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.

[Toxicological evaluation of nanosized colloidal silver, stabilized with polyvinylpyrrolidone, in 92-day experiment on rats. II. Internal organs morphology].

The aim of the study was to evaluate the safe doses of commercially available nanosized colloidal silver (NCS), stabilized with polyvinilpirrolidone (PVP, food additive E1201) when administered in gastrointestinal tract of rats in the 92-day experiment in terms of the morphological changes in the internals of animals. The sample studied contained non-aggregated nanoparticles (NPs) of silver belonging to size fractions with a diameter of less than 5 nm, 10-20 nm or 50-80 nm. 80% of NPs were inside the range of hydrodynamic diameters 10.6-61.8 nm. The preparation of NCS was administered to growing male Wistar rats. (initial body weight 80 ± 10 g) for 1 month by intragastric gavage and then consumed with food at doses of 0.1, 1.0 and 10 mg/kg of body weight based on silver. The control animals received water or vehicle of nanomaterial--water solution of PVP. After withdrawal of animals from the experiment by exsanguination under ether anesthesia organs (liver, spleen, kidney, ileum) were isolated and their slides were prepared by standard methods following 'by staining with hematoxylin-eosin. Analysis was performed in light optical microscope equipped with a digital camera at a magnification from 1 x 100 to 1 x 1000. It was shown that the experimental animals treated with the NCS developed series of morphological changes in the tissues of the internal organs (liver, spleen and kidney) with the elevation of the range and severity of structural changes with increasing doses of silver. The most sensitive target of NCS action was apparently liver, which has already shown at a dose of 0.1 mg of silver NP/kg of body weight marked eosinophilic infiltration of portal tracts, which was accompanied at doses of 1.0 and 10.0 mg/kg by the emergence of medium and large-drop fat vacuoles in the cytoplasm of hepatocytes, swelling and lympho-macrophage. infiltration of the portal tracts. Detectable changes can be regarded as symptoms of inflammation of hepatocytes, at least, at a dose nanomaterial of 1.0 mg/kg body weight or more. Relative intensity of morphological changes in the internal organs correlated with published data on the biodistribution of silver NP administered to the gastrointestinal tract. It is concluded that the threshold dose corresponding to the minimum adverse effect of NCS is, according to the study of the above, no more than 1.0 mg/kg of body weight based on silver.

[Experience of using subcutaneous interstitial fluid for biomonitoring a dose load in workers of metallurgic industry].

Comparative envaluation of metals content of serum and subcutaneous interstitial fluid revealed that levels of chromium, manganese, arsenic, cadmium in these biologic media have no reliable differences (p = 0.056-0.907) but strong correlation (r = 0.52-0.93). Findings are that sorlie metals (nickel, copper, zinc, lead) contents of subcutaneous interstitial fluid have reliable differences in contents of serum and subcutaneous interstitial fluid., but these metals have strong correlations between their contents in the biologic media (r=0.43-0.72). Chances are that subcutaneous interstitial fluid is a promising medium for biomonitoring of dose load in metallurgic industry workers and enables to increase efficiency of measures reducing risk of occupational hazards influence on the worker's health.

[Inhalation toxicity of nanodispersed manganese oxide aerosol].

The article presents results of experimental study concerning acute inhalation toxicity of nanodispersed manganese oxide widely used in novelty goods production. During the technology process, nano-dimensional manganese oxide aerosol could appear in workplace air--that causes danger of the workers' inhalation exposure. Findings are that the studied substance is a nanomaterial with prevalent fihiform particles measuring 15-29 nm. The substance is acutely toxic when inhaled as an aerosol. Cl50 for 4-hour exposure for Wistar rats equals 120 mg/m³ (1 jeopardy class). Clinical manifestations of acute intoxication are mostly irritative and neurotoxic effects supported by morphologic studies. Histopathologic changes in brain tissue are cerebellar ischemia with dilated perecellular spaces, faveolate appearance.

[Toxicological assessment of nanostructured silica. The acute oral toxicity].

Nanostructured amorphous silica (SiO2) is one of the priorities of nanomaterials, exposing human to the ever-increasing scale as a component of food additives, drugs and cosmetic products. According to numerous publications SiO2 nanoparticles (NPs) possess deleterious effects on animal and human cells in vitro and also exhibit inhalation toxicity. However, the biological effects in vivo of silica NPs taken orally are studied insufficiently. This article represents the first section of this study which aim is identification of silica preparation as nanomaterial and estimating of acute toxicity after oral administration in the form of aqueous suspension. Studies of size and shape of the particles in aqueous suspension of silica used in the study by electron and atomic force microscopy, spectroaqustic analysis and dynamic laser light scattering showed that the test substance is a nanomaterial. Estimation of acute toxicity of an aqueous suspension of nanostructured silica with a single intragastric gavage to male BALB/C mice allowed to conclude that the test material has LD50 by the oral route greater than 10 000 mg/kg and consequently belongs to class IV (low danger agents). Pathological changes in jejunum and colon of mice in the acute experiment (at a dose of 10 000 mg/kg) haven't been identified. Thus SiO2 NPs possess low toxicity when administered in the gastrointestinal tract. The available data, however, do not exclude possible presence of adverse effects under their long-term administration by oral way.

[Effect of nanodispersed manganese oxide (III, IV) on morphological properties of various tissues and body systems].

The study of morphological features of tissue organs of experimental animals after a single intragastric administration of an aqueous suspension nanodis-dispersed manganese (III, IV) at doses of 2000, 3500 and 5000 mg/kg showed complex of morphological changes in the form of circulatory disorders up to hemostasis and bleeding which lead to changes in the structure of organs, enhanced apoptosis, activation of macrophage system by proliferation and macrophage phagocytosis of degradation products of cells, hypertrophy of lymphoid tissue of immune system; development of histiocytic infiltrates in parenchymal organs and central nervous system. With the introduction of microsized analogues in the same doses of the morphological changes in the circulatory system, macrophage system and immune organsfound predominantly in a dose of 5000 mg/kg. Changes were less significant and were presented as activation of macrophages in the liver in the form of activation of Kupffer cells, proliferation of lymphoid tissue in the spleen, lymph histiocytic infiltrates in the liver, kidney, medium size.

[Hygienic and toxicological safety assessment of nano- and micro-dispersed manganese oxide (III, IV)].

The findings of the experimental study performed within the preliminary hygienic and toxicological assessment of nanodispersed manganese oxide (III, IV), which was endogastrics administered to white nonlinear mice once, showed that, under acute oral exposure to manganese oxide, nanodispersed particles were 2,6-fold more toxic and cumulative (class 3, according to the oral LD50 values) than dispersed solution with micro-sized particles (0.4-592 microm, class 4). Endogactric exposure to nanodispersed solution of manganese oxide (III, IV) in organisme produced such negative health effects as vasodilation and plethora, pathological inclusions and increased cell aggregation, which increased and became more prevalent with higher doses. The prediction of chronic toxicity parameters demonstrated that threshold and NOAEL doses for nanodispersed solution were 1.6 and 0.26 mg/kg, respectively, which are 2.3 times lower than for finely divided solution of manganese oxide (III, IV).

[Surgical treatment of cancer of the large intestine, complicated by intestinal obstruction]. / Khirurgicheskoe lechenie raka tol'stoi kishki, oslozhnennogo kishechnoi neprokhodimost'iu.

Retrospective analysis of the treatment of 3,080 patients with cancer of the large intestine made it possible to distinguish compensated, sub- and decompensated degrees of the neoplastic stenosis and place the two last named into the category of a complicated course of cancer. In subcompensated stenosis (complications in 16.3% of cases, mortality 5.9%), irrespective of the location of the tumor, the authors consider one-stage intervention with primary restoration of the continuity of the intestine to be the operation of choice. In decompensated stenosis (complications 28.9%, mortality 13.2%) caused by tumors of the right colon and the rectum they recommend a similar policy, while in tumors of the left colon they believe Hartmann's operation to be indicated.

[Radical and palliative combined surgery in the surgical treatment of cancer of the rectum]. / Radikal'nye i palliativnye kombinirovannye operatsii v khirurgicheskom lechenii raka priamoi kishki.

Analysis of 186 combined resections and extirpations of the rectum for primary carcinoma, among which 31 were palliative operations, showed that the indications for them are limited with age from 21.7% in the young to 12.8% in patients over 60 years of age. Combined interventions were carried out irrespective of the site of the tumor in the rectum. Morphological verification of growth of the tumor into other organs in 32.3% of patients who had been operated on merely points to the expediency of a combined operation excluding ungrounded refusal of treatment. Combined operations failed to lead to an increase in postoperative mortality, which was 5.4%, and produce favourable late-term results--54.5 +/- 4.3% of patients have a survival period of 5 years.

[Improvement of the effectiveness of the surgical treatment in cancer of the colon]. / Povyshenie éffektivnosti khirugicheskogo lecheniia raka obodochnoi kishki.

The authors studied the results of 585 resections of the colon and hemicolectomies which were performed for carcinoma; 105 of the operations were palliative. Among the radical operations 82 (17.1%) and among the palliative operations 28 (26.7%) were combined. 247 patients were 60 years of age and older. In the early postoperative period 5.0% of patients died after radical and 16.2% after palliative operations. Postoperative lethality was higher in combined operations and among elderly and old aged patients.

[Combined surgery in cancer of the colon]. / Kombinirovannye operatsii pri rake obodochnoi kishki.

The study was concerned with the short- and long-term results of 80 radical and 27 palliative combined resections and hemicolectomy for colonic cancer involving other organs or accompanied by solitary distant metastases. Tumor elements were morphologically identified in other resected organs (53.3%). 15.9% of surgical patients died within short periods after operation. Colon was anastomosed immediately in 97 patients (90.7%), cecostomy being performed in two of them shortly after operation. Five--year survival was 53.5% in radically--operated patients and 31.6%--in cases of palliative treatment.

[Combination primary restorative surgical interventions in cancer of the rectum in women]. / Kombinirovannye pervichno-vosstanovitel'nye khirurgicheskie vmeshatel'stva pri rake priamoi kishki u zhenshchin.

In the past 15 years, 136 combined operations in women with locally disseminated rectal cancer were performed. Eighty-two (60.3%) operations were primary-restorative. After them, the lethality was 2.9%, 3- and 5-year survival--64 and 40%, respectively. The performance of the combined sphincter-preserving operations is expedient, despite the spreading of the primary tumour.

[Expansion of the indications for surgery in neglected rectal cancer]. / O rasshirenii pokazanii k operatsii pri zapushchennoi rake priamoi kishki.

Radical surgery was performed in 971 (86.0%) cases of rectal cancer, while palliative resection or extirpation of the rectum--in 158 (14.0%). The postoperative lethality rates were 5.8 and 11.4%, respectively. Sixty-seven patients aged 60 and more underwent palliative resection or extirpation, with eight (11.9%) fatalities occurring shortly after treatment. The postoperative lethality rate was 8.8% in cases of exploratory laparotomy and symptomatic surgery, reaching 15.3% in a group of patients aged 60 years and more. It is inferred that palliative resection, extirpation and palliative combined surgery are justified in treatment advanced rectal cancer whatever patient's age.

[Surgical treatment of complicated cancer of the rectum]. / Khirurgicheskoe lechenie oslozhnennogo raka priamoi kishki.

The authors make an analysis of results of the surgical treatment of 246 patients with rectum carcinoma complicated by partial or complete ileus, pararectal abscess and perforation of the tumor. The data obtained show that primary resection of the tumor especially with a one-step reestablishment of the colon continuity is one of the main prerequisites for the elimination of postoperative complications and lethality.