Synergistic effects of calcium silicate/zinc silicate dual compounds andin-situinterconnected pores on promoting bone regeneration of composite scaffolds.

Rapid bone regeneration in implants is important for successful transplantation. In this regard, we report the development of calcium silicate/zinc silicate (CS/ZS) dual-compound-incorporated calcium phosphate cement (CPC) scaffolds with a three-dimensional poly (lactic-co-glycolic acid) network that synergistically promote bone regeneration.In vitroresults demonstrated that the incorporation of CS/ZS dual compounds into the CPC significantly promoted the osteogenic differentiation of stem cells compared to the addition of CS or ZS alone. Moreover, the bone-regeneration efficacy of the composite scaffolds was validated by filling in femur condyle defects in rabbits, which showed that the scaffolds with CS and ZS possessed a great bone repair effect, as evidenced by more new bone formation and a faster scaffold biodegradation compared to the scaffold with CS alone.

pH-Responsive nanoplatform synergistic gas/photothermal therapy to eliminate biofilms in poly(L-lactic acid) scaffolds.

To date, implant-associated infection is still a significant clinical challenge, which cannot be effectively eliminated by single therapies due to the formation of microbial biofilms. Herein, a pH-responsive nanoplatform was constructed via the in situ growth of zinc sulfide (ZnS) nanoparticles on the surface of Ti3C2 MXene nanosheets, which was subsequently introduced in poly(L-lactic acid) (PLLA) to prepare a composite bone scaffold via selective laser sintering technology. In the acidic biofilm microenvironment, the degradation of ZnS released hydrogen sulfide (H2S) gas to eliminate the biofilm extracellular DNA (eDNA), thus destroying the compactness of the biofilm. Then, the bacterial biofilm became sensitive to hyperthermia, which could be further destroyed under near-infrared light irradiation due to the excellent photothermal property of MXene, finally achieving gas/photothermal synergistic antibiofilm and efficient sterilization. The results showed that the synergistic gas/photothermal therapy for the composite scaffold not only evidently inhibited the formation of biofilms, but also effectively eradicated the eDNA of the already-formed biofilms and killed 90.4% of E. coli and 84.2% of S. aureus under near infrared light irradiation compared with single gas or photothermal therapy. In addition, the composite scaffold promoted the proliferation and osteogenic differentiation of mouse bone marrow mesenchymal stem cells. Thus, the designed scaffold with excellent biofilm elimination and osteogenesis ability has great potential as an alternative treatment for implant-associated bone infections.

[Preparation of berberine-naringin dual drug-loaded composite microspheres and evaluation of their antibacterial-osteogenic properties].

Objective: To develop a drug-loaded composite microsphere that can simultaneously release the berberine (BBR) and naringin (NG) to repair infectious bone defects. Methods: The NG was loaded on mesoporous microspheres (MBG) to obtain the drug-loaded microspheres (NG-MBG). Then the dual drug-loaded compound microspheres (NG-MBG@PDA-BBR) were obtained by wrapping NG-MBG with polydopamine (PDA) and modifying the coated PDA with BBR. The composite microspheres were characterized by scanning electron microscopy, X-ray diffraction, specific surface area and pore volume analyzer, and Fourier transform infrared spectroscopy; the drug loading rate and release of NG and BBR were measured; the colony number was counted and the bacterial inhibition rate was calculated after co-culture with Staphylococcus aureus and Escherichia coli for 12 hours to observe the antibacterial effect; the biocompatibility was evaluated by live/dead cell fluorescence staining and cell counting kit 8 assay after co-culture with rat's BMSCs for 24 and 72 hours, respectively, and the osteogenic property was evaluated by alkaline phosphatase (ALP) staining and alizarin red staining after 7 and 14 days, respectively. Results: NG-MBG@PDA-BBR and three control microspheres (MBG, MBG@PDA, and NG-MBG@PDA) were successfully constructed. Scanning electron microscopy showed that NG-MBG@PDA-BBR had a rough lamellar structure, while MBG had a smooth surface, and MBG@PDA and NG-MBG@PDA had a wrapped agglomeration structure. Specific surface area analysis showed that MBG had a mesoporous structure and had drug-loading potential. Low angle X-ray diffraction showed that NG was successfully loaded on MBG. The X-ray diffraction pattern contrast showed that all groups of microspheres were amorphous. Fourier transform infrared spectroscopy showed that NG and BBR peaks existed in NG-MBG@PDA-BBR. NG-MBG@PDA-BBR had good sustained drug release ability, and NG and BBR had early burst release and late sustained release. NG-MBG@PDA-BBR could inhibit the growth of Staphylococcus aureus and Escherichia coli, and the antibacterial ability was significantly higher than that of MBG, MBG@PDA, and NG-MBG@PDA ( P<0.05). But there was a significant difference in biocompatibility at 72 hours among microspheres ( P<0.05). ALP and alizarin red staining showed that the ALP positive area and the number of calcium nodules in NG-MBG@PDA-BBR were significantly higher than those of MBG and NG-MBG ( P<0.05), and there was no significant difference between NG-MBG@PDA and NG-MBG@PDA ( P>0.05). Conclusion: NG-MBG@PDA-BBR have sustained release effects on NG and BBR, indicating that it has ideal dual performance of osteogenesis and antibacterial property.

Direct osteogenesis and immunomodulation dual function via sustained release of naringin from the polymer scaffold.

Many traditional Chinese medicine monomers, such as naringin (NG), can regulate the local immune microenvironment to benefit osteogenesis. However, the rapid release of NG from scaffolds severely influences the osteogenesis-promoting effect. Herein, NG was loaded into mesoporous bioglass (MBG) to achieve sustained release through physical adsorption and the barrier role of mesoporous channels, then MBG loaded with NG was added to poly(L-lactic acid) (PLLA) to fabricate composite scaffolds by selective laser sintering (SLS) technology. The results showed that the NG-MBG/PLLA scaffolds could continuously and slowly release NG for 14 days compared with NG/PLLA scaffolds, and the cumulative release amount for the NG-MBG/PLLA scaffolds was 44.26%. In addition, the NG-MBG/PLLA scaffolds can promote the proliferation and osteogenesis differentiation of mouse bone marrow mesenchymal stem cells (mBMSCs). Meanwhile, the composite scaffolds decreased the reactive oxygen species (ROS) level of RAW264.7 under the stimulation of lipopolysaccharide (LPS) and significantly suppressed interleukin-6 (IL-6) and enhanced arginase-1 (Arg-1) protein expressions. Moreover, calcium nodule and alkaline phosphatase production of mBMSCs in a macrophage-conditioned medium for the NG-MBG/PLLA group also evidently increased compared with the PLLA and MBG/PLLA groups. These NG sustained-release composite scaffolds with osteo-immunomodulation function have great application prospects in the clinic.

A spatiotemporal drug release scaffold with antibiosis and bone regeneration for osteomyelitis.

INTRODUCTION: Scaffolds loaded with antibacterial agents and osteogenic drugs are considered essential tools for repairing bone defects caused by osteomyelitis. However, the simultaneous release of two drugs leads to premature osteogenesis and subsequent sequestrum formation in the pathological situation of unthorough antibiosis. OBJECTIVES: In this study, a spatiotemporal drug-release polydopamine-functionalized mesoporous silicon nanoparticle (MSN) core/shell drug delivery system loaded with antibacterial silver (Ag) nanoparticles and osteogenic dexamethasone (Dex) was constructed and introduced into a poly-l-lactic acid (PLLA) scaffold for osteomyelitis therapy. METHODS: MSNs formed the inner core and were loaded with Dex through electrostatic adsorption (MSNs@Dex), and then polydopamine was used to seal the core through the self-assembly of dopamine as the outer shell (pMSNs@Dex). Ag nanoparticles were embedded in the polydopamine shell via an in situ growth technique. Finally, the Ag-pMSNs@Dex nanoparticles were introduced into PLLA scaffolds (Ag-pMSNs@Dex/PLLA) constructed by selective laser sintering (SLS). RESULTS: The Ag-pMSNs@Dex/PLLA scaffold released Ag+ at the 12th hour, followed by the release of Dex starting on the fifth day. The experiments verified that the scaffold had excellent antibacterial performance against Escherichia coli and Staphylococcus aureus. Moreover, the scaffold significantly enhanced the osteogenic differentiation of mouse bone marrow mesenchymal stem cells. CONCLUSION: The findings suggested that this spatiotemporal drug release scaffold had promising potential for osteomyelitis therapy.

A bifunctional zoledronate sustained-release system in scaffold: Tumor therapy and bone repair.

It is of great challenges to repair bone defect and prevent tumor recurrence in bone tumors postoperative treatment. Bone scaffolds loaded with zoledronate (ZOL) are expected to solve these issues due to its osteogenesis and anti-tumor ability. Furthermore, ZOL needs to be sustained release to meet the requirement of long-term therapy. In this study, ZOL was loaded into amination functionalized mesoporous silicon (SBA15NH2), and then incorporated into poly (L-lactic acid) to prepare PLLA/SBA15NH2-ZOL scaffold via selective laser sintering technology. On one hand, ZOL of local release not only can inhibit growth and proliferation of bone tumor cells but also inhibit osteoclast differentiation through competitive binding of receptor activator of nuclear factor (NF)-kB (RANK) in osteoclast precursors. On the other hand, amination function could change the surface charge of mesoporous silica to positive charge to enhance the absorption of ZOL, mesoporous structure and abundant amino groups of SBA15NH2 play a barrier role and form hydrogen bond with phosphate groups of ZOL, respectively, thereby achieving its sustained release. The results showed that the loading amount of ZOL was 236.53 mg/g, and the scaffold could sustainedly release ZOL for more than 6 weeks. The scaffold inhibited proliferation of osteosarcoma cells through inducing apoptosis and cell cycle arrest. TRAP staining and F-actin ring formation experiment showed the scaffold inhibited differentiation and mature of osteoclast. Pit formation assay indicated that bone resorption activity was inhibited strongly.

Construction of magnetic nanochains to achieve magnetic energy coupling in scaffold.

BACKGROUND: Fe3O4 nanoparticles are highly desired for constructing endogenous magnetic microenvironment in scaffold to accelerate bone regeneration due to their superior magnetism. However, their random arrangement easily leads to mutual consumption of magnetic poles, thereby weakening the magnetic stimulation effect. METHODS: In this study, magnetic nanochains are synthesized by magnetic-field-guided interface co-assembly of Fe3O4 nanoparticles. In detail, multiple Fe3O4 nanoparticles are aligned along the direction of magnetic force lines and are connected in series to form nanochain structures under an external magnetic field. Subsequently, the nanochain structures are covered and fixed by depositing a thin layer of silica (SiO2), and consequently forming linear magnetic nanochains (Fe3O4@SiO2). The Fe3O4@SiO2 nanochains are then incorporated into poly l-lactic acid (PLLA) scaffold prepared by selective laser sintering technology. RESULTS: The results show that the Fe3O4@SiO2 nanochains with unique core-shell structure are successfully constructed. Meanwhile, the orderly assembly of nanoparticles in the Fe3O4@SiO2 nanochains enable to form magnetic energy coupling and obtain a highly magnetic micro-field. The in vitro tests indicate that the PLLA/Fe3O4@SiO2 scaffolds exhibit superior capacity in enhancing cell activity, improving osteogenesis-related gene expressions, and inducing cell mineralization compared with PLLA and PLLA/Fe3O4 scaffolds. CONCLUSION: In short, the Fe3O4@SiO2 nanochains endow scaffolds with good magnetism and cytocompatibility, which have great potential in accelerating bone repair.

Silver-decorated black phosphorus: a synergistic antibacterial strategy.

Black phosphorus (BP) exhibits great potential as antibacterial materials due to its unique photocatalytic activity. However, the unsatisfactory optical absorption and quick recombination of photoinduced electron-hole pairs restrain its photocatalytic antibacterial performance. In this work, silver nanoparticles (AgNPs) were decorated on BP to construct BP@AgNPs nanohybrids and then introduced into poly-l-lactic acid scaffold. Combining the tunable bandgap of BP and the LSPR effect of AgNPs, BP@AgNPs nanohybrids displayed the broaden visible light absorption. Furthermore, AgNPs acted as electron acceptors could accelerate charge transfer and suppress electron-hole recombination. Therefore, BP@AgNPs nanohybrids achieved synergistically enhanced photocatalytic antibacterial activity under visible light irradiation. Fluorescence probe experiment verified that BP@AgNPs promoted the generation of reactive oxygen species, which could disrupt bacteria membrane, damage DNA and oxide proteins, and finally lead to bacteria apoptosis. As a result, the scaffold possessed strong antibacterial efficiency with a bactericidal rate of 97% under light irradiation. Moreover, the scaffold also exhibited good cytocompatibility. This work highlighted a new strategy to develop photocatalytic antibacterial scaffold for bone implant application.

Silver-doped bioglass modified scaffolds: A sustained antibacterial efficacy.

Implant-related bacterial infection is a serious complication, which even causes implant failure. Silver (Ag) nanoparticles are broadly used antibacterial agents due to their excellent antibacterial ability and broad-spectrum bactericidal property. However, the significance of burst release cannot be entirely ignored. In this study, Ag doped mesoporous bioactive glasses (Ag-MBG) nanospheres were synthesized using modified Stöber method, then incorporated into poly L-lactic acid (PLLA) matrix to prepare the composite scaffolds via selective laser sintering (SLS) technology. Herein, Mesoporous bioactive glasses (MBG) sol had many negatively-charged silicon hydroxyl groups, which could adsorb positively-charged Ag ions by electrostatic interaction and eventually form Si-O-Ag bonds into MBG. Moreover, MBG promoted osteoblast colonization due to its continuous release of Si ions. The results showed the Ag-MBG/PLLA scaffold could sustainedly release Ag ions for 28 days, and exhibited significantly antibacterial ability against Escherichia coli, its bacterial inhibition rate was over 80%. In addition, the composite scaffold also showed good cytocompatibility. It may be concluded that the prepared Ag-MBG/PLLA scaffold has great potential to repair implant-associated bone infection.

Polydopamine-decorated black phosphorous to enhance stability in polymer scaffold.

Black phosphorous (BP) is recognized as an effective reinforcement for polymer scaffold because of its excellent mechanical property and biocompatibility. Nevertheless, its poor stability in physiological environment limits its application in bone repair. In this work, BP was modified with dopamine by self-polymerization approach (donated as BP@PDA) to improve its stability, and then was introduced into poly-L-lactic acid (PLLA) scaffold fabricated by selective laser sintering technology. Results showed the compressive and tensile strength of PLLA/BP@PDA scaffold were improved by 105% and 50%, respectively. The enhanced strength was ascribed to the increased stability of BP and the improved compatibility of BP@PDA with PLLA matrix after modifying with polydopamine. Simultaneously, the bioactivity of PLLA scaffold was significantly improved. It was attributed to that BP@PDA provided the sustained source ofPO43-ions which could capture Ca2+ions from physiological medium to facilitatein situbiomineralization, thereby promoting cell adhesion, proliferation and differentiation. This study demonstrated the great potential of BP@PDA in bone repair.

3D Printed Zn-doped Mesoporous Silica-incorporated Poly-L-lactic Acid Scaffolds for Bone Repair.

Poly-L-lactic acid (PLLA) lacks osteogenic activity, which limits its application in bone repair. Zinc (Zn) is widely applied to strengthen the biological properties of polymers due to its excellent osteogenic activity. In the present study, Zn-doped mesoporous silica (Zn-MS) particles were synthesized by one-pot hydrothermal method. Then, the particles were induced into PLLA scaffolds prepared by selective laser sintering technique, aiming to improve their osteogenic activity. Our results showed that the synthesized particles possessed rosette-like morphology and uniform mesoporous structure, and the composite scaffold displayed the sustained release of Zn ion in a low concentration range, which was attributed to the shield effect of the PLLA matrix and the strong bonding interaction of Si-O-Zn. The scaffold could evidently promote osteogenesis differentiation of mouse bone marrow mesenchymal stem cells by upregulating their osteogenesis-related gene expression. Besides, Zn-MS particles could significantly increase the compressive strength of the PLLA scaffold because of their rosette-like morphology and mesoporous structure, which can form micromechanical interlocking with the PLLA matrix. The Zn-MS particles possess great potential to improve various polymer scaffold properties due to their advantageous morphology and physicochemical properties.

Synthesis of a mace-like cellulose nanocrystal@Ag nanosystem via in-situ growth for antibacterial activities of poly-L-lactide scaffold.

Antibacterial property for scaffolds is an urgent problem to prevent infections in bone repair. Ag nanoparticles possess excellent bactericidal activities, whereas their agglomeration restricts the full play of antibacterial property in scaffold. Herein, a mace-like nanosystem was constructed to improve their dispersion by in-situ growth of Ag nanoparticles on cellulose nanocrystal (CNC), which was labeled CNC@Ag nanosystem. Subsequently, the CNC@Ag nanosystem was introduced into poly-L-lactide (PLLA) scaffolds. Results demonstrated that the nanosystem uniformly dispersed in scaffold. The antibacterial tests demonstrated that the scaffolds possessed robust antibacterial activities against E. coli, with bacterial inhibition rate over 95%. Moreover, ion release behavior corroborated the scaffolds continuously released Ag+ for more than 28 days, which benefited from the immobilization effect of CNC on Ag. Encouragingly, the mechanical properties of the scaffolds were remarkably higher than that of PLLA/CNC scaffolds, owing to the mace-like CNC@Ag nanosystem improved the load transfer efficiency in the scaffold.

Construction of Fe3O4-Loaded Mesoporous Carbon Systems for Controlled Drug Delivery.

Magnetite (Fe3O4) nanoparticles as drug carriers can achieve precise drug target due to their magnetic property. However, they are easy to aggregate in the physiological environment, which obviously limits their application in drug delivery. The development of the Fe-MIL-88B-derived method to construct the Fe3O4-loaded mesoporous carbon (Fe3O4/carbon) system is a feasible strategy to solve the issue. First, iron atoms evenly distribute in the organic links through coordination bonds in Fe-MIL-88B. After the carbonization of Fe-MIL-88B, mesoporous carbon acts as a barrier to prevent the aggregation of Fe3O4 nanoparticles. Herein, Fe-MIL-88B particles were fabricated by the hydrothermal method and then pyrolyzed to construct Fe3O4/carbon systems. Results showed that Fe3O4 nanoparticles uniformly in situ grew on mesoporous carbon generated by the carbonization of organic components. More encouragingly, the Fe3O4/carbon system loaded with DOX demonstrated pH-responsive DOX release, efficient delivery of DOX into cancer cells, and significant cancer cell killing ability. Therefore, the Fe3O4/carbon systems prepared by the Fe-MIL-88B-derived method might open up a way for targeted and controlled drug delivery.

Novel Strategy to Accelerate Bone Regeneration of Calcium Phosphate Cement by Incorporating 3D Plotted Poly(lactic-co-glycolic acid) Network and Bioactive Wollastonite.

Inefficient bone regeneration of self-hardening calcium phosphate cement (CPC) increases the demand for interconnected macropores and osteogenesis-stimulated substances. It remains a challenge to fabricate porous CPC with interconnected macropores while maintaining its advantages, such as plasticity. Herein, pastes containing CPC and wollastonite (WS) are infiltrated into a 3D plotted poly(lactic-co-glycolic acid) (PLGA) network to fabricate plastic CPC-based composite cement (PLGA/WS/CPC). The PLGA/WS/CPC recovers the plasticity of CPC after being heated above the glass transition temperature of PLGA. The presence of the 3D PLGA network significantly increases the flexibility of CPC in prophase and generates 3D interconnected macropores in situ upon its degradation. The addition of WS is helpful to improve the attachment, proliferation, and osteogenic differentiation of mouse bone marrow stromal cells in vitro. The in vivo experimental results indicate that PLGA/WS/CPC promotes rapid angiogenesis and bone formation. Therefore, the plastic CPC-based composite cement with a 3D PLGA network and wollastonite shows an obviously improved efficiency for repairing bone defects and is expected to facilitate the wider application of CPC in the clinic.

An immuno-potentiating vehicle made of mesoporous silica-zinc oxide micro-rosettes with enhanced doxorubicin loading for combined chemoimmunotherapy.

Herein, mesoporous silica-zinc oxide (MS-Zn) micro-rosettes with controllable petal thickness were synthesized by a facile one-pot hydrothermal method. MS-Zn loaded with doxorubicin and polyinosinic-polycytidylic acid sodium salt not only significantly inhibits tumor growth but also effectively rejects tumor metastasis in vivo.

Improving the anti-washout property of calcium phosphate cement by introducing konjac glucomannan/κ-carrageenan blend.

Anti-washout calcium phosphate cement (CPC) was prepared by dissolving water-soluble konjac glucomannan (KGM) and κ-carrageenan (KC) blend in the cement liquid. The anti-washout property, setting time, compressive strength and in vitro cytocompatibility of the CPC modified with KGM/KC blend were evaluated. The results indicated that the CPC pastes modified with KGM/KC blend exhibited excellent anti-washout property. The addition of KGM/KC blend shortened the setting time and increased the injectability of CPC. Although the introduction of KGM/KC blend reduced the compressive strength of CPC, the compressive strength still surpassed that of human cancellous bone. The optimal KGM/KC mass ratio was 2:8, with which the modified cement exhibited the most efficient washout resistance and the highest compressive strength. The introduction of KGM/KC blend obviously promoted the proliferation of mouse bone marrow mesenchymal stem cells. This anti-washout CPC modified by KGM/KC blend with excellent in vitro cytocompatibility will have good prospects for application in bone defect repair.

Improvement of anti-washout property of calcium phosphate cement by addition of konjac glucomannan and guar gum.

The inferior anti-washout property of injectable calcium phosphate cement (CPC) limits its wider application in clinic. In this study, the improvement of anti-washout performance of CPC by addition of konjac glucomannan or guar gum, which was dissolved in the CPC liquid, was first studied. The influence of KGM/GG blend with different mass ratios on the anti-washout property, compressive strength and in vitro cytocompatibility of CPC was estimated. The results revealed that small amount of KGM or GG could obviously enhance the anti-washout property of CPC. Moreover, the washout resistance efficiency of KGM/GG blend was better than KGM or GG alone. The addition of KGM/GG blend slightly shortened the final setting time of CPC. Although the introduction of KGM/GG blend reduced the compressive strength of CPC, the compressive strength still reached or surpassed that of human cancellous bone. The best KGM/GG mass ratio was 5:5, which was most efficient at not only reducing CPC disintegration, but also increasing compressive strength. The addition of KGM/GG blend obviously promoted the cells proliferation on the CPC. In short, the CPC modified by KGM/GG blend exhibited excellent anti-washout property, appropriate setting time, adequate compressive strength, and good cytocompatibility, and has the potential to be used in bone defect repair. The addition of KGM/GG blend significantly improved the anti-washout property of CPC. The best KGM/GG mass ratio was 5:5, which was most efficient in reducing the CPC disintegration.

Fabrication of ß-tricalcium phosphate composite ceramic sphere-based scaffolds with hierarchical pore structure for bone regeneration.

Polymer sphere-based scaffolds, which are prepared by bonding the adjacent spheres via sintering the randomly packed spheres, feature uniform pore structure, full three-dimensional (3D) interconnection, and considerable mechanical strength. However, bioceramic sphere-based scaffolds fabricated by this method have never been reported. Due to high melting temperature of bioceramic, only limited diffusion rate can be achieved when sintering the bioceramic spheres, which is far from enough to form robust bonding between spheres. In the present study, for the first time we fabricated 3D interconnected ß-tricalcium phosphate ceramic sphere-based (PG/TCP) scaffolds by introducing phosphate-based glass (PG) as sintering additive and placing uniaxial pressure during the sintering process. The sintering mechanism of PG/TCP scaffolds was unveiled. The PG/TCP scaffolds had hierarchical pore structure, which was composed by interconnected macropores (>200 µm) among spheres, pores (20­120 µm) in the interior of spheres, and micropores (1­3 µm) among the grains. During the sintering process, partial PG reacted with ß-TCP, forming ß-Ca2P2O7; metal ions from PG substituted to Ca2+ sites of ß-TCP. The mechanical properties (compressive strength 2.8­10.6 MPa; compressive modulus 190­620 MPa) and porosity (30%­50%) of scaffolds could be tailored by manipulating the sintering temperatures. The introduction of PG accelerated in vitro degradation of scaffolds, and the PG/TCP scaffolds showed good cytocompatibility. This work may offer a new strategy to prepare bioceramic scaffolds with satisfactory physicochemical properties for application in bone regeneration.

A tracheal scaffold of gelatin-chondroitin sulfate-hyaluronan-polyvinyl alcohol with orientated porous structure.

Scaffold of gelatin-chondroitin sulfate-hyaluronan-polyvinyl alcohol (GCH-PVA) with orientated microtubule structure and good hydrophilicity was fabricated by unidirectional freeze-drying method mimicking the composition and structure of tracheal cartilage extracellular matrix. PVA was incorporated to improve flexibility and viscoelasticity of GCH scaffold. All wet scaffolds showed similar compressive elastic modulus with native cartilage. GCH-PVA scaffolds showed high relative remaining stress during relaxation indicating good mechanical stability. The hysteresis ratio during cyclic compression increased gradually with PVA content and close to native cartilage. During multiple frequency compression, all scaffolds showed a low loss tangent close to native cartilage, and PVA incorporation enhanced the elasticity of scaffolds when they were stressed under high frequency. The incorporation of PVA promoted gene expression of adhesion related integrin α5ß1 and actin by mouse bone marrow mesenchymal stem cells (mBMSCs). With the orientated microtubule structure, cells ingrowth into scaffolds was facilitated by dynamic culture method.