Neovascularization is critical to improve the diabetic microenvironment,deliver abundant nutrients to the wound and promote wound closure.However,the excess of oxidative stress impedes the healing process.Herein,a sel...Neovascularization is critical to improve the diabetic microenvironment,deliver abundant nutrients to the wound and promote wound closure.However,the excess of oxidative stress impedes the healing process.Herein,a self-adaptive multifunctional hydrogel with self-healing property and injectability is fabricated through a boronic ester-based reaction between the phenylboronic acid groups of the 3-carboxyl-4-fluorophenylboronic acid-grafted quaternized chitosan and the hydroxyl groups of the polyvinyl alcohol,in which pro-angiogenic drug of desferrioxamine(DFO)is loaded in the form of gelatin microspheres(DFO@G).The boronic ester bonds of the hydrogel can self-adaptively react with hyperglycemic and hydrogen peroxide to alleviate oxidative stress and release DFO@G in the early phase of wound healing.A sustained release of DFO is then realized by responding to overexpressed matrix metalloproteinases.In a full-thickness diabetic wound model,the DFO@G loaded hydrogel accelerates angiogenesis by upregulating expression of hypoxia-inducible factor-1 and angiogenic growth factors,resulting in collagen deposition and rapid wound closure.This multifunctional hydrogel can not only self-adaptively change the microenvironment to a pro-healing state by decreasing oxidative stress,but also respond to matrix metalloproteinases to release DFO.The self-adaptive multifunctional hydrogel has a potential for treating diabetic wounds.展开更多
The homeostasis of the wound microenvironment is fundamental for scarless wound healing,while the excessive accumulation of transforming growth factor-beta(TGF-β)in the wound microenvironment always leads to hypertro...The homeostasis of the wound microenvironment is fundamental for scarless wound healing,while the excessive accumulation of transforming growth factor-beta(TGF-β)in the wound microenvironment always leads to hypertrophic scars(HS)formation by regulating cell fates and crosstalk among various types of cells,such as macrophages and fibroblasts.This study reports that an injectable,self-assembling LA-peptide hydrogel has the potential to facilitate scarless cutaneous wound healing through dynamically adsorbing TGF-β within the wound environment.We found that the released LA peptides led to the suppression of both the PI3K/Akt and TGF-β/Smad2/3 pathways in macrophages and fibroblasts.As expected,the application of LA-peptide hydrogel alleviated the M2 type polarization of macrophages and inhibited fibroblasts activation by adsorbing TGF-β both in vitro and in vivo.Furthermore,designated concentrations of the LA-peptide hydrogel achieved controlled release of LA peptides,enabling dynamic regulation of TGF-β for maintaining microenvironment homeostasis during different phases of wound healing.This contributed to the inhibition of HS formation without delaying wound healing in both a mouse full-thickness skin wound model and a rabbit ear scar model.Overall,the LA-peptide hydrogel provides promising avenues for promoting scarless healing of wounds,exemplifying precision medicine-guided targeting of specific pathogenic molecules,such as TGF-β,and highlighting the significance of dynamic regulation of TGF-β homeostasis in wound microenvironment.展开更多
Background:Diabetic foot ulcer(DFU)is one of the most common and complex complications of diabetes,but the underlying pathophysiology remains unclear.Single-cell RNA sequencing(scRNA-seq)has been conducted to explore ...Background:Diabetic foot ulcer(DFU)is one of the most common and complex complications of diabetes,but the underlying pathophysiology remains unclear.Single-cell RNA sequencing(scRNA-seq)has been conducted to explore novel cell types or molecular profiles of DFU from various perspectives.This study aimed to comprehensively analyze the potential mechanisms underlying impaired re-epithelization of DFU in a single-cell perspective.Methods:We conducted scRNA-seq on tissues from human normal skin,acute wound,and DFU to investigate the potential mechanisms underlying impaired epidermal differentiation and the pathological microenvironment.Pseudo-time and lineage inference analyses revealed the distinct states and transition trajectories of epidermal cells under different conditions.Transcription factor analysis revealed the potential regulatory mechanism of key subtypes of keratinocytes.Cell-cell interaction analysis revealed the regulatory network between the proinflammatory microenvironment and epidermal cells.Laser-capture microscopy coupled with RNA sequencing(LCM-seq)and multiplex immunohistochemistry were used to validate the expression and location of key subtypes of keratinocytes.Results:Our research provided a comprehensive map of the phenotypic and dynamic changes that occur during epidermal differentiation,alongside the corresponding regulatory networks in DFU.Importantly,we identified two subtypes of keratinocytes:basal cells(BC-2)and diabetes-associated keratinocytes(DAK)that might play crucial roles in the impairment of epidermal homeostasis.BC-2 and DAK showed a marked increase in DFU,with an inactive state and insufficient motivation for epidermal differentiation.BC-2 was involved in the cellular response and apoptosis processes,with high expression of TXNIP,IFITM1,and IL1R2.Additionally,the pro-differentiation transcription factors were downregulated in BC-2 in DFU,indicating that the differentiation process might be inhibited in BC-2 in DFU.DAK was associated with cellular glucose homeostasis.Furthermore,increased CCL2+CXCL2+fibroblasts,VWA1+vascular endothelial cells,and GZMA+CD8+T cells were detected in DFU.These changes in the wound microenvironment could regulate the fate of epidermal cells through the TNFSF12-TNFRSF12A,IFNG-IFNGR1/2,and IL-1B-IL1R2 pathways,which might result in persistent inflammation and impaired epidermal differentiation in DFU.Conclusions:Our findings offer novel insights into the pathophysiology of DFU and present potential therapeutic targets that could improve wound care and treatment outcomes for DFU patients.展开更多
Stem cell-laden hydrogel patches are promising candidates to treat chronic ulcers due to cells’long-lasting and dynamic responses to wound microenvironment.However,their clinical translations are prohibited by the cr...Stem cell-laden hydrogel patches are promising candidates to treat chronic ulcers due to cells’long-lasting and dynamic responses to wound microenvironment.However,their clinical translations are prohibited by the cryopreservation difficulty due to their weak mechanical strength and slow biotransport capability,and by the morphological mismatch between clinical ulcers and pre-fabricated patches.Here we report a stem cell-laden alginate-dopamine hydrogel patch that can be readily cryopreserved,processed,and scaled toward clinical usages.This cell-hydrogel patch not only maintains cell viability and structure integrity during cryo-preservation,but also can be directly utilized without centrifugation or incubation post cryopreservation.In addition,this tissue-adhesive hydrogel patch enables close wound contact and fast cellular response,and its scalable and flexible structure enables assembly for large or irregularly shaped ulcers.Therefore,it accelerates ulcer healing and reduces scar formation via continuous,versatile,self-adjusting paracrine of imbedded,cryopreserved stem cells.These findings highlight its po-tential for scalable clinical applications in chronic wound management and pave the way for broader adoption of ready-to-use regenerative therapies.展开更多
基金supported by the National Natural Science Foundation of China(Nos.52103186,51725303,52033007)the Fundamental Research Funds for the Central Universities(Nos.2682020ZT84,2682021ZTPY008).
文摘Neovascularization is critical to improve the diabetic microenvironment,deliver abundant nutrients to the wound and promote wound closure.However,the excess of oxidative stress impedes the healing process.Herein,a self-adaptive multifunctional hydrogel with self-healing property and injectability is fabricated through a boronic ester-based reaction between the phenylboronic acid groups of the 3-carboxyl-4-fluorophenylboronic acid-grafted quaternized chitosan and the hydroxyl groups of the polyvinyl alcohol,in which pro-angiogenic drug of desferrioxamine(DFO)is loaded in the form of gelatin microspheres(DFO@G).The boronic ester bonds of the hydrogel can self-adaptively react with hyperglycemic and hydrogen peroxide to alleviate oxidative stress and release DFO@G in the early phase of wound healing.A sustained release of DFO is then realized by responding to overexpressed matrix metalloproteinases.In a full-thickness diabetic wound model,the DFO@G loaded hydrogel accelerates angiogenesis by upregulating expression of hypoxia-inducible factor-1 and angiogenic growth factors,resulting in collagen deposition and rapid wound closure.This multifunctional hydrogel can not only self-adaptively change the microenvironment to a pro-healing state by decreasing oxidative stress,but also respond to matrix metalloproteinases to release DFO.The self-adaptive multifunctional hydrogel has a potential for treating diabetic wounds.
基金financially sponsored by grants from the National Natural Science Foundation of China(No.82372530 and 82072182)“Young doctor training program”of Xijing Hospital(No.XJZT24QN60,J.Yuan)+1 种基金Special topic of health care in Air Force Military Medical University(No.23KYBJ02,X.Li)Scientific and technological research of major problems in military medicine and aviation medicine(No.2023JSYX09,K.Tao and Y.Wang).
文摘The homeostasis of the wound microenvironment is fundamental for scarless wound healing,while the excessive accumulation of transforming growth factor-beta(TGF-β)in the wound microenvironment always leads to hypertrophic scars(HS)formation by regulating cell fates and crosstalk among various types of cells,such as macrophages and fibroblasts.This study reports that an injectable,self-assembling LA-peptide hydrogel has the potential to facilitate scarless cutaneous wound healing through dynamically adsorbing TGF-β within the wound environment.We found that the released LA peptides led to the suppression of both the PI3K/Akt and TGF-β/Smad2/3 pathways in macrophages and fibroblasts.As expected,the application of LA-peptide hydrogel alleviated the M2 type polarization of macrophages and inhibited fibroblasts activation by adsorbing TGF-β both in vitro and in vivo.Furthermore,designated concentrations of the LA-peptide hydrogel achieved controlled release of LA peptides,enabling dynamic regulation of TGF-β for maintaining microenvironment homeostasis during different phases of wound healing.This contributed to the inhibition of HS formation without delaying wound healing in both a mouse full-thickness skin wound model and a rabbit ear scar model.Overall,the LA-peptide hydrogel provides promising avenues for promoting scarless healing of wounds,exemplifying precision medicine-guided targeting of specific pathogenic molecules,such as TGF-β,and highlighting the significance of dynamic regulation of TGF-β homeostasis in wound microenvironment.
基金supported by the National Natural Science Foundation of China(Nos 82256147,82072178)Support Program for Growth Factor Research(No.SZYZ-TR-06)GuangDong Basic and Applied Basic Research Foundation(No.2023A1515111087).
文摘Background:Diabetic foot ulcer(DFU)is one of the most common and complex complications of diabetes,but the underlying pathophysiology remains unclear.Single-cell RNA sequencing(scRNA-seq)has been conducted to explore novel cell types or molecular profiles of DFU from various perspectives.This study aimed to comprehensively analyze the potential mechanisms underlying impaired re-epithelization of DFU in a single-cell perspective.Methods:We conducted scRNA-seq on tissues from human normal skin,acute wound,and DFU to investigate the potential mechanisms underlying impaired epidermal differentiation and the pathological microenvironment.Pseudo-time and lineage inference analyses revealed the distinct states and transition trajectories of epidermal cells under different conditions.Transcription factor analysis revealed the potential regulatory mechanism of key subtypes of keratinocytes.Cell-cell interaction analysis revealed the regulatory network between the proinflammatory microenvironment and epidermal cells.Laser-capture microscopy coupled with RNA sequencing(LCM-seq)and multiplex immunohistochemistry were used to validate the expression and location of key subtypes of keratinocytes.Results:Our research provided a comprehensive map of the phenotypic and dynamic changes that occur during epidermal differentiation,alongside the corresponding regulatory networks in DFU.Importantly,we identified two subtypes of keratinocytes:basal cells(BC-2)and diabetes-associated keratinocytes(DAK)that might play crucial roles in the impairment of epidermal homeostasis.BC-2 and DAK showed a marked increase in DFU,with an inactive state and insufficient motivation for epidermal differentiation.BC-2 was involved in the cellular response and apoptosis processes,with high expression of TXNIP,IFITM1,and IL1R2.Additionally,the pro-differentiation transcription factors were downregulated in BC-2 in DFU,indicating that the differentiation process might be inhibited in BC-2 in DFU.DAK was associated with cellular glucose homeostasis.Furthermore,increased CCL2+CXCL2+fibroblasts,VWA1+vascular endothelial cells,and GZMA+CD8+T cells were detected in DFU.These changes in the wound microenvironment could regulate the fate of epidermal cells through the TNFSF12-TNFRSF12A,IFNG-IFNGR1/2,and IL-1B-IL1R2 pathways,which might result in persistent inflammation and impaired epidermal differentiation in DFU.Conclusions:Our findings offer novel insights into the pathophysiology of DFU and present potential therapeutic targets that could improve wound care and treatment outcomes for DFU patients.
基金the National Natural Science Foundation of China (Grant No. GYKP019, 52076157)Xian Jiaotong University (Young Talent Support Program).
文摘Stem cell-laden hydrogel patches are promising candidates to treat chronic ulcers due to cells’long-lasting and dynamic responses to wound microenvironment.However,their clinical translations are prohibited by the cryopreservation difficulty due to their weak mechanical strength and slow biotransport capability,and by the morphological mismatch between clinical ulcers and pre-fabricated patches.Here we report a stem cell-laden alginate-dopamine hydrogel patch that can be readily cryopreserved,processed,and scaled toward clinical usages.This cell-hydrogel patch not only maintains cell viability and structure integrity during cryo-preservation,but also can be directly utilized without centrifugation or incubation post cryopreservation.In addition,this tissue-adhesive hydrogel patch enables close wound contact and fast cellular response,and its scalable and flexible structure enables assembly for large or irregularly shaped ulcers.Therefore,it accelerates ulcer healing and reduces scar formation via continuous,versatile,self-adjusting paracrine of imbedded,cryopreserved stem cells.These findings highlight its po-tential for scalable clinical applications in chronic wound management and pave the way for broader adoption of ready-to-use regenerative therapies.
