Journal: Journal of Cell Communication and Signaling

Article Title: Hypoxic adipose‐derived stem cell exosomes as carriers of miR‐100‐5p to enhance angiogenesis and suppress inflammation in diabetic foot ulcers

doi: 10.1002/ccs3.70018

Figure Lengend Snippet: Therapeutic effects of ADSCs‐Exo in a DFU rat model. (A) Schematic diagram illustrating the treatment process of the DFU rat model with ADSCs‐Exo; (B) visual observation and digital image recording of ulcer area changes in the DFU rat model; (C) the wound area and skin thickness were measured using a vernier caliper; (D) hematoxylin and eosin staining of skin tissue morphology in the DFU rat model; (E) IHC analysis of angiogenesis‐related factors VEGF and CD31, and inflammatory factors TNF‐α and IL‐6; (F) quantification of VEGF, CD31, TNF‐α, and IL‐6 expression through IHC. All data are presented as mean ± standard error, 10 rats per group ( n = 10). Statistical analysis was performed using ANOVA followed by Tukey's post hoc test. ADSCs‐Exo, adipose‐derived stem cell exosomes; DFU, diabetic foot ulcer; IHC, immunohistochemistry; IL‐6, interleukin‐6; TNF‐α, tumor necrosis factor alpha; VEGF, vascular endothelial growth factor. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: In vitro experiments were conducted using human umbilical vein endothelial cells (HUVEC, CRL‐1730, ATCC, USA) and rat skin fibroblasts (FR, CRL‐1213, ATCC, USA).

Techniques: Staining, Expressing, Derivative Assay, Immunohistochemistry

Journal: Journal of Cell Communication and Signaling

Article Title: Hypoxic adipose‐derived stem cell exosomes as carriers of miR‐100‐5p to enhance angiogenesis and suppress inflammation in diabetic foot ulcers

doi: 10.1002/ccs3.70018

Figure Lengend Snippet: The role of H‐ADSCs‐Exo‐mediated miR‐100‐5p delivery in cell proliferation and angiogenesis. (A) Schematic diagram of H‐ADSCs‐Exo delivering miR‐100‐5p to treat HUVEC and FR cells; (B) RT‐qPCR analysis of miR‐100‐5p expression levels in H‐ADSCs‐Exo and N‐ADSCs‐Exo; (C) confocal microscopy observation of fluorescently labeled H‐ADSCs‐Exo and N‐ADSCs‐Exo uptake in HUVEC and FR cells; (D) RT‐qPCR analysis of miR‐100‐5p expression in HUVEC and FR cells after treatment with H‐ADSCs‐Exo and N‐ADSCs‐Exo; (E) CCK‐8 assay to determine the proliferation rates of HUVEC and FR cells treated with PBS, N‐ADSCs‐Exo, and H‐ADSCs‐Exo; (F) Transwell assay to assess the migration abilities of HUVEC and FR cells treated with PBS, N‐ADSCs‐Exo, and H‐ADSCs‐Exo; (G) Western blot analysis of the expression levels of vascular endothelial growth factor, angiopoietin 1, collagen I, and fibronectin proteins in HUVEC treated with PBS, N‐ADSCs‐Exo, and H‐ADSCs‐Exo; (H) tube formation assay to evaluate the number, length, and branching points of tubular structures formed by HUVEC treated with PBS, N‐ADSCs‐Exo, and H‐ADSCs‐Exo. All data are presented as mean ± standard error, with experiments repeated in triplicate ( n = 3 biological replicates). Statistical analysis was performed using ANOVA followed by Tukey's post hoc test. FR, fibroblasts; H‐ADSCs‐Exo, hypoxic adipose‐derived stem cell exosomes; HUVEC, human umbilical vein endothelial cells; N‐ADSCs‐Exo, normoxic adipose‐derived stem cell exosomes; RT‐qPCR, quantitative real‐time polymerase chain reaction. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: In vitro experiments were conducted using human umbilical vein endothelial cells (HUVEC, CRL‐1730, ATCC, USA) and rat skin fibroblasts (FR, CRL‐1213, ATCC, USA).

Techniques: Quantitative RT-PCR, Expressing, Confocal Microscopy, Labeling, CCK-8 Assay, Transwell Assay, Migration, Western Blot, Tube Formation Assay, Derivative Assay, Real-time Polymerase Chain Reaction

Journal: Journal of Cell Communication and Signaling

Article Title: Hypoxic adipose‐derived stem cell exosomes as carriers of miR‐100‐5p to enhance angiogenesis and suppress inflammation in diabetic foot ulcers

doi: 10.1002/ccs3.70018

Figure Lengend Snippet: The role of H‐ADSCs‐Exo overexpressing miR‐100‐5p in promoting angiogenesis and cell proliferation. (A) Schematic representation of the procedure for treating HUVEC with H‐ADSCs‐Exo overexpressing miR‐100‐5p; (B) quantitative real‐time polymerase chain reaction analysis of miR‐100‐5p expression levels in H‐ADSCs‐Exo overexpressing miR‐100‐5p mimic and NC mimic; (C) transmission electron microscopy and nanoparticle tracking analysis to characterize the morphology and size distribution of H‐ADSCs‐Exo overexpressing miR‐100‐5p mimic and NC mimic; (D) cell counting kit‐8 assay to measure the proliferation rates of HUVEC and fibroblasts cells treated with Exo overexpressing miR‐100‐5p mimic and NC mimic; (E) Transwell assay to evaluate the migration ability of cells treated with Exo overexpressing miR‐100‐5p mimic and NC mimic; (F) Western blot analysis of the expression levels of vascular endothelial growth factor, angiopoietin 1, collagen I, and fibronectin proteins in HUVEC treated with miR‐100‐5p mimic and NC mimic Exo; (G) tube formation assay to assess the number of tubular structures formed by HUVEC treated with Exo overexpressing miR‐100‐5p mimic and NC mimic; (H) immunofluorescence staining to detect F‐actin and VE‐cadherin expression in HUVEC treated with Exo overexpressing miR‐100‐5p mimic and NC mimic. All data are presented as mean ± standard error, with experiments repeated three times ( n = 3 biological replicates). Statistical analysis was performed using ANOVA followed by Tukey's post hoc test. H‐ADSCs‐Exo, hypoxic adipose‐derived stem cell exosomes; HUVEC, human umbilical vein endothelial cells. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: In vitro experiments were conducted using human umbilical vein endothelial cells (HUVEC, CRL‐1730, ATCC, USA) and rat skin fibroblasts (FR, CRL‐1213, ATCC, USA).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Transmission Assay, Electron Microscopy, Cell Counting, Transwell Assay, Migration, Western Blot, Tube Formation Assay, Immunofluorescence, Staining, Derivative Assay

Journal: Journal of Cell Communication and Signaling

Article Title: Hypoxic adipose‐derived stem cell exosomes as carriers of miR‐100‐5p to enhance angiogenesis and suppress inflammation in diabetic foot ulcers

doi: 10.1002/ccs3.70018

Figure Lengend Snippet: miR‐100‐5p Exo promote healing and reduce inflammatory response in DFU rat models. (A) Schematic diagram illustrating the treatment process of DFU rat models using hypoxic adipose‐derived stem cell exosomes with miR‐100‐5p overexpression; (B) weekly observation and recording of ulcer size changes in DFU rat models; (C) analysis of ulcer area and skin thickness changes; (D) quantitative real‐time polymerase chain reaction detection of miR‐100‐5p expression levels in rat skin tissue; (E) hematoxylin and eosin staining to assess skin tissue morphology; (F) immunohistochemistry detection of angiogenesis‐related factors vascular endothelial growth factor and CD31, and inflammatory factors TNF‐α and IL‐6; (G) measurement of blood glucose and insulin levels; (H) ELISA detection of inflammatory factors IL‐6 and TNF‐α. All data are presented as mean ± standard error, 10 rats per group ( n = 10). Statistical analysis was performed using ANOVA and Tukey's post hoc test. DFU, diabetic foot ulcer; IL‐6, interleukin‐6; TNF‐α, tumor necrosis factor alpha. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: In vitro experiments were conducted using human umbilical vein endothelial cells (HUVEC, CRL‐1730, ATCC, USA) and rat skin fibroblasts (FR, CRL‐1213, ATCC, USA).

Techniques: Derivative Assay, Over Expression, Real-time Polymerase Chain Reaction, Expressing, Staining, Immunohistochemistry, Enzyme-linked Immunosorbent Assay