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Effects of CM@MOF on HaCaT cells in vitro. (A) Migration ability of HaCaT cells evaluated by scratch assay, violet staining and immunofluorescence staining of Ki67. Green: Ki67; Blue: nuclei (DAPI). Scale bars: 100 μm. (B) Quantitative analysis of cell migration using ImageJ software. (C) Quantitative analysis of cell migrating rate. (D) Quantification of Ki67-positive cells percentage of HaCaT cells. (E) Live (Green)/Dead (Red) staining of HaCaT cells. Scale bars: 100 μm. (F) ROS(Green) in HaCaT cells observed by fluorescence microscopy. Scale bars: 50 μm. (G) Quantitative analysis of the percentage of live HaCaT cells. (H) Quantitative analysis of ROS fluorescence intensity (mean gray value) in HaCaT cells. (I) mRNA levels of IL6, IL17a, IL22, IL23a and Tnfα in HaCaT cells assessed by RT-qPCR. HaCaT: human <t>keratinocyte</t> cells; DAPI: 4′,6-diamidino-2-phenylindole; LPS: Lipopolysaccharide; M2: M2 membrane; CM: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage); MOF: Metal-organic framework loaded Cu/Zn; CM@MOF: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage) loaded Cu/Zn metal-organic framework (Cu/Zn-MOF); S. aureus : Staphylococcus aureus ; ROS: reactive oxygen species; RT-qPCR: quantitative Real-Time PCR. Results were presented as the mean ± SD, one way ANOVA was performed for multiple comparison tests, while Student's t -test was performed for two independent groups (ns, no significance, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗P < 0.0001).
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Effects of CM@MOF on HaCaT cells in vitro. (A) Migration ability of HaCaT cells evaluated by scratch assay, violet staining and immunofluorescence staining of Ki67. Green: Ki67; Blue: nuclei (DAPI). Scale bars: 100 μm. (B) Quantitative analysis of cell migration using ImageJ software. (C) Quantitative analysis of cell migrating rate. (D) Quantification of Ki67-positive cells percentage of HaCaT cells. (E) Live (Green)/Dead (Red) staining of HaCaT cells. Scale bars: 100 μm. (F) ROS(Green) in HaCaT cells observed by fluorescence microscopy. Scale bars: 50 μm. (G) Quantitative analysis of the percentage of live HaCaT cells. (H) Quantitative analysis of ROS fluorescence intensity (mean gray value) in HaCaT cells. (I) mRNA levels of IL6, IL17a, IL22, IL23a and Tnfα in HaCaT cells assessed by RT-qPCR. HaCaT: human <t>keratinocyte</t> cells; DAPI: 4′,6-diamidino-2-phenylindole; LPS: Lipopolysaccharide; M2: M2 membrane; CM: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage); MOF: Metal-organic framework loaded Cu/Zn; CM@MOF: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage) loaded Cu/Zn metal-organic framework (Cu/Zn-MOF); S. aureus : Staphylococcus aureus ; ROS: reactive oxygen species; RT-qPCR: quantitative Real-Time PCR. Results were presented as the mean ± SD, one way ANOVA was performed for multiple comparison tests, while Student's t -test was performed for two independent groups (ns, no significance, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗P < 0.0001).
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Effects of CM@MOF on HaCaT cells in vitro. (A) Migration ability of HaCaT cells evaluated by scratch assay, violet staining and immunofluorescence staining of Ki67. Green: Ki67; Blue: nuclei (DAPI). Scale bars: 100 μm. (B) Quantitative analysis of cell migration using ImageJ software. (C) Quantitative analysis of cell migrating rate. (D) Quantification of Ki67-positive cells percentage of HaCaT cells. (E) Live (Green)/Dead (Red) staining of HaCaT cells. Scale bars: 100 μm. (F) ROS(Green) in HaCaT cells observed by fluorescence microscopy. Scale bars: 50 μm. (G) Quantitative analysis of the percentage of live HaCaT cells. (H) Quantitative analysis of ROS fluorescence intensity (mean gray value) in HaCaT cells. (I) mRNA levels of IL6, IL17a, IL22, IL23a and Tnfα in HaCaT cells assessed by RT-qPCR. HaCaT: human <t>keratinocyte</t> cells; DAPI: 4′,6-diamidino-2-phenylindole; LPS: Lipopolysaccharide; M2: M2 membrane; CM: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage); MOF: Metal-organic framework loaded Cu/Zn; CM@MOF: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage) loaded Cu/Zn metal-organic framework (Cu/Zn-MOF); S. aureus : Staphylococcus aureus ; ROS: reactive oxygen species; RT-qPCR: quantitative Real-Time PCR. Results were presented as the mean ± SD, one way ANOVA was performed for multiple comparison tests, while Student's t -test was performed for two independent groups (ns, no significance, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗P < 0.0001).
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Effects of CM@MOF on HaCaT cells in vitro. (A) Migration ability of HaCaT cells evaluated by scratch assay, violet staining and immunofluorescence staining of Ki67. Green: Ki67; Blue: nuclei (DAPI). Scale bars: 100 μm. (B) Quantitative analysis of cell migration using ImageJ software. (C) Quantitative analysis of cell migrating rate. (D) Quantification of Ki67-positive cells percentage of HaCaT cells. (E) Live (Green)/Dead (Red) staining of HaCaT cells. Scale bars: 100 μm. (F) ROS(Green) in HaCaT cells observed by fluorescence microscopy. Scale bars: 50 μm. (G) Quantitative analysis of the percentage of live HaCaT cells. (H) Quantitative analysis of ROS fluorescence intensity (mean gray value) in HaCaT cells. (I) mRNA levels of IL6, IL17a, IL22, IL23a and Tnfα in HaCaT cells assessed by RT-qPCR. HaCaT: human <t>keratinocyte</t> cells; DAPI: 4′,6-diamidino-2-phenylindole; LPS: Lipopolysaccharide; M2: M2 membrane; CM: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage); MOF: Metal-organic framework loaded Cu/Zn; CM@MOF: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage) loaded Cu/Zn metal-organic framework (Cu/Zn-MOF); S. aureus : Staphylococcus aureus ; ROS: reactive oxygen species; RT-qPCR: quantitative Real-Time PCR. Results were presented as the mean ± SD, one way ANOVA was performed for multiple comparison tests, while Student's t -test was performed for two independent groups (ns, no significance, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗P < 0.0001).
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Effects of CM@MOF on HaCaT cells in vitro. (A) Migration ability of HaCaT cells evaluated by scratch assay, violet staining and immunofluorescence staining of Ki67. Green: Ki67; Blue: nuclei (DAPI). Scale bars: 100 μm. (B) Quantitative analysis of cell migration using ImageJ software. (C) Quantitative analysis of cell migrating rate. (D) Quantification of Ki67-positive cells percentage of HaCaT cells. (E) Live (Green)/Dead (Red) staining of HaCaT cells. Scale bars: 100 μm. (F) ROS(Green) in HaCaT cells observed by fluorescence microscopy. Scale bars: 50 μm. (G) Quantitative analysis of the percentage of live HaCaT cells. (H) Quantitative analysis of ROS fluorescence intensity (mean gray value) in HaCaT cells. (I) mRNA levels of IL6, IL17a, IL22, IL23a and Tnfα in HaCaT cells assessed by RT-qPCR. HaCaT: human <t>keratinocyte</t> cells; DAPI: 4′,6-diamidino-2-phenylindole; LPS: Lipopolysaccharide; M2: M2 membrane; CM: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage); MOF: Metal-organic framework loaded Cu/Zn; CM@MOF: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage) loaded Cu/Zn metal-organic framework (Cu/Zn-MOF); S. aureus : Staphylococcus aureus ; ROS: reactive oxygen species; RT-qPCR: quantitative Real-Time PCR. Results were presented as the mean ± SD, one way ANOVA was performed for multiple comparison tests, while Student's t -test was performed for two independent groups (ns, no significance, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗P < 0.0001).
Keratinocyte Growth Kit, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Effects of CM@MOF on HaCaT cells in vitro. (A) Migration ability of HaCaT cells evaluated by scratch assay, violet staining and immunofluorescence staining of Ki67. Green: Ki67; Blue: nuclei (DAPI). Scale bars: 100 μm. (B) Quantitative analysis of cell migration using ImageJ software. (C) Quantitative analysis of cell migrating rate. (D) Quantification of Ki67-positive cells percentage of HaCaT cells. (E) Live (Green)/Dead (Red) staining of HaCaT cells. Scale bars: 100 μm. (F) ROS(Green) in HaCaT cells observed by fluorescence microscopy. Scale bars: 50 μm. (G) Quantitative analysis of the percentage of live HaCaT cells. (H) Quantitative analysis of ROS fluorescence intensity (mean gray value) in HaCaT cells. (I) mRNA levels of IL6, IL17a, IL22, IL23a and Tnfα in HaCaT cells assessed by RT-qPCR. HaCaT: human <t>keratinocyte</t> cells; DAPI: 4′,6-diamidino-2-phenylindole; LPS: Lipopolysaccharide; M2: M2 membrane; CM: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage); MOF: Metal-organic framework loaded Cu/Zn; CM@MOF: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage) loaded Cu/Zn metal-organic framework (Cu/Zn-MOF); S. aureus : Staphylococcus aureus ; ROS: reactive oxygen species; RT-qPCR: quantitative Real-Time PCR. Results were presented as the mean ± SD, one way ANOVA was performed for multiple comparison tests, while Student's t -test was performed for two independent groups (ns, no significance, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗P < 0.0001).
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low passage hacat human keratinocytes  (ATCC)
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Effects of CM@MOF on HaCaT cells in vitro. (A) Migration ability of HaCaT cells evaluated by scratch assay, violet staining and immunofluorescence staining of Ki67. Green: Ki67; Blue: nuclei (DAPI). Scale bars: 100 μm. (B) Quantitative analysis of cell migration using ImageJ software. (C) Quantitative analysis of cell migrating rate. (D) Quantification of Ki67-positive cells percentage of HaCaT cells. (E) Live (Green)/Dead (Red) staining of HaCaT cells. Scale bars: 100 μm. (F) ROS(Green) in HaCaT cells observed by fluorescence microscopy. Scale bars: 50 μm. (G) Quantitative analysis of the percentage of live HaCaT cells. (H) Quantitative analysis of ROS fluorescence intensity (mean gray value) in HaCaT cells. (I) mRNA levels of IL6, IL17a, IL22, IL23a and Tnfα in HaCaT cells assessed by RT-qPCR. HaCaT: human <t>keratinocyte</t> cells; DAPI: 4′,6-diamidino-2-phenylindole; LPS: Lipopolysaccharide; M2: M2 membrane; CM: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage); MOF: Metal-organic framework loaded Cu/Zn; CM@MOF: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage) loaded Cu/Zn metal-organic framework (Cu/Zn-MOF); S. aureus : Staphylococcus aureus ; ROS: reactive oxygen species; RT-qPCR: quantitative Real-Time PCR. Results were presented as the mean ± SD, one way ANOVA was performed for multiple comparison tests, while Student's t -test was performed for two independent groups (ns, no significance, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗P < 0.0001).
Low Passage Hacat Human Keratinocytes, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Effects of CM@MOF on HaCaT cells in vitro. (A) Migration ability of HaCaT cells evaluated by scratch assay, violet staining and immunofluorescence staining of Ki67. Green: Ki67; Blue: nuclei (DAPI). Scale bars: 100 μm. (B) Quantitative analysis of cell migration using ImageJ software. (C) Quantitative analysis of cell migrating rate. (D) Quantification of Ki67-positive cells percentage of HaCaT cells. (E) Live (Green)/Dead (Red) staining of HaCaT cells. Scale bars: 100 μm. (F) ROS(Green) in HaCaT cells observed by fluorescence microscopy. Scale bars: 50 μm. (G) Quantitative analysis of the percentage of live HaCaT cells. (H) Quantitative analysis of ROS fluorescence intensity (mean gray value) in HaCaT cells. (I) mRNA levels of IL6, IL17a, IL22, IL23a and Tnfα in HaCaT cells assessed by RT-qPCR. HaCaT: human keratinocyte cells; DAPI: 4′,6-diamidino-2-phenylindole; LPS: Lipopolysaccharide; M2: M2 membrane; CM: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage); MOF: Metal-organic framework loaded Cu/Zn; CM@MOF: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage) loaded Cu/Zn metal-organic framework (Cu/Zn-MOF); S. aureus : Staphylococcus aureus ; ROS: reactive oxygen species; RT-qPCR: quantitative Real-Time PCR. Results were presented as the mean ± SD, one way ANOVA was performed for multiple comparison tests, while Student's t -test was performed for two independent groups (ns, no significance, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗P < 0.0001).

Journal: Materials Today Bio

Article Title: Cutibacterium acnes -macrophage fusion membrane coating Cu/Zn-MOF for psoriasis treatment via pathological innate lymphoid cells inhibition

doi: 10.1016/j.mtbio.2025.102731

Figure Lengend Snippet: Effects of CM@MOF on HaCaT cells in vitro. (A) Migration ability of HaCaT cells evaluated by scratch assay, violet staining and immunofluorescence staining of Ki67. Green: Ki67; Blue: nuclei (DAPI). Scale bars: 100 μm. (B) Quantitative analysis of cell migration using ImageJ software. (C) Quantitative analysis of cell migrating rate. (D) Quantification of Ki67-positive cells percentage of HaCaT cells. (E) Live (Green)/Dead (Red) staining of HaCaT cells. Scale bars: 100 μm. (F) ROS(Green) in HaCaT cells observed by fluorescence microscopy. Scale bars: 50 μm. (G) Quantitative analysis of the percentage of live HaCaT cells. (H) Quantitative analysis of ROS fluorescence intensity (mean gray value) in HaCaT cells. (I) mRNA levels of IL6, IL17a, IL22, IL23a and Tnfα in HaCaT cells assessed by RT-qPCR. HaCaT: human keratinocyte cells; DAPI: 4′,6-diamidino-2-phenylindole; LPS: Lipopolysaccharide; M2: M2 membrane; CM: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage); MOF: Metal-organic framework loaded Cu/Zn; CM@MOF: Fusion membrane combined with cell wall ( C.acnes ) and membrane (M2-Macrophage) loaded Cu/Zn metal-organic framework (Cu/Zn-MOF); S. aureus : Staphylococcus aureus ; ROS: reactive oxygen species; RT-qPCR: quantitative Real-Time PCR. Results were presented as the mean ± SD, one way ANOVA was performed for multiple comparison tests, while Student's t -test was performed for two independent groups (ns, no significance, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗P < 0.0001).

Article Snippet: Human immortalized keratinocytes (HaCaT; obtained from Cell Lines Service, Eppelheim, #300493) were cultured at 37 °C in a humidified atmosphere with 5 % CO 2 in DMEM supplemented with 10 % FBS.

Techniques: In Vitro, Migration, Wound Healing Assay, Staining, Immunofluorescence, Software, Fluorescence, Microscopy, Quantitative RT-PCR, Membrane, Real-time Polymerase Chain Reaction, Comparison