Development of Safirinium dyes for new applications : fluorescent staining of bacteria, human kidney cells, and the horny layer of the epidermis

Low-molecular synthetic fluorophores are convenient tools in bioimaging applications. Several derivatives of Safirinium dyes as well as their reactive N-hydroxysuccinimide (NHS) esters bearing diverse substituents were synthesized and evaluated experimentally in terms of their lipophilicity by means of reverse-phase and immobilized artificial membrane high-performance liquid chromatography. Subsequently, the selected compounds were employed as novel cellular imaging agents for staining Gram-positive and Gram-negative bacteria, human kidney cell line, as well as human skin tissue. The analyzed dyes allowed for visualization of cellular structures such as mitochondria, endoplasmic reticulum, and cellular nuclei. They proved to be useful in fluorescent staining of stratum corneum, especially in the aspect of xenobiotic exposure and its penetration into the skin. The best results were obtained with the use of moderately lipophilic NHS esters of Safirinium Q. The development of Safirinium dyes is a promising alternative for commercially available dyes since the reported molecules have low molecular masses and exhibit efficient staining and remarkable water solubility. Moreover, they are relatively simple and low-cost in synthesis.Peer reviewe

Wpływ rodzaju nośnika oraz właściwości fizykochemicznych terpenów na ich przenikanie do warstw skóry ludzkiej

Zawiera prace stanowiące podstawę habilitacji.

DEVELOPMENT OF HYDROGEL CAPSULES FOR USE IN DUAL-PHASE TOPICAL PREPARATIONS CONTAINING DIHYDROXYACETONE AND AMINO ACIDS – PRELIMINARY RESULTS

The aim of the following work was a preliminary assessment of the possibility of producing a topical preparation containing dihydroxyacetone (DHA) and amino acids, encapsulated separately in polysaccharide capsules. The colour reaction between the DHA and amino acids would be possible after mixing of the two components triggered with mechanical disruption of the capsules on application. Such preparation could be used in patients suffering from vitiligo, by masking the symptoms of the disease. Several capsule compositions were evaluated in terms of appearance, dimensions and mechanical parameters. In addition, the reactions of various amino acids with DHA were carried out and obtained colours were analysed. The results indicate that formation of the dual-phase formulation of encapsulated DHA and amino acids is possible. It was also found that certain amino acids vary in colour resulting from reaction with DHA. This feature can be potentially utilized to better design the colour resulting from application of the prepared composition

The effect of mangiferin on skin: Penetration, permeation and inhibition of ECM enzymes.

Mangiferin (2-C-β-D-glucopyranosyl-1,3,6,7-tetrahydroxyxanthone) is a polyphenol with strong antioxidant properties. Mangiferin is obtained from the mango tree (Mangifera indica L., Anacardiaceae). It has been proven that mangiferin exhibits many pharmacological activities. The aim of this study was to analyze the penetration of mangiferin into the human skin and through the skin. According to our knowledge, skin penetration and permeation studies of mangiferin have not been analyzed so far. Additionally, the influence of mangiferin on two Extracellular Matrix Enzymes (ECM): collagenase and elastase, was evaluated for the first time. It has been indicated that mangiferin is able to permeate the stratum corneum and penetrate into the epidermis and dermis in comparable amounts. For confirmation of the obtained results, fluorescence microscopy was successfully utilized. The analysis revealed the capability of mangiferin to reversibly inhibit elastase and collagenase activity. The mechanism of mangiferin interaction with both enzymes was estimated as a noncompetitive inhibition

Mangiferin and Hesperidin Transdermal Distribution and Permeability through the Skin from Solutions and Honeybush Extracts (Cyclopia sp.)—A Comparison Ex Vivo Study

Polyphenolic compounds—mangiferin and hesperidin—are, among others, the most important secondary metabolites of African shrub Cyclopia sp. (honeybush). The aim of this study was to compare the percutaneous absorption of mangiferin and hesperidin from solutions (water, ethanol 50%, (v/v)) and extracts obtained from green and fermented honeybush (water, ethanol 50%, (v/v)). Research was performed with the Bronaugh cells, on human dorsal skin. The mangiferin and hesperidin distributions in skin layers (stratum corneum, epidermis, and dermis) and in acceptor fluid (in every 2, 4, 6, and 24 h) were evaluated by HPLC–Photodiode Array Coulometric and Coulometric Electrochemical Array Detection. The transdermal distribution of hesperidin was also demonstrated by fluorescence microscopy. Results indicated that mangiferin and hesperidin were able to cross the stratum corneum and penetrate into the epidermis and dermis. An advantage of hesperidin penetration into the skin from the water over ethanol solution was observed (451.02 ± 14.50 vs. 357.39 ± 4.51 ng/cm2), as well as in the mangiferin study (127.56 ± 9.49 vs. 97.23 ± 2.92 ng/cm2). Furthermore, mangiferin penetration was more evident from nonfermented honeybush ethanol extract (189.85 ± 4.11 ng/cm2) than from solutions. The permeation of mangiferin and hesperidin through the skin to the acceptor fluid was observed regardless of whether the solution or the honeybush extract was applied. The highest ability to permeate the skin was demonstrated for the water solution of hesperidin (250.92 ± 16.01 ng/cm2), while the hesperidin occurring in the extracts permeated in a very low capacity. Mangiferin from nonfermented honeybush ethanol extract had the highest ability to permeate to the acceptor fluid within 24 h (152.36 ± 8.57 ng/cm2)

Mangiferin (C₁₉H₁₈O₁₁).

<p>Mangiferin (C₁₉H₁₈O₁₁).</p

Changes in the activity of elastase (%) in the presence of increasing concentrations of mangiferin solution (μM).

<p>Changes in the activity of elastase (%) in the presence of increasing concentrations of mangiferin solution (μM).</p

Permeation of mangiferin through the skin.

<p>The aqueous and ethanol mangiferin solutions were applied on the skin and after 2, 4, 6 and 24 h the amount of mangiferin in the acceptor fluid was measured by HPLC. The obtained HPLC data (ng/ml) were used to calculate the amount of mangiferin that permeates through the skin surface area of 1 cm². Error bars represent standard deviations. Significant differences between the results are marked with an “*” (p<0.05, two-way ANOVA with post-hoc Tukey test).</p

Activity of elastase (mM/min) vs elastase concentration (μg/ml) in the presence of different concentrations of mangiferin solution (0, 59, 118 and 177 μM).

<p>Activity of elastase (mM/min) vs elastase concentration (μg/ml) in the presence of different concentrations of mangiferin solution (0, 59, 118 and 177 μM).</p

Activity of collagenase (mM/min) vs collagenase concentration (μg/ml) in the presence of different concentrations of mangiferin solution (0, 200, 270 and 350 μM).

<p>Activity of collagenase (mM/min) vs collagenase concentration (μg/ml) in the presence of different concentrations of mangiferin solution (0, 200, 270 and 350 μM).</p