Characterisation and Assay of Five Enzymatic Activities in the Stratum Corneum Using Tape-Strippings

Redoulès, Daniel; Tarroux, R.; Assalit, M.F.; Périé, Jacques

doi:10.1159/000066242

Cited by 61 publications

(68 citation statements)

References 26 publications

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“…The acidic pH is also required for normal permeability barrier function 19,8 , and several other key SC functions. For example, the activity of acidic, lysosomal type, lipid processing enzymes is required to fully establish the permeability barrier 13,12,16,36 , and an acidic pH controls the activity of proteases necessary for SC integrity and desquamation 9,38,33,17 . We recently demonstrated that the Sodium-Hydrogen-Exchanger 1 (NHE1) plays a key role in establishing this critical acidic SC milieu 2 .…”

Section: Introductionmentioning

confidence: 99%

Fluorescence lifetime to image epidermal ionic concentrations

Behne

Barry

Moll³

et al. 2004

SPIE Proceedings

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Measurements of ionic concentrations in skin have traditionally been performed with an array of methods which either did not reveal detailed localization information, or only provided qualitative, not quantitative information. FLIM combines a number of advantages into a method ideally suited to visualize concentrations of ions such as H + in intact, unperturbed epidermis and stratum corneum (SC). Fluorescence lifetime is dye concentration-independent, the method requires only low light intensities and is therefore not prone to photobleaching or phototoxic artifacts, and because multiphoton lasers of IR wavelength are used, light penetrates deep into intact tissue. The standard method to measure SC pH is the flat pH electrode, which provides reliable information only about surface pH changes, without further vertical or subcellular spatial resolution; i.e., specific microdomains such as the corneocyte interstices are not resolved, and the deeper SC is inaccessible without resorting to inherently disruptive stripping methods. Furthermore, the concept of a gradient of pH through the SC stems from such stripping experiments, but other confirmation for this concept is lacking. Our investigations into the SC pH distribution so far have revealed the crucial role of the Sodium/Hydrogen Antiporter NHE1 in generation of SC acidity, the colocalization of enzymatic lipid processing activity in the SC with acidic domains of the SC, and the timing and localization of emerging acidity in the SC of newborns. Together, these results have led to an improved understanding of the SC pH, its distribution, origin, and regulation. Future uses for this method include measurements of other ions important for epidermal processes, such as Ca 2+ , and a quantitative approach to topical drug penetration.

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Section: Introductionmentioning

confidence: 99%

Fluorescence lifetime to image epidermal ionic concentrations

Behne

Barry

Moll³

et al. 2004

SPIE Proceedings

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“…In previous studies [3][4] we described and analyzed the enzymes involved in generating this protective envelope and showed that in dry, non-eczematous atopic dermatitis, the phosphatase-like and trypsin-like activities are altered. The present study pursues these investigations in the same type of pathology but in cases involving an inflammatory component.…”

Section: Introductionmentioning

confidence: 99%

Variability of Enzyme Markers during Clinical Regression of Atopic Dermatitis

Tarroux

Assalit²,

Licu³

et al. 2001

Skin Pharmacol Physiol

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Skin surface enzyme activities were found to be significantly different in healthy and in skin with atopic dermatitis and, following appropriate treatment, a close correlation was observed between the clinical staging of the atopic dermatitis and the levels of the assayed marker enzymes. Samples were taken, by stripping with simple adhesive tapes, from a group of subjects on cure in a spa. The corneocytes were recovered from the first layers of the stratum corneum. Aqueous extracts of the strips were tested for their activity on chromophoric substrates which allow fluorescence spectrometry to be used to assay the trypsin-like, acid-phosphatase-like and phospholipase-A2-like activities. We show that the restoration of return to activities close to those of healthy subjects is related to the general condition of the patients, who showed a clearly improved SCORAD. Recovery of the trypsin-like activity and attenuation of the phospholipase-like activity, paralleled the regression of the dermatitis as assessed by a decrease in clinically evaluated parameters of xerosis and inflammation.

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“…And the PTD group compared with the PSD group showed more irregular granular keratin in granular cells and immature keratin formation. It is well documented that ACP activity is increased in damaged tissues, [44][45][46] and ACP activity in PTD group was significantly higher than that of PSD group. Therefore, these results suggest that PPD skin toxicity in a tungstate supplemented diet is enhanced.…”

Section: Discussionmentioning

confidence: 84%

The Effects of Tungstate on Skin Lesions Caused by PPD in Rats

Lee¹,

Cho

S³

2008

Biological & Pharmaceutical Bulletin

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Red brownish p-phenylenediamine (PPD) has been widely used as one of the ingredients in hair dyes. The use by consumers of hair dye results in exposure of the hair, scalp skin, eyes, nail, and hands to PPD. PPD studies have found that women using dye with a formulation that included PPD suffered acute sensitive responses such as headaches, dizziness, and systemic anaphylaxis for the long time after using the product.1) And in patch tests, PPD caused erythema, rash, and a burning sensation on human skin. Furthermore, workers occupationally exposed to PPD in the field suffered from the bladder cancer.2,3) Previous studies have shown that PPD application induces injury to rat skin. 4,5) It is reported that PPD is metabolized to N-acetylated adducts as the major metabolites by N-acetyltransferase in human skin and liver, 6) and rats. 7) Moreover, it is reported that N-oxidation of arylamines like PPD occurs in the liver and is generally considered to involve cytochrome P450. 8) Furthermore, N-acetylated PPD metabolites are less toxic and they are excreted in urine.9) And, lipid peroxidation was increased while glutathione was decreased in skin by PPD application.5,10) In addition, PPD toxicity is increased by MnSOD polymorphism 11) while PPD toxicity is alleviated by thiol antioxidants.12) These reports suggest that production of ROS can be related to PPD skin toxicity. However, there are ongoing controversies whether PPD induced organ toxicity or whether it is safe in animal and clinical experiment. 13,14) It is well known that xenobiotic-induced tissue damage is related to reactive metabolites as well as produced reactive oxygen species (ROS) such as superoxide, hydroxyl radical, single oxygen and hydrogen peroxide under the metabolism. 15,16) Many studies have reported an imbalance between ROS generating systems and scavenging systems such as the increasing of ROS generating enzymes and/or decreasing of scavenging enzymes and nonenzymatic antioxidants. 17,18) It is accepted that xanthine oxidase (XO), one of the ROS generating systems, is increased in the injured cells.19) It has been revealed that inhibition of XO activity by allopurinol 20) or tungstate 21) alleviates experimental tissue damage. This indicates therefore a relationship of XO to skin cell injury, which suggests that the XO plays a role in the toxicity by the exposure of xenobiotics such as cosmetics and chemicals due to relatively higher XO activity in skin tissue compared with other tissues except liver and intestine. 19,22) It is reported that oxidized PPD, imine derivatives by hydrogen peroxide 23) could induce malignant and benign tumors in the mammary gland, uterus, and soft tissues of rats.24) Moreover, it is reported that XO could generate ROS such as superoxide and hydrogen peroxide.25,26) So, we hypothesized hydrogen peroxide derived from the XO system in skin involved with PPD oxidation and produced oxidized PPD is associated with dermal tissue damage.It is well established that sodium tungstate has relatively lower toxicity than that o...

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Characterisation and Assay of Five Enzymatic Activities in the Stratum Corneum Using Tape-Strippings

Cited by 61 publications

References 26 publications

Fluorescence lifetime to image epidermal ionic concentrations

Fluorescence lifetime to image epidermal ionic concentrations

Variability of Enzyme Markers during Clinical Regression of Atopic Dermatitis

The Effects of Tungstate on Skin Lesions Caused by PPD in Rats

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