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

Journal: International Journal of Molecular Sciences

Article Title: HB-EGF Improves the Hair Regenerative Potential of Adipose-Derived Stem Cells via ROS Generation and Hck Phosphorylation

doi: 10.3390/ijms21010122

Figure Lengend Snippet: Thrombopoietin (THPO) injection promotes hair growth in vivo, stimulating dermal papilla cells (DPCs). ( A ) Recombinant human THPO was injected into the dorsal skin of shaved mice. The hair weights were measured, skin sections were analyzed by HE staining, and numbers of mature hair follicles and hair follicles with Ki67 + cells were measured 16–17 days later. Asterisks indicate hair follicles with Ki67 + cells in the matrix amplifying zone. n = 4 per group. Three independent experiments were conducted. ( B ) THPO increased the length of mouse vibrissal hair follicles compared with the control. ( C ) THPO increased DPC proliferation dose- and time-dependently. ( D ) THPO upregulated the mRNA expression of DPC marker genes in DPC. * p < 0.05, ** p < 0.01, *** p < 0.001. Three independent experiments were conducted per data point. All error bars indicate the S.E.M.

Article Snippet: Human DPCs were purchased from PromoCell (#C-12071) and cultured in follicle DPC medium with supplement mix (PromoCell, Heidelberg, Germany) and 0.1% anti-antibiotics (Gibco).

Techniques: Injection, In Vivo, Recombinant, Staining, Expressing, Marker