ABSTRACT
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. AIMS: 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. MATERIALS AND 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. CONCLUSION: A 60 °C, heat shock stimulates the human dermal fibroblasts in ex vivo skin to upregulate their procollagen type I and type III expression.
Subject(s)
Collagen Type III/genetics , Collagen Type I/genetics , Fibroblasts/physiology , Heat-Shock Response/physiology , Skin Aging/physiology , Apoptosis/physiology , Collagen Type I/metabolism , Collagen Type III/metabolism , Dermis/pathology , Dermis/physiopathology , Epidermis/pathology , Epidermis/physiopathology , Fibroblasts/pathology , Gene Expression Regulation/physiology , HSP27 Heat-Shock Proteins/genetics , HSP27 Heat-Shock Proteins/metabolism , HSP47 Heat-Shock Proteins/genetics , HSP47 Heat-Shock Proteins/metabolism , HSP70 Heat-Shock Proteins/genetics , HSP70 Heat-Shock Proteins/metabolism , Heat-Shock Proteins , Hot Temperature/adverse effects , Humans , In Vitro Techniques , Models, Biological , Molecular Chaperones , Rejuvenation/physiologyABSTRACT
BACKGROUND: The formation of wrinkles is associated with degeneration of the collagen matrix. For regeneration of the matrix, fibroblasts need to be stimulated in producing new collagen. AIMS: In this study, the effect of short-pulsed heat shocks on gene expression of procollagen type I, procollagen type III, heat shock protein (hsp)27, hsp47 and hsp70 and on the expression of remodeling markers, procollagen type I carboxy-terminal peptide (P1P) and carboxy-terminal telopeptide of type I (ICTP), of human dermal fibroblasts in vitro, is investigated. MATERIALS AND METHODS: Temperatures of 45 degrees C and 60 degrees C were used for the heat shocks. The proliferation rates, viability and metabolic activity were measured directly after the pulsed heat shocks and quantitative PCR was performed at five different time points after the heat shocks. Enzyme Immuno Assays were performed to determine the concentrations of P1P and ICTP. RESULTS: A decreased proliferation rate of the 60 degrees C heat shocked cells was shown, whereas the viability and metabolic activity did not differ. Furthermore, gene expressions were upregulated in both 45 degrees C and 60 degrees C heat-shocked cells. However, remodeling marker analyses showed a larger amount of collagen produced by 60 degrees C heat-shocked cells. CONCLUSION: It can be concluded that these findings, together with upregulation in gene expression, show that it is possible to stimulate the cells to produce more collagen with short-pulsed heat shocks.
