human hair follicle dermal papilla cells  (PromoCell)

Journal: International Journal of Molecular Sciences

Article Title: Poly-D,L-Lactic Acid Filler Restores Hair Thickness and Shine by Ameliorating Age-Associated Follicular Decline

doi: 10.3390/ijms27052098

Figure Lengend Snippet: PDLLA filler restores cell-cycle activity and paracrine function in senescent hDPCs, thereby promoting keratin synthesis in senescent hHFKs. ( A ) Schematic overview of the hDPC experimental design. hDPCs (1 × 10 6 cells) were treated with H 2 O 2 (150 µM) for 1.5 h, cultured in fresh growth medium for 3 days, and subsequently treated with PDLLA filler (300 µg/mL). Cells and CM were harvested 2 days after PDLLA filler treatment. ( B – D ) Cell-cycle distribution of senescent hDPCs analyzed by flow cytometry using PI staining. Percentages of cells in the G0/G1 ( B ), S ( C ), and G2/M ( D ) phases are shown. ( E ) Proliferation of senescent hDPCs assessed by proliferation assay after PDLLA filler treatment and expressed as fold change relative to PBS-treated senescent controls. ( F ) IGF-1 secretion level in CM from senescent hDPCs was quantified by ELISA and expressed as fold change relative to PBS-treated controls. ( G ) Schematic overview of the hHFK experimental design. Senescent hHFKs were cultured with CM derived from PBS-treated or PDLLA filler-treated senescent hDPCs. ( H ) Proliferation of hHFKs assessed after exposure to CM from PBS-treated (CM PBS ) or PDLLA filler-treated hDPCs (CM PDLLA filler ). ( I ) Western blot analysis of pan-keratin expression in senescent hHFKs. Molecular weight markers are indicated; pan-keratin bands were expected at 46–58 kDa. GAPDH served as the loading control. ( J ) Quantitative densitometric analysis of pan-keratin protein levels shown in ( I ), normalized to GAPDH and expressed as fold change relative to CM PBS . Data are presented as mean ± standard deviation. Differences among groups were analyzed using the Kruskal-Wallis test, with Mann–Whitney U tests used for post hoc pairwise comparisons. *, p < 0.05 and **, p < 0.01 vs. PBS or CM PBS . CM, conditioned medium; d, days; ELISA, enzyme-linked immunosorbent assay; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; h, hours; hDPCs, human dermal papilla cells; hHFKs, human hair follicular keratinocytes; H 2 O 2, hydrogen peroxide; IGF-1, Insulin-like growth factor-1; PBS, phosphate-buffered saline; PDLLA, poly-D,L-lactic acid; PI, propidium iodide.

Article Snippet: Human DPCs (hDPCs) were purchased from PromoCell GmbH (Heidelberg, Germany) and cultured in Follicle Dermal Papilla Cell Growth Medium (PromoCell) supplemented with the provided growth supplement mix and 1% penicillin/streptomycin, in accordance with the manufacturer’s instructions.

Techniques: Activity Assay, Cell Culture, Flow Cytometry, Staining, Proliferation Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Western Blot, Expressing, Molecular Weight, Control, Standard Deviation, MANN-WHITNEY, Saline

Journal: International Journal of Molecular Sciences

Article Title: Poly-D,L-Lactic Acid Filler Restores Hair Thickness and Shine by Ameliorating Age-Associated Follicular Decline

doi: 10.3390/ijms27052098

Figure Lengend Snippet: PDLLA filler restores dermal papilla cell proliferation and promotes HMK proliferation in the hair matrix, as well as hair shaft keratin formation, in vivo. ( A ) Schematic illustration indicating the dermal papilla region within the hair follicle, where DPCs are densely localized and were analyzed for proliferation. Darker shaded areas indicate analyzed regions (dermal papilla). ( B ) Representative immunofluorescence images showing PCNA (green) expression in the dermal papilla region of hair follicles from saline- and PDLLA filler-treated mice. Nuclei were counterstained with DAPI (blue). Dashed boxes indicate the dermal papilla region, shown at higher magnification in the bottom panels. Proliferating cells were quantified by counting PCNA-positive nuclei-colocalized with DAPI within defined regions of interest. Scale bar = 100 μm. ( C ) Quantification of PCNA-positive cells in the dermal papilla of each hair follicle, expressed as the number of PCNA-positive cells per dermal papilla. ( D ) IGF-1 protein levels in whole skin tissue, measured by ELISA and expressed as fold change relative to the saline-treated control group. ( E ) Schematic illustration indicating hair matrix regions analyzed for HMK proliferation. Darker shaded areas represented the analyzed regions (hair matrix). ( F ) Representative immunofluorescence images showing PCNA expression (green) in the hair matrix region of hair follicles from saline- and PDLLA filler-treated mice. Nuclei were counterstained with DAPI (blue). Dashed boxes indicate the hair matrix region, shown at higher magnification in the bottom panels. Quantification was performed by counting PCNA-positive nuclei co-localized with DAPI within standardized regions of interest. Scale bar = 100 μm. ( G ) Quantification of PCNA-positive cells within the hair matrix region, expressed as the number of PCNA-positive cells per hair follicle. ( H ) Schematic illustration indicating hair shaft cortex regions analyzed for keratin expression. Darker shaded areas indicate analyzed regions (hair cortex). ( I ) Representative immunofluorescence images showing the expression of hair shaft cortex-specific hard keratins K35 (type I; green) and K85 (type II; red) in hair follicles from saline- and PDLLA filler-treated mice. Nuclei were counterstained with DAPI (blue). Scale bar = 100 μm. ( J , K ) Quantitative analysis of K35-positive ( J ) and K85-positive ( K ) fluorescence intensity within the hair shaft cortex region, expressed as fold change relative to the saline-treated control group. Data are presented as mean ± standard deviation (n = 5 per group). Group comparisons were performed using the Kruskal–Wallis test and Mann–Whitney U test. **, p < 0.01 vs. Saline. DAPI, 4′,6-diamidino-2-phenylindole; DPCs, dermal papilla cells; ELISA, enzyme-linked immunosorbent assay; HMK, hair matrix keratinocytes; IGF-1, Insulin-like growth factor-1; PCNA, proliferating cell nuclear antigen; PDLLA, poly-D,L-lactic acid.

Article Snippet: Human DPCs (hDPCs) were purchased from PromoCell GmbH (Heidelberg, Germany) and cultured in Follicle Dermal Papilla Cell Growth Medium (PromoCell) supplemented with the provided growth supplement mix and 1% penicillin/streptomycin, in accordance with the manufacturer’s instructions.

Techniques: In Vivo, Immunofluorescence, Expressing, Saline, Enzyme-linked Immunosorbent Assay, Control, Fluorescence, Standard Deviation, MANN-WHITNEY