Osteoarthritis(OA),a common degenerative disease,is characterized by high disability and imposes substantial economic impacts on individuals and society.Current clinical treatments remain inadequate for effectively ma...Osteoarthritis(OA),a common degenerative disease,is characterized by high disability and imposes substantial economic impacts on individuals and society.Current clinical treatments remain inadequate for effectively managing OA.Organoids,miniature 3D tissue structures from directed differentiation of stem or progenitor cells,mimic native organ structures and functions.They are useful for drug testing and serve as active grafts for organ repair.However,organoid construction requires extracellular matrix-like 3D scaffolds for cellular growth.Hydrogel microspheres,with tunable physical and chemical properties,show promise in cartilage tissue engineering by replicating the natural microenvironment.Building on prior work on SF-DNA dual-network hydrogels for cartilage regeneration,we developed a novel RGD-SF-DNA hydrogel microsphere(RSD-MS)via a microfluidic system by integrating photopolymerization with self-assembly techniques and then modified with Pep-RGDfKA.The RSD-MSs exhibited uniform size,porous surface,and optimal swelling and degradation properties.In vitro studies demonstrated that RSD-MSs enhanced bone marrow mesenchymal stem cells(BMSCs)proliferation,adhesion,and chondrogenic differentiation.Transcriptomic analysis showed RSD-MSs induced chondrogenesis mainly through integrin-mediated adhesion pathways and glycosaminoglycan biosynthesis.Moreover,in vivo studies showed that seeding BMSCs onto RSD-MSs to create cartilage organoid precursors(COPs)significantly enhanced cartilage regeneration.In conclusion,RSD-MS was an ideal candidate for the construction and long-term cultivation of cartilage organoids,offering an innovative strategy and material choice for cartilage regeneration and tissue engineering.展开更多
To provide materials used in investigating the relationship between amino acid compositions of silk-like protein, structure, and functions, especially the biological functions, the motif genes encoding the silk fibroi...To provide materials used in investigating the relationship between amino acid compositions of silk-like protein, structure, and functions, especially the biological functions, the motif genes encoding the silk fibroin amorphous domain, SGFGPVANGGSGEASSESDFGSSGFGPVANASSGEASSESDFAG(F) were designed and extended using a "head-to-tail" construction strategy. The designed genes were cloned into PSLFA1180FA and multimerized to form structures containing a two-timer, a four-timer, an eight-timer, and a twelve-timer. All the resulting plasmids were digested using the restriction enzyme BamHI and the double-enzymes BglII/HindIII. Restriction enzyme analysis and DNA sequencing revealed the motif was successfully cloned into PSLFA1180FA and multimerized to form a twelve-timer without gene deletion or mutation.展开更多
基金supported by the National Key Research and Development Program of China(No.2022YFB3804300)Integrated Project of Major Research Plan of National Natural Science Foundation of China(92249303)+1 种基金Key Project of the National Natural Science Foundation of China(82230071)National Natural Science Foundation of China(32101084).
文摘Osteoarthritis(OA),a common degenerative disease,is characterized by high disability and imposes substantial economic impacts on individuals and society.Current clinical treatments remain inadequate for effectively managing OA.Organoids,miniature 3D tissue structures from directed differentiation of stem or progenitor cells,mimic native organ structures and functions.They are useful for drug testing and serve as active grafts for organ repair.However,organoid construction requires extracellular matrix-like 3D scaffolds for cellular growth.Hydrogel microspheres,with tunable physical and chemical properties,show promise in cartilage tissue engineering by replicating the natural microenvironment.Building on prior work on SF-DNA dual-network hydrogels for cartilage regeneration,we developed a novel RGD-SF-DNA hydrogel microsphere(RSD-MS)via a microfluidic system by integrating photopolymerization with self-assembly techniques and then modified with Pep-RGDfKA.The RSD-MSs exhibited uniform size,porous surface,and optimal swelling and degradation properties.In vitro studies demonstrated that RSD-MSs enhanced bone marrow mesenchymal stem cells(BMSCs)proliferation,adhesion,and chondrogenic differentiation.Transcriptomic analysis showed RSD-MSs induced chondrogenesis mainly through integrin-mediated adhesion pathways and glycosaminoglycan biosynthesis.Moreover,in vivo studies showed that seeding BMSCs onto RSD-MSs to create cartilage organoid precursors(COPs)significantly enhanced cartilage regeneration.In conclusion,RSD-MS was an ideal candidate for the construction and long-term cultivation of cartilage organoids,offering an innovative strategy and material choice for cartilage regeneration and tissue engineering.
基金National Natural Science Foundation of China(No. 51173125)Natural Science Foundations of Jiangsu Province of China(No. BK2010253,No. BK2012633)+2 种基金College Natural Science Research Project of Jiangsu Province of China(No. 12KJA43004)Science and Technology Plan Foundation of Suzhou of China(No. ZXS2012002)Priority Academic Program Development of Jiangsu Higher Education Institutions,China
文摘To provide materials used in investigating the relationship between amino acid compositions of silk-like protein, structure, and functions, especially the biological functions, the motif genes encoding the silk fibroin amorphous domain, SGFGPVANGGSGEASSESDFGSSGFGPVANASSGEASSESDFAG(F) were designed and extended using a "head-to-tail" construction strategy. The designed genes were cloned into PSLFA1180FA and multimerized to form structures containing a two-timer, a four-timer, an eight-timer, and a twelve-timer. All the resulting plasmids were digested using the restriction enzyme BamHI and the double-enzymes BglII/HindIII. Restriction enzyme analysis and DNA sequencing revealed the motif was successfully cloned into PSLFA1180FA and multimerized to form a twelve-timer without gene deletion or mutation.
