Heat shocks enhance procollagen type I and III expression in fibroblasts in ex vivo human skin.
BACKGROUND
The well-known characteristics of aging skin are the development of fine lines and wrinkles, but changes in skin tone, skin texture, thickness and moisture content are also aspects of aging. Rejuvenation of the skin aims at reversing the signs of aging and can be established in the epidermis as well as in the dermis. Aged dermis, in fact, has a degenerated collagen matrix. To regenerate this matrix, fibroblasts need to be stimulated into synthesizing new collagen.


OBJECTIVE
In this study, the effects of heat shocks of different temperatures on human dermal fibroblasts in ex vivo skin on the expression of procollagen 1, procollagen 3, heat shock protein (hsp)27, hsp47, and hsp70 are investigated.


METHODS
The heat shocks were applied on ex vivo skin samples by immersing the samples in heated phosphate-buffered saline of 45 °C or 60 °C. Metabolic activity was measured and at similar time points propidium-iodide-calceine staining was performed to establish cell viability. Quantitative polymerase chain reaction (qPCR) was performed after the heat shock to determine gene expression levels relative to the reference temperature. Furthermore, PicroSirius Red and hematoxylin stainings were performed to visualize the collagen network and the cells.


RESULTS
The skin samples were shown to be viable and metabolically active. Histology indicated that the heat shocks did not influence the structure of the collagen network or cell appearance. qPCR results showed that in contrast to the 45 °C heat shock the 60 °C heat shock resulted in significant upregulations of procollagen type I and III, hsp70 and hsp47.


CONCLUSIONS
A 60 °C, heat shock stimulates the human dermal fibroblasts in ex vivo skin to upregulate their procollagen type I and type III expression.
Philips Research Eindhoven, Eindhoven, The Netherlands. susanne.van.den.berg@philips.com
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D017209:Apoptosis / Q000502:physiology; D024042:Collagen Type I / Q000235:genetics* / Q000378:metabolism; D024061:Collagen Type III / Q000235:genetics* / Q000378:metabolism; D020405:Dermis / Q000473:pathology / Q000503:physiopathology; D004817:Epidermis / Q000473:pathology / Q000503:physiopathology; D005347:Fibroblasts / Q000473:pathology / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology; D055551:HSP27 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D050958:HSP47 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins; D018869:Heat-Shock Response / Q000502:physiology*; D006358:Hot Temperature / Q000009:adverse effects; D006801:Humans; D066298:In Vitro Techniques; D008954:Models, Biological; D018832:Molecular Chaperones; D012060:Rejuvenation / Q000502:physiology; D015595:Skin Aging / Q000502:physiology*
D024042:Collagen Type I; D024061:Collagen Type III; D055551:HSP27 Heat-Shock Proteins; D050958:HSP47 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; C483693:Hspa14 protein, human; D018832:Molecular Chaperones; C494660:SERPINH1 protein, human