Tendon-bone healing is essential for an effective rotator cuff tendon repair surgery,however,this remains a significant challenge due to the lack of biomaterials with high strength and bioactivity.Inspired by the high...Tendon-bone healing is essential for an effective rotator cuff tendon repair surgery,however,this remains a significant challenge due to the lack of biomaterials with high strength and bioactivity.Inspired by the high-performance exoskeleton of natural organisms,we set out to apply natural fish scale(FS)modified by calcium silicate nanoparticles(CS NPs)as a new biomaterial(CS-FS)to overcome the challenge.Benefit from its“Bouligand”microstructure,such FS-based scaffold maintained excellent tensile strength(125.05 MPa)and toughness(14.16 MJ/m^(3)),which are 1.93 and 2.72 times that of natural tendon respectively,allowing it to well meet the requirements for rotator cuff tendon repair.Additionally,CS-FS showed diverse bioactivities by stimulating the differentiation and phenotypic maintenance of multiple types of cells participated into the composition of tendon-bone junction,(e.g.bone marrow mesenchymal stem cells(BMSCs),chondrocyte,and tendon stem/progenitor cells(TSPCs)).In both rat and rabbit rotator cuff tear(RCT)models,CS-FS played a key role in the tendon-bone interface regeneration and biomechanical function,which may be achieved by activating BMP-2/Smad/Runx2 pathway in BMSCs.Therefore,natural fish scale-based biomaterials are the promising candidate for clinical tendon repair due to their outstanding strength and bioactivity.展开更多
The treatment of rotator cuff tear is one of the major challenges for orthopedic surgeons.The key to treatment is the reconstruction of the tendon-bone interface(TBI).Autologous platelet-rich plasma(PRP)is used as a t...The treatment of rotator cuff tear is one of the major challenges for orthopedic surgeons.The key to treatment is the reconstruction of the tendon-bone interface(TBI).Autologous platelet-rich plasma(PRP)is used as a therapeutic agent to accelerate the healing of tendons,as it contains a variety of growth factors and is easy to prepare.Graphene oxide(GO)is known to improve the physical properties of biomaterials and promote tissue repair.In this study,PRP gels containing various concentrations of GO were prepared to promote TBI healing and supraspinatus tendon reconstruction in a rabbit model.The incorporation of GO improved the ultrastructure and mechanical properties of the PRP gels.The gels containing 0.5 mg/ml GO(0.5 GO/PRP)continuously released transforming growth factor-b1(TGF-b1)and platelet-derived growth factor(PDGF)-AB,and the released TGF-b1 and PDGF-AB were still at high concentrations,1063.451 pg/ml and814.217 pg/ml,respectively,on the 14th day.In vitro assays showed that the 0.5 GO/PRP gels had good biocompatibility and promoted bone marrow mesenchymal stem cells proliferation and osteogenic and chondrogenic differentiation.After 12 weeks of implantation,the magnetic resonance imaging,microcomputed tomography and histological results indicated that the newly regenerated tendons in the 0.5 GO/PRP group had a similar structure to natural tendons.Moreover,the biomechanical results showed that the newly formed tendons in the 0.5 GO/PRP group had better biomechanical properties compared to those in the other groups,and had more stable TBI tissue.Therefore,the combination of PRP and GO has the potential to be a powerful advancement in the treatment of rotator cuff injuries.展开更多
Healing of an anterior cruciate ligament(ACL)autologous graft in a bone tunnel occurs through the formation of fibrovascular scar tissue,which is structurally and compositionally inferior to normal fibrocartilaginous ...Healing of an anterior cruciate ligament(ACL)autologous graft in a bone tunnel occurs through the formation of fibrovascular scar tissue,which is structurally and compositionally inferior to normal fibrocartilaginous insertion and thus may increase the reconstruction failure and the rate of failure recurrence.In this study,an injectable hydroxyapatite/type I collagen(HAp/Col I)paste was developed to construct a suitable local microenvironment to accelerate the healing of bone-tendon interface.Physicochemical characterization demonstrated that the HAp/Col I paste was injectable,uniform and stable.The in vitro cell culture illustrated that the paste could promote MC3T3-E1 cells proliferation and osteogenic expression.The results of a canine ACL reconstruction study showed that the reconstructive ACL had similar texture and color as the native ACL.The average width of the tunnel,total bone volume,bone volume/tissue volume and trabecular number acquired from micro-CT analysis suggested that the healing of tendon-bone interface in experimental group was better than that in control group.The biomechanical test showed the maximal loads in experimental group achieved approximately half of native ACL’s maximal load at 24 weeks.According to histological examination,Sharpey fibers could be observed as early as 12 weeks postoperatively while a typical four-layer transitional structure of insertion site was regenerated at 48 weeks in the experimental group.The injectable HAp/Col I paste provided a biomimetic scaffold and microenvironment for early cell attachment and proliferation,further osteogenic expression and extracellular matrix deposition,and in vivo structural and functional regeneration of the tendon-bone interface.展开更多
Despite transosseous rotator cuff tear repair using sutures is widely accepted for tendon-bone fixation,the fibrocartilaginous enthesis regeneration is still hardly achieved with the traditional sutures.In the present...Despite transosseous rotator cuff tear repair using sutures is widely accepted for tendon-bone fixation,the fibrocartilaginous enthesis regeneration is still hardly achieved with the traditional sutures.In the present work,degradable magnesium(Mg)alloy wire was applied to suture supraspinatus tendon in a rat acute rotator cuff tear model with Vicryl Plus 4±0 absorbable suture as control.The shoulder joint humerus-supraspinatus tendon complex specimens were retrieved at 4,8,and 12 weeks after operation.The Mg alloy suture groups showed better biomechanical properties in terms of ultimate load to failure.Gross observation showed that hyperplastic response of the scar tissue at the tendon-bone interface is progressively alleviated over time in the both Mg alloy suture and Vicryl suture groups.In the histological analysis,for Mg alloy suture groups,chondrocytes appear to proliferate at 4 weeks postoperatively,and the tendon-bone interface showed an orderly structural transition zone at 8 weeks postoperatively.The collagenous fiber tended to be aligned and the tendon-bone interlocking structures apparently formed,where transitional structure from unmineralized fibrocartilage to mineralized fibrocartilage was closer to the native fibrocartilaginous enthesis.In vivo degradation of the magnesium alloy wire was completed within 12 weeks.The results indicated that Mg alloy wire was promising as degradable suture with the potential to promotes fibrocartilaginous interface regeneration in rotator cuff repair.展开更多
BACKGROUND Fibroblast plays a major role in tendon-bone healing.Exosomes derived from bone marrow mesenchymal stem cells(BMSCs)can activate fibroblasts and promote tendon-bone healing via the contained microRNAs(miRNA...BACKGROUND Fibroblast plays a major role in tendon-bone healing.Exosomes derived from bone marrow mesenchymal stem cells(BMSCs)can activate fibroblasts and promote tendon-bone healing via the contained microRNAs(miRNAs).However,the underlying mechanism is not comprehensively understood.Herein,this study aimed to identify overlapped BMSC-derived exosomal miRNAs in three GSE datasets,and to verify their effects as well as mechanisms on fibroblasts.AIM To identify overlapped BMSC-derived exosomal miRNAs in three GSE datasets and verify their effects as well as mechanisms on fibroblasts.METHODS BMSC-derived exosomal miRNAs data(GSE71241,GSE153752,and GSE85341)were downloaded from the Gene Expression Omnibus(GEO)database.The candidate miRNAs were obtained by the intersection of three data sets.TargetScan was used to predict potential target genes for the candidate miRNAs.Functional and pathway analyses were conducted using the Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases,respectively,by processing data with the Metascape.Highly interconnected genes in the protein-protein interaction(PPI)network were analyzed using Cytoscape software.Bromodeoxyuridine,wound healing assay,collagen contraction assay and the expression of COL I andα-smooth muscle actin positive were applied to investigate the cell proliferation,migration and collagen synthesis.Quantitative real-time reverse transcription polymerase chain reaction was applied to determine the cell fibroblastic,tenogenic,and chondrogenic potential.RESULTS Bioinformatics analyses found two BMSC-derived exosomal miRNAs,has-miR-144-3p and hasmiR-23b-3p,were overlapped in three GSE datasets.PPI network analysis and functional enrichment analyses in the GO and KEGG databases indicated that both miRNAs regulated the PI3K/Akt signaling pathway by targeting phosphatase and tensin homolog(PTEN).In vitro experiments confirmed that miR-144-3p and miR-23b-3p stimulated proliferation,migration and collagen synthesis of NIH3T3 fibroblasts.Interfering with PTEN affected the phosphorylation of Akt and thus activated fibroblasts.Inhibition of PTEN also promoted the fibroblastic,tenogenic,and chondrogenic potential of NIH3T3 fibroblasts.CONCLUSION BMSC-derived exosomes promote fibroblast activation possibly through the PTEN and PI3K/Akt signaling pathways,which may serve as potential targets to further promote tendon-bone healing.展开更多
Magnesium(Mg)screws perform clinical potential in anterior cruciate ligament(ACL)reconstruction,and promote fibrocartilaginous entheses regeneration at the femoral entrance.We aim to prove that high-purity Magnesium(H...Magnesium(Mg)screws perform clinical potential in anterior cruciate ligament(ACL)reconstruction,and promote fibrocartilaginous entheses regeneration at the femoral entrance.We aim to prove that high-purity Magnesium(HP Mg)screws modulate macrophage polarization in fibrocartilage interface regeneration both in vitro and in vivo.HP Mg extracts performed good cytocompatibility and significantly promoted M2 macrophage polarization in the flow cytometry and ELISA assays.M2 macrophages stimulated fibrochondrocyte differentiation of cocultured hBMSCs,and HP Mg extracts had synergistic effect on the process.Then we applied HP Mg screws,with Ti screws as control,in the ACL reconstruction rabbit model.In the histological and immunofluorescence analysis,HP Mg screws inhibited M1 polarization at 2 weeks and highly promoted M2 polarization at 2 and 4 weeks at the tendon–bone interface.Furthermore,regeneration of fibrocartilaginous entheses,rather than the fibrovascular scar interface,was detected in the HP Mg group at 12 weeks.For further mechanism study via RNA-seq detection and WB assays,we found that AKT1 was highly activated in M2 polarization,and HP Mg could stimulate AKT1 expression,rather than AKT2,in the early phase of tendon–bone healing.Our study elucidated macrophage polarization during tendon–bone healing process and emphasized HP Mg on M2 polarization and fibrocartilage interface regeneration via the selective activation of AKT1 and PI3K/AKT pathway.展开更多
基金supported by the National Basic Research Program of China(grant No.2021YFA0715700)the Natural Science Foundation of China(32130062)+1 种基金Innovation Cross Team of Chinese Academy of Sciences(JCTD-2018-13)Science and Technology Commission of Shanghai Municipality(20442420300,21DZ1205600).
文摘Tendon-bone healing is essential for an effective rotator cuff tendon repair surgery,however,this remains a significant challenge due to the lack of biomaterials with high strength and bioactivity.Inspired by the high-performance exoskeleton of natural organisms,we set out to apply natural fish scale(FS)modified by calcium silicate nanoparticles(CS NPs)as a new biomaterial(CS-FS)to overcome the challenge.Benefit from its“Bouligand”microstructure,such FS-based scaffold maintained excellent tensile strength(125.05 MPa)and toughness(14.16 MJ/m^(3)),which are 1.93 and 2.72 times that of natural tendon respectively,allowing it to well meet the requirements for rotator cuff tendon repair.Additionally,CS-FS showed diverse bioactivities by stimulating the differentiation and phenotypic maintenance of multiple types of cells participated into the composition of tendon-bone junction,(e.g.bone marrow mesenchymal stem cells(BMSCs),chondrocyte,and tendon stem/progenitor cells(TSPCs)).In both rat and rabbit rotator cuff tear(RCT)models,CS-FS played a key role in the tendon-bone interface regeneration and biomechanical function,which may be achieved by activating BMP-2/Smad/Runx2 pathway in BMSCs.Therefore,natural fish scale-based biomaterials are the promising candidate for clinical tendon repair due to their outstanding strength and bioactivity.
基金supported by the Luzhou Municipal People’s Government-SouthwestMedical University Science and Technology Cooperation Achievements Transformation Project(2019LZXNYDJ20C01).
文摘The treatment of rotator cuff tear is one of the major challenges for orthopedic surgeons.The key to treatment is the reconstruction of the tendon-bone interface(TBI).Autologous platelet-rich plasma(PRP)is used as a therapeutic agent to accelerate the healing of tendons,as it contains a variety of growth factors and is easy to prepare.Graphene oxide(GO)is known to improve the physical properties of biomaterials and promote tissue repair.In this study,PRP gels containing various concentrations of GO were prepared to promote TBI healing and supraspinatus tendon reconstruction in a rabbit model.The incorporation of GO improved the ultrastructure and mechanical properties of the PRP gels.The gels containing 0.5 mg/ml GO(0.5 GO/PRP)continuously released transforming growth factor-b1(TGF-b1)and platelet-derived growth factor(PDGF)-AB,and the released TGF-b1 and PDGF-AB were still at high concentrations,1063.451 pg/ml and814.217 pg/ml,respectively,on the 14th day.In vitro assays showed that the 0.5 GO/PRP gels had good biocompatibility and promoted bone marrow mesenchymal stem cells proliferation and osteogenic and chondrogenic differentiation.After 12 weeks of implantation,the magnetic resonance imaging,microcomputed tomography and histological results indicated that the newly regenerated tendons in the 0.5 GO/PRP group had a similar structure to natural tendons.Moreover,the biomechanical results showed that the newly formed tendons in the 0.5 GO/PRP group had better biomechanical properties compared to those in the other groups,and had more stable TBI tissue.Therefore,the combination of PRP and GO has the potential to be a powerful advancement in the treatment of rotator cuff injuries.
基金supported by the National Key Research and Development Program of China(2018YFC1106200,2018YFC1106203)the National Natural Science Foundation of China(32071330)+1 种基金the Sichuan Science and Technology Innovation Team(2019JDTD0008)the Key Science and Technology Program of Guangxi Province(AA17204085-2).
文摘Healing of an anterior cruciate ligament(ACL)autologous graft in a bone tunnel occurs through the formation of fibrovascular scar tissue,which is structurally and compositionally inferior to normal fibrocartilaginous insertion and thus may increase the reconstruction failure and the rate of failure recurrence.In this study,an injectable hydroxyapatite/type I collagen(HAp/Col I)paste was developed to construct a suitable local microenvironment to accelerate the healing of bone-tendon interface.Physicochemical characterization demonstrated that the HAp/Col I paste was injectable,uniform and stable.The in vitro cell culture illustrated that the paste could promote MC3T3-E1 cells proliferation and osteogenic expression.The results of a canine ACL reconstruction study showed that the reconstructive ACL had similar texture and color as the native ACL.The average width of the tunnel,total bone volume,bone volume/tissue volume and trabecular number acquired from micro-CT analysis suggested that the healing of tendon-bone interface in experimental group was better than that in control group.The biomechanical test showed the maximal loads in experimental group achieved approximately half of native ACL’s maximal load at 24 weeks.According to histological examination,Sharpey fibers could be observed as early as 12 weeks postoperatively while a typical four-layer transitional structure of insertion site was regenerated at 48 weeks in the experimental group.The injectable HAp/Col I paste provided a biomimetic scaffold and microenvironment for early cell attachment and proliferation,further osteogenic expression and extracellular matrix deposition,and in vivo structural and functional regeneration of the tendon-bone interface.
基金the National Key Research and Development Program of China(No.2020YFC1107501)the National Natural Science Foundation of China(No.51971222,51801220)+4 种基金the Natural Science Foundation of Liaoning Province of China(No.2020-MS-001)the Dong Guan Innovative Research Team Program(No.2020607134012)the Military Translational Medicine Fund of Chinese PLA General Hospital(ZH19008)Capital’s Funds for Health Improvement and Research(CFH 2022-2-5051)the Dong Guan Science and Technology Service Network Initiative(20201600200042)。
文摘Despite transosseous rotator cuff tear repair using sutures is widely accepted for tendon-bone fixation,the fibrocartilaginous enthesis regeneration is still hardly achieved with the traditional sutures.In the present work,degradable magnesium(Mg)alloy wire was applied to suture supraspinatus tendon in a rat acute rotator cuff tear model with Vicryl Plus 4±0 absorbable suture as control.The shoulder joint humerus-supraspinatus tendon complex specimens were retrieved at 4,8,and 12 weeks after operation.The Mg alloy suture groups showed better biomechanical properties in terms of ultimate load to failure.Gross observation showed that hyperplastic response of the scar tissue at the tendon-bone interface is progressively alleviated over time in the both Mg alloy suture and Vicryl suture groups.In the histological analysis,for Mg alloy suture groups,chondrocytes appear to proliferate at 4 weeks postoperatively,and the tendon-bone interface showed an orderly structural transition zone at 8 weeks postoperatively.The collagenous fiber tended to be aligned and the tendon-bone interlocking structures apparently formed,where transitional structure from unmineralized fibrocartilage to mineralized fibrocartilage was closer to the native fibrocartilaginous enthesis.In vivo degradation of the magnesium alloy wire was completed within 12 weeks.The results indicated that Mg alloy wire was promising as degradable suture with the potential to promotes fibrocartilaginous interface regeneration in rotator cuff repair.
基金Supported by Sanming Project of Medicine in Shenzhen,No.SZSM201612078Health Shanghai Initiative Special Fund(Medical-Sports Integration,Creating a New Model of Exercise for Health),No.JKSHZX-2022-02.
文摘BACKGROUND Fibroblast plays a major role in tendon-bone healing.Exosomes derived from bone marrow mesenchymal stem cells(BMSCs)can activate fibroblasts and promote tendon-bone healing via the contained microRNAs(miRNAs).However,the underlying mechanism is not comprehensively understood.Herein,this study aimed to identify overlapped BMSC-derived exosomal miRNAs in three GSE datasets,and to verify their effects as well as mechanisms on fibroblasts.AIM To identify overlapped BMSC-derived exosomal miRNAs in three GSE datasets and verify their effects as well as mechanisms on fibroblasts.METHODS BMSC-derived exosomal miRNAs data(GSE71241,GSE153752,and GSE85341)were downloaded from the Gene Expression Omnibus(GEO)database.The candidate miRNAs were obtained by the intersection of three data sets.TargetScan was used to predict potential target genes for the candidate miRNAs.Functional and pathway analyses were conducted using the Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases,respectively,by processing data with the Metascape.Highly interconnected genes in the protein-protein interaction(PPI)network were analyzed using Cytoscape software.Bromodeoxyuridine,wound healing assay,collagen contraction assay and the expression of COL I andα-smooth muscle actin positive were applied to investigate the cell proliferation,migration and collagen synthesis.Quantitative real-time reverse transcription polymerase chain reaction was applied to determine the cell fibroblastic,tenogenic,and chondrogenic potential.RESULTS Bioinformatics analyses found two BMSC-derived exosomal miRNAs,has-miR-144-3p and hasmiR-23b-3p,were overlapped in three GSE datasets.PPI network analysis and functional enrichment analyses in the GO and KEGG databases indicated that both miRNAs regulated the PI3K/Akt signaling pathway by targeting phosphatase and tensin homolog(PTEN).In vitro experiments confirmed that miR-144-3p and miR-23b-3p stimulated proliferation,migration and collagen synthesis of NIH3T3 fibroblasts.Interfering with PTEN affected the phosphorylation of Akt and thus activated fibroblasts.Inhibition of PTEN also promoted the fibroblastic,tenogenic,and chondrogenic potential of NIH3T3 fibroblasts.CONCLUSION BMSC-derived exosomes promote fibroblast activation possibly through the PTEN and PI3K/Akt signaling pathways,which may serve as potential targets to further promote tendon-bone healing.
基金sponsored by the National Natural Science Foundation of China(No.81702183,No.81974325)Shanghai Sailing Program(No.17YF1414100).
文摘Magnesium(Mg)screws perform clinical potential in anterior cruciate ligament(ACL)reconstruction,and promote fibrocartilaginous entheses regeneration at the femoral entrance.We aim to prove that high-purity Magnesium(HP Mg)screws modulate macrophage polarization in fibrocartilage interface regeneration both in vitro and in vivo.HP Mg extracts performed good cytocompatibility and significantly promoted M2 macrophage polarization in the flow cytometry and ELISA assays.M2 macrophages stimulated fibrochondrocyte differentiation of cocultured hBMSCs,and HP Mg extracts had synergistic effect on the process.Then we applied HP Mg screws,with Ti screws as control,in the ACL reconstruction rabbit model.In the histological and immunofluorescence analysis,HP Mg screws inhibited M1 polarization at 2 weeks and highly promoted M2 polarization at 2 and 4 weeks at the tendon–bone interface.Furthermore,regeneration of fibrocartilaginous entheses,rather than the fibrovascular scar interface,was detected in the HP Mg group at 12 weeks.For further mechanism study via RNA-seq detection and WB assays,we found that AKT1 was highly activated in M2 polarization,and HP Mg could stimulate AKT1 expression,rather than AKT2,in the early phase of tendon–bone healing.Our study elucidated macrophage polarization during tendon–bone healing process and emphasized HP Mg on M2 polarization and fibrocartilage interface regeneration via the selective activation of AKT1 and PI3K/AKT pathway.
