Chronic diabetic wounds result from a disrupted microenvironment where oxidative stress,impaired angiogenesis,and persistent infection create a vicious cycle that delays healing.Unfortunately,existing treatments often...Chronic diabetic wounds result from a disrupted microenvironment where oxidative stress,impaired angiogenesis,and persistent infection create a vicious cycle that delays healing.Unfortunately,existing treatments often fail to address these interrelated issues,resulting in suboptimal healing.Here,we propose a base-tip dual-component hydrogel microneedle(MN)system(GBEVs-pVEGF/AgNPs@MNs),consisting of a tip loaded with plant-bacterial hybrid extracellular vesicles(GBEVs-pVEGF)and a base containing silver nanoparticles(AgNPs).Upon penetrating the necrotic tissue of diabetic wounds,our multifunctional MNs could effectively deliver GBEVs-pVEGF,thereby alleviating oxidative stress,promoting cell migration,and facilitating angiogenesis.Additionally,the physical barrier formed by the basal layer synergistically mitigates persistent bacterial infections during wound healing in conjunction with the antimicrobial agent AgNPs.This multifunctional MN system,integrating antioxidant,angiogenic,and antimicrobial properties,effectively restores the disrupted wound microenvironment,offering significant potential for accelerating diabetic wound healing.展开更多
Bone organoids are emerging as powerful tools for studying bone development and related diseases.However,the simplified design of current methods somewhat limits their application potential,as these methods produce si...Bone organoids are emerging as powerful tools for studying bone development and related diseases.However,the simplified design of current methods somewhat limits their application potential,as these methods produce single-tissue organoids that fail to replicate the bone microarchitecture or achieve effective mineralization.To address this issue,we propose a three-dimensional(3D)construction strategy for generating mineralized bone structures using bone marrow-derived mesenchymal stem cells(BMSCs).By mixing BMSCs with hydrogel to create a bone matrix-mimicking bioink and employing projection-based light-curing 3D printing technology,we constructed 3D-printed structures,which were then implanted subcutaneously into nude mice,away from the native bone microenvironment.Even without external stimulation,these implants spontaneously formed mineralized bone domains.With long-term culture,these structures gradually matured into fully differentiated bone tissue,completing both mineralization and vascularization.This in vivo bone organoid model offers a novel platform for studying bone development,exploring congenital diseases,testing drugs,and developing thera-peutic applications.展开更多
Rheumatoid arthritis(RA)is a chronic autoimmune disease that leads to joint deformities and functional impairments.Traditional treatment approaches,such as nonsteroidal anti-inflammatory drugs,disease-modifying antirh...Rheumatoid arthritis(RA)is a chronic autoimmune disease that leads to joint deformities and functional impairments.Traditional treatment approaches,such as nonsteroidal anti-inflammatory drugs,disease-modifying antirheumatic drugs,and molecular targeted therapies,often fail to simultaneously achieve efficient inflammation relief and cartilage tissue repair.DNA hydrogels,derived from nucleic acid nanotechnology,have demonstrated potential in RA therapy due to their programmability,high biocompatibility,and tunable degradation properties.However,their application is still hindered by challenges including high synthesis costs,immunogenicity risks,and uncontrolled degradation rates.To address these limitations,this study proposes a dual-action strategy involving a polymer-modified DNA hydrogel co-delivering nanozymes and living mitochondria to overcome the constraints of traditional therapies and comprehensively optimize RA treatment outcomes.The incorporation of functionalized polymers significantly reduces synthesis costs and immunogenicity while fine-tuning the degradation rate of the hydrogel,enabling sustained support during bone and cartilage repair.The hydrogel is loaded with Prussian blue nanozymes to scavenge excessive reactive oxygen species(ROS)within the RA microenvironment,alleviating inflammation,and facilitates intracellular delivery of living mitochondria to inhibit ROS production at its source,promoting tissue repair.By integrating endogenous ROS reduction with exogenous ROS clearance,this strategy markedly enhances therapeutic efficacy,offering a novel approach for precise RA treatment and advancing the clinical translation of biomaterials.展开更多
The healing of large skin defects remains a significant challenge in clinical settings.The lack of epidermal sources,such as autologous skin grafting,limits full-thickness skin defect repair and leads to excessive sca...The healing of large skin defects remains a significant challenge in clinical settings.The lack of epidermal sources,such as autologous skin grafting,limits full-thickness skin defect repair and leads to excessive scar formation.Skin organoids have the potential to generate a complete skin layer,supporting in-situ skin regeneration in the defect area.In this study,skin organoid spheres,created with human keratinocytes,fibroblasts,and endothelial cells,showed a specific structure with a stromal core surrounded by surface keratinocytes.We selected an appropriate bioink and innovatively combined an extrusion-based bioprinting technique with dual-photo source cross-linking technology to ensure the overall mechanical properties of the 3D bioprinted skin organoid.Moreover,the 3D bioprinted skin organoid was customized to match the size and shape of the wound site,facilitating convenient implantation.When applied to full-thickness skin defects in immunodeficient mice,the 3D bioprinted human-derived skin organoid significantly accelerated wound healing through in-situ regeneration,epithelialization,vascularization,and inhibition of excessive inflammation.The combination of skin organoid and 3D bioprinting technology can overcome the limitations of current skin substitutes,offering a novel treatment strategy to address large-area skin defects.展开更多
Posttraumatic osteoarthritis(PTOA)patients are often diagnosed by X-ray imaging at a middle-late stage when drug interventions are less effective.Early PTOA is characterized by overexpressed matrix metalloprotease 13(...Posttraumatic osteoarthritis(PTOA)patients are often diagnosed by X-ray imaging at a middle-late stage when drug interventions are less effective.Early PTOA is characterized by overexpressed matrix metalloprotease 13(MMP13).Herein,we constructed an integrated diagnosis and treatment micelle modified with MMP13 enzyme-detachable,cyanine 5(Cy5)-containing PEG,black hole quencher-3(BHQ3),and cRGD ligands and loaded with siRNA silencing MMP13(siM13),namely ERMs@siM13.ERMs@siM13 could be cleaved by MMP13 in the diseased cartilage tissues to detach the PEG shell,causing cRGD exposure.Accordingly,the ligand exposure promoted micelle uptake by the diseased chondrocytes by binding to cell surfaceαvβ3 integrin,increasing intracellular siM13 delivery for on-demand MMP13 downregulation.Meanwhile,the Cy5 fluorescence was restored by detaching from the BHQ3-containing micelle,precisely reflecting the diseased cartilage state.In particular,the intensity of Cy5 fluorescence generated by ERMs@siM13 that hinged on the MMP13 levels could reflect the PTOA severity,enabling the physicians to adjust the therapeutic regimen.Finally,in the murine PTOA model,ERMs@siM13 could diagnose the early-stage PTOA,perform timely interventions,and monitor the OA progression level during treatment through a real-time detection of MMP13.Therefore,ERMs@siM13 represents an appealing approach for early-stage PTOA theranostics.展开更多
Infected wounds pose a significant clinical challenge due to bacterial resistance, recurrent infections, and impaired healing. Reactive oxygen species (ROS)-based strategies have shown promise in eradicating bacterial...Infected wounds pose a significant clinical challenge due to bacterial resistance, recurrent infections, and impaired healing. Reactive oxygen species (ROS)-based strategies have shown promise in eradicating bacterial infections. However, the excess ROS in the infection site after treatments may cause irreversible damage to healthy tissues. To address this issue, we developed bovine serum albumin-iridium oxide nanoclusters (BSA-IrOx NCs) which enable photo-regulated ROS generation and scavenging using near infrared (NIR) laser. Upon NIR laser irradiation, BSA-IrOx NCs exhibit enhanced photodynamic therapy, destroying biofilms and killing bacteria. When the NIR laser is off, the nanoclusters' antioxidant enzyme-like activities prevent inflammation and repair damaged tissue through ROS clearance. Transcriptomic and metabolomic analyses revealed that BSA-IrOx NCs inhibit bacterial nitric oxide synthase, blocking bacterial growth and biofilm formation. Furthermore, the nanoclusters repair impaired skin by strengthening cell junctions and reducing mitochondrial damage in a fibroblast model. In vivo studies using rat infected wound models confirmed the efficacy of BSA-IrOx NCs. This study presents a promising strategy for treating biofilm-induced infected wounds by regulating the ROS microenvironment, addressing the challenges associated with current ROS-based antibacterial approaches.展开更多
基金support from the National Natural Science Foundation of China(No.82472444).
文摘Chronic diabetic wounds result from a disrupted microenvironment where oxidative stress,impaired angiogenesis,and persistent infection create a vicious cycle that delays healing.Unfortunately,existing treatments often fail to address these interrelated issues,resulting in suboptimal healing.Here,we propose a base-tip dual-component hydrogel microneedle(MN)system(GBEVs-pVEGF/AgNPs@MNs),consisting of a tip loaded with plant-bacterial hybrid extracellular vesicles(GBEVs-pVEGF)and a base containing silver nanoparticles(AgNPs).Upon penetrating the necrotic tissue of diabetic wounds,our multifunctional MNs could effectively deliver GBEVs-pVEGF,thereby alleviating oxidative stress,promoting cell migration,and facilitating angiogenesis.Additionally,the physical barrier formed by the basal layer synergistically mitigates persistent bacterial infections during wound healing in conjunction with the antimicrobial agent AgNPs.This multifunctional MN system,integrating antioxidant,angiogenic,and antimicrobial properties,effectively restores the disrupted wound microenvironment,offering significant potential for accelerating diabetic wound healing.
基金support from National Natural Science Foundation of China(82230071,82172098,82472444)Shanghai Committee of Science and Technology(23141900600,Laboratory Animal Research Project)+1 种基金Shanghai Clinical Research Plan of SHDC2023CRT01Generaral Project of Natural Science Foundation of Jiangsu Province(BK20231218).
文摘Bone organoids are emerging as powerful tools for studying bone development and related diseases.However,the simplified design of current methods somewhat limits their application potential,as these methods produce single-tissue organoids that fail to replicate the bone microarchitecture or achieve effective mineralization.To address this issue,we propose a three-dimensional(3D)construction strategy for generating mineralized bone structures using bone marrow-derived mesenchymal stem cells(BMSCs).By mixing BMSCs with hydrogel to create a bone matrix-mimicking bioink and employing projection-based light-curing 3D printing technology,we constructed 3D-printed structures,which were then implanted subcutaneously into nude mice,away from the native bone microenvironment.Even without external stimulation,these implants spontaneously formed mineralized bone domains.With long-term culture,these structures gradually matured into fully differentiated bone tissue,completing both mineralization and vascularization.This in vivo bone organoid model offers a novel platform for studying bone development,exploring congenital diseases,testing drugs,and developing thera-peutic applications.
基金financially supported by National Natural Science Foundation of China(32471396,82230071,82172098)Shanghai Committee of Science and Technology(23141900600,Laboratory Animal Research Project)+3 种基金Shanghai Clinical Research Plan of SHDC2023CRT01Shanghai Municipal Demonstration Project for Innovative Medical Device Applications(23SHS05700)Young Elite Scientist Sponsorship Program by China Association for Science and Technology(YESS20230049)Key Project of the Seed Program for Medical New Technology Research and Translation of the Shanghai Municipal Health Commission(2024ZZ1001).
文摘Rheumatoid arthritis(RA)is a chronic autoimmune disease that leads to joint deformities and functional impairments.Traditional treatment approaches,such as nonsteroidal anti-inflammatory drugs,disease-modifying antirheumatic drugs,and molecular targeted therapies,often fail to simultaneously achieve efficient inflammation relief and cartilage tissue repair.DNA hydrogels,derived from nucleic acid nanotechnology,have demonstrated potential in RA therapy due to their programmability,high biocompatibility,and tunable degradation properties.However,their application is still hindered by challenges including high synthesis costs,immunogenicity risks,and uncontrolled degradation rates.To address these limitations,this study proposes a dual-action strategy involving a polymer-modified DNA hydrogel co-delivering nanozymes and living mitochondria to overcome the constraints of traditional therapies and comprehensively optimize RA treatment outcomes.The incorporation of functionalized polymers significantly reduces synthesis costs and immunogenicity while fine-tuning the degradation rate of the hydrogel,enabling sustained support during bone and cartilage repair.The hydrogel is loaded with Prussian blue nanozymes to scavenge excessive reactive oxygen species(ROS)within the RA microenvironment,alleviating inflammation,and facilitates intracellular delivery of living mitochondria to inhibit ROS production at its source,promoting tissue repair.By integrating endogenous ROS reduction with exogenous ROS clearance,this strategy markedly enhances therapeutic efficacy,offering a novel approach for precise RA treatment and advancing the clinical translation of biomaterials.
基金Science Foundation of China(NO.92249303,NO.82230071,No.82172098,No.82371603)Jiangsu Province Natural Science and Technological Project(No.BK20231218)+4 种基金Experimental Animal Research Field Project Shanghai Science and Technology Commission(No.23141900600)Research Physician Innovation and Transformation Ability Training Project from Shanghai Health Commission(No.SHDC2023CRT013)Basic Medical Innovation Project of Changhai Hospital(No.20237Y38)Deep Blue Talent Project of Naval Medical University(Jin Cui)Shanghai Oriental Talent Program(Xiao Chen,Yuanyuan Liu).
文摘The healing of large skin defects remains a significant challenge in clinical settings.The lack of epidermal sources,such as autologous skin grafting,limits full-thickness skin defect repair and leads to excessive scar formation.Skin organoids have the potential to generate a complete skin layer,supporting in-situ skin regeneration in the defect area.In this study,skin organoid spheres,created with human keratinocytes,fibroblasts,and endothelial cells,showed a specific structure with a stromal core surrounded by surface keratinocytes.We selected an appropriate bioink and innovatively combined an extrusion-based bioprinting technique with dual-photo source cross-linking technology to ensure the overall mechanical properties of the 3D bioprinted skin organoid.Moreover,the 3D bioprinted skin organoid was customized to match the size and shape of the wound site,facilitating convenient implantation.When applied to full-thickness skin defects in immunodeficient mice,the 3D bioprinted human-derived skin organoid significantly accelerated wound healing through in-situ regeneration,epithelialization,vascularization,and inhibition of excessive inflammation.The combination of skin organoid and 3D bioprinting technology can overcome the limitations of current skin substitutes,offering a novel treatment strategy to address large-area skin defects.
基金supported by Integrated Project of Major Research Plan of National Natural Science Foundation of China (92249303)National Natural Science Foundation of China (82230071,82172098,82371603,82102217,81872428,and 81703010)+7 种基金the Shanghai Rising Star Program (21QA1412000)Shanghai Hospital Development Center (SHDC2023CRT013)Shanghai Committee of Science and Technology (23141900600,Laboratory Animal Research Project)Shanghai Baoshan District Medical Health Project (21-E-14)the Construction of Key Medical Disciplines of Baoshan District of Shanghai (BSZK-2023-Z07)the Shanghai Municipal Natural Science Foundation (23ZR1463300)Postdoctoral Fellowship Program of CPSF (GZB20230397)General Funding for China Postdoctoral Science Foundation (2023M732179).
文摘Posttraumatic osteoarthritis(PTOA)patients are often diagnosed by X-ray imaging at a middle-late stage when drug interventions are less effective.Early PTOA is characterized by overexpressed matrix metalloprotease 13(MMP13).Herein,we constructed an integrated diagnosis and treatment micelle modified with MMP13 enzyme-detachable,cyanine 5(Cy5)-containing PEG,black hole quencher-3(BHQ3),and cRGD ligands and loaded with siRNA silencing MMP13(siM13),namely ERMs@siM13.ERMs@siM13 could be cleaved by MMP13 in the diseased cartilage tissues to detach the PEG shell,causing cRGD exposure.Accordingly,the ligand exposure promoted micelle uptake by the diseased chondrocytes by binding to cell surfaceαvβ3 integrin,increasing intracellular siM13 delivery for on-demand MMP13 downregulation.Meanwhile,the Cy5 fluorescence was restored by detaching from the BHQ3-containing micelle,precisely reflecting the diseased cartilage state.In particular,the intensity of Cy5 fluorescence generated by ERMs@siM13 that hinged on the MMP13 levels could reflect the PTOA severity,enabling the physicians to adjust the therapeutic regimen.Finally,in the murine PTOA model,ERMs@siM13 could diagnose the early-stage PTOA,perform timely interventions,and monitor the OA progression level during treatment through a real-time detection of MMP13.Therefore,ERMs@siM13 represents an appealing approach for early-stage PTOA theranostics.
基金Institutional Research Project of Shanghai Sixth People's Hospital(LY33.X-4020).
文摘Infected wounds pose a significant clinical challenge due to bacterial resistance, recurrent infections, and impaired healing. Reactive oxygen species (ROS)-based strategies have shown promise in eradicating bacterial infections. However, the excess ROS in the infection site after treatments may cause irreversible damage to healthy tissues. To address this issue, we developed bovine serum albumin-iridium oxide nanoclusters (BSA-IrOx NCs) which enable photo-regulated ROS generation and scavenging using near infrared (NIR) laser. Upon NIR laser irradiation, BSA-IrOx NCs exhibit enhanced photodynamic therapy, destroying biofilms and killing bacteria. When the NIR laser is off, the nanoclusters' antioxidant enzyme-like activities prevent inflammation and repair damaged tissue through ROS clearance. Transcriptomic and metabolomic analyses revealed that BSA-IrOx NCs inhibit bacterial nitric oxide synthase, blocking bacterial growth and biofilm formation. Furthermore, the nanoclusters repair impaired skin by strengthening cell junctions and reducing mitochondrial damage in a fibroblast model. In vivo studies using rat infected wound models confirmed the efficacy of BSA-IrOx NCs. This study presents a promising strategy for treating biofilm-induced infected wounds by regulating the ROS microenvironment, addressing the challenges associated with current ROS-based antibacterial approaches.
