In regenerative medicine,extracellular vesicles(EVs)possess the potential to repair injured cells by delivering modulatory factors.However,the therapeutic effect of EVs in large-scale tissue defects,which are subject ...In regenerative medicine,extracellular vesicles(EVs)possess the potential to repair injured cells by delivering modulatory factors.However,the therapeutic effect of EVs in large-scale tissue defects,which are subject to prolonged timelines for tissue architecture and functional restoration,remains poorly understood.In this study,we introduce EVs and cell-tethering hybrid hydrogels composed of tyramine-conjugated gelatin(GelTA)that can be in-situ crosslinked with EVs derived from human induced pluripotent stem cell-derived myofibers(hiPSC-myofibers)and hiPSC-muscle precursor cells.This hybrid hydrogel sustains the release of EVs and provides a beneficial nano-topography and mechanical properties for creating a favorable extracellular matrix.Secreted EVs from the hiPSC-myofibers contain specific microRNAs,potentially improving myogenesis and angiogenesis.Herein,we demonstrate increased myogenic markers and fusion/differentiation indexes through the combina-tory effects of EVs and integrin-mediated adhesions in the 3D matrix.Furthermore,we observe a unique impact of EVs,which aid in maintaining the viability and phenotype of myofibers under harsh environments.The hybrid hydrogel in-situ crosslinked with hiPSCs and EVs is facilely used to fabricate large-scale muscle constructs by the stacking of micro-patterned hydrogel domains.Later,we confirmed a combinational effect,whereby muscle tissue regeneration and functional restoration were improved,via an in vivo murine volumetric muscle loss model.展开更多
Cellular reprogramming technologies have been developed with different physicochemical factors to improve the reprogramming efficiencies of induced pluripotent stem cells(iPSCs).Ultrasound is a clinically applied nonc...Cellular reprogramming technologies have been developed with different physicochemical factors to improve the reprogramming efficiencies of induced pluripotent stem cells(iPSCs).Ultrasound is a clinically applied noncontact biophysical factor known for regulating various cellular behaviors but remains uninvestigated for cellular reprogramming.Here,we present a new reprogramming strategy using low-intensity ultrasound(LIUS)to improve cellular reprogramming of iPSCs in vitro and in vivo.Under 3D microenvironment conditions,increased LIUS stimulation shows enhanced cellular reprogramming of the iPSCs.The cellular reprogramming process facilitated by LIUS is accompanied by increased mesenchymal to epithelial transition and histone modification.LIUS stimulation transiently modulates the cytoskeletal rearrangement,along with increased membrane fluidity and mobility to increase HA/CD44 interactions.Furthermore,LIUS stimulation with HA hydrogel can be utilized in application of both human cells and in vivo environment,for enhanced reprogrammed cells into iPSCs.Thus,LIUS stimulation with a combinatorial 3D microenvironment system can improve cellular reprogramming in vitro and in vivo environments,which can be applied in various biomedical fields.展开更多
基金funded by the National Institutes of Health(R01AR077132)AHA collaborative award(944227)+3 种基金the Gillian Reny Stepping Strong Center for Trauma Innovationpartially supported by Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education(NRF-2021R1A6A3A14039720)supporting Jiseong Kim and Jieun Jeon by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute(KHIDI),funded by the Minstry of Health&Welfare,Republic of Korea(HI19C0757)partially funded by the Deanship of Scientific Research(DSR)at King Abdulaziz University,Jeddah,under Grant No.RG-22-135-39.
文摘In regenerative medicine,extracellular vesicles(EVs)possess the potential to repair injured cells by delivering modulatory factors.However,the therapeutic effect of EVs in large-scale tissue defects,which are subject to prolonged timelines for tissue architecture and functional restoration,remains poorly understood.In this study,we introduce EVs and cell-tethering hybrid hydrogels composed of tyramine-conjugated gelatin(GelTA)that can be in-situ crosslinked with EVs derived from human induced pluripotent stem cell-derived myofibers(hiPSC-myofibers)and hiPSC-muscle precursor cells.This hybrid hydrogel sustains the release of EVs and provides a beneficial nano-topography and mechanical properties for creating a favorable extracellular matrix.Secreted EVs from the hiPSC-myofibers contain specific microRNAs,potentially improving myogenesis and angiogenesis.Herein,we demonstrate increased myogenic markers and fusion/differentiation indexes through the combina-tory effects of EVs and integrin-mediated adhesions in the 3D matrix.Furthermore,we observe a unique impact of EVs,which aid in maintaining the viability and phenotype of myofibers under harsh environments.The hybrid hydrogel in-situ crosslinked with hiPSCs and EVs is facilely used to fabricate large-scale muscle constructs by the stacking of micro-patterned hydrogel domains.Later,we confirmed a combinational effect,whereby muscle tissue regeneration and functional restoration were improved,via an in vivo murine volumetric muscle loss model.
基金funding from the National Research Foundation of Korea(NRF)grant funded by the Korea government(MSIT,MOE)(NRF-2019M3A9H1032376,NRF-2022R1A2C3004850,RS-2023-00214410,RS-2023-00257290,RS-2023-00246418,and RS-2023-00275407).
文摘Cellular reprogramming technologies have been developed with different physicochemical factors to improve the reprogramming efficiencies of induced pluripotent stem cells(iPSCs).Ultrasound is a clinically applied noncontact biophysical factor known for regulating various cellular behaviors but remains uninvestigated for cellular reprogramming.Here,we present a new reprogramming strategy using low-intensity ultrasound(LIUS)to improve cellular reprogramming of iPSCs in vitro and in vivo.Under 3D microenvironment conditions,increased LIUS stimulation shows enhanced cellular reprogramming of the iPSCs.The cellular reprogramming process facilitated by LIUS is accompanied by increased mesenchymal to epithelial transition and histone modification.LIUS stimulation transiently modulates the cytoskeletal rearrangement,along with increased membrane fluidity and mobility to increase HA/CD44 interactions.Furthermore,LIUS stimulation with HA hydrogel can be utilized in application of both human cells and in vivo environment,for enhanced reprogrammed cells into iPSCs.Thus,LIUS stimulation with a combinatorial 3D microenvironment system can improve cellular reprogramming in vitro and in vivo environments,which can be applied in various biomedical fields.
