Abstract:Background: Using human keratinocyte HaCaT cell line model, we screened for proteins that changed their content due to SDS exposure in non-toxic dose (25 μg/ml, as determined by the MTT assay and microscopic examination) during 48 h. Methods: The altered level of proteins from HaCaT keratinocytes exposed to SDS was analyzed by LC-MS/MS approach and quantified using Progenesis LC software. Results: The Pathview map of 131 upregulated proteins was built, and enhancement of glycolysis/gluconeogenesis was found.
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“…Finally, both esculentin peptides displayed a non-toxic profile on different types of mammalian cells, e.g., erythrocytes, keratinocytes, alveolar epithelial cells, and macrophages, either after a short-or long-term treatment. Importantly, HaCaT cells are a reliable and helpful in vitro model to determine the toxicity of various agents on the skin layer because keratinocytes represent 95% of the epidermal cells [61,62]. In comparison, the evaluation of the peptides' effect against lung epithelial cells, i.e., the A549 alveolar cell line, and immune cells, like macrophages, is of great interest for their crucial role in orchestrating both immune defense and inflammatory responses [63][64][65].…”
Corynebacterium jeikeium is a commensal bacterium that colonizes human skin, and it is part of the normal bacterial flora. In non-risk subjects, it can be the cause of bad body smell due to the generation of volatile odorous metabolites, especially in the wet parts of the body that this bacterium often colonizes (i.e., groin and axillary regions). Importantly, in the last few decades, there have been increasing cases of serious infections provoked by this bacterium, especially in immunocompromised or hospitalized patients who have undergone installation of prostheses or catheters. The ease in developing resistance to commonly-used antibiotics (i.e., glycopeptides) has made the search for new antimicrobial compounds of clinical importance. Here, for the first time, we characterize the antimicrobial activity of some selected frog skin-derived antimicrobial peptides (AMPs) against C. jeikeium by determining their minimum inhibitory and bactericidal concentrations (MIC and MBC) by a microdilution method. The results highlight esculentin-1b(1-18) [Esc(1-18)] and esculentin-1a(1-21) [Esc(1-21)] as the most active AMPs with MIC and MBC of 4–8 and 0.125–0.25 µM, respectively, along with a non-toxic profile after a short- and long-term (40 min and 24 h) treatment of mammalian cells. Overall, these findings indicate the high potentiality of Esc(1-18) and Esc(1-21) as (i) alternative antimicrobials against C. jeikeium infections and/or as (ii) additives in cosmetic products (creams, deodorants) to reduce the production of bad body odor.
“…Finally, both esculentin peptides displayed a non-toxic profile on different types of mammalian cells, e.g., erythrocytes, keratinocytes, alveolar epithelial cells, and macrophages, either after a short-or long-term treatment. Importantly, HaCaT cells are a reliable and helpful in vitro model to determine the toxicity of various agents on the skin layer because keratinocytes represent 95% of the epidermal cells [61,62]. In comparison, the evaluation of the peptides' effect against lung epithelial cells, i.e., the A549 alveolar cell line, and immune cells, like macrophages, is of great interest for their crucial role in orchestrating both immune defense and inflammatory responses [63][64][65].…”
Corynebacterium jeikeium is a commensal bacterium that colonizes human skin, and it is part of the normal bacterial flora. In non-risk subjects, it can be the cause of bad body smell due to the generation of volatile odorous metabolites, especially in the wet parts of the body that this bacterium often colonizes (i.e., groin and axillary regions). Importantly, in the last few decades, there have been increasing cases of serious infections provoked by this bacterium, especially in immunocompromised or hospitalized patients who have undergone installation of prostheses or catheters. The ease in developing resistance to commonly-used antibiotics (i.e., glycopeptides) has made the search for new antimicrobial compounds of clinical importance. Here, for the first time, we characterize the antimicrobial activity of some selected frog skin-derived antimicrobial peptides (AMPs) against C. jeikeium by determining their minimum inhibitory and bactericidal concentrations (MIC and MBC) by a microdilution method. The results highlight esculentin-1b(1-18) [Esc(1-18)] and esculentin-1a(1-21) [Esc(1-21)] as the most active AMPs with MIC and MBC of 4–8 and 0.125–0.25 µM, respectively, along with a non-toxic profile after a short- and long-term (40 min and 24 h) treatment of mammalian cells. Overall, these findings indicate the high potentiality of Esc(1-18) and Esc(1-21) as (i) alternative antimicrobials against C. jeikeium infections and/or as (ii) additives in cosmetic products (creams, deodorants) to reduce the production of bad body odor.
“…In contrast, several metabolic pathways, such as negative regulation of apoptotic process and epidermis development, were downregulated after treatment of HaCaT cells with SDS. Nevertheless, by conventional criteria, all the differences were considered to be not statistically significant, apparently, because doses of SDS were non-toxic (25 μg/mL) and did not lead to cell death during 48 h of exposure 6 . Figure 4 GO enrichment analysis of the proteins identified by ≥ 2 unique validated peptides in ( a ) biological process and ( b ) cellular component.…”
Section: Resultsmentioning
confidence: 99%
“…Direct contact with SDS (≤ 20%) may cause moderate inflammation, irritation of the skin, eyes, mouth, lungs, and this surfactant may slowly build up in skin cells over long-term use. Although there is no direct evidence supporting that SDS is a carcinogen 5 , we have previously demonstrated that SDS exposure resulted in the upregulation of proteins that control glycolysis 6 . There was a correlation between enhanced glycolytic ATP production and tumor malignancy 7 .…”
There is no direct evidence supporting that SDS is a carcinogen, so to investigate this fact, we used HaCaT keratinocytes as a model of human epidermal cells. To reveal the candidate proteins and/or pathways characterizing the SDS impact on HaCaT, we proposed comparative proteoinformatics pipeline. For protein extraction, the performance of two sample preparation protocols was assessed: 0.2% SDS-based solubilization combined with the 1DE-gel concentration (Protocol 1) and osmotic shock (Protocol 2). As a result, in SDS-exposed HaCaT cells, Protocol 1 revealed 54 differentially expressed proteins (DEPs) involved in the disease of cellular proliferation (DOID:14566), whereas Protocol 2 found 45 DEPs of the same disease ID. The ‘skin cancer’ term was a single significant COSMIC term for Protocol 1 DEPs, including those involved in double-strand break repair pathway (BIR, GO:0000727). Considerable upregulation of BIR-associated proteins MCM3, MCM6, and MCM7 was detected. The eightfold increase in MCM6 level was verified by reverse transcription qPCR. Thus, Protocol 1 demonstrated high effectiveness in terms of the total number and sensitivity of MS identifications in HaCaT cell line proteomic analysis. The utility of Protocol 1 was confirmed by the revealed upregulation of cancer-associated MCM6 in HaCaT keratinocytes induced by non-toxic concentration of SDS. Data are available via ProteomeXchange with identifier PXD035202.
“…The HaCaT was cultured in Dulbecco’s Modified Eagle Medium (DMEM) along with 10% FBS and 1% penicillin-streptomycin [ 19 ]. A total of 5000 cells per well were seeded in a 96-well plate.…”
Hidradenitis suppurativa (HS) has been considered an orphan disease with limited treatments available. The available topical treatment for this condition is clindamycin lotion; however, short retention and frequent application are the main setbacks. Thus, the present study aimed to attain an optimized antibacterial in situ spray formulation for the hidradenitis suppurativa skin condition, which gels once in contact with the skin surface at around 37 °C and possesses bioadhesion as well as sustained-release properties of the incorporated drug. Different concentrations of thermo-reversible gelling polymer, Pluronic F-127, were investigated along with the selected bioadhesive polymers, HPMC and SA. The optimized formulation F3 consisting of 18% Pluronic F-127 with 0.2% HPMC and 0.2% SA was characterized based on various physicochemical properties. The gelation temperature of F3 was found to be 29.0 ± 0.50 °C with a gelation time of 1.35 ± 0.40 min and a pH of 5.8. F3 had the viscosity of 178.50 ± 5.50 cP at 25 °C and 7800 ± 200 cP at 37 °C as the gel set. The optimized formulation was found to be bioadhesive and cytocompatible. Cumulative drug release was 65.05% within the time-frame of 8 h; the release pattern of the drug followed zero-order kinetics with the Higuchi release mechanism. The average zone of inhibition was found to be 43.44 ± 1.34 mm. The properties of F3 formulation reflect to improve residence time at the site of application and can enhance sustained drug release. Therefore, it could be concluded that optimized formulation has better retention and enhanced antimicrobial activity for superior efficacy against HS.
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