Fig. 5

Improvements in the skin ageing phenotype promoted by MNRF treatment and its association with senescent fibroblasts. (a) Representative images of immunofluorescence staining for procollagen-1 (red) and DAPI (blue) after MN and MNRF treatments. The graph on the right indicates the number of procollagen-1-positive cells after MN and MNRF treatments (n = 25). Original magnification ×400. (b) Representative images of dermal collagen deposition using Masson’s trichrome staining in the skin treated with MN and MNRF. The graphs on the right represent the results of the quantitative assessment of dermal collagen density in the papillary and reticular dermis after MN and MNRF treatments (n = 26). Scale bar = 50 μm. (c) Representative images of immunofluorescence staining for elastin (green) and DAPI (blue) after MN and MNRF treatments. The graphs on the right represent the results of the quantitative assessment of dermal elastic fibres in the papillary and reticular dermis after MN and MNRF treatments (n = 25). Scale bar = 50 μm. (d) The graphs indicate the evaluation of collagen density of papillary dermis (left) and hydration (right) in the skin treated with MN and MNRF of MNRF- responders (n = 12) and non-responders (n = 13). MNRF- responders and non-responders were distinguished based on differences in the number of senescent fibroblasts. Statistical significance: *p < 0.05, **p < 0.01, ****p < 0.0001. ns indicates no significance.