrictions of these therapies for cartilage regeneration has expanded the field of cartilage tissue engineering (CTE), in which novel engineering and biological approaches are introduced to accelerate the development of new biomimetic cartilage to replace the injured tissue. Until now, a wide range of hydrogels and cell sources have been employed for CTE to either recapitulate microenvironmental cues during a new tissue growth or to compel the recovery of cartilaginous structures via manipulating biochemical and biomechanical properties of the original tissue. Towards modifying current cartilage treatments, advanced hydrogels have been designed and synthesized in recent years to improve network crosslinking and self-recovery of implanted scaffolds after damage in vivo. This review focused on the recent advances in CTE, especially self-healing hydrogels. The article firstly presents the cartilage tissue, its defects, and treatments. Subsequently, introduces CTE and summarizes the polymeric hydrogels and their advances. Furthermore, characterizations, the advantages, and disadvantages of advanced hydrogels such as multi-materials, IPNs, nanomaterials, and supramolecular are discussed. Afterward, the self-healing hydrogels in CTE, mechanisms, and the physical and chemical methods for the synthesis of such hydrogels for improving the reformation of CTE are introduced. The article then briefly describes the fabrication methods in CTE. Finally, this review presents a conclusion of prevalent challenges and future outlooks for self-healing hydrogels in CTE applications.

Sequential Enzymatic Digestion of Different Cartilage Tissues: A Rapid and High-Efficiency Protocol for Chondrocyte Isolation, and Its Application in Cartilage Tissue Engineering.

The classic chondrocyte isolation protocol is a 1-step enzymatic digestion protocol in which cartilage samples are digested in collagenase solution for a single, long period. However, this method usually results in incomplete cartilage dissociation and low chondrocyte quality. In this study, we aimed to develop a rapid, high-efficiency, and flexible chondrocyte isolation protocol for cartilage tissue engineering. Cartilage tissues harvested from rabbit ear, rib, septum, and articulation were minced and subjected to enzymatic digestion using the classic protocol or the newly developed sequential protocol. In the classic protocol, cartilage fragments were subjected to one 12-hour digestion. In the sequential protocol, cartilage fragments were sequentially subjected to 2-hour first digestion, followed by two 3-hour digestions. The collected cells were then subjected to analyses of cell-yield efficiency, viability, proliferation, phenotype, and cartilage matrix synthesis capacity. Overall, the sequential protocol exhibited higher cell-yield efficiency than the classic protocol for the 4 cartilage types. The cells harvested from the second and third digestions demonstrated higher cell viability, more proliferative activity, a better chondrocyte phenotype, and a higher cartilage-specific matrix synthesis ability than those harvested from the first digestion and after the classic 1-step protocol. The sequential protocol is a rapid, flexible, high-efficiency chondrocyte isolation protocol for different cartilage tissues. We recommend using this protocol for chondrocyte isolation, and in particular, the cells obtained after the subsequent 3-hour sequential digestions should be used for chondrocyte-based therapy.

Dual crosslinked pullulan–gelatin cryogel scaffold for chondrocyte-mediated cartilage repair: synthesis, characterization and in vitro evaluation

Cryogels, a subset of hydrogels, have recently drawn attention for cartilage tissue engineering due to its inherent microporous architecture and good mechanical properties. In this study a dual crosslinked pullulan–gelatin cryogel (PDAG) scaffold was synthesized by crosslinking gelatin with oxidized pullulan by Schiff’s base reaction followed by cryogelation. Chondrocytes seeded within the PDAG scaffolds and cultured for 21 d in vitro demonstrated enhanced cell proliferation, enhanced production of cartilage-specific extracellular matrix and up-regulated sulfated glycosaminoglycan without altering the articular chondrocyte phenotype. Quantitative reverse transcription-polymerase chain reaction-based gene expression studies, immunofluorescence, and histological studies demonstrated that the PDAG scaffold significantly enhanced the expression of chondrogenic marker genes such as type II collagen, aggrecan, and SOX9. Taken together, these results demonstrated that PDAG scaffold prepared by sequential Schiff’s base reaction and cryogelation would be a promising cell-responsive scaffold for cartilage tissue engineering applications.

The Application of Cartilage Tissue Engineering with Cell-Laden Hydrogel in Plastic Surgery: A Systematic Review.

As a contour-supporting material, the cartilage has a significant application value in plastic surgery. Since the development of hydrogel scaffolds with sufficient biomechanical strength and high biocompatibility, cell-laden hydrogels have been widely studied for application in cartilage bioengineering. This systematic review summarizes the latest research on engineered cartilage constructed using cell-laden hydrogel scaffolds in plastic surgery. A systematic review was performed by searching the PubMed and Web of Science databases using selected keywords and Medical Subject Headings search terms. Forty-two studies were identified based on the search criteria. After full-text screening for inclusion and exclusion criteria, 18 studies were included. Data collected from each study included culturing form, seed cell types and sources, concentration of cells and gels, scaffold materials and bio-printing structures, and biomechanical properties of cartilage constructs. These cell-laden hydrogel scaffolds were reported to show some feasibility of cartilage engineering, including better cell proliferation, enhanced deposition of glycosaminoglycans and collagen type II in the extracellular matrix, and better biomechanical properties close to the natural state. Cell-laden hydrogels have been widely used in cartilage bioengineering research. Through 3-dimensional (3D) printing, the cell-laden hydrogel can form a bionic contour structure. Extracellular matrix expression was observed in vivo and in vitro, and the elastic modulus was reported to be similar to that of natural cartilage. The future direction of cartilage tissue engineering in plastic surgery involves the use of novel hydrogel materials and more advanced 3D printing technology combined with biochemistry and biomechanical stimulation.

Fabrication and characterization ofchitosan‐gelatin/single‐walledcarbon nanotubes electrospun composite scaffolds for cartilage tissue engineering applications

AbstractCartilage is a connective tissue with a slow healing rate due to lack in blood circulation and slow metabolism. Designing tissue engineering scaffolds modified based on its specific features can assist its natural regeneration process. In this study, the chitosan‐gelatin/single‐walled carbon nanotubes functionalized by COOH (SWNTs‐COOH) nanocomposite scaffolds were fabricated through electrospinning. The effect of each component and different duration of cross‐linking were assessed in terms of morphology, porosity, chemical structure, thermal behavior, mechanical properties, wettability, biodegradability, and in vitro cell culture study. Adding SWNTs‐COOH decreased fiber diameter, water contact angle and degradation rate while increased tensile strength, hydrophilicity, stability and cell viability, due to their high intrinsic electrical conductivity, and mechanical properties and the presence of COOH functional groups in its structure. All the sample presented a porosity percentage of more than 80%, which is essential for tissue engineering scaffolds. The presence SWNTs‐COOH did not have any adverse effect on cytocompatibility. The optimal cross‐linking time increased the stability of the scaffolds in PBS. It can be concluded that the chitosan‐gelatin/1wt% SWNTs‐COOH scaffold can be appropriate for cartilage tissue engineering applications.

3D Bioprinting of Hydrogels for Cartilage Tissue Engineering.

Three-dimensional (3D) bioprinting is an emerging technology based on 3D digital imaging technology and multi-level continuous printing. The precise positioning of biological materials, seed cells, and biological factors, known as “additive biomanufacturing”, can provide personalized therapy strategies in regenerative medicine. Over the last two decades, 3D bioprinting hydrogels have significantly advanced the field of cartilage and bone tissue engineering. This article reviews the development of 3D bioprinting and its application in cartilage tissue engineering, followed by a discussion of the current challenges and prospects for 3D bioprinting. This review presents foundational information on the future optimization of the design and manufacturing process of 3D additive biomanufacturing.

Enhanced efficacy of transforming growth factor-β1 loaded an injectable cross-linked thiolated chitosan and carboxymethyl cellulose-based hydrogels for cartilage tissue engineering

Growth factors (GFs) are soluble proteins extracellular that control a wide range of cellular processes as well as tissue regeneration. While transforming growth factor beta-1 (TGF-β1) promotes chondrogenesis, its medical use is restricted by its potential protein instability, which necessitates high doses of the protein, which can result in adverse side effects such as inefficient cartilage formation. In this work, we have developed a novel hydrogel composite based on the polymer, cross-linked thiolated chitosan; TCS and carboxymethyl cellulose; CMC (TCS/CMC) hydrogel system was utilized as injectable TGF-β1 carriers for cartilage tissue engineering applications. Rheological measurements showed that the elastic modulus of TCS/CMC hydrogels with an optimized CMC concentration could reach around 2.5 kPa or higher than their respective viscous modulus, indicating that they behaved like strong hydrogels. Crosslinking significantly alters the overall network distribution, surface morphology, pore size, porosity, gelation time, swelling ratio, water content, and in vitro degradation of the TCS/CMC hydrogels. TCS/CMC hydrogels maintain more than 90% of their weight and retain their original form after 21 days. TGF-β1 released marginally from TCS/CMC hydrogels as incubation time increased, up to 21 days, with around 18.6 ± 0.9% of the drug stored inside the TCS/CMC hydrogels. On day 21, BMSC treated with TGF-β1 in medium or TGF-β1-loaded TCS/CMC hydrogels grew faster than the other groups. For in vivo cartilage repair, full-thickness cartilage defects were induced on rat knees for 8 weeks. The optimal ability of this novel TGF-β1-loaded TCS/CMC hydrogel system was further demonstrated by histological analysis, resulting in a novel therapeutic strategy for repairing articular cartilage defects.

Allogeneic mesenchymal stromal cells for cartilage regeneration: A review of in vitro evaluation, clinical experience, and translational opportunities.

The paracrine signaling, immunogenic properties and possible applications of mesenchymal stromal cells (MSCs) for cartilage tissue engineering and regenerative medicine therapies have been investigated through numerous in vitro, animal model and clinical studies. The emerging knowledge largely supports the concept of MSCs as signaling and modulatory cells, exerting their influence through trophic and immune mediation rather than as a cell replacement therapy. The virtues of allogeneic cells as a ready‐to‐use product with well‐defined characteristics of cell surface marker expression, proliferative ability, and differentiation capacity are well established. With clinical applications in mind, a greater focus on allogeneic cell sources is evident, and this review summarizes the latest published and upcoming clinical trials focused on cartilage regeneration adopting allogeneic and autologous cell sources. Moreover, we review the current understanding of immune modulatory mechanisms and the role of trophic factors in articular chondrocyte‐MSC interactions that offer feasible targets for evaluating MSC activity in vivo within the intra‐articular environment. Furthermore, bringing labeling and tracking techniques to the clinical setting, while inherently challenging, will be extremely informative as clinicians and researchers seek to bolster the case for the safety and efficacy of allogeneic MSCs. We therefore review multiple promising approaches for cell tracking and labeling, including both chimerism studies and imaging‐based techniques, that have been widely explored in vitro and in animal models. Understanding the distribution and persistence of transplanted MSCs is necessary to fully realize their potential in cartilage regeneration techniques and tissue engineering applications.

Chondro-inductive hyaluronic acid/chitosan coacervate-based scaffolds for cartilage tissue engineering

Injuries related to articular cartilage are among the most challenging musculoskeletal problems because of poor repair capacity of this tissue. The lack of efficient treatments for chondral defects has stimulated research on cartilage tissue engineering applications combining porous biocompatible scaffolds with stem cells in the presence of external stimuli. This work presents the role of rat bone marrow mesenchymal stem cell (BMSC) encapsulated-novel three-dimensional (3D) coacervate scaffolds prepared through complex coacervation between different chitosan salts (CHI) and sodium hyaluronate (HA). The 3D architecture of BMSC encapsulated scaffolds (HA/CHI) was shown by scanning electron microscopy (SEM) to have an interconnected structure to allow cell-cell and cell-matrix interactions. Chondrogenic induction of encapsulated BMSCs within HA/CHI coacervates demonstrated remarkable cellular viability in addition to the elevated expression levels of chondrogenic markers such as sex determining region Y-box 9 protein (SOX9), aggrecan (ACAN), cartilage oligomeric matrix protein (COMP) and collagen type II (COL2A1) by immunofluorescence staining, qPCR and ELISA test. Collectively, HA/CHI coacervates are promising candidates for future use of these scaffolds in cartilage tissue engineering applications.

Blending with Poly(l-lactic acid) Improves the Printability of Poly(l-lactide-co-caprolactone) and Enhances the Potential Application in Cartilage Tissue Engineering.

Poly(l-lactide-co-caprolactone) (PLCL, 50:50) has been used in cartilage tissue engineering because of its high elasticity. However, its mechanical properties, including its rigidity and viscoelasticity, must be improved for compatibility with native cartilage. In this study, a set of PLCL/poly(l-lactic acid) (PLLA) blends was prepared by blending with different mass ratios of PLLA that range from 10 to 50%, using thermoplastic techniques. After testing the properties of these PLCL/PLLA blends, they were used to fabricate scaffolds by the 3D printing technology. The structures and viscoelastic behavior of the PLCL/PLLA scaffolds were determined, and then, the potential application of the scaffolds in cartilage tissue engineering was evaluated by chondrocytes culture. All blends demonstrate good thermal stability for the 3D printing technology. All blends show good toughness, while the rigidity of PLCL is increased through PLLA blending, and Young’s modulus of blends with 10–20% PLLA is similar to that of native cartilage. Furthermore, blending with PLLA improves the processability of PLCL for 3D printing, and the compression modulus and viscoelasticity of 3D-printed PLCL/PLLA scaffolds are different from that of PLCL. Additionally, the stress relaxation time (t1/2) of the PLCL/PLLA scaffolds, which is important for chondrogenesis, is dramatically shortened compared with the pure PLCL scaffold at the same 3D-printing filling rate. Consistently, the PLCL90PLLA10 scaffold at a 70% filling rate with much shorter t1/2 is more conducive to the proliferation and chondrogenesis of in vitro seeded chondrocytes accompanied by upregulated expression of SOX9 than the PLCL scaffold. Taken together, these results demonstrate that blending with PLLA improves the printability of PLCL and enhances its potential application, particularly PLCL/PLLA scaffolds with a low ratio of PLLA, in cartilage tissue engineering.

Digital Light Processing Bioprinted Human Chondrocyte-Laden Poly (γ-Glutamic Acid)/Hyaluronic Acid Bio-Ink towards Cartilage Tissue Engineering.

Cartilage injury is the main cause of disability in the United States, and it has been projected that cartilage injury caused by osteoarthritis will affect 30% of the entire United States population by the year 2030. In this study, we modified hyaluronic acid (HA) with γ-poly(glutamic) acid (γ-PGA), both of which are common biomaterials used in cartilage engineering, in an attempt to evaluate them for their potential in promoting cartilage regeneration. As seen from the results, γ-PGA-GMA and HA, with glycidyl methacrylate (GMA) as the photo-crosslinker, could be successfully fabricated while retaining the structural characteristics of γ-PGA and HA. In addition, the storage moduli and loss moduli of the hydrogels were consistent throughout the curing durations. However, it was noted that the modification enhanced the mechanical properties, the swelling equilibrium rate, and cellular proliferation, and significantly improved secretion of cartilage regeneration-related proteins such as glycosaminoglycan (GAG) and type II collagen (Col II). The cartilage tissue proof with Alcian blue further demonstrated that the modification of γ-PGA with HA exhibited suitability for cartilage tissue regeneration and displayed potential for future cartilage tissue engineering applications. This study built on the previous works involving HA and further showed that there are unlimited ways to modify various biomaterials in order to further bring cartilage tissue engineering to the next level.

Porcine fibrin sealant combined with autologous chondrocytes successfully promotes full-thickness cartilage regeneration in a rabbit model.

Xenogeneic porcine fibrin sealant (PFS), derived from porcine blood, was used as a scaffold for cartilage tissue engineering. PFS has a porous microstructure, biocompatibility and degradation, and it provides a perfect extracellular matrix environment for the adhesion and proliferation of chondrocytes. Recently, PFS in combination with autologous chondrocytes (ACs) were used to study the microstructure of PFS scaffolds and promotion effect on the proliferation and migration of ACs. In this study, we investigated the effects of PFS in combination with ACs on the healing of cartilage defects in rabbits. A full‐thickness cartilage defect was made in the femoral trochlear in rabbits, subsequently, three surgical procedures were used to repair the defect, namely: the defect was treated with microfracture (MF group); the defect was filled with PFS alone (PFS group) or in combination with ACs (PFS + ACs group); the unrepaired cartilage defects served as the control group (CD group). Three and 6 months after the operation, the reparative effect was evaluated using medical imaging, gross scoring, pathological staining, biomechanical testing and biochemical examination. The PFS group showed a limited effect on defect repair, this result was significantly worse than the MF group. The best reparative effect was observed in the PFS + ACs group. These results hinted that PFS in combination with autologous chondrocytes has broad prospects for clinical applications in cartilage tissue engineering.

Bioorthogonal hydroxyethyl cellulose-based scaffold crosslinked via click chemistry for cartilage tissue engineering applications

In this study, azide and alkyne moieties were introduced to the structure of citric acid-modified hydroxyethyl cellulose (HEC) and then through a bioorthogonal click chemistry method: Strain-promoted azide-alkyne cycloaddition, a novel crosslinked HEC scaffold (click sample) was obtained. Chemical modifications and successful crosslinking of the samples were assessed with FTIR and 1H NMR spectroscopy. Lyophilized samples exhibited a porous interconnected microarchitecture with desirable features for commensurate cartilage tissue engineering applications. As the stability of scaffolds improved upon crosslinking, considerable water uptake and swelling degree of ~650% could still be measured for the click sample. Offering Young's modulus of ~10 MPa and tensile strength of ~0.43 MPa, the mechanical characteristics of click sample were comparable with those of normal cartilage tissue. Various in vitro biological assays, including MTT analysis, cellular attachment, histological staining with safranin O, and real-time PCR decisively approved significant biocompatibility, chondrogenic ability, and bioorthogonal features of click sample.

Non-mulberry silk fiber-based scaffolds reinforced by PLLA porous microspheres for auricular cartilage: An in vitro study

Designing clinical applicable polymeric composite scaffolds for auricular cartilage tissue engineering requires appropriate mechanical strength and biological characteristics. In this study, silk fiber-based scaffolds co-reinforced with poly-L-lactic acid porous microspheres (PLLA PMs) combined with either Bombyx mori (Bm) or Antheraea pernyi (Ap) silk fibers were fabricated as inspired by the “steel bars reinforced concrete” structure in architecture and their chondrogenic functions were also investigated. We found that the Ap silk fiber-based scaffolds reinforced by PLLA PMs (MAF) exhibited superior physical properties (the mechanical properties in particular) as compared to the Bm silk fiber-based scaffolds reinforced by PLLA PMs (MBF). Furthermore, in vitro evaluation of chondrogenic potential showed that the MAF provided better cell adhesion, viability, proliferation and GAG secretion than the MBF. Therefore, the MAF are promising in auricular cartilage tissue engineering and relevant plastic surgery-related applications.

Review of Hybrid Materials Based on Polyhydroxyalkanoates for Tissue Engineering Applications.

This review is focused on hybrid polyhydroxyalkanoate-based (PHA) biomaterials with improved physico-mechanical, chemical, and piezoelectric properties and controlled biodegradation rate for applications in bone, cartilage, nerve and skin tissue engineering. PHAs are polyesters produced by a wide range of bacteria under unbalanced growth conditions. They are biodegradable, biocompatible, and piezoelectric polymers, which make them very attractive biomaterials for various biomedical applications. As naturally derived materials, PHAs have been used for multiple cell and tissue engineering applications; however, their widespread biomedical applications are limited due to their lack of toughness, elasticity, hydrophilicity and bioactivity. The chemical structure of PHAs allows them to combine with other polymers or inorganic materials to form hybrid composites with improved structural and functional properties. Their type (films, fibers, and 3D printed scaffolds) and properties can be tailored with fabrication methods and materials used as fillers. Here, we are aiming to fill in a gap in literature, revealing an up-to-date overview of ongoing research strategies that make use of PHAs as versatile and prospective biomaterials. In this work, a systematic and detailed review of works investigating PHA-based hybrid materials with tailored properties and performance for use in tissue engineering applications is carried out. A literature survey revealed that PHA-based composites have better performance for use in tissue regeneration applications than pure PHA.

Copper-based biomaterials for bone and cartilage tissue engineering.

BackgroudTissue engineering using cells, scaffolds, and bioactive molecules can promote the repair and regeneration of injured tissues. Copper is an essential element for the human body that is involved in many physiological activities and in recent years, copper has been used increasingly in tissue engineering.MethodsThe current advances of copper-based biomaterial for bone and cartilage tissue engineering were searched on PubMed and Web of Science.ResultsVarious forms of copper-based biomaterials, including pure copper, copper ions, copper nanoparticles, copper oxides, and copper alloy are introduced. The incorporation of copper into base materials provides unique properties, resulting in tuneable porosity, mechanical strength, degradation, and crosslinking of scaffolds. Copper also shows promising biological performance in cell migration, cell adhesion, osteogenesis, chondrogenesis, angiogenesis, and antibacterial activities. In vivo applications of copper for bone and cartilage tissue engineering are discussed.ConclusionThis review focuses on copper’s physiochemical and biological effects, and its applications in bone and cartilage tissue engineering. The potential limitations and future perspectives are also discussed.Translational potential of this articleThis review introduces the recent advances in copper-based biomaterial for bone and cartilage tissue engineering. This revie could guide researchers to apply copper in biomaterials, improving the generation of bone and cartilages, decrease the possibility of infection and shorten the recovery time so as to decrease medical costs.

The Role of Histone Demethylase in Osteogenic and Chondrogenic Differentiation of Mesenchymal Stem Cells: A Literature Review

The proliferation and multi-directional differentiation potential of mesenchymal stem cells (MSCs) enabled its wide use in the development of new therapies for bone and cartilage repair. Although preliminary work has been done to verify the gene expression profile of MSCs osteogenic and chondrogenic differentiation, it is still unclear what key factors initiate the differentiation of MSCs, resulting in its limited application in bone and cartilage tissue engineering. The epigenetic mechanism mediated by histone demethylases (lysine [K]-specific histone demethylases, KDMs) is the key link in regulating MSCs lineage differentiation. The lysine-specific histone demethylase (LSD) family containing Tower domain and the histone demethylase family containing Jumonji C (JmjC) domain regulate the expression of various osteogenic-related genes, including Runt-related transcription factor 2 ( RUNX2), osterix ( OSX), osteocalcin ( OCN), to mediate MSCs osteogenic differentiation. The KDM2/4/6 subfamilies regulate the chondrogenic differentiation of MSCs through multiple pathways centered on SRY-related high-mobility-group-box gene 9 ( SOX9). In addition, nanotopology, mircoRNAs, etc. regulate the expression of a variety of osteogenic and chondrogenic transcription factors through up- and down-regulation of KDMs. In summary, the role of histone demethylase in the osteogenic and chondrogenic differentiation of mesenchymal stem cells will help us better understand the pathogenesis of bone and cartilage damage diseases, and establish the foundation of future clinical applications for bone and cartilage tissue engineering.

Novel three‐dimensional printing of poly(ester urethane) scaffolds for biomedical applications

AbstractTissue engineering requires highly porous complex structures with interconnected pores and tightly controlled pore sizes, added to customized production. Additive manufacturing (AM) is nowadays one of the most suitable technologies for the preparation of structures with strictly defined complex three‐dimensional architectures. Moreover, computed tomography and magnetic resonance imaging can be employed to obtain personalized medical devices. Fused deposition modeling (FDM) is one of the most common, easiest, and cost‐effective AM techniques to obtain 3D objects from a wide variety of materials. However, there is a lack of bioresorbable materials that can fit the increasing demand for biomedical applications requiring stiff elastomers. In this work, a series of bioresorbable segmented poly(ester urethanes) (SPEU) with high hard segment content was synthesized and physicochemically characterized. The materials with higher thermal stability were processed into homogeneous filaments by hot‐melt extrusion with no need for additives nor plasticizers. These filaments were evaluated for FDM, and structures with controlled porosity, filament diameter, and in‐plane pore size could be easily obtained by modifying the printing parameters. Regarding SPEU mechanical properties, the obtained scaffolds could be promising for cartilage tissue engineering applications.

A Review of Tissue‐Engineered Cartilage Utilizing Fibrin and Its Composite

Suitable alternatives are made for damaged or diseased organs and tissues in tissue engineering by combining cellular and molecular biology with materials and mechanical engineering. Fibrin is a critical blood component responsible for homeostasis, used extensively as a biopolymer scaffold in tissue engineering. This study summarizes the latest developments in organ and tissue regeneration using fibrin as a scaffold material. The combination of active peptides and growth factors through a heparin-bound delivery system improves fibrin function as a scaffold. Besides, the development of fibrin precursors as recombinant proteins solves multiple or single donor fibrin adhesives. Its composite allows biomolecules to be combined with fibrin and can significantly enhance fibrin efficacy in cartilage tissue engineering applications.

In vitro anti-inflammatory potential of marine macromolecules cross-linked bio-composite scaffold on LPS stimulated RAW 264.7 macrophage cells for cartilage tissue engineering applications

Biomaterials serve as an integral component of tissue engineering. They are designed to provide architectural framework of native extracellular matrix so as to encourage cell growth and eventual tissue regeneration. Naturally occurring biopolymers as scaffolds offer options for cartilage tissue engineering due to anti-inflammatory, biocompatibility, biodegradability, low toxicity of degradation by-products and plasticity in processing into a variety of material formats. Here we studied in vitro anti-inflammatory potential of marine macromolecules cross-linked bio-composite scaffold composed of hydroxyapatite, alginate, chitosan and fucoidan named as HACF on LPS stimulated RAW 264.7 macrophage cells. The effects of HACF on the viability of RAW264.7 cells, nitrite level, intracellular ROS as well as the mRNA levels of NF-κB, iNOS, COX-2, TNF-α, IL-1β and IL-6 were examined in LPS induced RAW264.7 macrophage cells. The results revealed that HACF hydrogel scaffold exerts anti-inflammatory effect by inhibiting the production of ROS, suppress NF-kB translocation to the nucleus and thereby inhibiting the production of inflammatory mediators. Hence, our results confirm that HACF has a strong anti-oxidant capacity to inhibit inflammation associated gene expression by suppressing NF-kB signaling pathway. It clearly reveals the anti-oxidant and anti-inflammatory effect of HACF hydrogel scaffold on LPS induced RAW 264.7 cells.