Application of cytogenetic model Allium cepa for screening potential cytogenotoxicity of herbal-based hair dyes.

Plant models may be useful as test organisms for initial screening of potential toxicity of personal care products. The objective of the present study was to assess the efficacy of the Allium cepa (common onion) test system as a bioanalytical tool for screening potential cytotoxicity and genotoxicity of herbal-based hair dye formulations. Exposure of black hair dye formulations for 48 hours resulted in root growth retardation and mitosis suppression in the root meristems of A. cepa bulbs indicating concentration dependent cytotoxicity. At the 72 hour post exposure, cytotoxic effects on the roots were reduced but not recovered completely signifying prolong toxic action of the hair dyes. The condensed nuclei was the most frequent nuclear abnormality found in the dye exposed root meristematic cells indicating the cell death process. Induction of micronuclei and chromosomal aberrations in the root meristematic cells even at the post exposure stage indicates persistent genotoxicity of the hair dyes which may be attributed to the interactive effects of chemical mixtures present in the commercial hair dye formulations. The results revealed that A. cepa test system is an effective bioanalytical tool for screening cytogenotoxicity of commercial hair dye formulations.

Scanning electron microscopy approach for evaluation of hair dyed with Lawsonia inermis powder: in vitro Study / Método de microscopía electrónica de barrido para la evaluación de cabello teñido con extracto de Lawsonia inermis: estudio in vitro

During aging, usually graying of the hair occurs as a result of oxidative stress. Driven by social acceptance and self-perception of the exterior appearance, both men and women rely on hair dyeing products, in order to mask the graying hair. At the same time, a frequent use of synthetic products and treatment can damage the hair shaft; for this reason, this study aimed to evaluate the morphological effect of the herbal dye derived from Lawsonia inermis (henna), on hair. Dyed hairs were evaluated by means of SEM. Subsequently, they were compared, qualitatively and quantitatively, with undyed hairs. Results showed a positive impact on the cuticula pattern and on the diameters of the examined samples, after henna application. Different results, about the degree and the type of morphological changes occurring on pigmented hairs, may depend on the phenotype and on the health condition of hair, before dye treatment.

Durante el envejecimiento, generalmente se produce el envejecimiento del cabello como resultado del estrés oxidativo. Motivados por la aceptación social y la autopercepción de la apariencia, tanto hombres como mujeres confían en productos para teñir el cabello para enmascarar las canas. Al mismo tiempo, el uso frecuente de productos y tratamientos sintéticos puede dañar el tallo del cabello. Por esta razón, este estudio tuvo como objetivo evaluar el efecto morfológico del tinte derivado de Lawsonia inermis (henna) en el cabello. Los cabellos teñidos se evaluaron mediante SEM. Posteriormente, se compararon, cualitativa y cuantitativamente, con cabellos sin teñir. Los resultados mostraron un impacto positivo en el patrón de la cutícula y en los diámetros de las muestras examinadas, después de la aplicación de henna. Los diferentes resultados, sobre el grado y el tipo de cambios morfológicos que ocurren en los cabellos pigmentados, pueden depender del fenotipo y del estado de salud del cabello, antes del tratamiento con tinte.

Investigation on the 2D-Distribution of Metallic Elements after Hair Dyeing.

Long-term use of hair dyes has potential effects on metal content in hair. However, little research dissects the specific distribution and composition variations of the metal after dyeing. In this study, we investigated the morphological change and metallic elements content variation after dyeing. The results showed that the concentration of essential metal elements decreased, among which the Ca, K, and Na decreased sharply even above 50%. As for the heavy metal, the most significant observation is that Pb increased almost by five times after dyeing. Besides, it revealed, using scanning electron microscope coupled with energy-dispersive X-ray spectroscopy (SEM-EDS), that Pb concentrated at the outer layer of the hair. In addition, two-dimensional proton-induced X-ray emission (2D-PIXE) was applied to analyze the distribution of metallic elements along the longitudinal and cross section of the hair. The results showed that Ca and Zn distributed evenly in the hair along the longitudinal and cross section. It is the first time that 2D-PIXE is applied to analyze the metallic distribution in the hair. This method exhibits high sensitivity and can be widely used in the environmental and medical field to analyze the distribution of metallic elements.

The effects of para-phenylenediamine (PPD) on the skin fibroblast cells.

1. Para-phenylenediamine (PPD) is the commonest and most well-known component of hair dyes. PPD is found in more than 1000 hair dye formulations and is the most frequently used permanent hair dye component in Europe, North America and East Asia. PPD containing hair dyes have been associated with cancer and mutagenicity. Apart from that, PPD has potential toxicity which includes acute toxicity such as allergic contact dermatitis and subacute toxicity. 2. In this study, we examined the effects of the PPD composition on the skin-isolated fibroblast cells. Fibroblast cells were isolated from the skin and cell viability, reactive oxygen species (ROS) production, the collapse of mitochondrial membrane potential (MMP), lipid peroxidation (LPO), damage to the lysosome release of lactate dehydrogenase (LDH) and finally release of cytochrome c were examined following the exposure to various concentrations of PPD. 3. Our results showed that exposure to PPD increased ROS generation, LPO, the collapse of MMP, LDH release and cytochrome c release. Our results suggest that PPD can induce damage to the lysosomal membrane. 4. These results showed that PPD composition has a selective toxicity on skin fibroblasts cell and mitochondria are considered one of the goals of its toxicity.

The role of chelating agents and amino acids in preventing free radical formation in bleaching systems.

The control of bleaching reaction is important in hair bleaching and laundry detergents to ensure quality of the final product. A better understanding of the reaction mechanisms is needed to minimize product failures. 31P NMR-spectroscopy-based spin trap technique was employed to detect and quantify the free radical species that were generated in different bleaching solutions. These solutions contained the key actives in an alkaline hair colorant/bleaching product, an ammonium salt and hydrogen peroxide at pH = 10. Generally, the main radical species detected in hair oxidative coloring or bleaching processes, were hydroperoxyl/superoxide radicals HO2·/O2.-, amino radicals ·NH2 and hydroxyl radicals ·OH. Their amounts showed a variation based on the chemical composition of the bleaching systems and the metal ion content. The generation of free radicals from reactions between transition metal ions, such as copper, and hydrogen peroxide at pH = 10 was evaluated. In the absence of chelating agents, the copper ions generated a significant level of hydroxyl radicals in a Fenton-like reaction with hydrogen peroxide at pH = 10. Besides that, an increase in copper ion content led to an increase of amino radical ·NH2, whereas the concentration of superoxide radical O2·- decreased which was not yet well reported in the previous literature. The effect of chelating agents like ethylenediaminetetraacetic acid (EDTA), tetrasodium-iminodisuccinate (IDS), a mixture of basic amino acids and dicarboxylic acid on free radical formation was investigated in the presence of binary Cu2+-Ca2+ bleaching systems. As expected, in the binary Cu2+-Ca2+ ion system EDTA did not suppress hydroxyl radical formation effectively, but the mixture containing sodium succinate, lysine and arginine reduced hydroxyl radical formation, whereas IDS (nearly) completely inhibited hydroxyl radical formation. The results indicated that each bleaching solution has its characteristic performance and damage profile. Whereas the reactivity can be controlled by the usage of chelating agents.

The Late Stages of Melanogenesis: Exploring the Chemical Facets and the Application Opportunities.

In the last decade, the late stages of melanin biosynthesis involving the oxidative polymerization of 5,6-dihydroxyindole (DHI) and 5,6-dihydroxyindole-2-carboxylic acid (DHICA) have been extensively investigated. Most of the information derived from a biomimetic approach in which the oxidation of melanogenic indoles was carried out under conditions mimicking those occurring in the biological environment. Characterization of the early oligomers allowed for drawing a structural picture of DHI and DHICA melanins, providing also an interpretative basis for the different properties exhibited by these pigments, e.g., the chromophore and the antioxidant ability. The improved knowledge has opened new perspectives toward the exploitation of the unique chemistry of melanins and its precursors in cosmetic and health care applications. A noticeable example is the development of an innovative hair dyeing system that is based on the marked ease of DHI to give rise to black melanin on air oxidation under slightly alkaline conditions. The advantage of this method for a step-wise coverage of gray hair with a natural shade pigmentation on repeated treatment with a DHI-based formulation with respect to traditional dyes is presented. A variant of DHICA melanin combining solubility in water-miscible organic solvents, an intense chromophore in the UltraViolet-A UV-A region, and a marked antioxidant potency was evaluated as an ingredient for cosmetic formulations.

Protective effect of conditioner agents on hair treated with oxidative hair dye.

BACKGROUND: Hair coloring is broadly used by women and men either to change their natural hair color or to delay the onset of gray hair. Oxidative dyes may damage the hair, as chemical and physical processes are required to convert the fiber structure and, consequently, alterations in its mechanical and surface properties. OBJECTIVES: The aim of this study was to evaluate the protective effect of silanetriol (and) Panthenol, PEG-12 dimethicone, and hydrolyzed silk (and) hydrolyzed milk protein (and) lactose as conditioner agents on hair treated with oxidative hair dye by protein loss, combability, and breaking strength. METHODS: In this research, we analyzed the untreated hair (sample I) and the effect of oxidative hair dye emulsions, with or without conditioner agents (sample II) silanetriol (and) Panthenol (sample III), PEG-12 dimethicone (sample IV), and hydrolyzed silk (and) hydrolyzed milk protein (and) lactose (sample V) on Caucasian hair. The hair samples were submitted to protein loss quantification, breaking strength, and combing analysis. RESULTS: For protein loss, the results were: IIa  = Va  > IVb  > IIIc  > Id . For the breaking strength: Ie  = IIe  = IIIe  = IVe  = Ve . For the combing analysis for wet and dry hair, the results were, respectively: IIa  > IIIb  = IVb  > Vc  > Id and IIA  > IIIb  = Vb  > IV c = Ic . Data classified by different letters presented statistically significant alterations, α = 5, P ≤ . 05, n = 15. CONCLUSIONS: Based on these results, the incorporation of conditioner agents into emulsion blond color decreased the damage caused by the coloring process.

Influence of bleaching and coloring on ethyl glucuronide content in human hair.

Ethyl glucuronide (EtG) is increasingly used in forensic toxicology as a marker for alcohol use in analyses of hair samples, especially in abstinence control. Some cosmetic treatments are considered to markedly reduce the EtG content. In view of especially many women with coloured hair the present study was performed to further investigate the effect of a variety of colouring procedures (bleaching, tinting, permanent and semi-permanent dyeing, henna) on the EtG content. Untreated hair samples (n = 12, EtG 13.9-64.7 pg/mg) were re-analyzed (gas chromatography- negative chemical ionization mass spectrometry, 0.8 pg/mg quantification limit) after different treatment procedures. A decrease of the EtG content of at least 10% occurred in every case. The reduction in comparison to the untreated hair was expectedly high for permanent dyeing and bleaching with 18.1% of the initial content (median, range 0.0-50.9%) and 18.4% (0.0-46.7%), respectively. For henna this was 38.3% (0.0-83.0%), for tinting 70.4% (29.0-90.8%), for semi-permanent dyeing 41.9% (0.0-77.4%). With permanent hair dye the EtG content was decreased to below 7 pg/mg in 10 of 12 cases, in 3 cases even below the LOD (0.2 pg/mg). Surprisingly henna treatment without oxidative component had a marked influence, EtG was below 2 pg/mg in 2 of 12 samples. The study showed that all tested coloration procedures markedly affected the deposited EtG content. Even temporary or henna coloration may have a marked effect. The present data support the recommendation to exclude hair samples with colour manipulations for analysis on the EtG content as a precaution in alcohol abstinence programs. Copyright © 2017 John Wiley & Sons, Ltd.

Investigation of hair dye deposition, hair color loss, and hair damage during multiple oxidative dyeing and shampooing cycles.

Color fastness is a major concern for consumers and manufacturers of oxidative hair dye products. Hair dye loss results from multiple wash cycles in which the hair dye is dissolved by water and leaches from the hair shaft. In this study, we carried out a series of measurements to help us better understand the kinetics of the leaching process and pathways associated with its escape from the fiber. Hair dye leaching kinetics was measured by suspending hair in a dissolution apparatus and monitoring the dye concentration in solution (leached dye) with an ultraviolet-visible spectrophotometer. The physical state of dye deposited in hair fibers was evaluated by a reflectance light microscopy technique, based on image stacking, allowing enhanced depth of field imaging. The dye distribution within the fiber was monitored by infrared spectroscopic imaging of hair fiber cross sections. Damage to the ultrafine structure of the hair cuticle (surface, endocuticle, and cell membrane complex) and cortex (cell membrane complex) was determined in hair cross sections and on the hair fiber surface with atomic force microscopy. Using differential scanning calorimetry, we investigated how consecutive coloring and leaching processes affect the internal proteins of hair. Further, to probe the surface properties of hair we utilized contact angle measurements. This study was conducted on both pigmented and nonpigmented hair to gain insight into the influence of melanin on the hair dye deposition and leaching processes. Both types of hair were colored utilizing a commercial oxidative hair dye product based on pyrazole chemistry.

Protein loss in human hair from combination straightening and coloring treatments.

BACKGROUND: Hair chemical treatments, such as dyeing and straightening products, are known to cause damage that can be assessed by protein loss. OBJECTIVES: The aim of this study was to evaluate the hair protein loss caused by combined chemical treatments (dye and relaxer) using the validated bicinchoninic acid (BCA) method. Three kinds of straighteners, based on ammonium thioglycolate, guanidine hydroxide and sodium hydroxide, were evaluated and the least harmful combination indicated. METHODS: Caucasian virgin dark brown hair tresses were treated with developed natural brown color oxidative hair dyeing and/or straightening commercial products based on ammonium thioglycolate, sodium hydroxide, or guanidine hydroxide. Protein loss quantification was assessed by the validated BCA method which has several advantages for quantifying protein loss in chemically treated hair. RESULTS: When both treatments (straightening and dyeing) were combined, a higher negative effect was observed, particularly for dyed hair treated with sodium hydroxide. In this case, a 356% increase in protein loss relative to virgin hair was observed and 208% in relation to only dyed hair. The combination of dying and relaxers based on ammonium thioglycolate or guanidine hydroxide caused a small increase in protein loss, suggesting that these straightening products could be the best alternatives for individuals wishing to combine both treatments. CONCLUSIONS: These results indicated that when application of both types of products is desired, ammonium thioglycolate or guanidine hydroxide should be chosen for the straightening process.

Coloring, bleaching, and perming: influence on EtG content in hair.

BACKGROUND: Hair analysis of ethyl glucuronide (EtG) has become, beside fatty acid ethyl ester, a valuable marker for the detection of moderate and chronic excessive alcohol consumption. So far, only few studies exist about the influence of cosmetic treatment on EtG content in hair. The aim of this study was to evaluate the effect of coloring, bleaching, and perming on the concentration of this alcohol marker in hair. Studies were also performed to evaluate the chemical stability of EtG in the presence of hydrogen peroxide and ammonium thioglycolate. METHODS: Six air samples were treated in vitro by the different commercial cosmetics following the suppliers' instructions. After washing, pulverization, incubation in ultrasonic bath, and solid phase extraction, EtG was determined by GC/MS-NICI after solid phase extraction and heptafluorobutyric anhydride derivatization. RESULTS: The results showed that samples (n = 10) treated with the coloring product did not show any important change in the EtG results. In the bleaching study (n = 23), a mean decrease of 73.5% was observed. After incubation of a solution of EtG with hydrogen peroxide (15%), a decrease of 45% was shown supporting the hypothesis of a chemical degradation of EtG and a leaching out effect from the hair matrix. In the perm treatment study (n = 23), a mean decrease of 95.7% of EtG was found. Incubation of a solution of EtG with ammonium thioglycolate (5%) showed a total decrease of EtG supporting the hypothesis of a chemical degradation. CONCLUSIONS: Coloring treatment did not importantly influence EtG content in hair. However, an important decrease of EtG in hair could be found after bleaching and permanent wave treatment. This decrease seems to be because of a chemical degradation of EtG, after bleaching, and a leaching out effect from the matrix. After perming, it seems to be more of a chemical degradation of EtG. These data have to be considered for the correct interpretation of EtG amounts in hair.

Permanent hair dye-incorporated hyaluronic acid nanoparticles.

We prepared p-phenylenediamine (PDA)-incorporated nanoparticles using hyaluronic acid (HA). PDA-incorporated HA nanoparticles have spherical shapes and sizes were less than 300 nm. The results of FT-IR spectra indicated that PDA-incorporated HA nanoparticles were formed by ion-complex formation between amine group of PDA and carboxyl group of HA. Furthermore, powder-X-ray diffractogram (XRD) measurement showed that intrinsic crystalline peak of PDA disappeared by formation of nanoparticle with HA at XRD measurement. These results indicated that PDA-incorporated HA nanoparticles were formed by ion-complex formation. At drug release study, the higher PDA contents induced faster release rate from nanoparticles. PDA-incorporated nanoparticles showed reduced intrinsic toxicity against HaCaT human keratinocyte cells at MTT assay and apoptosis assay. We suggest that PDA-incorporated HA nanoparticles are promising candidates for novel permanent hair dye.

Preparation and evaluation of semi-permanent hair dyes liposome.

This study used phospholipids from fresh egg yolks to prepare liposome-encapsulated semi-permanent hair dyes in different pH buffer solutions and evaluated the functions and colour fastness to washing of the dyes. The extraction ratio of egg yolk phospholipids was 5%, and the purity was 91.8%. Empty liposome solutions were then prepared using high-speed homogenizer with particle size 219-848 nm. After being stored at 4 °C for 28 days, the average particle size of the liposome-encapsulated dye formulas increased from 1.36-1.92 µm to 1.99-2.38 µm. The ΔE colour difference values of the five hair extension sets dyed with the control group and hair dyes on the market were of the range 6.56-13.39 after eight times of washing, whereas the ΔE values of the four hair extension sets dyed with the liposome-encapsulated dyes were of the range 3.56-5.21 after eight times of washing. The liposome-encapsulated dye at pH 3 showed the best result.

Elicitation of the immune response to p-phenylenediamine in allergic patients: the role of dose and exposure time.

BACKGROUND: Usage of hair dye products containing p-phenylenediamine (PPD) is a concern for PPD-allergic individuals. OBJECTIVES: The present study investigates the role of dose and exposure time on elicitation of allergic contact dermatitis under conditions of permanent hair dyeing. METHODS: Elicitation responses after application of a typical hair dye product containing 2% PPD for 30 min followed by rinsing were analysed in 38 PPD-allergic individuals with a documented history of hair dye-related allergy. Skin binding experiments in vitro were performed to distinguish the dose available for elicitation from the dose applied. RESULTS: A positive reaction was elicited in 20 of 20 patients with grades ++ to +++ and 12 of 18 with grade + according to the classification of the International Contact Dermatitis Research Group. Under conditions of diagnostic patch testing (48 h exposure), the dose available for elicitation is more than 10-fold higher compared with the dose available for hair dyeing (30-min exposure, rinsing of product). CONCLUSIONS: This investigation demonstrates that under simulated hair dye use conditions the actual exposure to PPD is more than an order of magnitude lower than under diagnostic patch testing, although sufficient to elicit a clearly noticeable reaction in 84% of PPD patch test-positive individuals.

Repeated exposure to hair dye induces regulatory T cells in mice.

BACKGROUND: We have recently shown that commercial p-phenylenediamine (PPD)-containing hair dyes are potent immune activators that lead to severe contact hypersensitivity in an animal model. However, only a minority of people exposed to permanent hair dyes develops symptomatic contact hypersensitivity. This suggests that the majority of people exposed to hair dyes does not become sensitized or develop immunological tolerance. OBJECTIVES: To study the immune response in mice repeatedly exposed to PPD-containing hair dye in a consumer-like manner. METHODS: A commercial hair dye containing PPD was tested in C57BL/6 mice. The local immune response was measured by ear swelling and by histological examinations. The immune response in the draining lymph nodes was analysed by flow cytometry. RESULTS: The hair dye induced local inflammation as seen by swelling and cell infiltration of the treated ears. In addition, exposure to hair dye caused T-cell activation as seen by T-cell proliferation and production of interferon-γ and interleukin (IL)-17 within the draining lymph nodes. The inflammatory response peaked at the fourth exposure to hair dye. From this point on, an upregulation of regulatory T cells and IL-10-producing cells was seen. CONCLUSIONS: This study shows that PPD-containing hair dyes strongly affect the immune system. In addition to being potent skin sensitizers that activate inflammatory T cells, hair dyes also induce anti-inflammatory mechanisms. This might explain why many consumers can use hair dyes repeatedly without developing noticeable allergies, but it also raises the question whether the immune modulatory effects of hair dyes might influence the development of autoimmune diseases and cancers.

A novel LCD (coal tar) solution for psoriasis does not discolor naturally light or color-processed hair in an exaggerated exposure test model.

BACKGROUND: Scalp psoriasis is reported to occur in 50-80% of psoriasis sufferers. Treatment of scalp psoriasis requires special consideration of product esthetics and staining potential due to the presence of hair. AIM: To evaluate the potential of a new, marketed liquor carbonis distillate (LCD; coal tar) solution to discolor naturally light or color-processed hair under exaggerated exposure conditions. METHODS: Samples of naturally light and color-processed hair from a single donor were exposed to LCD solution repeatedly over 14 days and via submergence for 24 h. Color of LCD-treated hair samples was compared with untreated control hair samples. RESULTS: LCD solution did not discolor naturally light or color-processed hair following repeated exposures and 24 h submergence. CONCLUSION: The marketed LCD solution does not appear to discolor naturally light or color-processed hair.

A new oxidant for hair coloring.

Coloring hair using a level 3 permanent colorant involves two processes, lightening the underlying melanin and information of the colored chromophores inside the hair. In a typical in-market products the oxidant used to achieve these changes is hydrogen peroxide buffered at pH 10 with an alkalizer such as ammonium hydroxide. A new oxidant has been developed based on the combination of ammonium carbonate, hydrogen peroxide and glycine at pH 9 that can match the lightening and color performance of the current oxidant. It has the advantage that both the carbonate and hydrogen peroxide concentrations can be changed to alter the lightening performance making it a more flexible oxidant. This allows the capability to lighten the hair in a shorter time, or with lower hydrogen peroxide levels. This paper discusses the key oxidizing species that are present in both systems and the mechanisms of melanin lightening. In addition, the lightening performance will be assessed as a function of time, pH, hydrogen peroxide concentration and carbonate concentration. The importance of glycine to the oxidant is also described along with a proposal for its mechanism of action. It has been demonstrated that the addition of glycine can control the undesired formation of carbonate radicals that can be generated from the oxidant. The control of these radicals enables the oxidant to deliver excellent lightening with no negatives in fiber damage bs. conventional oxidants.

An in vitro assay to screen for the sensitizing potential of xenobiotics.

We are developing a new, animal-free assay for determination of the sensitizing potential of a substance. The design of this assay is based on current immunological knowledge of the pathogenesis of allergic contact dermatitis. It integrates human dendritic cells and keratinocytes, which are both known to be critically involved in vivo. The read-out system uses molecular responses typically occurring after an encounter of the skin with contact allergens. The assay provides concentration-response information, by which the relative ability of a chemical to induce sensitization can be predicted. Additionally, the assay defines the border-concentration of general toxicity of a substance. Weak allergens and even prohaptens are detectable. We have called the assay LCSA, loose-fit coculture-based sensitization assay.