Microneedle patch based on dissolving, detachable microneedle technology for improved skin quality of the periorbital region. Part 2: Clinical Evaluation.

OBJECTIVE: Microneedle patches based on dissolving, detachable microneedle technology (Russian patent No. 2652567; US patent EFS No. 32735812; WO/2019/231360) are novel dermatological products that allow safe, painless and effective reduction of epidermal wrinkles after six procedures. The purpose of this study was two-fold (a) to assess the safety and efficacy of microneedle patches comprising 650 microneedles containing hyaluronic and ferulic acids after 6 procedures of applying the applicator to the skin for 25 min; and (b) to correlate our previously reported ex vivo findings. METHODS: The microneedle applicator contains 650 microneedles, which dissolve in 25 min of exposure. The effectiveness of microneedle applicators was confirmed by a randomized split-phase study involving 82 subjects. Applications of microneedle patches were performed at intervals of two times per week, and the effectiveness was assessed at 6 weeks after the start of the study. RESULTS: The results of the profilometric evaluation of skin smoothness demonstrated a significant reduction in the average roughness index by 65.32 ± 2.99% and in the mean skin waviness by 66.84 ± 1.6% compared with these indicators in the control group (P < 0.05). The therapeutic effect of the procedure was confirmed by an ultrasound examination that registered a 72.2 ± 5.4% and 25 ± 1.4% increase in echo-density of the epidermis and dermis, respectively. An independent blinded skin evaluation by dermatologists revealed steady decrease in puffiness of the application area in 89.9% of patients, increased elasticity in 78.3% of the patients, and reduced severity of epidermal wrinkles in 89.9% of the patients. CONCLUSIONS: The applicator with 650 soluble microneedles on its surface containing hyaluronic and ferulic acids is considered a safe, effective and convenient way to improve the skin quality in the periorbital region after six procedures. Nevertheless, additional studies of soluble microneedles are required to fully assess the amount and distribution area of the injected hyaluronic acid and other active components, as well as to detail the mechanism of action of soluble microneedles to improve skin quality.

OBJECTIF: Les patchs Microneedle basés sur la technologie de micro-aiguille dissolvante et détachable (brevet russe n ° 2652567; brevet américain EFS n ° 32735812; WO / 2019/231360) sont de nouveaux produits dermatologiques qui permettent une réduction sûre, indolore et efficace des rides épidermiques après 6 procédures. Le but de cette étude était double: a) évaluer l'innocuité et l'efficacité des patchs de micro-aiguilles comprenant 650 micro-aiguilles contenant des acides hyaluronique et férulique après 6 procédures d'application de l'applicateur sur la peau pendant 25 minutes; et b) pour corréler nos résultats ex-vivo précédemment rapportés. MÉTHODES: L'applicateur de micro-aiguilles contient 650 micro-aiguilles, qui se dissolvent en 25 minutes d'exposition. L'efficacité des applicateurs de micro-aiguilles a été confirmée par une étude randomisée en hémi-visage portant sur 82 sujets. Des applications de patchs de micro-aiguilles ont été effectuées à des intervalles de 2 fois par semaine, et l'efficacité a été évaluée 6 semaines après le début de l'étude. RÉSULTATS: Les résultats de l'évaluation profilométrique de la douceur de la peau ont démontré une réduction significative de l'indice de rugosité moyen de 65,32 ± 2,99% et de l'ondulation cutanée moyenne de 66,84 ± 1,6% par rapport à ces indicateurs dans le groupe témoin (p <0,05). L'effet thérapeutique de la procédure a été confirmé par un examen échographique qui a enregistré une augmentation de 72,2 ± 5,4% et 25 ± 1,4% de l'écho-densité de l'épiderme et du derme, respectivement. Une évaluation indépendante de la peau en aveugle par des dermatologues a révélé une diminution constante des poches de la zone d'application chez 89,9% des patients, une augmentation de l'élasticité chez 78,3% des patients et une réduction de la gravité des rides épidermiques chez 89,9% des patients. CONCLUSIONS: L'applicateur avec 650 micro-aiguilles solubles à sa surface contenant des acides hyaluronique et férulique est considéré comme un moyen sûr, efficace et pratique d'améliorer la qualité de la peau dans la région périorbitaire après 6 procédures. Néanmoins, des études supplémentaires sur les micro-aiguilles solubles sont nécessaires pour évaluer pleinement la quantité et la zone de distribution de l'acide hyaluronique injecté et d'autres composants actifs, ainsi que pour détailler le mécanisme d'action des micro-aiguilles solubles pour améliorer la qualité de la peau.

Microneedle patch based on dissolving, detachable microneedle technology for improved skin quality - Part 1: ex vivo safety evaluation.

OBJECTIVE: The aim of the paper presented herein is the description and safety evaluation of the process of dissolution of an 86-microneedle patch composed of hyaluronic acid, when applied topically to human abdominal skin explants. Such explants were chosen to replace the inability of obtaining periorbital skin. In order to evaluate penetration and dissolution of the microneedles, we employed histochemical methods and a fluorescent dye FITC (fluorescein isothiocyanate). METHODS: Abdominoplasty human skin explants were treated with square microneedle patches with a 1.5-cm2 surface area, containing 86 microneedles and having 450 ± 23.5 µm in height with 1 mm interspacing between nearest neighbouring microneedles. Histological processing and staining for cell viability, FITC distributions and glycosaminoglycans were performed. The stained surface percentage for each treatment was compared to control untreated samples at given time points. A Mann-Whitney test was used to identify the difference between two populations (sites of skin samples punctured with stained and clear microneedles, respectively) at the given level of statistical significance (P < 0.05). RESULTS: The application of the MN patch to excised skin explants showed these microneedles to be non-invasive into the dermis of the skin. Skin puncturing with MN patches revealed 17 different sites of microneedle penetration immediately afterwards and 4 sites, 2 h later. Although there were some variances in the epidermal depth of penetration, these variances did not impact on cell viability. The hyaluronic acid-based microneedles having 450 µm in length penetrated the epidermis at an averaged depth by 26 µm without disrupting skin cell viability and without causing an inflammatory response. Hyaluronic acid could be detected in most of these penetration sites, with no diffusion into the dermis, which is important for cosmetic applications. FITC analysis uncovered fluorescein isothiocyanate distribution within microneedle insertion site, which remained steady after 2 and 6 h of experimentation. CONCLUSION: Using ex vivo tracer staining studies, we have shown that the evaluated microneedle applicator is capable of penetrating the skin epidermis and delivering substances embedded in the needle polymer matrix. In addition, the tested product was shown to be safe, which provides a broad perspective for delivering cosmetic and pharmaceutic agents.

OBJECTIF: Le but de cet article est de décrire et évaluer l'innocuité du processus de dissolution d'un patch de 86 micro-aiguilles composé d'acide hyaluronique, lorsqu'il est appliqué par voie topique sur des explants de peau abdominale humaine. De tels explants ont été choisis pour palier à l'impossibilité d'obtenir une peau périorbitaire. Afin d'évaluer la pénétration et la dissolution des micro-aiguilles, nous avons utilisé des méthodes histochimiques et un marqueur fluorescent FITC (isothiocyanate de fluorescéine). MÉTHODES: Les explants de la peau humaine de l'abdominoplastie ont été traités avec des patchs à microaiguilles, carrés d'une surface de 1,5 cm2, contenant 86 micro-aiguilles et ayant 450 ± 23,5 µm de hauteur avec 1 mm d'espacement entre les micro-aiguilles. Un traitement histologique et des colorations ont été réalisée pour observer la viabilité cellulaire et les glycosaminoglycanes. La diffusion de FITC a été observée en épifluorescence. Le pourcentage de surface colorée pour chaque traitement a été comparé à des échantillons témoins non traités à différents temps de cinétique. Un test de Mann-Whitney a été utilisé pour identifier la différence entre deux populations (sites de pénétration des micro-aiguilles et peau normale) avec une limite de significativité statistique de P < 0.05. RÉSULTATS: L'application du patch MN sur les explants de peau a montré que ces micro-aiguilles ne Pénétraient pas dans le derme de la peau. L'application cutanée des patchs MN a révélé 17 sites différents de pénétration de micro-aiguille immédiatement après l'application et 4 sites, 2 heures plus tard. Bien qu'il y ait eu quelques variations dans la profondeur de pénétration épidermique, ces variations n'ont pas eu d'impact sur la viabilité cellulaire. Les micro-aiguilles à base d'acide hyaluronique d'une longueur de 450 µm ont pénétré l'épiderme à une profondeur moyenne de 26 µm sans perturber la viabilité des cellules de la peau et sans provoquer de réponse inflammatoire. L'acide hyaluronique composant les micro-aiguilles a été détecté sur la plupart de ces sites de pénétration, sans diffusion dans le derme, ce qui est important pour les applications cosmétiques. L'analyse FTIC a révélé une distribution d'isothiocyanate de fluorescéine dans le site d'insertion de micro-aiguille qui est restée stable après 2 et 6 heures d'expérimentation. CONCLUSION: En utilisant des études de coloration de traceurs sur explant de peau humaine ex vivo, nous avons montré que l'applicateur de micro-aiguille est capable de pénétrer l'épiderme et de délivrer des substances incorporées dans la matrice polymère de l'aiguille. De plus, le produit testé s'est avéré sûr et bien toléré, ce qui ouvre une large perspective pour l'administration d'agents cosmétiques et pharmaceutique.

[Biosorption ability of mutants of Rhodotorula mucilaginosa UCM Y-1776].

Twenty stable mutants with various coloration intensity have been allocated in carotene-synthesizing natural strain Rhodotorula mucilaginosa UCM Y-1776 (wild type) after nitrosoguanidine action. Two brightly orange mutants 4L and 11 and one non-pigmented mutant 2 were chosen for the further researches. The ultraviolet was inefficient as a mutagen. Resistance to high concentration of copper ions (up to 200 mg/g), high sorption ability (Qmax = 9.1 mmol/g) was characteristic of R. mucilaginosa UCM Y-1776. Concentration of copper ions 50 mg/l was toxic for mutants 4L, 11 and 2, which sorption ability was lower in comparison with carotene pigmented R. mucilaginosa UCM Y-1776. It was shown, for the first time that there was a direct dependence between the presence of carotenoid pigments, resistance to high concentration of copper ions and sorption ability for yeast R. mucilaginosa UCM Y-1776.

[Intensification of extraction by yeast Saccharamyces cerevisiae 1968 of copper ions from a solution in magnetic field].

It was determined whether it is possible to intensify the biosorption of copper ions from a copper sulfate solution with yeast Saccharomyces cerevisiae 1968 by introducing a metal headpiece into the solution and by applying an external magnetic field. The study was carried out in a magnetic field oriented both parallel and perpendicular to the axes of the rods (with parallel and perpendicular geometry of the system) that make up the headpiece. It was shown that the extent of intensification of the extraction of copper ions at different geometries of the system differs insignificantly and that the extraction of copper ions from the solution occurs by biosorption and cementation onto the metal headpiece.

[Effect of chromium (VI) on growth physiology and sorptional capacity of yeast]. / Vliianie khroma (VI) na fiziologiiu rosta i sorbtsionnuiu sposobnost' drozzhzhei.

Resistance of 7 yeast species (7 strains) to various chrome concentrations in the medium and their sorptional activity have been studied. It is shown that the biomass of the yeast Williopsis californica UKM-248, Candida krusei UKM-61t under their concentration in the medium of 200 and 500 mg/l of Cr6+ is 0.74 and 0.45 g/l DWB, respectively. For the rest of strains these chrome concentrations are toxical and the concentration of 100 mg/l of Cr6+ is the bordering threshold for them. Under periodical cultivation in the medium with 30 mg/l of Cr6+ one can observe the delay of cultures growth displayed in the increase of the lag-phase and exponential growth phase as well as the decrease of medium pH (to 2). The resistance of the studied yeast to high concentrations of Cr6+ correlates with their sorptional activity.

[Search of heavy metals biosorbents among yeasts of different taxonomic groups]. / Poisk biosorbentov tiazhelykh metallov sredi drozhzhei razlichnykh taksonomicheskikh grupp.

The use of microbial cells as heavy metal biosorbents is a potential alternative for the existing methods of the environment treatment for pollution by these metals. Yeast may be successfully used in the metals sorption. An agar-plate screening method was used for rapid isolation of metal-accumulating strains of yeast and preliminary estimation of their biosorption capacity. The capacity to accumulate heavy metals, such as copper, zinc, lead, chrome and cobalt was estimated in 68 pigmented and pigmentless yeast strains isolated from different sources. The best capacity of copper and zinc accumulation was shown by pink-coloured Rhodotorula mucilaginosa, Rhodotorula aurantiaca, Rhodotorula glutinis and pigmentless strains of the yeast Candida krusei, Williopsis california.

[Yeasts--biosorbents of heavy metals]. / Drozhzhi--biosorbenty tiazhelykh metallov.

The sharp increase of the level of environment pollution by heavy metals caused the increase of interest to the problem of live organisms (including microorganisms) resistance to these metals. Biosorption is one of the mechanisms of microorganisms resistance to heavy metals. Yeasts as biosorbents are of special interest. An analysis of the data from literature have shown that the yeast biomass may be used successfully as biosorption material for such metals as Ag, Au, Cd, Co, Cr, Cu, Ni, Pb, U, Th, Zn. Yeasts of genera Saccharomyces, Candida, Pichia are efficient biosorbents of metals. The sorptional system estimation is based on the classic sorption isotherm obtained in the course of equilibrium experiments and depends on pH, properties of metal ions, biomass concentration, preliminary physical or chemical treatment of the biomass, presence of various organic and inorganic ions and on temperature. The yeast biomass may be obtained using numerous industrial processes, that decreases considerably the biosorbent cost. Most yeasts can sorb a wide range of metals or be strictly specific in respect of only one metal. Special attention would be paid to the cell wall which structure determines sorption proceeding mechanisms. Problems of mechanisms of heavy metal biosorption by microorganisms at molecular level are discussed. The review also deals with the newest developments on improving the biosorption processes in microorganisms, yeast in particular.

[Bioethics of research on humans and animals in aviation, space and marine medicine]. / Bioéticheskie pravila provedeniia issledovanii na cheloveke i zhivotnykh v aviatsionnoi, kosmicheskoi i morskoi meditsine.

The article includes excerpts from The Bioethic Rules of Research With Humans and Animals that have established on analysis of national and international bioethic guidelines for biomedical research with the use of humans and animals and are a part of the ISS human use guidelines.

[Ethics of space life science research with animals]. / Etika éksperimental'nykh issledovanii na zhivotnykh v kosmicheskoi biologii i meditsine.

The authors present a historical sketch on the ethics of use of animals in life sciences experiments. Experiments with animals are necessary to expand knowledge in the field of life sciences including the space ones, and health service. It is emphasized that these experiments must be performed with observance of certain moral rules and utilize minimum animals suffice to obtain statistically significant data. The first draft of SRC-IBMP Guidelines on the use of laboratory animals has been published.

[The biology of bacteria used to treat industrial sewage to remove heavy metals]. / Biologiia bakterii, ispol'zuemykh pri ochistke promyshlennykh stochnykh vod ot tiazhelykh metallov.

The content of heavy metals in the water bodies increases as a result of anthropogenic effects. Heavy metals exert toxical effect on hydrobionts. Involved into the trophic chain they can be the cause of grave human diseases. Microorganisms possess a capacity to accumulate heavy metals in their body, evoke the process of their enzymatic transformation into nontoxical compounds and adsorb them on the cell surface. When breeding bacterial strains for the biological treatment of sewage from the metal-working plants the authors recommend to choose the cultures from econiches containing heavy metal compounds and to establish their resistance to high concentrations of metals. This resistance correlates with high biological activity. The bacterial strains of Pseudomonas genus possessing a capacity to sulphate reduction have been selected. As a result of their activity the high-solvable toxical sulphates are restored to practically unsolvable precipitating forms. A new species Aeromonas dechromatica transforming high-toxical six-valence chromium into low-toxical three-valent ones is described. The grounds of biotechnologies of the use of the breeded cultures for treatment of industrial sewage are stated.

[The sulfate-reducing capacity of bacteria in the genus Pseudomonas]. / Sul'fatvosstanavlivaiushchaia sposobnost' bakterii roda Pseudomonas.

Active sulphate-reducing microorganism which belongs to the genus Pseudomonas has been distinguished and described. The culture is a facultative aerobe, optimum Eh is -170-180 mV. Pseudomonas sp. being cultivated under strictly anaerobic conditions sulphate-reduction proceeds more intensively than under aerobic conditions. This fact should be taken into account under treatment of industrial sewage.

[Use of metabolic and pathogenetic criteria in group standardization of organically active chlortriazine dyes]. / Ob ispol'zovanii metabolicheskikh i patogeneticheskikh kriteriev dlia gruppovogo normirovaniia krasitelei organicheskikh aktivnykh khlortriazinovykh.

The article presents research data related to the biotransformation, intoxication mechanisms and pathogenic properties of chlortriazine dye-stuffs. All the dye-stuffs studies were characterized by similar intoxication properties and pathogenesis, and were related to the 3rd class of hazardous substances. The MAC for the organic chlortriazine dye-stuffs constituted 2 mg/m3.

[Bacteria reducing chromium in nature and in effluents of industrial plants]. / Bakterii, vosstanavlivaiushchie khrom v prirode i v stokakh proizvodstvennykh predpriiatii.

Bacteria capable of hexavalent chromium (Cr6+) reduction can be found in Cr6+-containing sewage and sediments of purification tanks of industrial plants. They cannot be detected in water and soil samples containing no chromium compounds. Bacteria reducing chromium belong to the genera Aeromonas, Escherichia, Pseudomonas and Enterobacter. Their activity of Cr6+ reduction correlates with the high resistance to the elevated content of this ion in the medium. The fine cell structure of these bacteria is described.

[Use of Robolact and Linolac dry milk mixtures in the overall therapy of infants with acute intestinal infections]. / Ispol'zovanie sukhikh molochnykh smesei "Robolakt" i "Linolak" v kompleksnoi terapii detei, bol'nykh ostrymi kishechnymi infektsiiami.

The clinical efficacy of the dry milk mixtures "Robolact" and "Linolac" manufactured in Hungary was studied comprehensively in young children with acute intestinal infections. It was shown that "Robolact" has a good palatability and produces a satiation sensation. The children suffering from coli-infection, staphylococcal enterocolitis and mixed intestinal infection received different types of feeding. In children on "Robolact" feeding, the stool returned to normal over a shorter period of time which was equal to 10 (+/-2) days versus 16 (+/-3) days in children on natural feeding, and 12 (+/-5) days on feeding with kefir and its mixtures. The mixture "Linolac" given during the amelioration of acute clinical symptoms of the intestinal infection was tolerated more poorly that was manifested in regurgitations in part of children and in food refusals in 15% of the cases. Examination of the microflora revealed dysbacteriosis in all the patients with intestinal infections regardless the type of feeding. Substantial differences in the microflora composition concerned the anaerobic flora in both experimental and control groups. The children on natural feeding showed the decreased bifidoflora half as frequently as those on formula feeding.