Journal: The American Journal of Pathology

Article Title: Release of miR-29 Target Laminin C2 Improves Skin Repair

doi: 10.1016/j.ajpath.2023.11.002

Figure Lengend Snippet: miR-29a is expressed in suprabasal neoepidermis and in the granulation tissue of healing wounds in mice. A: The role of miR-29 in cell adhesion in the epidermis was tested using the skin wound model. Two wounds ( dashed lines ) are shown in the mouse dorsum at day 0 after biopsy (middle), followed by the experimental plan (bottom). A decrease in miR-29 may affect cutaneous repair through the up-regulation of mRNA targets of miR-29. B and C: miR-29a in sections of wounds harvested at days 1, 3, and 5 after biopsy [fluorescence in situ hybridization (FISH); nuclei counterstained with DAPI]. The inset corresponds to the boxed region at greater magnification. Dashed lines demarcate the border between the dermis and the epidermis. Arrowhead shows suprabasal keratinocytes positive for miR-29a at the edge of the wound; double arrowheads point to the basal layer of regenerating epidermis void of miR-29. Scrambled probe was used to control the background and is shown at day 3 for representation; the image at day 5 is representative of the undetected expression of miR-29b. D: Quantification of positively stained pixels in day-5 wound sections from WT and KO mice; representative images are shown in Figure 2 H. E: Wound contraction, assessed as the percentage of the original wound diameter, in WT and KO wounds, as shown in Figure 2 A. F: Area of newly formed wound epidermis, as shown in Figure 2 A. Data are expressed as actual values and means ± SD. n = 12 wounds per genotype. Scale bars: 5 mm ( A ); 10 μm ( B, inset); 50 μm ( B and C ). E, epidermis; D, dermis; H&E, hematoxylin and eosin; HF, hair follicle; NE, neoepidermis; W, wound bed.

Article Snippet: Primary human keratinocytes (PeloBiotech, Planegg, Germany) were maintained in serum-free keratinocyte growth medium (PeloBiotech) at 37°C, 5% CO 2 on collagen I (Σ) or Coating Matrix Kit Protein (Gibco/Thermo Fisher Scientific, Schwerte, Germany) coated flasks.

Techniques: Fluorescence, In Situ Hybridization, Control, Expressing, Staining

Journal: The American Journal of Pathology

Article Title: Release of miR-29 Target Laminin C2 Improves Skin Repair

doi: 10.1016/j.ajpath.2023.11.002

Figure Lengend Snippet: LAMC2 deposition in the matrix of human traumatic wounds. A: Section of a human wound biopsy specimen, stained with anti-LAMC2 antibody and counterstained with hematoxylin. Note the brown signal for LAMC2 between the epidermis (Epi) and dermis on the wound (W) edge ( double arrowheads ) and around the small blood vessels (BV, single arrowheads ). B: Basal keratinocytes of the wound epidermal (W Epi) tongue deposit LAMC2 near blood vessels (BV; single arrowheads ). C and D: Blood vessels in maturing wound matrix retain LAMC2 ( arrows ). Blue staining is immature wound matrix, which turns golden brown as more collagen is formed. Epithelial and endothelial cells appear deep purple, along with lighter purple staining of other cell types in the dermis (fibroblasts and immune cells). Erythrocytes are in red. E: Suggested mechanism of action of miR-29 and LAMC2 in wound repair. Deposition LAMC2 into wound bed by keratinocytes at the leading edge contributes to angiogenesis, which, in turn, feeds epidermal advancement and wound repair. Scale bars: 200 μm ( A ); 100 μm ( B , top; C ); 20 μm ( B , bottom; D ). GT, granulation tissue.

Article Snippet: Primary human keratinocytes (PeloBiotech, Planegg, Germany) were maintained in serum-free keratinocyte growth medium (PeloBiotech) at 37°C, 5% CO 2 on collagen I (Σ) or Coating Matrix Kit Protein (Gibco/Thermo Fisher Scientific, Schwerte, Germany) coated flasks.

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: Experimental and Clinical Evidence Suggests That Treatment with Betacellulin Can Alleviate Th2-Type Cytokine-Mediated Impairment of Skin Barrier Function

doi: 10.3390/ijms231911520

Figure Lengend Snippet: Cutaneous expression and distribution of BTC and effects of its knockdown on human keratinocytes. ( a ). Representative immunohistochemistry images ( left ) and quantification ( right ) of BTC in healthy normal skin tissues and AD lesional skin tissues. Scale bar: 200 μm, n = 5. The mRNA expression levels of BTC ( b ), FLG , LOR , CLDN1 , TJP1 ( c ), DEFB1 , DEFB4A , DEFB103 , DEFB104 , and CAMP ( d ) in keratinocytes in the presence or absence of BTC siRNA were detected. n = 3. * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Article Snippet: Primary normal human epidermal keratinocytes (FC-0007, Kurabo Industries, Osaka, Japan) isolated from neonatal foreskins were cultured in serum-free HuMedia-KG2 keratinocyte growth medium (KK-2150S, Kurabo Industries) containing human epidermal growth factor, insulin, hydrocortisone, gentamicin, amphotericin B, and bovine brain pituitary extract at 37 °C in a humidified atmosphere consisting of 95% air and 5%, as previously described [ ].

Techniques: Expressing, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: Experimental and Clinical Evidence Suggests That Treatment with Betacellulin Can Alleviate Th2-Type Cytokine-Mediated Impairment of Skin Barrier Function

doi: 10.3390/ijms231911520

Figure Lengend Snippet: BTC increases the expression of TJ components and skin-derived antimicrobial peptides and improves the TER in keratinocytes. The mRNA expression levels of FLG , LOR , CLDN1 , and TJP1 ( a ); the protein levels of filaggrin, loricrin, claudin-1, ZO-1 ( b ), and TER ( c ); and the mRNA expression levels of DEFB1 , DEFB4A , DEFB103 , DEFB104 , and CAMP ( d ) in keratinocytes with or without BTC administration were detected. n = 3. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Primary normal human epidermal keratinocytes (FC-0007, Kurabo Industries, Osaka, Japan) isolated from neonatal foreskins were cultured in serum-free HuMedia-KG2 keratinocyte growth medium (KK-2150S, Kurabo Industries) containing human epidermal growth factor, insulin, hydrocortisone, gentamicin, amphotericin B, and bovine brain pituitary extract at 37 °C in a humidified atmosphere consisting of 95% air and 5%, as previously described [ ].

Techniques: Expressing, Derivative Assay