Infectious wound healing is complicated with and limited by infection and oxidative stress at the wound site.In recent years,various evidences suggest that nanozymes with multiple enzymatic activities have enabled the...Infectious wound healing is complicated with and limited by infection and oxidative stress at the wound site.In recent years,various evidences suggest that nanozymes with multiple enzymatic activities have enabled the development of novel strategies for infectious wound healing.In this study,epigallocatechin gallate loaded polydopamine(P@E)was developed to act as a potent reactive oxygen species(ROS)scavenger for scavenging ROS,alleviating inflammatory responses,and promoting infectious wound healing.Combining with near infrared(NIR)irradiation,P@E presented excellent antibacterial ability of Escherichia coli(E.coli,93.6%)and methicillin-resistant Staphylococcus aureus(MRSA,87.6%).Specifically,P@E+NIR exhibited the most potent antioxidant,anti-inflammatory and cell proliferation behaviors through downregulating intracellular ROS levels(81.9%and 94.3%for NIH3T3 and RAW264.7 respectively)and inducible nitric oxide synthase(iNOS)expression level(55.7%),and up-regulating the expression levels of arginase-1(Arg-1,71.4%),heat shock protein 70(HSP70,48.6%)and platelet endothelial cell adhesion molecule(CD31,35.3%)compared to control group.Meanwhile,it also efficiently induced M2 directional polarization of lipopolysaccharide induced murine macrophages to achieve anti-inflammation,indicated by the down-regulation of CD86(86.2%),and up-regulation of CD206(85.6%).Significantly,it was also observed that P@E+NIR presented the excellent behaviors of inhibiting wound infection,alleviating wound inflammation,as well as promoting skin tissue repairing.Altogether,it has developed the strategy of using P@E combining with NIR irradiation for the synergistic enhanced healing of infectious skin wound,which can serve as a promising therapeutic strategy for its clinical treatment.展开更多
Background:Bacterial infections pose a considerable threat to skin wounds,particularly in the case of challenging-to-treat diabetic wounds.Systemic antibiotics often struggle to penetrate deep wound tissues and topica...Background:Bacterial infections pose a considerable threat to skin wounds,particularly in the case of challenging-to-treat diabetic wounds.Systemic antibiotics often struggle to penetrate deep wound tissues and topically applied antibiotics may lead to sensitization,necessitating the development of novel approaches for effectively treating germs in deep wound tissues.Neutrophils,the predominant immune cells in the bloodstream,rapidly release an abundance of molecules via degranulation upon activation,which possess the ability to directly eliminate pathogens.This study was designed to develop novel neutrophil cell engineered nanovesicles(NVs)with high production and explore their bactericidal properties and application in promoting infectious wound healing.Methods:Neutrophils were isolated from peripheral blood and activated in vitro via phorbol myristate acetate(PMA)stimulation.Engineered NVs were prepared by sequentially extruding activated neutrophils followed by ultracentrifugation and were compared with neutrophil-derived exosomes in terms of morphology,size distribution and protein contents.The bactericidal effect of NVs in vitro was evaluated using the spread plate technique,LIVE/DEAD backlight bacteria assay and observation of bacterial morphology.The therapeutic effects of NVs in vivo were evaluated using wound contraction area measurements,histopathological examinations,assessments of inflammatory factors and immunochemical staining.Results:Activated neutrophils stimulated with PMA in vitro promptly release a substantial amount of bactericidal proteins.NVs are similar to exosomes in terms of morphology and particle size,but they exhibit a significantly higher enrichment of bactericidal proteins.In vitro,NVs demonstrated a significant bactericidal effect,presumably mediated by the enrichment of bactericidal proteins such as lysozyme.These NVs significantly accelerated wound healing,leading to a marked reduction in bacterial load,downregulation of inflammatory factors and enhanced collagen deposition in a fullthickness infectious skin defect model.Conclusions:We developed engineered NVs derived from activated neutrophils to serve as a novel debridement method targeting bacteria in deep tissues,ultimately promoting infectious wound healing.展开更多
Diabetic infectious wound is usually in the environment of high glucose levels and immune disorder.It always results in high reactive oxygen species(ROS)levels,serious immune dysfunction,and extreme susceptibility to ...Diabetic infectious wound is usually in the environment of high glucose levels and immune disorder.It always results in high reactive oxygen species(ROS)levels,serious immune dysfunction,and extreme susceptibility to infection,delaying wound healing.Herein,we prepared an immunomodulatory polyphenol metal organic framework nanozyme(CE)combined with near infrared(NIR)irradiation for diabetic infectious wound healing.This nanozyme was formed by the self-assembly of Cu ions and ellagic acid.In vitro experiments confirmed that CE+NIR could efficiently lower the inflammatory factors(IL-6(50.72%),IL-1β(61.44%)and iNOS(57.33%))expression,and upregulate anti-inflammatory factor(IL-10(111.44%))expression of lipopolysaccharide induced RAW264.7,decreasing the ROS levels,promoting cellular migration(527.91%),and accelerating angiogenesis(91.81%)of high glucose treated L929,together with inhibiting the bacterial growth of E.coli(92.21%)and MRSA(95.89%).Transcriptome sequencing results and their related validation experiments demonstrated that CE+NIR achieved antioxidant and anti-inflammation through ferroptosis inhibition.And transcriptomics and metabolomics analysis proved the efficient antibacterial activity of CE+NIR through NIR amplified cuproptosis.Markedly,it also confirmed that the strategy of CE+NIR was helpful to immunoregu-lation activation via inducing macrophage M2 directional polarization,and immune activating T cells number,thereby restoring immune homeostasis,and accelerating tissue repair.It might offer an efficient strategy of other infectious diseases’immunotherapy with high efficacy and biosafety.展开更多
Background:Photodynamic therapy(PDT)is a widely used therapeutic approach for eradicating bacterial biofilms in infected wound,but its effectiveness is limited by the hypoxic environment within the biofilm.This study ...Background:Photodynamic therapy(PDT)is a widely used therapeutic approach for eradicating bacterial biofilms in infected wound,but its effectiveness is limited by the hypoxic environment within the biofilm.This study aimed to investigate whether the efficiency of photodynamic removing biofilm is improving by providing oxygen(O_(2)),as well as the expression of cytokines involved in infected wound healing.Methods:Manganese dioxide(MnO_(2))nanoparticles with catalase-like activity were grown in situ on graphitic phase carbon nitride(g-C_(3)N_(4),CN)nanosheets to construct an all-in-one CN-MnO2 nanozyme,which was then incorporated into poly-L-lactic acid(PLLA)to prepare CN-MnO2/PLLA wound dressing by electrospinning.Subsequently,the in vitro antibacterial biofilm ratio and antibacterial ratio of CN-MnO_(2)/PLLA wound dressing were examined by spread plate and crystal violet staining under irradiation with 808 nm near-infrared light and 660 nm visible light.Meanwhile,the rat skin injury model was established,and hematoxylin and eosin(H&E),Masson’s,tumor necrosis factor-α(TNF-α),Arginase 1(Arg-1),vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(BFGF)were evaluated in vivo to assess the effect of CN-MnO_(2)/PLLA wound dressing on wound healing.Results:Biofilm density caused by Staphylococcus aureus and Pseudomonas aeruginosa had elimination rates of 83 and 62%,respectively,when treated with CN-MnO_(2)/PLLA dressing.Additionally,the dressing exhibited high antibacterial efficacy against both bacteria,achieving 99 and 98.7%elimination of Staphylococcus aureus and Pseudomonas aeruginosa,respectively.Furthermore,in vivo experiments showed that the CN-MnO_(2)/PLLA wound dressing achieved complete healing of infected wounds on Day 14,with a wound healing rate of>99%by increasing collagen deposition,expression of anti-inflammatory cytokine Arg-1,vascularization cytokine VEGF,and epithelial cell BFGF,and inhibiting the expression of inflammatory cytokine TNF-α.Conclusions:The CN-MnO_(2)/PLLA wound dressing exhibited excellent antibacterial properties in vitro and in vivo.In addition,CN-MnO_(2)/PLLA wound dressing accelerated rapid wound healing through an anti-inflammatory,pro-vascular regeneration and skin tissue remodeling mechanism.展开更多
基金financially supported by the Natural Science Foundation of Guangxi(Nos.2022GXNSFAA035505 and 2024GXNSFAA010164)the Key Research&Development Program of Guangxi(No.GuiKe2024AB08057)+1 种基金the Guangxi Medical and Health Suitable Technology Development and Popularization Applications Project(Nos.S2023093 and S2021084)the Major Talent Project of Guangxi Autonomous Region.
文摘Infectious wound healing is complicated with and limited by infection and oxidative stress at the wound site.In recent years,various evidences suggest that nanozymes with multiple enzymatic activities have enabled the development of novel strategies for infectious wound healing.In this study,epigallocatechin gallate loaded polydopamine(P@E)was developed to act as a potent reactive oxygen species(ROS)scavenger for scavenging ROS,alleviating inflammatory responses,and promoting infectious wound healing.Combining with near infrared(NIR)irradiation,P@E presented excellent antibacterial ability of Escherichia coli(E.coli,93.6%)and methicillin-resistant Staphylococcus aureus(MRSA,87.6%).Specifically,P@E+NIR exhibited the most potent antioxidant,anti-inflammatory and cell proliferation behaviors through downregulating intracellular ROS levels(81.9%and 94.3%for NIH3T3 and RAW264.7 respectively)and inducible nitric oxide synthase(iNOS)expression level(55.7%),and up-regulating the expression levels of arginase-1(Arg-1,71.4%),heat shock protein 70(HSP70,48.6%)and platelet endothelial cell adhesion molecule(CD31,35.3%)compared to control group.Meanwhile,it also efficiently induced M2 directional polarization of lipopolysaccharide induced murine macrophages to achieve anti-inflammation,indicated by the down-regulation of CD86(86.2%),and up-regulation of CD206(85.6%).Significantly,it was also observed that P@E+NIR presented the excellent behaviors of inhibiting wound infection,alleviating wound inflammation,as well as promoting skin tissue repairing.Altogether,it has developed the strategy of using P@E combining with NIR irradiation for the synergistic enhanced healing of infectious skin wound,which can serve as a promising therapeutic strategy for its clinical treatment.
基金supported by National Natural Science Foundation of China(No.82072217,81772135 and U21A20370).
文摘Background:Bacterial infections pose a considerable threat to skin wounds,particularly in the case of challenging-to-treat diabetic wounds.Systemic antibiotics often struggle to penetrate deep wound tissues and topically applied antibiotics may lead to sensitization,necessitating the development of novel approaches for effectively treating germs in deep wound tissues.Neutrophils,the predominant immune cells in the bloodstream,rapidly release an abundance of molecules via degranulation upon activation,which possess the ability to directly eliminate pathogens.This study was designed to develop novel neutrophil cell engineered nanovesicles(NVs)with high production and explore their bactericidal properties and application in promoting infectious wound healing.Methods:Neutrophils were isolated from peripheral blood and activated in vitro via phorbol myristate acetate(PMA)stimulation.Engineered NVs were prepared by sequentially extruding activated neutrophils followed by ultracentrifugation and were compared with neutrophil-derived exosomes in terms of morphology,size distribution and protein contents.The bactericidal effect of NVs in vitro was evaluated using the spread plate technique,LIVE/DEAD backlight bacteria assay and observation of bacterial morphology.The therapeutic effects of NVs in vivo were evaluated using wound contraction area measurements,histopathological examinations,assessments of inflammatory factors and immunochemical staining.Results:Activated neutrophils stimulated with PMA in vitro promptly release a substantial amount of bactericidal proteins.NVs are similar to exosomes in terms of morphology and particle size,but they exhibit a significantly higher enrichment of bactericidal proteins.In vitro,NVs demonstrated a significant bactericidal effect,presumably mediated by the enrichment of bactericidal proteins such as lysozyme.These NVs significantly accelerated wound healing,leading to a marked reduction in bacterial load,downregulation of inflammatory factors and enhanced collagen deposition in a fullthickness infectious skin defect model.Conclusions:We developed engineered NVs derived from activated neutrophils to serve as a novel debridement method targeting bacteria in deep tissues,ultimately promoting infectious wound healing.
基金financially supported by the Key Research&Development Program of Guangxi(Grant No.GuiKeAB22080088)the National Natural Science Foundation of China(Grant No.82360372)+2 种基金the Key Project of Hunan Provincial Administration of Traditional Chinese Medicine(Grant No.C2024004)the First-class Discipline Innovation-driven Talent Program of Guangxi Medical Universitythe Major Talent Project of Guangxi Autonomous Region.
文摘Diabetic infectious wound is usually in the environment of high glucose levels and immune disorder.It always results in high reactive oxygen species(ROS)levels,serious immune dysfunction,and extreme susceptibility to infection,delaying wound healing.Herein,we prepared an immunomodulatory polyphenol metal organic framework nanozyme(CE)combined with near infrared(NIR)irradiation for diabetic infectious wound healing.This nanozyme was formed by the self-assembly of Cu ions and ellagic acid.In vitro experiments confirmed that CE+NIR could efficiently lower the inflammatory factors(IL-6(50.72%),IL-1β(61.44%)and iNOS(57.33%))expression,and upregulate anti-inflammatory factor(IL-10(111.44%))expression of lipopolysaccharide induced RAW264.7,decreasing the ROS levels,promoting cellular migration(527.91%),and accelerating angiogenesis(91.81%)of high glucose treated L929,together with inhibiting the bacterial growth of E.coli(92.21%)and MRSA(95.89%).Transcriptome sequencing results and their related validation experiments demonstrated that CE+NIR achieved antioxidant and anti-inflammation through ferroptosis inhibition.And transcriptomics and metabolomics analysis proved the efficient antibacterial activity of CE+NIR through NIR amplified cuproptosis.Markedly,it also confirmed that the strategy of CE+NIR was helpful to immunoregu-lation activation via inducing macrophage M2 directional polarization,and immune activating T cells number,thereby restoring immune homeostasis,and accelerating tissue repair.It might offer an efficient strategy of other infectious diseases’immunotherapy with high efficacy and biosafety.
基金supported by the National Science Foundation of China under Grant No.32360232,32060222 and 82160354Jiangxi Provincial Natural Science Foundation under Grant No.20232BAB216052 and 20212BAB204005.
文摘Background:Photodynamic therapy(PDT)is a widely used therapeutic approach for eradicating bacterial biofilms in infected wound,but its effectiveness is limited by the hypoxic environment within the biofilm.This study aimed to investigate whether the efficiency of photodynamic removing biofilm is improving by providing oxygen(O_(2)),as well as the expression of cytokines involved in infected wound healing.Methods:Manganese dioxide(MnO_(2))nanoparticles with catalase-like activity were grown in situ on graphitic phase carbon nitride(g-C_(3)N_(4),CN)nanosheets to construct an all-in-one CN-MnO2 nanozyme,which was then incorporated into poly-L-lactic acid(PLLA)to prepare CN-MnO2/PLLA wound dressing by electrospinning.Subsequently,the in vitro antibacterial biofilm ratio and antibacterial ratio of CN-MnO_(2)/PLLA wound dressing were examined by spread plate and crystal violet staining under irradiation with 808 nm near-infrared light and 660 nm visible light.Meanwhile,the rat skin injury model was established,and hematoxylin and eosin(H&E),Masson’s,tumor necrosis factor-α(TNF-α),Arginase 1(Arg-1),vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(BFGF)were evaluated in vivo to assess the effect of CN-MnO_(2)/PLLA wound dressing on wound healing.Results:Biofilm density caused by Staphylococcus aureus and Pseudomonas aeruginosa had elimination rates of 83 and 62%,respectively,when treated with CN-MnO_(2)/PLLA dressing.Additionally,the dressing exhibited high antibacterial efficacy against both bacteria,achieving 99 and 98.7%elimination of Staphylococcus aureus and Pseudomonas aeruginosa,respectively.Furthermore,in vivo experiments showed that the CN-MnO_(2)/PLLA wound dressing achieved complete healing of infected wounds on Day 14,with a wound healing rate of>99%by increasing collagen deposition,expression of anti-inflammatory cytokine Arg-1,vascularization cytokine VEGF,and epithelial cell BFGF,and inhibiting the expression of inflammatory cytokine TNF-α.Conclusions:The CN-MnO_(2)/PLLA wound dressing exhibited excellent antibacterial properties in vitro and in vivo.In addition,CN-MnO_(2)/PLLA wound dressing accelerated rapid wound healing through an anti-inflammatory,pro-vascular regeneration and skin tissue remodeling mechanism.
