Intra-articular injection of mesenchymal stem cells(MSCs)is a promising strategy for osteoarthritis(OA)treatment.However,more and more studies reveal that the injected MSCs have poor adhesion,migration,and survival in...Intra-articular injection of mesenchymal stem cells(MSCs)is a promising strategy for osteoarthritis(OA)treatment.However,more and more studies reveal that the injected MSCs have poor adhesion,migration,and survival in the joint cavity.A recent study shows that tropoelastin(TE)regulates adhesion,proliferation and phenotypic maintenance of MSCs as a soluble additive,indicating that TE could promote MSCs-homing in regenerative medicine.In this study,we used TE as injection medium,and compared it with classic media in MSCs intra-articular injection such as normal saline(NS),hyaluronic acid(HA),and platelet-rich plasma(PRP).We found that TE could effectively improve adhesion,migration,chondrogenic differentiation of infrapatellar fat pad MSCs(IPFP-MSCs)and enhance matrix synthesis of osteoarthritic chondrocytes(OACs)in indirect-coculture system.Moreover,TE could significantly enhance IPFP-MSCs adhesion via activation of integrin β1,ERK1/2 and vinculin(VCL)in vitro.In addition,intra-articular injection of TE-IPFP MSCs suspension resulted in a short-term increase in survival rate of IPFP-MSCs and better histology scores of rat joint tissues.Inhibition of integrin β1 or ERK1/2 attenuated the protective effect of TE-IPFP MSCs suspension in vivo.In conclusion,TE promotes performance of IPFP-MSCs and protects knee cartilage from damage in OA through enhancement of cell adhesion and activation of integrin β1/ERK/VCL pathway.Our findings may provide new insights in MSCs intra-articular injection for OA treatment.展开更多
The highly organized extracellular matrix(ECM)of musculoskeletal tissues,encompassing tendons,ligaments and muscles,is structurally anisotropic,hierarchical and multi-compartmental.These features collectively contribu...The highly organized extracellular matrix(ECM)of musculoskeletal tissues,encompassing tendons,ligaments and muscles,is structurally anisotropic,hierarchical and multi-compartmental.These features collectively contribute to their unique function.Previous studies have investigated the effect of tissue-engineered scaffold anisotropy on cell morphology and organization for musculoskeletal tissue repair and regeneration,but the hierarchical arrangement of ECM and compartmentalization are not typically replicated.Here,we present a method for multi-compartmental scaffold design that allows for physical mimicry of the spatial architecture of musculoskeletal tissue in regenerative medicine.This design is based on an ECM-inspired macromolecule scaffold.Polycaprolactone(PCL)scaffolds were fabricated with aligned fibers by electrospinning and mechanical stretching,and then surface-functionalized with the cell-supporting ECM protein molecule,tropoelastin(TE).TE was attached using two alternative methods that allowed for either physisorption or covalent attachment,where the latter was achieved by plasma ion immersion implantation(PIII).Aligned fibers stimulated cell elongation and improved cell alignment,in contrast to randomly oriented fibers.TE coatings bound by physisorption or covalently following 200 s PIII treatment promoted fibroblast proliferation.This represents the first cytocompatibility assessment of novel PIII-treated TE-coated PCL scaffolds.To demonstrate their versatility,these 2D anisotropic PCL scaffolds were assembled into 3D hierarchical constructs with an internally compartmentalized structure to mimic the structure of musculoskeletal tissue.展开更多
基金supported by CHONGQING TALENTS PROJECT(4246ZJ1)Science and technology projects of Chongqing Education Commission(KJQN202000427).
文摘Intra-articular injection of mesenchymal stem cells(MSCs)is a promising strategy for osteoarthritis(OA)treatment.However,more and more studies reveal that the injected MSCs have poor adhesion,migration,and survival in the joint cavity.A recent study shows that tropoelastin(TE)regulates adhesion,proliferation and phenotypic maintenance of MSCs as a soluble additive,indicating that TE could promote MSCs-homing in regenerative medicine.In this study,we used TE as injection medium,and compared it with classic media in MSCs intra-articular injection such as normal saline(NS),hyaluronic acid(HA),and platelet-rich plasma(PRP).We found that TE could effectively improve adhesion,migration,chondrogenic differentiation of infrapatellar fat pad MSCs(IPFP-MSCs)and enhance matrix synthesis of osteoarthritic chondrocytes(OACs)in indirect-coculture system.Moreover,TE could significantly enhance IPFP-MSCs adhesion via activation of integrin β1,ERK1/2 and vinculin(VCL)in vitro.In addition,intra-articular injection of TE-IPFP MSCs suspension resulted in a short-term increase in survival rate of IPFP-MSCs and better histology scores of rat joint tissues.Inhibition of integrin β1 or ERK1/2 attenuated the protective effect of TE-IPFP MSCs suspension in vivo.In conclusion,TE promotes performance of IPFP-MSCs and protects knee cartilage from damage in OA through enhancement of cell adhesion and activation of integrin β1/ERK/VCL pathway.Our findings may provide new insights in MSCs intra-articular injection for OA treatment.
基金supported by an Australian Commonwealth Government Research Training Program Tuition Fee Offset and Stipend Scholarship.A.S.W.acknowledges funding from the National Health and Medical Research Council(APP1195827)M.M.M.B.acknowledges funding from the Australian Research Council(FL190100216).
文摘The highly organized extracellular matrix(ECM)of musculoskeletal tissues,encompassing tendons,ligaments and muscles,is structurally anisotropic,hierarchical and multi-compartmental.These features collectively contribute to their unique function.Previous studies have investigated the effect of tissue-engineered scaffold anisotropy on cell morphology and organization for musculoskeletal tissue repair and regeneration,but the hierarchical arrangement of ECM and compartmentalization are not typically replicated.Here,we present a method for multi-compartmental scaffold design that allows for physical mimicry of the spatial architecture of musculoskeletal tissue in regenerative medicine.This design is based on an ECM-inspired macromolecule scaffold.Polycaprolactone(PCL)scaffolds were fabricated with aligned fibers by electrospinning and mechanical stretching,and then surface-functionalized with the cell-supporting ECM protein molecule,tropoelastin(TE).TE was attached using two alternative methods that allowed for either physisorption or covalent attachment,where the latter was achieved by plasma ion immersion implantation(PIII).Aligned fibers stimulated cell elongation and improved cell alignment,in contrast to randomly oriented fibers.TE coatings bound by physisorption or covalently following 200 s PIII treatment promoted fibroblast proliferation.This represents the first cytocompatibility assessment of novel PIII-treated TE-coated PCL scaffolds.To demonstrate their versatility,these 2D anisotropic PCL scaffolds were assembled into 3D hierarchical constructs with an internally compartmentalized structure to mimic the structure of musculoskeletal tissue.
文摘目的:通过检测弹力蛋白原(tropoelastin)基因单核苷酸多态性(single nucleotide polymorphisms,SNPs)位点表达、tropoelastin mRNA和弹性蛋白与弹力纤维水平,探讨tropoelastin基因SNPs与主动脉夹层(aortic dissection,AD)的关系。方法:留取96例AD患者(AD组)和95例对照组患者的外周血和主动脉组织标本,提取外周血DNA,应用MassARRAY技术进行tropoelastin基因SNPs检测,并构建单体型;采用real-time PCR和蛋白质印迹法分别检测主动脉中膜层tropoelastin mRNA和弹性蛋白的表达;采用EVG(Elastin Van Gieson)染色观察主动脉管形态和壁弹力纤维的分布。比较tropoelastin基因各SNPs位点基因型和单体型在两组间频率的分布,以及tropoelastin m RNA、弹性蛋白和弹力纤维在两组中表达的差异。结果:两组共检测tropoelastin基因SNPs位点7个,5个位点呈多态性,其中rs2071307(G/A)、rs34945509(C/T)和rs17855988(G/C)位点频率分布在两组之间差异均有统计学意义(均P<0.05)。3个SNP位点[rs2071307(G/A)、rs34945509(C/T)和rs17855988(G/C)]的单体型频率分布,在两组之间差异均有统计学意义(均P<0.01)。Real-time PCR和蛋白质印迹法结果显示AD组主动脉中膜层tropoelastin m RNA和弹性蛋白的表达量均显著低于对照组(均P<0.05)。EVG染色可见AD组主动脉中膜层撕裂,弹力纤维破碎、断裂,且排列不规则;而对照组主动脉中膜层结构完整、层次分明,弹力纤维排列紧密、有序。结论:Tropoelastin基因rs2071307(G/A)、rs34945509(C/T)、rs17855988(G/C)位点的多态性改变可能最终会影响弹力纤维的合成,并有可能在AD的发生中起重要作用。
