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Effect of SCFF on HFDPC and HaCaT cell viability and proliferation. <t>HFDPCs</t> and HaCaT cells were treated with different concentrations of SCFF as indicated for 24 h. ( a ) HFDPC and ( b ) HaCaT viability and proliferation were assessed at various SCFF concentrations (50, 100, 250, 500, 1000, 2500, 5000 μg/mL) using a CCK-8 assay. Values are mean ± SD of three independent experiments. ( a – c ) *** p < 0.001, ** p < 0.01 and * p < 0.05 versus NC group (100%). ( c ) Representative images of cellular proliferation activity in HFDPCs and HaCaT cells after 48 h of incubation ( n = 3; Scale bar = 200 μm). SCFF, Staphylococcus capitis KMH304 ferment filtrate; HFDPCs, <t>human</t> <t>follicle</t> dermal papilla cells; NC, negative control.
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Screening process to determine peptides with hair-restoring properties. ( A ) <t>Human</t> <t>Follicle</t> <t>Dermal</t> <t>Papilla</t> <t>Cell</t> (HFDPCs) viability of over 200 cell-penetrating peptides, with EGF as the positive control. The red dots mark the peptides with the highest cell viability. ( B ) Western Blot results showing the upregulation of β-catenin and p-ERK proteins of 22 chosen peptides compared to the control. Peptide No.218 (in red rectangle) showed the highest expression level of β-catenin and p-ERK proteins, and was chosen for subsequent analyses. ( C ) Cell-penetrating capability of DualPep-ALO was evaluated using FITC-conjugated peptides in HFDPCs. Cells were treated with FITC-DualPep-ALO or FITC-TAT (2.5 µM, 2 h), followed by extensive washing and trypsinization to remove membrane-bound peptides. Intracellular fluorescence intensity was quantified by flow cytometry. Each experiment was performed three times
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Screening process to determine peptides with hair-restoring properties. ( A ) <t>Human</t> <t>Follicle</t> <t>Dermal</t> <t>Papilla</t> <t>Cell</t> (HFDPCs) viability of over 200 cell-penetrating peptides, with EGF as the positive control. The red dots mark the peptides with the highest cell viability. ( B ) Western Blot results showing the upregulation of β-catenin and p-ERK proteins of 22 chosen peptides compared to the control. Peptide No.218 (in red rectangle) showed the highest expression level of β-catenin and p-ERK proteins, and was chosen for subsequent analyses. ( C ) Cell-penetrating capability of DualPep-ALO was evaluated using FITC-conjugated peptides in HFDPCs. Cells were treated with FITC-DualPep-ALO or FITC-TAT (2.5 µM, 2 h), followed by extensive washing and trypsinization to remove membrane-bound peptides. Intracellular fluorescence intensity was quantified by flow cytometry. Each experiment was performed three times
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Effect of SCFF on HFDPC and HaCaT cell viability and proliferation. HFDPCs and HaCaT cells were treated with different concentrations of SCFF as indicated for 24 h. ( a ) HFDPC and ( b ) HaCaT viability and proliferation were assessed at various SCFF concentrations (50, 100, 250, 500, 1000, 2500, 5000 μg/mL) using a CCK-8 assay. Values are mean ± SD of three independent experiments. ( a – c ) *** p < 0.001, ** p < 0.01 and * p < 0.05 versus NC group (100%). ( c ) Representative images of cellular proliferation activity in HFDPCs and HaCaT cells after 48 h of incubation ( n = 3; Scale bar = 200 μm). SCFF, Staphylococcus capitis KMH304 ferment filtrate; HFDPCs, human follicle dermal papilla cells; NC, negative control.

Journal: Microorganisms

Article Title: Anti-Hair Loss Activity of Healthy Human Scalp-Derived Staphylococcus capitis KMH304 Ferment Filtrate in Human Hair-Follicle Dermal Papilla and Keratinocyte Cells

doi: 10.3390/microorganisms14040929

Figure Lengend Snippet: Effect of SCFF on HFDPC and HaCaT cell viability and proliferation. HFDPCs and HaCaT cells were treated with different concentrations of SCFF as indicated for 24 h. ( a ) HFDPC and ( b ) HaCaT viability and proliferation were assessed at various SCFF concentrations (50, 100, 250, 500, 1000, 2500, 5000 μg/mL) using a CCK-8 assay. Values are mean ± SD of three independent experiments. ( a – c ) *** p < 0.001, ** p < 0.01 and * p < 0.05 versus NC group (100%). ( c ) Representative images of cellular proliferation activity in HFDPCs and HaCaT cells after 48 h of incubation ( n = 3; Scale bar = 200 μm). SCFF, Staphylococcus capitis KMH304 ferment filtrate; HFDPCs, human follicle dermal papilla cells; NC, negative control.

Article Snippet: HFDPCs (C-12071; Promocell, Heidelberg, Germany) were cultured in Follicle Dermal Papilla Cell Growth Medium (C-26501; Promocell) supplemented with 1% antibiotic–antimycotic (Gibco; Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: CCK-8 Assay, Activity Assay, Incubation, Negative Control

Hair growth-related and AGA-inducing signature gene expression following SCFF treatment in HFDPCs. HFDPCs were treated with different SCFF concentrations for 24 h, and gene expression was evaluated using qPCR. ( a ) Hair growth-related factors (KGF, IGF-1, and HGF). ( b ) AGA-inducing factors (AR and TGF-β2). MXD at 5 μg/mL was used as PC for TGF-β2, and 2.5 μg/mL MXD was used as PC for other markers. Data are mean ± SD of three independent experiments. *** p < 0.001, ** p < 0.01 and * p < 0.05 versus NC group (100%). SCFF, Staphylococcus capitis KMH304 ferment filtrate; HFDPCs, human follicle dermal papilla cells; MXD, minoxidil; PC, positive control.

Journal: Microorganisms

Article Title: Anti-Hair Loss Activity of Healthy Human Scalp-Derived Staphylococcus capitis KMH304 Ferment Filtrate in Human Hair-Follicle Dermal Papilla and Keratinocyte Cells

doi: 10.3390/microorganisms14040929

Figure Lengend Snippet: Hair growth-related and AGA-inducing signature gene expression following SCFF treatment in HFDPCs. HFDPCs were treated with different SCFF concentrations for 24 h, and gene expression was evaluated using qPCR. ( a ) Hair growth-related factors (KGF, IGF-1, and HGF). ( b ) AGA-inducing factors (AR and TGF-β2). MXD at 5 μg/mL was used as PC for TGF-β2, and 2.5 μg/mL MXD was used as PC for other markers. Data are mean ± SD of three independent experiments. *** p < 0.001, ** p < 0.01 and * p < 0.05 versus NC group (100%). SCFF, Staphylococcus capitis KMH304 ferment filtrate; HFDPCs, human follicle dermal papilla cells; MXD, minoxidil; PC, positive control.

Article Snippet: HFDPCs (C-12071; Promocell, Heidelberg, Germany) were cultured in Follicle Dermal Papilla Cell Growth Medium (C-26501; Promocell) supplemented with 1% antibiotic–antimycotic (Gibco; Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Gene Expression, Positive Control

The regulation of cellular senescence by SCFF in HFDPCs. HFDPCs were treated with different concentrations of SCFF for 24 h, followed by analysis using qPCR and SA-β-gal staining. ( a , b ) Anti-aging factors (SIRT1, SIRT7, COL13A1, and p21). ( c ) SA-β-gal staining in young (P6) and senescent (P16) passage cells. Quantitative and representative images of SA-β-gal-positive cells compared with senescent cells (Scale bar = 200 μm). For SIRT, 1 μg/mL fisetin (Fis) was used as a positive control (PC), and for COL13 and p21, 1 μg/mL Fis was used as PC. Data are the mean ± SD of three independent experiments. **** p < 0.0001, *** p < 0.001, ** p < 0.01, and * p < 0.05 versus the negative control (NC) and the senescent group (100%). SCFF, Staphylococcus capitis KMH304 ferment filtrate; HFDPCs, human follicle dermal papilla cells.

Journal: Microorganisms

Article Title: Anti-Hair Loss Activity of Healthy Human Scalp-Derived Staphylococcus capitis KMH304 Ferment Filtrate in Human Hair-Follicle Dermal Papilla and Keratinocyte Cells

doi: 10.3390/microorganisms14040929

Figure Lengend Snippet: The regulation of cellular senescence by SCFF in HFDPCs. HFDPCs were treated with different concentrations of SCFF for 24 h, followed by analysis using qPCR and SA-β-gal staining. ( a , b ) Anti-aging factors (SIRT1, SIRT7, COL13A1, and p21). ( c ) SA-β-gal staining in young (P6) and senescent (P16) passage cells. Quantitative and representative images of SA-β-gal-positive cells compared with senescent cells (Scale bar = 200 μm). For SIRT, 1 μg/mL fisetin (Fis) was used as a positive control (PC), and for COL13 and p21, 1 μg/mL Fis was used as PC. Data are the mean ± SD of three independent experiments. **** p < 0.0001, *** p < 0.001, ** p < 0.01, and * p < 0.05 versus the negative control (NC) and the senescent group (100%). SCFF, Staphylococcus capitis KMH304 ferment filtrate; HFDPCs, human follicle dermal papilla cells.

Article Snippet: HFDPCs (C-12071; Promocell, Heidelberg, Germany) were cultured in Follicle Dermal Papilla Cell Growth Medium (C-26501; Promocell) supplemented with 1% antibiotic–antimycotic (Gibco; Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Staining, Positive Control, Negative Control

Overall effect of SCFF in HFDPCs and HaCaT cells. In keratinocytes, SCFF enhanced barrier function and antioxidant activity. SCFF not only regulated cellular aging through longevity-, collagen-, and p21-related genes, but also promoted hair growth by increasing the expression of hair-growth–associated factors and suppressing hair loss-inducing factors in HFDPCs. SCFF, Staphylococcus capitis KMH304 ferment filtrate; HFDPCs, human follicle dermal papilla cells.

Journal: Microorganisms

Article Title: Anti-Hair Loss Activity of Healthy Human Scalp-Derived Staphylococcus capitis KMH304 Ferment Filtrate in Human Hair-Follicle Dermal Papilla and Keratinocyte Cells

doi: 10.3390/microorganisms14040929

Figure Lengend Snippet: Overall effect of SCFF in HFDPCs and HaCaT cells. In keratinocytes, SCFF enhanced barrier function and antioxidant activity. SCFF not only regulated cellular aging through longevity-, collagen-, and p21-related genes, but also promoted hair growth by increasing the expression of hair-growth–associated factors and suppressing hair loss-inducing factors in HFDPCs. SCFF, Staphylococcus capitis KMH304 ferment filtrate; HFDPCs, human follicle dermal papilla cells.

Article Snippet: HFDPCs (C-12071; Promocell, Heidelberg, Germany) were cultured in Follicle Dermal Papilla Cell Growth Medium (C-26501; Promocell) supplemented with 1% antibiotic–antimycotic (Gibco; Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Antioxidant Activity Assay, Expressing

Screening process to determine peptides with hair-restoring properties. ( A ) Human Follicle Dermal Papilla Cell (HFDPCs) viability of over 200 cell-penetrating peptides, with EGF as the positive control. The red dots mark the peptides with the highest cell viability. ( B ) Western Blot results showing the upregulation of β-catenin and p-ERK proteins of 22 chosen peptides compared to the control. Peptide No.218 (in red rectangle) showed the highest expression level of β-catenin and p-ERK proteins, and was chosen for subsequent analyses. ( C ) Cell-penetrating capability of DualPep-ALO was evaluated using FITC-conjugated peptides in HFDPCs. Cells were treated with FITC-DualPep-ALO or FITC-TAT (2.5 µM, 2 h), followed by extensive washing and trypsinization to remove membrane-bound peptides. Intracellular fluorescence intensity was quantified by flow cytometry. Each experiment was performed three times

Journal: BMC Biotechnology

Article Title: A novel cell-penetrating peptide supports hair follicle growth through anti-inflammatory and growth factor–associated mechanisms in preclinical models

doi: 10.1186/s12896-026-01130-4

Figure Lengend Snippet: Screening process to determine peptides with hair-restoring properties. ( A ) Human Follicle Dermal Papilla Cell (HFDPCs) viability of over 200 cell-penetrating peptides, with EGF as the positive control. The red dots mark the peptides with the highest cell viability. ( B ) Western Blot results showing the upregulation of β-catenin and p-ERK proteins of 22 chosen peptides compared to the control. Peptide No.218 (in red rectangle) showed the highest expression level of β-catenin and p-ERK proteins, and was chosen for subsequent analyses. ( C ) Cell-penetrating capability of DualPep-ALO was evaluated using FITC-conjugated peptides in HFDPCs. Cells were treated with FITC-DualPep-ALO or FITC-TAT (2.5 µM, 2 h), followed by extensive washing and trypsinization to remove membrane-bound peptides. Intracellular fluorescence intensity was quantified by flow cytometry. Each experiment was performed three times

Article Snippet: HFDPCs were cultured in Human Follicle Dermal Papilla Cell Growth Medium (PromoCell, Heidelberg, Germany) supplemented with fetal calf serum, bovine pituitary extract, basic fibroblast growth factor, and insulin.

Techniques: Positive Control, Western Blot, Control, Expressing, Membrane, Fluorescence, Flow Cytometry

Effects of DualPep-ALO on cell viability. ( A ) Mouse macrophage viability after 24-hour treatment with various concentrations of DualPep-ALO (0.64, 3.2, 16, 80 µM), compared with L-NMMA (25 µM). ( B ) Human Follicle Dermal Papilla Cells viability after treatment with DualPep-ALO (3.2, 16, 80 µM) for 24, 48, and 72 hours, compared with minoxidil (20 µM). Data are presented as mean ± SD. *p < 0.05 vs. negative control at the same time point. Each experiment was performed three times

Journal: BMC Biotechnology

Article Title: A novel cell-penetrating peptide supports hair follicle growth through anti-inflammatory and growth factor–associated mechanisms in preclinical models

doi: 10.1186/s12896-026-01130-4

Figure Lengend Snippet: Effects of DualPep-ALO on cell viability. ( A ) Mouse macrophage viability after 24-hour treatment with various concentrations of DualPep-ALO (0.64, 3.2, 16, 80 µM), compared with L-NMMA (25 µM). ( B ) Human Follicle Dermal Papilla Cells viability after treatment with DualPep-ALO (3.2, 16, 80 µM) for 24, 48, and 72 hours, compared with minoxidil (20 µM). Data are presented as mean ± SD. *p < 0.05 vs. negative control at the same time point. Each experiment was performed three times

Article Snippet: HFDPCs were cultured in Human Follicle Dermal Papilla Cell Growth Medium (PromoCell, Heidelberg, Germany) supplemented with fetal calf serum, bovine pituitary extract, basic fibroblast growth factor, and insulin.

Techniques: Negative Control

Antioxidant enzyme activities in Human Follicle Dermal Papilla Cells (HFPDCs) treated with DualPep-ALO. DualPep-ALO (3.2, 16, 80 µM) enhanced the activities of ( A ) Superoxide dismutase (SOD) and ( B ) catalase (CAT) in HFPDCs after oxidative stress induction with H 2 O 2 (400 µM). L-Ascorbic acid (100 µM) was used as a positive control. Data are expressed as mean ± SD. *p < 0.05 vs. negative control; #p < 0.05 vs. H 2 O 2 -treated group. Each experiment was performed three times

Journal: BMC Biotechnology

Article Title: A novel cell-penetrating peptide supports hair follicle growth through anti-inflammatory and growth factor–associated mechanisms in preclinical models

doi: 10.1186/s12896-026-01130-4

Figure Lengend Snippet: Antioxidant enzyme activities in Human Follicle Dermal Papilla Cells (HFPDCs) treated with DualPep-ALO. DualPep-ALO (3.2, 16, 80 µM) enhanced the activities of ( A ) Superoxide dismutase (SOD) and ( B ) catalase (CAT) in HFPDCs after oxidative stress induction with H 2 O 2 (400 µM). L-Ascorbic acid (100 µM) was used as a positive control. Data are expressed as mean ± SD. *p < 0.05 vs. negative control; #p < 0.05 vs. H 2 O 2 -treated group. Each experiment was performed three times

Article Snippet: HFDPCs were cultured in Human Follicle Dermal Papilla Cell Growth Medium (PromoCell, Heidelberg, Germany) supplemented with fetal calf serum, bovine pituitary extract, basic fibroblast growth factor, and insulin.

Techniques: Positive Control, Negative Control