The Protective Effects of Cath-MH With Anti-Propionibacterium Acnes and Anti-Inflammation Functions on Acne Vulgaris

Acne vulgaris is a common adolescent skin condition which is mainly caused by Propionibacterium acnes overcolonization and subsequent inflammation. Our previous studies have demonstrated that Cath-MH, an antimicrobial peptide from the skin of the frog Microhyla heymonsivogt, possesses potential antimicrobial, LPS-binding, and anti-septicemic properties. However, its protective effects and potential mechanisms against acne vulgaris are still unclear. In the present study, its anti-P. acnes effects were measured by two-fold broth dilution method, agglutination assay, scanning electron microscopy and confocal laser scanning microscopy experiments. Its treatment potential for acne vulgaris was further evaluated in mice ear inoculated by P. acnes. In addition, the binding ability between Cath-MH and LTA was measured by the Circular Dichroism and antibacterial assay. Moreover, the anti-inflammatory efficiency of Cath-MH was evaluated in LTA- and LPS-induced RAW 264.7 macrophage cells. Cath-MH was found to kill P. acnes with a MIC value of about 1.56 μM by membrane disruption mechanism. It also exhibited agglutination activity against P. acnes. Cath-MH was able to bind LTA as well as LPS, inhibit LTA/LPS-stimulated TLR2/4 expression, and subsequently decreased the inflammatory response in RAW 264.7 cells. As expected, Cath-MH alleviated the formation of edema and the infiltration of inflammatory cells in acne mouse model with concurrent suppression of P. acnes growth and inflammatory cytokines expression in vivo. The potent P. acnes inhibition activity combined with powerful anti-inflammatory effect of Cath-MH indicates its potential as a novel therapeutic option for acne vulgaris.

Download Full-text

Studies on the Mechanisms of Anti-Inflammatory Activity of Heparin- and Hyaluronan-Containing Multilayer Coatings—Targeting NF-κB Signalling Pathway

International Journal of Molecular Sciences ◽

10.3390/ijms21103724 ◽

2020 ◽

Vol 21 (10) ◽

pp. 3724 ◽

Cited By ~ 6

Author(s):

Hala Alkhoury ◽

Adrian Hautmann ◽

Bodo Fuhrmann ◽

Frank Syrowatka ◽

Frank Erdmann ◽

...

Keyword(s):

Signal Transduction ◽

Laser Scanning ◽

Adhesion Formation ◽

Laser Scanning Microscopy ◽

Inflammatory Activity ◽

Confocal Laser ◽

Layer By Layer ◽

Ethylene Imine ◽

Inflammatory Reactions ◽

Anti Inflammatory

The use of implants can be hampered by chronic inflammatory reactions, which may result in failure of the implanted device. To prevent such an outcome, the present study examines the anti-inflammatory properties of surface coatings made of either hyaluronic acid (HA) or heparin (Hep) in combination with chitosan (Chi) prepared as multilayers through the layer-by-layer (LbL) technique. The properties of glycosaminoglycan (GAG)-modified surfaces were characterized in terms of surface topography, thickness and wettability. Results showed a higher thickness and hydrophilicity after multilayer formation compared to poly (ethylene imine) control samples. Moreover, multilayers containing either HA or Hep dampened the inflammatory response visible by reduced adhesion, formation of multinucleated giant cells (MNGCs) and IL-1β release, which was studied using THP-1 derived macrophages. Furthermore, investigations regarding the mechanism of anti-inflammatory activity of GAG were focused on nuclear transcription factor-кB (NF-κB)-related signal transduction. Immunofluorescence staining of the p65 subunit of NF-κB and immunoblotting were performed that showed a significant decrease in NF-κB level in macrophages on GAG-based multilayers. Additionally, the association of FITC-labelled GAG was evaluated by confocal laser scanning microscopy and flow cytometry showing that macrophages were able to associate with and take up HA and Hep. Overall, the Hep-based multilayers demonstrated the most suppressive effect making this system most promising to control macrophage activation after implantation of medical devices. The results provide an insight on the anti-inflammatory effects of GAG not only based on their physicochemical properties, but also related to their mechanism of action toward NF-κB signal transduction.

Download Full-text

A Novel Hybrid Peptide Composed of LfcinB6 and KR-12-a4 With Enhanced Antimicrobial, Anti-Inflammatory and Anti-Biofilm Activities

10.21203/rs.3.rs-1109468/v1 ◽

2021 ◽

Author(s):

Chelladurai Ajish ◽

Sungtae Yang ◽

S. Dinesh Kumar ◽

Eun Young Kim ◽

Hye Jung Min ◽

...

Keyword(s):

Antimicrobial Activity ◽

Laser Scanning ◽

Antimicrobial Agents ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Bacterial Cells ◽

Bacterial Strains ◽

Hybrid Peptide ◽

Cell Selectivity ◽

Anti Inflammatory

Abstract Hybridizing two known antimicrobial peptides (AMPs) is a simple and effective strategy for designing antimicrobial agents with enhanced cell selectivity against bacterial cells. Here, we generated a hybrid peptide Lf-KR in which LfcinB6 and KR-12-a4 were linked with a Pro hinge to obtain a novel AMP with potent antimicrobial, anti-inflammatory, and anti-biofilm activities. Lf-KR exerted superior cell selectivity for bacterial cells over sheep red blood cells. Lf-KR showed broad-spectrum antimicrobial activities (MIC: 4–8 mM) against tested 12 bacterial strains and retained its antimicrobial activity in the presence of salts at physiological concentrations. Membrane depolarization and dye leakage assays showed that the enhanced antimicrobial activity of Lf-KR was due to increased permeabilization and depolarization of microbial membranes. Lf-KR significantly inhibited the expression and production of pro-inflammatory cytokines (NO and TNF-a) in LPS-stimulated mouse macrophage RAW264.7 cells. In addition, Lf-KR showed a powerful eradication effect on preformed multidrug-resistant Pseudomonas aeruginosa (MDRPA) biofilms. We confirmed using confocal laser scanning microscopy that a large portion of the preformed MDRPA biofilm structure was perturbed by the addition of Lf-KR. Collectively, our results suggest that Lf-KR can be an antimicrobial, anti-inflammatory, and anti-biofilm candidate as a pharmaceutical agent.

Download Full-text

Bioprocessed Brewers’ Spent Grain Improves Nutritional and Antioxidant Properties of Pasta

Antioxidants ◽

10.3390/antiox10050742 ◽

2021 ◽

Vol 10 (5) ◽

pp. 742

Author(s):

Rosa Schettino ◽

Michela Verni ◽

Marta Acin-Albiac ◽

Olimpia Vincentini ◽

Annika Krona ◽

...

Keyword(s):

Laser Scanning ◽

Antioxidant Properties ◽

Laser Scanning Microscopy ◽

Sensory Profile ◽

Confocal Laser ◽

Protective Effects ◽

Food Ingredient ◽

Cell Wall Disruption ◽

Spent Grain

Brewers’ spent grain (BSG), the by-product of brewing, was subjected to a xylanase treatment followed by fermentation with Lactiplantibacillus plantarum PU1. Bioprocessed BSG has been used as ingredient to obtain a fortified semolina pasta which can be labeled as “high fiber” and “source of protein” according to the European Community Regulation No. 1924/2006. Compared to native BSG, the use of bioprocessed BSG led to higher protein digestibility and quality indices (essential amino acid index, biological value, protein efficiency ratio, nutritional index), as well as lower predicted glycemic index. Bioprocessing also improved the technological properties of fortified pasta. Indeed, brightfield and confocal laser scanning microscopy revealed the formation of a more homogeneous protein network, resulting from the degradation of the arabinoxylan structure of BSG, and the release of the components entrapped into the cellular compartments. The extensive cell wall disruption contributed to the release of phenols, and conferred enhanced antioxidant activity to the fortified pasta. The persistence of the activity was demonstrated after in vitro-mimicked digestion, evaluating the protective effects of the digested pasta towards induced oxidative stress in Caco-2 cells cultures. The fortified pasta showed a peculiar sensory profile, markedly improved by the pre-treatment, thus confirming the great potential of bioprocessed BSG as health-promoting food ingredient.

Download Full-text

Metal Chelating, Inhibitory DNA Damage, and Anti-Inflammatory Activities of Phenolics from Rambutan (Nephelium lappaceum) Peel and the Quantifications of Geraniin and Corilagin

Molecules ◽

10.3390/molecules23092263 ◽

2018 ◽

Vol 23 (9) ◽

pp. 2263 ◽

Cited By ~ 9

Author(s):

Yujing Li ◽

Zhaojie Li ◽

Hu Hou ◽

Yongliang Zhuang ◽

Liping Sun

Keyword(s):

Dna Damage ◽

Cell Model ◽

Biological Properties ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dependent Manner ◽

Protective Effects ◽

Metal Chelating ◽

Study Results ◽

Anti Inflammatory

Whereas the preparation and biological properties of rambutan peel phenolics (RPP) were explored in our previous studies, the metal chelating, inhibitory DNA damage, and anti-inflammatory activities of RPP were evaluated and the important phenolics of RPP quantified in this study. Results showed that RPP had high Fe2+ and Cu2+-chelating activities with EC50 of 0.80 mg/mL and 0.13 mg/mL, respectively. RPP effectively decreased the production of hydroxyl radical with IC50 of 62.4 μg/mL. The protective effects of RPP against AAPH-induced DNA damage were also explored. RPP efficiently inhibited peroxyl radical-induced plasmid DNA strand breakage. The anti-inflammatory effects of RPP were determined using a lipopolysaccharide (LPS)-induced RAW 264.7 cell model. RPP significantly inhibited the production of nitric oxide (NO) and controlled the levels of inducible NO synthase mRNA in LPS-induced RAW 264.7 cells. The inhibitory activity increased in a dose-dependent manner. The above bioactivity of RPP was associated with its phenolic content and phenolic profiles. Furthermore, the contents of geraniin and corilagin in RPP were determined by an ultra-high performance liquid chromatography coupled with triple quadruple mass spectrometry (UPLC-QQQ-MS), showing 140.02 and 7.87 mg/g extract dry weight. Thus, RPP has potential applications as a novel nutraceutical and functional food in health promotion.

Download Full-text

Inhibition of Cronobacter Sakazakii Biofilm Formation and Expression of Virulence Factors by Coenzyme Q0

10.21203/rs.3.rs-58153/v1 ◽

2020 ◽

Author(s):

Ning Guan ◽

Yiqi Shi ◽

Haoyu Tong ◽

Yanpeng Yang ◽

Jiahui Li ◽

...

Keyword(s):

Biofilm Formation ◽

Laser Scanning ◽

Coenzyme Q ◽

Polymerase Chain Reaction Analysis ◽

Laser Scanning Microscopy ◽

Raw 264.7 Cells ◽

Confocal Laser ◽

Dependent Manner ◽

Cronobacter Sakazakii ◽

And Control

Abstract Objectives：Here, we investigated the inhibitory effects of coenzyme Q0 (CoQ0) on biofilm formation and the expression of virulence genes by Cronobacter sakazakii. Results：We found that the minimum inhibitory concentration of CoQ0 against C. sakazakii strains ATCC29544 and ATCC29004 was 100 μg/mL, while growth curve assays showed that sub-inhibitory concentrations (SICs) of CoQ0 for both strains were 6.4, 3.2, 1.6 and 0.8 μg/mL. Assays exploring the inhibition of specific biofilm formation showed that SICs of CoQ0 inhibited biofilm formation by C. sakazakii in a dose-dependent manner, which was confirmed by scanning electron microscopy and confocal laser scanning microscopy analyses. CoQ0 inhibited the swimming and swarming motility of C. sakazakii and reduced its ability to adhere to and invade HT-29 cells. In addition, CoQ0 impeded the ability of C. sakazakii to survive and replicate within RAW 264.7 cells. Finally, real time polymerase chain reaction analysis confirmed that nine C. sakazakii genes associated with biofilm formation and virulence were down-regulated in response to CoQ0 treatment. Conclusion：Overall, our findings suggest that CoQ0 is a promising antibiofilm agent and provide new insights for the prevention and control of infections caused by C. sakazakii.

Download Full-text

Size-Dependent Internalization Efficiency of Macrophages from Adsorbed Nanoparticle-Based Monolayers

Nanomaterials ◽

10.3390/nano11081963 ◽

2021 ◽

Vol 11 (8) ◽

pp. 1963

Author(s):

Tatiana Petithory ◽

Laurent Pieuchot ◽

Ludovic Josien ◽

Arnaud Ponche ◽

Karine Anselme ◽

...

Keyword(s):

Electron Microscopy ◽

Laser Scanning ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Raw 264.7 ◽

Functional Coatings ◽

Size Dependent ◽

Cell Internalization ◽

Biomedical Field ◽

Scanning Electron

Functional coatings based on the assembly of submicrometric or nanoparticles are found in many applications in the biomedical field. However, these nanoparticle-based coatings are particularly fragile since they could be exposed to cells that are able to internalize nanoparticles. Here, we studied the efficiency of RAW 264.7 murine macrophages to internalize physisorbed silica nanoparticles as a function of time and particle size. This cell internalization efficiency was evaluated from the damages induced by the cells in the nanoparticle-based monolayer on the basis of scanning electron microscopy and confocal laser scanning microscopy observations. The internalization efficiency in terms of the percentage of nanoparticles cleared from the substrate is characterized by two size-dependent regimes. Additionally, we highlighted that a delay before internalization occurs, which increases with decreasing adsorbed nanoparticle size. This internalization is characterized by a minimal threshold that corresponds to 35 nm nanoparticles that are not internalized during the 12-h incubation considered in this work.

Download Full-text

FEATURES OF THE APPLICATION OF ENZYME PREPARATIONS FOR THE TREATMENT AND PREVENTION OF POSTACNE

Russian Journal of Skin and Venereal Diseases ◽

10.18821/1560-9588-2017-20-4-209-213 ◽

2017 ◽

Vol 20 (4) ◽

pp. 209-213

Author(s):

Mariya N. Ostretsova ◽

E. S Snarskaya ◽

S. B Tkachenko ◽

A. S Allenova

Keyword(s):

Placebo Group ◽

Laser Scanning ◽

Acne Vulgaris ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Double Blind ◽

Inflammatory Infiltration ◽

Doppler Flowmetry ◽

Enzyme Preparations ◽

Gluteus Muscle

Postacne is a frequent consequence of vulgar acne. In our research the possibility of using hyaluronidase-based medication with prolonged action was considered. Materials and methods. Type of study: randomized, double-blind, placebo-controlled, comparative in parallel groups. 78 patients aged 18 to 35 years with moderate and severe acne vulgaris and postacne scarring were observed, patients were divided into 2 groups of 39 each by randomization. The first group received Bovhyaluronidase + Azoximer (HA) 3000 ME, the second group received a placebo. In accordance with the method of administration within the groups, patients were divided into subgroups. The drug or placebo was administered by intramuscular injection into the gluteus muscle (N = 10, 1 per 3 days) or by ultraphoonophoresis (N = 15, daily). Evaluation of the skin was performed by confocal laser scanning microscopy (CLSM) in vivo (VivaScope 1500, “Lucid Inc.”, USA) and laser Doppler flowmetry (LDF) (LAKK-01 laser capillary blood flow analyzer, “LAZMA”, Russia). The effectiveness was evaluated by the Qualitative scaring grading system and Goodman’s quantitative global acne scarring system. Results. With the use of Bovhyaluronidase + Azoximer 3000 ME improvement was in 84.6%, 5.13% of patients had a significant improvement. In the placebo group the reduction of indices was insignificant. In the drug group a statistically significant change in the microcirculation index (flux) was established. In the placebo group the dynamics of flux was less pronounced. CLSM results showed a significant decrease in fibrosis and inflammatory infiltration in the scar area in the BGA group. In inflammatory areas was reduction of fibrosis and vasodilation. In the placebo group these changes were less pronounced.

Download Full-text

Antibiofilm and Immune Response of Engineered Bioactive Nanoparticles for Endodontic Disinfection

Journal of Clinical Medicine ◽

10.3390/jcm9030730 ◽

2020 ◽

Vol 9 (3) ◽

pp. 730 ◽

Cited By ~ 2

Author(s):

Hebatullah Hussein ◽

Anil Kishen

Keyword(s):

Root Canal ◽

Laser Scanning ◽

Bacterial Load ◽

Chitosan Nanoparticles ◽

Bacterial Biofilm ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Scanning Electron Micrographs ◽

Anti Inflammatory ◽

Scanning Electron

The biological aim of root canal treatment is to facilitate periapical tissue healing following endodontic therapy. This study aimed to develop an organotypic infected root canal model to understand the interaction of bacterial biofilm with macrophages and study the therapeutic effect of engineered bioactive chitosan nanoparticles (CSnp) on macrophages. Ex-vivo experiments were conducted in two phases; Phase-1: Enterococcus faecalis biofilms (two and six weeks old) developed in organotypic root canal model were used to characterize residual biofilm after conventional chemical treatment alone and combined with CSnp utilizing Confocal Laser Scanning Microscopy, Scanning Electron Microscopy and colony-forming units from pulverized dentin. Phase-2: The interaction of post-treatment biofilm and RAW macrophages was evaluated regarding pro/anti-inflammatory markers, cell viability and spreading at 24, 48 and 72 h. Compared to conventionally disinfected six-week-old biofilm, CSnp resulted in less viable bacteria (p < 0.01). Scanning electron micrographs demonstrated disruption of the biofilm. CSnp exhibited less residual bacterial load in pulverized dentin (p < 0.001). Macrophage interaction with CSnp-treated biofilm reduced proinflammatory markers (nitric oxide, TNF-α, IL-1β, and IL-6), increased anti-inflammatory marker (TGF-β1) and enhanced cell survival and spreading over time (p < 0.01 at 72 h). Engineered chitosan nanoparticles concurrently inactivated biofilm and altered the inflammatory response of macrophages that would promote healing.

Download Full-text

Antibacterial therapy of vulgar acne

Vestnik dermatologii i venerologii ◽

10.25208/0042-4609-2019-95-5-58-67 ◽

2019 ◽

Vol 95 (5) ◽

pp. 58-67

Author(s):

E. S. Snarskaya

Keyword(s):

Quality Of Life ◽

Conflict Of Interest ◽

Laser Scanning ◽

Acne Vulgaris ◽

Blood Analysis ◽

Laser Scanning Microscopy ◽

Scanning Microscopy ◽

Confocal Laser ◽

Index Of Quality

Purpose: to evaluate the efficacy and safety of minocycline in the therapy of medium-heavy and severe forms of acne vulgaris according to modern ideas about pathogenesis of the disease.Materials and methods. Long-term multi-purpose observation which included 28 patients with the established diagnosis of acne vulgaris, medium-weight and severe forms is made. Patients received monotherapy by Minoleksin® medicine, depending on severity — 50–100 mg a day. Prior to the research and after the end of treatment inspection of the patient with calculation the rash elements, clinical and biochemical blood analysis, a research of function of sebaceous glands, the laser scanning microscopy, a laser Doppler floumetry, definition of the dermatological index of quality of life, photography (was performed at the consent of the patient). In the course of therapy inspection of the patient and assessment of tolerance of drug was performed.Results. The evidence of efficiency of minocycline in treatment of vulgar acnes of average and heavy severity is obtained that is confirmed by visual dermatological indicators, non-invasive methods of a research (results of a sebumetriya, the confocal laser scanning microscopy, a Doppler floumetriya), indicators of the dermatological index of quality of life. When studying clinical and laboratory indicators security concerns are not revealed. The undesirable phenomena in the course of the research are not registered.Conclusion. Minolexin® (minocycline) is highly effective and safe drug for treatment of vulgar acnes of average and heavy severity and it can be recommended for practical application in out-patient practice.Conflict of interest: the author states that there is no potential conflict of interest requiring disclosure in this article.

Download Full-text

Fucoxanthin-Rich Brown Algae Extract Improves Male Reproductive Function on Streptozotocin-Nicotinamide-Induced Diabetic Rat Model

International Journal of Molecular Sciences ◽

10.3390/ijms20184485 ◽

2019 ◽

Vol 20 (18) ◽

pp. 4485 ◽

Cited By ~ 6

Author(s):

Zwe-Ling Kong ◽

Sabri Sudirman ◽

Yu-Chun Hsu ◽

Chieh-Yu Su ◽

Hsiang-Ping Kuo

Keyword(s):

Reproductive Function ◽

Male Reproduction ◽

Raw 264.7 Cells ◽

Diabetic Rat ◽

Luteinizing Hormone Level ◽

Raw 264.7 ◽

Protective Effects ◽

Sprague Dawley ◽

Male Reproductive Function ◽

Anti Inflammatory

Hypogonadism and oxidative stress are occurring commonly in men with diabetes and associated male infertility. This study aimed to investigate the capability of anti-oxidative and anti-inflammatory properties of fucoxanthin as well as to evaluate its protective effects on male reproduction in diabetic rats. The RAW 264.7 macrophage cells were used to evaluate the anti-oxidative and anti-inflammatory activity. Thirty male Sprague-Dawley rats were induced by streptozotocin-nicotinamide for a diabetes model and fed either with three different doses of fucoxanthin (13, 26, and 65 mg/kg) or rosiglitazone (0.571 mg/kg) for four weeks. The fucoxanthin significantly inhibited nitric oxide production and reduced reactive oxygen species level in lipopolysaccharide-induced RAW 264.7 cells. In the animal study, fucoxanthin administration improved insulin resistance, restored sperm motility, decreased abnormal sperm number, and inhibited lipid peroxidation. Moreover, it restored GPR54 and SOCS-3 mRNA expression in the hypothalamus and recovered luteinizing hormone level, as well as the testosterone level. In conclusion, fucoxanthin not only possessed antioxidant and anti-inflammatory properties but also decreased the diabetes signs and symptoms as well as improved spermatogenesis and male reproductive function.

Download Full-text