Quantitative evaluation of chronological ageing and photoageing in vivo: studies on skin echogenicity and thickness

Skin ageing is divided into chronological ageing and photoageing due to the cumulative effects of solar ultraviolet radiation. It is, however, difficult to measure the degree of photoageing and chronological ageing in humans in vivo . Here, we have evaluated the usefulness of ultrasonography for measurement of chronological ageing and photoageing in vivo . Twenty megahertz ultrasonography was performed in 90 individuals (29 men, 61 women, age 18–94) to describe age-related changes in sun-exposed regions with different levels of sun exposure (dorsal and ventral forearm, forehead, ankle) and non-exposed buttock skin. Skin thickness and skin echogenicity in different layers of the dermis were measured in ultrasound images. Additionally, cutaneous photodamage was scored clinically. Age-related changes were dependent on body site as well as layer of the dermis. A progressive, age-related decrease in echogenicity of the upper dermis was found in sun-exposed regions (dorsal forearm, forehead), but not in moderately exposed regions (ventral forearm, ankle). In the buttock an increase in echogenicity was observed. The echogenicity of the lower dermis increased in all examined sites. Skin thickness increased with age in the forehead and buttock, but decreased in the extremity skin. Our findings show that photoageing causes a decrease in echogenicity in the upper dermis. In contrast, chronological ageing is associated with an increase in echogenicity in the lower dermis. Although both increases and decreases in skin thickness were observed in different anatomical regions, there was no general relationship between skin thickness and age. Dermal echogenicity was deemed valuable for in vivo study of chronological ageing and photoageing.     

The role of retinoids in the prevention and repair of aged and photoaged skin

Retinoids, either naturally occurring or synthetic, are defined by their ability to bind nuclear retinoid receptors of the steroid/thyroid superfamily. Their protean but key function in physiology is control of cellular proliferation and differentiation. Topical retinoids, namely tretinoin, have been proven to prevent and repair clinical features of photoageing; these processes are facilitated by an ability to prevent loss of collagen from, and stimulate new collagen formation in, the papillary dermis of sun-exposed skin. Emerging evidence indicates that intrinsic, chronological ageing of the skin shares several mechanistic features with photoageing. Indeed aged skin is characterized by retinoid sensitivity and is probably reparable by application of topical retinoids.     

A clinicopathological study of the effects of topical retinyl propionate cream in skin photoageing

The clinical and histological effects of retinyl propionate cream (a retinyl ester) on extrinsic skin ageing (photoageing) in man were assessed in a double-blind randomized placebo-controlled study of 80 subjects, individual parameters of this being assessed for each treatment site (face, dorsal right forearm and hand, dorsal left forearm and hand) at intervals throughout the study, while skin surface replicas from sites of fine wrinkling around the eye and skin biopsies from the dorsal right forearm were also regularly reviewed throughout. Seventy-five subjects completed an initial 24-week study period, following which 60 elected to continue for a further 24 weeks. Although minimal trends towards improvement occurred, no statistically significant differences between the effects of the retinyl propionate cream and the placebo preparation were apparent for any of the clinical, histological or profilometric parameters of skin photoageing; however, in the very few subjects affected, actinic keratoses in the active group were reduced virtually to zero by week 48.     

Dowling Oration delivered at the Royal College of Physicians, London, Friday 5 June 1998. Retinoids: renaissance and reformation

There is confusion as to the cutaneous signs of ageing. For the most part the features of photoageing, namely actinic lentigines and wrinkling, are misinterpreted as features of ageing. Wrinkling is associated with a loss of collagens from the papillary dermis resulting from imperfect remodelling of the dermal extracellular matrix following sun exposure. Retinoids are either derivatives of vitamin A or synthetic ligands of nuclear retinoid receptors. Retinoid receptors, notably retinoic acid receptor gamma and retinoid X receptor alpha are present in human skin. Topically applied all-trans retinoic acid can repair and probably prevent photoageing of the skin by modulation of collagen synthesis in the dermis. There is emerging evidence that intrinsic ageing of the skin is also amenable to reversal by topical retinoids.     

Pigmentation in Koreans: study of the differences from caucasians in age, gender and seasonal variations

BACKGROUND: Human skin colour shows variations throughout life, and many extrinsic and intrinsic factors influence melanogenesis. Facultative pigmentation of sun-exposed skin has been suggested to reflect cumulative lifetime ultraviolet (UV) exposure in caucasians. However, pigmentary changes due to various regulatory factors may be different in dark-skinned peoples. OBJECTIVES: To observe the variations in skin colour due to ageing, gender differences and seasonal changes in Koreans with skin type IV or V. METHODS: Skin pigmentation was measured at five body sites (buttock, glabella, the V of the neck area, inner arm and dorsal forearm) using skin reflectance spectroscopy in 497 subjects (age range 0-87 years) in winter and 311 subjects (age range 0-84 years) in summer. Among these subjects, 110 were assessed in both seasons. Three independent measurements at each site were done and the average value was used as the pigmentation level. RESULTS: Constitutive pigmentation of the buttock was highest in the first decade of life. It then decreased during the second decade and this decreased level was maintained after the third decade. In contrast to caucasians, facultative pigmentation and sun exposure index did not increase with ageing. Gender differences were significant at all body sites after the first decade. Seasonal changes were apparent in dorsal forearm pigmentation. Little difference was seen in forehead pigmentation between summer and winter. CONCLUSIONS: Basal melanogenic regulation might not be different between Asians and caucasians. However, the sun exposure index may not represent lifelong cumulative UV exposure in Koreans. Age-, gender- and season-related characteristics of skin pigmentation in Koreans imply that genetically determined basal skin colour plays an important part in characterizing later responsiveness to UV radiation and sex hormones. Understanding differences between races will be helpful in studying the regulatory mechanisms of melanogenesis.     

Skin ageing

Similar to the entire organism, skin is subject to an unpreventable intrinsic ageing process. Additionally, skin ageing is also influenced by exogenous factors. Ultraviolet radiation in particular results in premature skin ageing, also referred to as extrinsic skin ageing or photoageing, which is the main cause of the changes associated with the ageing process in sun‐exposed areas. Despite their morphological and pathophysiological differences, intrinsic and extrinsic ageing share several molecular similarities. The formation of reactive oxygen species and the induction of matrix metalloproteinases reflect the central aspects of skin ageing. Accumulation of fragmented collagen fibrils prevents neocollagenesis and accounts for the further degradation of the extracellular matrix by means of positive feedback regulation. The importance of extrinsic factors in skin ageing and the detection of its mechanisms have furthered the development of various therapeutic and preventive strategies.     

Effects of ageing on dermal echogenicity

Background/aims: Changes in the dermis associated with ageing can be detected by high-frequency skin ultrasonography. In photoaged skin, this technique shows a subepidermal low echogenic band (SLEB) that is probably an ultrasound manifestation of elastosis and oedema in the papillary dermis. Since some authors found an association between age and SLEB thickness or its echogenicity on exposed sites, it has been proposed to use these parameters to quantify skin photoageing.
Methods: To determine whether SLEB can be used as a quantitative marker of ageing, its prevalence was determined on forearm skin in a group of 55 individuals (age 18–57 years). The size of SLEB has been measured by quantifying the number of low echogenic pixels in the subepidermal area, which is an accurate method for assessing SLEB severity.
Results: The prevalence of SLEB increased with age, but SLEB was also present in young subjects. The echogenicity of the subepidermal area did not show any age dependence. However, when a ratio of echogenicity between upper and lower dermis was calculated, a linear dependence on age was found.
Conclusions: This study indicates that skin echogenicity measured as a ratio between the upper and lower dermis may be used to objectively estimate photoageing.     

Photoageing: mechanism, prevention and therapy

Photoageing is the superposition of chronic ultraviolet (UV)-induced damage on intrinsic ageing and accounts for most age-associated changes in skin appearance. It is triggered by receptor-initiated signalling, mitochondrial damage, protein oxidation and telomere-based DNA damage responses. Photodamaged skin displays variable epidermal thickness, dermal elastosis, decreased/fragmented collagen, increased matrix-degrading metalloproteinases, inflammatory infiltrates and vessel ectasia. The development of cosmetically pleasing sunscreens that protect against both UVA and UVB irradiation as well as products such as tretinoin that antagonize the UV signalling pathways leading to photoageing are major steps forward in preventing and reversing photoageing. Improved understanding of the skin's innate UV protective mechanisms has also given rise to several novel treatment concepts that promise to revolutionize this field within the coming decade. Such advances should not only allow for the improved appearance of skin in middle age and beyond, but also greatly reduce the accompanying burden of skin cancer.     

Aspects of cutaneous ageing

'Ageing is a multistep, multifaceted, time-dependent phenomenon characterized by the decreased ability of a system to respond to exogenous and endogenous stress from either physical, chemical or biologic agents'. Cutaneous ageing provides a visible model of the interaction between endogenous (intrinsic) factors and exogenous (extrinsic) factors. In skin, the principal extrinsic-factor is ultraviolet light (UV) which is responsible for the constellation of changes termed photoageing. In recent years, much interest has been directed towards defining the ageing processes in skin and excellent comprehensive reviews have been compiled. This review aims to highlight several areas of developing knowledge, and focuses on the potential importance of environmental changes as they influence skin ageing and carcinogenesis. Repeated reference to the effects of UV on the skin are inevitable in any review of skin ageing and this is scarcely surprising as the skin contains many cells as well as subcellular and extracellular chromophores which are capable of absorbing energy within the UV spectrum. Cellular chromophores include among others keratinocytes, melanocytes, Langerhans cells, dermal fibroblasts and mast cells. Subcellular chromophores include keratin, melanin, collagen, elastin and a number of proteins, lipids and steroids (such as vitamin D). Urocanic acid, a photoisomerization product of the amino-acid histidine, may provide some limited photoprotection and some believe it to be important in UV induced immunosuppression. Understanding events at the molecular and biochemical level has unfortunately not been paralleled by clinical advances and the common, troublesome skin-problems of old age such as cancer, xerosis and pruritus remain a major cause of morbidity and yet are poorly explained.     

The SCINEXA: A novel,validated score to simultaneously assess and differentiate between intrinsic and extrinsic skin ageing

BackgroundStudies on the pathogenesis of skin ageing as well as efficacy testing of cosmetic and aesthetic measures to prevent or reverse skin ageing require – as an easy to use method – a validated non-invasive clinical score, which allows to simultaneously assess and differentiate between intrinsic (=chronological) and extrinsic (=photo-) skin ageing. Such an ideal score, however, does currently not exist.ObjectivesWe developed a novel skin ageing score ‘SCINEXA’ comprising 5 items indicative of intrinsic and 18 items highly characteristic of extrinsic skin ageing. These items were used to define an index (index_discr) that allowed differentiating between intrinsic versus extrinsic skin ageing. In order to validate the ‘SCINEXA’, we asked whether it can be used to discriminate regular sunbed users, which have been chronically exposed to ultraviolet radiation and thus are prone to photoageing, from non-sunbed users, which were considered paradigmatic for intrinsic skin ageing.MethodsFor this purpose, 58 non-sunbed users and 16 regularly sunbed users were assessed. In addition to the clinical examination of the 23 score items potential confounders were considered by questionnaire.ResultsBy employing the index_discr, we were able to classify 92% of all study subjects correctly as sunbed or non-sunbed users. Specifically, an index above 2 was associated with sunbed use and thus extrinsic skin ageing, whereas an index below 2 indicated intrinsic skin ageing.ConclusionThe novel ‘SCINEXA’ is suitable for the simultaneous assessment of intrinsic and extrinsic skin ageing.     

Increased expression of TRPV1 channel in intrinsically aged and photoaged human skin in vivo

Abstract:  Transient receptor potential vanilloid type 1 (TRPV1) is activated by various stimuli including capsaicin, heat and acid. While TRPV1 has been localized in the epidermis, little is known about the physiological role of TRPV1 in the skin, especially in skin ageing. In this study, we investigated the effect of acute UV irradiation on TRPV1 expression in human skin and the changes in TRPV1 mRNA and protein in intrinsic ageing and photoageing using human sun-protected (upper inner arm) and sun-exposed (forearm) skin of young and elderly subjects.
Western blot analysis of UV-irradiated young buttock skin revealed that the expression of TRPV1 protein was increased at 24 h (2.3-fold) and 48 h (2.4-fold) after UV irradiation. Real-time PCR analysis also showed that the mRNA level of TRPV1 was augmented by 2.4-fold at 4 h after UV irradiation. TRPV1 protein was expressed at higher levels by 2.6-fold in the sun-protected skin of the elderly subjects than in that of young people according to western blotting, real-time PCR analysis and immunohistochemical staining. In addition, the photoaged skin of elderly showed increased expression of TRPV1 mRNA and protein compared with that of the sun-protected skin of the same individuals. Also, we found increased expression of TRPV1 in nerve fibres of elderly persons using double staining of TRPV1 and nerve fibres.
Based on the above results, our data suggest that the expression of TRPV1 is affected by both the intrinsic ageing and photoageing processes.     

In vivo quantification of epidermis pigmentation and dermis papilla density with reflectance confocal microscopy: variations with age and skin phototype

Reflectance confocal microscopy (RCM) may help to quantify variations of skin pigmentation induced by different stimuli such as UV radiation or therapeutic intervention. The objective of our work was to identify RCM parameters able to quantify in vivo dermis papilla density and epidermis pigmentation potentially applicable in clinical studies. The study included 111 healthy female volunteers with phototypes I-VI. Photo-exposed and photo-protected anatomical sites were imaged. The effect of age was also assessed. Four epidermis components were specifically investigated: stratum corneum, stratum spinosum, basal epidermal layer and dermo-epidermal junction. Laser power, diameter of corneocytes and upper spinous keratinocytes, brightness of upper spinous and interpapillary spinous keratinocytes, number of dermal papillae and papillary contrast were systematically assessed. Papillary contrast measured at the dermo-epidermal junction appeared to be a reliable marker of epidermis pigmentation and showed a strong correlation with skin pigmentation assessed clinically using the Fitzpatrick's classification. Brightness of upper spinous and interpapillary spinous keratinocytes was not influenced by the skin phototype. The number of dermal papillae was significantly lower in subjects with phototypes I-II as compared with darker skin subjects. A dramatic reduction in the number of dermal papillae was noticed with age, particularly in subjects with fair skin. The method presented here provides a new in vivo investigation tool for quantification of dermis papilla density and epidermal pigmentation. Papillary contrast measured at the dermo-epidermal junction may be selected as a marker of skin pigmentation for evaluation in clinical studies.     

Distribution and expression of type VI collagen in photoaged skin

BACKGROUND: Several of the characteristic clinical features of photoaged skin, including wrinkling, are thought to be dependent on changes in the dermal matrix brought about by chronic sun exposure. Such changes include reductions in collagens I, III and VII, an increase in elastotic material in the reticular dermis and a marked reduction in the microfibrillar glycoprotein fibrillin. OBJECTIVES: To examine whether type VI collagen, a microfibrillar collagen necessary for cell-cell and cell-matrix communication, is affected by the photoageing process. METHODS: Six healthy volunteers with moderate to severe photoageing were enrolled into the study. Immunohistochemistry and in situ hybridization histochemistry were used to examine the levels of type VI collagen in photoprotected and photoaged sites. RESULTS: In photoprotected skin, type VI collagen was concentrated in the papillary dermis immediately below the dermal-epidermal junction, around blood vessels, hair follicles and glandular structures. The distribution of type VI collagen was unchanged in photoaged skin, although we observed an increase in the abundance of the alpha3 chain of collagen VI in the upper papillary dermis, at its junction with the dermal-epidermal junction (P < 0.05). No alterations were observed for any alpha chain at the mRNA level. CONCLUSIONS: These studies suggest that chronic sun exposure (photoageing) has little or no effect on either the distribution, abundance or levels of expression of type VI collagen in human skin. Thus, type VI collagen, unlike other matrix components so far studied, appears to be relatively unaffected by the photoageing process.     

Noninvasive methods for the assessment of photoageing

Although histopathological dermal elastosis is the current gold standard for the diagnosis of photoageing, noninvasive methods for quantifying the amount of photodamage to skin are clearly preferable. This study is the first to survey five noninvasive methods of assessing photoageing (clinical examination, spectrophotometry, skin surface topography, reflectance confocal microscopy and fluorescence lifetime imaging microscopy) in the same individual. Measurements for each noninvasive method were compared across nine individuals from three participant groups (‘younger’, ‘older’ and ‘photodamaged’) in UV‐protected volar and UV‐exposed dorsal forearm skin. Overall, participants in the younger group had the lowest measures of photodamage, while those in the photodamaged group had the highest, as indicated by each modality. The five noninvasive strategies surveyed in this study may demonstrate potential as a suitable methodology for the quantification of photoageing. The advantage of such noninvasive methods is that they allow for skin visualisation in vivo and repeated assessments of the same site. The main limitation of this study was its small sample size, which may have precluded many findings of statistical significance.     

Computational characterization of reflectance confocal microscopy features reveals potential for automated photoageing assessment

Skin photoageing results from a combination of factors including ultraviolet (sun) exposure, leading to significant changes in skin morphology and composition. Conventional methods assessing the degree of photoageing, in particular histopathological assessment involve an invasive multistep process. Advances in microscopy have enabled a shift towards non‐invasive in vivo microscopy techniques such as reflectance confocal microscopy (RCM) in this context. Computational image analysis of RCM images has the potential to be of use in the non‐invasive assessment of photoageing. In this report, we computationally characterized a clinical RCM data set from younger and older Caucasians with varying levels of photoageing. We identified several mathematical relationships that related to the degree of photoageing as assessed by conventional scoring approaches (clinical photography, SCINEXA and RCM). Furthermore, by combining the mathematical features into a single computational assessment score, we observed significant correlations with conventional RCM (P < 0.0001) and the other clinical assessment techniques.     

The effect of ultraviolet radiation exposure on the prevalence of mast cells in human skin

BACKGROUND: Dermal mast cells have been implicated as important effector cells in innate immunity, hypersensitivity responses and ultraviolet (UV)B-induced suppression of cell-mediated immune responses to contact allergens. Humans, like mouse strains, display variations in dermal mast cell prevalence. The factors determining these differences are yet to be fully elucidated. In mice, expression of the receptor for stem cell factor, c-kit, on dermal mast cells correlates with prevalence. OBJECTIVES: To evaluate dermal mast cell prevalence and mast cell c-kit expression in non-sun-exposed and sun-exposed skin in the same donor. METHODS: In 14 subjects, biopsies of skin (4 mm) were sampled from the skin sites of buttock, inner arm, shoulder and back of hand skin and dermal mast cell prevalence quantified. Non-sun-exposed buttock and chronically sun-exposed hand skin were evaluated for mast cell expression of c-kit and elastin content, a feature of photoageing and surrogate marker of UV exposure. RESULTS: The prevalence of dermal mast cells was significantly higher in hand skin than in the three other anatomically different skin sites. Significant correlations were observed in hand but not buttock skin between increasing dermal mast cell densities, extent of elastin content in the papillary dermis and age of the subject. Cellular expression of c-kit correlated with mast cell prevalence in hand skin. However, no relationship was observed in hand skin between c-kit expression, elastin content and age. CONCLUSIONS: The prevalence of mast cells in human skin is altered by factors that are intrinsic (mechanisms regulating c-kit expression) and extrinsic (chronic sun exposure), and the fact that the associations of mast cell prevalence with age is explained by the latter being a correlate of cumulative sun exposure.     

Chronic sun exposure alters both the content and distribution of dermal glycosaminoglycans

Summary Chronic sun exposure leads to structural and functional alterations in exposed skin. Photoageing is a process distinct from the changes taking place due to chronological ageing. Unique alterations in the dermal extracellular matrix occur as a result of photoageing and are responsible for many of these physiological changes taking place in sun-damaged skin. Accompanying the deposition of abnormal elastic tissue, or solar elastosis, are significant alterations in dermal glycosaminoglycans (GAGs). Accumulation of GAGs as a result of photoageing as demonstrated in both humans and animal models of photoageing seems almost paradoxical in view of the large amounts of GAGs present in the skin of newborns, making their skin well hydrated and supple, in sharp contrast to the weathered appearance of photoaged skin. We investigate the relative GAG content of photoaged skin using immunoperoxidase stains specific for hyaluronic acid and chondroitin sulphate, and determine the location of these GAGs using confocal laser scanning microscopy. Our results demonstrate significant increases in GAG staining in sun-damaged vs. sun-protected skin from the same individuals, as measured by computer-based image analysis. Furthermore, confocal laser scanning microscopy reveals that the increased dermal GAGs in sun-damaged skin are deposited on the elastotic material of the superficial dermis of photodamaged skin, and not between collagen and elastic fibres as in normal skin. The abnormal location of GAGs on these fibres may explain the apparent paradoxical weathered appearance of photodamaged skin despite increased GAGs.     

Mechanisms of photodamage of the skin and its functional consequences for skin ageing

Chronic photodamage of the skin manifests itself as extrinsic skin ageing (photoageing) and photocarcinogenesis. DNA photodamage and UV-generated reactive oxygen species are the initial molecular events that lead to most of the typical histological and clinical manifestations of chronic photodamage of the skin. Knowledge of the UV-absorbing chromophores in the skin and of the molecular mechanisms leading to the unwanted effects of sun exposure provide a basis for the development of novel strategies for the prevention and repair of photoageing. This review provides an overview of the photochemistry of the major skin chromophores and their relationship to chronic photodamage.