title: Phenotypic and functional changes in dermal primary fibroblasts isolated from intrinsically aged human skin.
abstract: Dermal fibroblasts play a key role in maintaining skin homoeostasis by synthesizing and degrading extracellular matrix components. During ageing, they are subjected to changes, such as the loss of type I collagen expression and an increased synthesis of metalloproteinase I, leading to fragmentation of collagen fibrils with consequent reduction of the mechanical tension and defects of skin wound healing. Most information about fibroblast ageing was obtained from experiments performed on replicative-senescent dermal fibroblasts in vitro. However, the senescence status of fibroblasts isolated from intrinsically aged skins and its consequences on functionality need to be deeper investigated. Herein, we studied age-related phenotypic and functional alteration of fibroblasts from 'young' (<35 years) and 'old' (>50 years) donors. Our results brought evidence of the senescent status of 'old' fibroblasts by senescence associated β-galactosidase (SA-βgal) positive staining and p16 expression. A PCR array focusing on senescence highlighted a subset of downregulated genes including cell cycle progression and ECM genes in 'old' fibroblasts as well as a subset of upregulated genes involved in senescence features. In 'old' fibroblasts, we measured a downregulation of proliferative and contractile capacities of migratory potential under PDGF stimulation and activation into myofibroblasts under TGFβ. Old fibroblasts were also more sensitive to oxidative stress than 'young' ones. Of interest, downregulation of p16 expression partially reversed the senescent phenotype of 'old' fibroblasts but failed to restore their functional properties. In conclusion, our data brought evidence of phenotypic and functional differences between fibroblasts from young and intrinsically aged skin that may contribute to the alterations observed with ageing.
authors.affiliation: INSERM U976, Centre de Recherche en Dermatologie, Paris, France.|Université Paris Diderot, Sorbonne Paris Cité, Paris, France.|Centre de Recherche, Johnson & Johnson Santé Beauté France, Val de Reuil, France.
authors.forename: Cécilia
authors.initials: C
authors.lastname: Brun
authors.affiliation: INSERM U976, Centre de Recherche en Dermatologie, Paris, France.|Université Paris Diderot, Sorbonne Paris Cité, Paris, France.
authors.forename: Francette
authors.initials: F
authors.lastname: Jean-Louis
authors.affiliation: Centre de Recherche, Johnson & Johnson Santé Beauté France, Val de Reuil, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Oddos
authors.affiliation: INSERM U976, Centre de Recherche en Dermatologie, Paris, France.|Université Paris Diderot, Sorbonne Paris Cité, Paris, France.|AP-HP, Service de Dermatologie, Hôpital Saint-Louis, Paris, France.
authors.forename: Martine
authors.initials: M
authors.lastname: Bagot
authors.affiliation: INSERM U976, Centre de Recherche en Dermatologie, Paris, France.|Université Paris Diderot, Sorbonne Paris Cité, Paris, France.
authors.forename: Armand
authors.initials: A
authors.lastname: Bensussan
authors.affiliation: INSERM U976, Centre de Recherche en Dermatologie, Paris, France.|Université Paris Diderot, Sorbonne Paris Cité, Paris, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Michel
mesh_terms: D000328:Adult; D002453:Cell Cycle; D002455:Cell Division; D002478:Cells, Cultured; D016922:Cellular Senescence; D003094:Collagen / Q000096:biosynthesis / Q000235:genetics; D019941:Cyclin-Dependent Kinase Inhibitor p16; D020405:Dermis / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D019942:Genes, p16; D006801:Humans; D008875:Middle Aged; D058628:Myofibroblasts / Q000166:cytology; D009363:Neoplasm Proteins / Q000096:biosynthesis; D010641:Phenotype; D034622:RNA Interference; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D015595:Skin Aging; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
chemical_list: C000614131:CDKN2A protein, human; D019941:Cyclin-Dependent Kinase Inhibitor p16; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016212:Transforming Growth Factor beta; D003094:Collagen; D001616:beta-Galactosidase
keywords: ageing; dermal fibroblasts; senescence; skin