Normal Human Skin Fibroblasts Research Articles

Silica's silent threat: Contributing to skin fibrosis in systemic sclerosis by targeting the HDAC4/Smad2/3 pathway

Nowadays, silica products are widely used in daily life, especially in skin applications, which inevitably increases the risk of silica exposure in general population. However, inadequate awareness of silica's potential hazards and lack of self-protection are of concern. Systemic sclerosis (SSc) is characterized by progressive tissue fibrosis under environmental and genetic interactions. Silica exposure is considered an important causative factor for SSc, but its pathogenesis remains unclear. Within this study, we showed that lower doses of silica significantly promoted the proliferation, migration, and activation of human skin fibroblasts (HSFs) within 24 h. Silica injected subcutaneously into mice induced and exacerbated skin fibrosis. Notably, silica increased histone deacetylase-4 (HDAC4) expression by inducing its DNA hypomethylation in normal HSFs. The elevated HDAC4 expression was also confirmed in SSc HSFs. Furthermore, HDAC4 was positively correlated with Smad2/3 phosphorylation and COL1, α-SMA, and CTGF expression. The HDAC4 inhibitor LMK235 mitigated silica-induced upregulation of these factors and alleviated skin fibrosis in SSc mice. Taken together, silica induces and exacerbates skin fibrosis in SSc patients by targeting the HDAC4/Smad2/3 pathway. Our findings provide new insights for evaluating the health hazards of silica exposure and identify HDAC4 as a potential interventional target for silica-induced SSc skin fibrosis.

Senescence-Associated Heterochromatin Foci Suppress γ-H2AX Focus Formation Induced by Radiation Exposure.

DNA damage is induced by both endogenous and exogenous factors. Repair of DNA double-strand break (DSB), a serious damage that threatens genome stability, decreases with senescence. However, the molecular mechanisms underlying the decline in DNA repair capacity during senescence remain unclear. We performed immunofluorescence staining for phosphorylated histone H2AX (γ-H2AX) in normal human fetal lung fibroblasts and human skin fibroblasts of different ages after chronic irradiation (total dose, 1 Gy; dose rate, 1 Gy/day) to investigate the effect of cellular senescence and organismal aging on DSB repair. Accumulation of DSBs was observed with cellular senescence and organismal aging, probably caused by delayed DSB repair. Importantly, the formation of γ-H2AX foci, an early event in DSB repair, is delayed with cellular senescence and organismal aging. These results suggest that the delay in γ-H2AX focus formation might delay the overall DSB repair. Interestingly, immediate γ-H2AX foci formation was suppressed in cells with senescence-associated heterochromatin foci (SAHF). To investigate the relationship between the γ-H2AX focus formation and SAHF, we used LiCl to relax the SAHFs, followed by irradiation. We demonstrated that LiCl rescued the delayed γ-H2AX foci formation associated with cellular senescence. This indicates that SAHF interferes with γ-H2AX focus formation and inhibits DSB repair in radiation-induced DSB. Our results suggest that therapeutic targeting of SAHFs have potential to resolve DSB repair dysfunction associated with cellular senescence.

Development of propolis, hyaluronic acid, and vitamin K nano-emulsion for the treatment of second-degree burns in albino rats

Burns are the fourth most common type of injury worldwide. Many patients also suffer numerous infections and complications that impair the burn healing process, which makes the treatment of burns a challenge. This study aimed to prepare and characterize nano-emulsion (NE) of propolis, hyaluronic acid, and vitamin K for treatment of second-degree burns. High-Pressure Liquid Chromatography (HPLC) was used for the qualitative assessment of the phenolic and flavonoid contents in crude propolis. The structural, optical, and morphological characterization, besides the antimicrobial, antioxidant, cytotoxicity, in-vitro, and in-vivo wound healing activities were evaluated. For in-vivo study, 30 adult male albino rats were divided randomly into control and treated groups, which were treated with normal saline (0.9%), and NE, respectively. The wounds were examined clinicopathologically on the 3rd, 7th, and 14th days. The NE revealed the formation of a mesh-like structure with a size range of 80–180 nm and a 21.6 ± 6.22 mV zeta potential. The IC50 of NE was 22.29 μg/ml. Also, the NE showed antioxidant and antimicrobial activity against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. The in-vitro investigation of the NE on normal human skin fibroblasts using scratch assay proved an acceleration for wound healing. The treated rats showed improved wound healing clinically and pathologically and wound contraction percent (WC %) was 98.13% at 14th day, also increased epithelization, fibrous tissue formation, collagen deposition, and angiogenesis compared to the control. It could be concluded that the prepared NE possesses antimicrobial, antioxidant, and healing effect in the treatment of second-degree burns.

Functionalized Carbon Nanotubes for Delivery of Ferulic Acid and Diosgenin Anticancer Natural Agents.

It was investigated whether loading multi-wall carbon nanotubes (CNTs) with two natural anticancer agents: ferulic acid (FUA) and diosgenin (DGN), may enhance the anticancer effect of these drugs. The CNTs were functionalized with carboxylic acid (CNTCOOH) or amine (CNTNH2), loaded with the above pro-drugs, as well as both combined and coated with chitosan or chitosan-stearic acid. Following physicochemical characterization, the drug-loading properties and kinetics of the drug's release were investigated. Their effects on normal human skin fibroblasts and MCF-7 breast carcinoma cells, HepG2 hepatocellular carcinoma cells, and A549 non-small-cell lung cancer cells were evaluated in vitro. Their actions at the molecular level were evaluated by assessing the expression of lncRNAs (HULC, HOTAIR, CCAT-2, H19, and HOTTIP), microRNAs (mir-21, mir-92, mir-145, and mir-181a), and proteins (TGF-β and E-cadherin) in HepG2 cells. The release of both pro-drugs depended on the glutathione concentration, coating, and functionalization. Release occurred in two stages: a no-burst/zero-order release followed by a sustained release best fitted to Korsmeyer-Peppas kinetics. The combined nanoformulation cancer inhibition effect on HepG2 cancer cells was more pronounced than for A549 and MCF7 cells. The combined nanoformulations had an additive impact followed by a synergistic effect, with antagonism demonstrated at high concentrations. The nanoformulation coated with chitosan and stearic acid was particularly successful in targeting HepG2 cells and inducing apoptosis. The CNT functionalized with carboxylic acid (CNTCOOH), loaded with both FUA and DGN, and coated with chitosan-stearic acid inhibited the expression of lncRNAs and modulated both microRNAs and proteins. Thus, nanoformulations composed of functionalized CNTs dual-loaded with FUA and DGN and coated with chitosan-stearic acid are a promising drug delivery system that enhances the activity of natural pro-drugs.

CLC-3 regulates TGF-β/smad signaling pathway to inhibit the process of fibrosis in hypertrophic scar

ObjectiveTo study the role and mechanism of chloride channel-3 (ClC-3) in the formation of hypertrophic scar by constructing ClC-3 interference vectors and examining their effects on human hypertrophic scar fibroblasts (HSFB). MethodsHuman HSFB and human normal skin fibroblasts (NSFB) were used in this study, and ClC-3 interference vectors were constructed to transfect cells. ClC-3 inhibitors NPPB and Tamoxifen were used to treat cells. Cell migration and the expression of TGF-β/Smad, CollagenⅠ,CollagenⅢ were examined to explore the role of ClC-3 in the formation of hypertrophic scar. ResultsCompared with the normal skin tissue, the positive expression of ClC-3 and TGF–β in the scar tissue was significantly increased. The relative expression of ClC-3 and TGF-β1 in HSFB cells was higher than that in NSFB cells. Interfering with the expression of CLC-3 can inhibit the migration of HSFB cells and the expression of TGF- β/Smad, CollagenⅠ/Ⅲ. The experiment of HSFB cells treated by CLC-3 inhibitors can also obtain similar results. ConclusionInhibiting CLC-3 can reduce the formation of hypertrophic scars.

Electrospun Tamarindus indica-loaded antimicrobial PMMA/cellulose acetate/PEO nanofibrous scaffolds for accelerated wound healing: In-vitro and in-vivo assessments

In this work, Tamarindus indica (T. indica)-loaded crosslinked poly(methyl methacrylate) (PMMA)/cellulose acetate (CA)/poly(ethylene oxide) (PEO) electrospun nanofibers were designed and fabricated for wound healing applications. T. indica is a plant extract that possesses antidiabetic, antimicrobial, antioxidant, antimalarial and wound healing properties. T. indica leaves extract of different concentrations were blended with a tuned composition of a matrix comprised of PMMA (10 %), CA (2 %) and PEO (1.5 %), and were electrospun to form smooth, dense and continuous nanofibers as illustrated by SEM investigation. In vitro evaluation of T. indica-loaded nanofibers on normal human skin fibroblasts (HBF4) revealed a high compatibility and low cytotoxicity. T. indica-loaded nanofibers significantly increased the healing activity of scratched HBF4 cells, as compared to the free plant extract, and the healing activity was significantly enhanced upon increasing the plant extract concentration. Moreover, T. indica-loaded nanofibers demonstrated significant antimicrobial activity in vitro against the tested microbes. In vivo, nanofibers resulted in a superior wound healing efficiency compared to the control untreated animals. Hence, engineered nanofibers loaded with potent phytochemicals could be exploited as an effective biocompatible and eco-friendly antimicrobial biomaterials and wound healing composites.

Copper(II) Complexes with 1-(Isoquinolin-3-yl)heteroalkyl-2-ones: Synthesis, Structure and Evaluation of Anticancer, Antimicrobial and Antioxidant Potential.

Four copper(II) complexes, C1-4, derived from 1-(isoquinolin-3-yl)heteroalkyl-2-one ligands L1-4 were synthesized and characterized using an elemental analysis, IR spectroscopic data as well as single crystal X-ray diffraction data for complex C1. The stability of complexes C1-4 under conditions mimicking the physiological environment was estimated using UV-Vis spectrophotometry. The antiproliferative activity of both ligands L1-4 and copper(II) compounds C1-4 were evaluated using an MTT assay on four human cancer cell lines, A375 (melanoma), HepG2 (hepatoma), LS-180 (colon cancer) and T98G (glioblastoma), and a non-cancerous cell line, CCD-1059Sk (human normal skin fibroblasts). Complexes C1-4 showed greater potency against HepG2, LS180 and T98G cancer cell lines than etoposide (IC50 = 5.04-14.89 μg/mL vs. IC50 = 43.21->100 μg/mL), while free ligands L1-4 remained inactive in all cell lines. The prominent copper(II) compound C2 appeared to be more selective towards cancer cells compared with normal cells than compounds C1, C3 and C4. The treatment of HepG2 and T98G cells with complex C2 resulted in sub-G1 and G2/M cell cycle arrest, respectively, which was accompanied by DNA degradation. Moreover, the non-cytotoxic doses of C2 synergistically enhanced the cytotoxic effects of chemotherapeutic drugs, including etoposide, 5-fluorouracil and temozolomide, in HepG2 and T98G cells. The antimicrobial activities of ligands L2-4 and their copper(II) complexes C2-4 were evaluated using different types of Gram-positive bacteria, Gram-negative bacteria and yeast species. No correlation was found between the results of the antiproliferative and antimicrobial experiments. The antioxidant activities of all compounds were determined using the DPPH and ABTS radical scavenging methods. Antiradical tests revealed that among the investigated compounds, copper(II) complex C4 possessed the strongest antioxidant properties. Finally, the ADME technique was used to determine the physicochemical and drug-likeness properties of the obtained complexes.

Phytochemical Profile, Antioxidant Potential, Antimicrobial Activity, and Cytotoxicity of Dry Extract from Rosa damascena Mill.

Dry rose extract (DRE) obtained industrially by aqueous ethanol extraction from R. damascena flowers and its phenolic-enriched fraction, obtained by re-extraction with ethyl acetate (EAE) were the subject of this study. 1H NMR of DRE allowed the identification and quantitation of fructose and glucose, while the combined use of HPLC-DAD-ESIMS and HPLC-HRMS showed the presence of 14 kaempferol glycosides, 12 quercetin glycosides, 4 phenolic acids and their esters, 4 galloyl glycosides, 7 ellagitannins, and quinic acid. In addition, the structures of 13 of the flavonoid glycosides were further confirmed by NMR. EAE was found to be richer in TPC and TFC and showed better antioxidant activity (DPPH, ABTS, and FRAP) compared to DRE. Both extracts displayed significant activity against Propionibacterium acnes, Staphylococcus aureus, and S. epidermidis, but showed no activity against Candida albicans. Toxicity tests on normal human skin fibroblasts revealed low toxicity for both extracts with stronger effects observed at 24 hours of treatment that were compensated for over the following two days. Human hepatocarcinoma (HepG2) cells exhibited an opposite response after treatment with a concentration above 350 µg/mL for EAE and 500 µg/mL for DRE, showing increased toxicity after the third day of treatment. Lower concentrations were non-toxic and did not significantly affect the cell cycle parameters of either of the cell lines.

Antibacterial and antibiofilm effects of Camellia oleifera seed dreg extract and its application in cosmetics.

Cosmetic care products contain a high proportion of water and nutrients. Therefore, preventing bacterial growth is an important issue to ensure product quality and safety. The application of antibacterial natural ingredients derived from plants is considered to have the potential to maintain product quality and reduce the use of chemicals in formulations. Additionally, chemically synthesized antiseptic and antibacterial agents are widely used in the industry at present. However, some preservative ingredients have been reported that may cause skin irritation, redness, allergies, and even dermatitis. This study aimed to prepare extract from Camellia oleifera tea seed dregs (CTSD), investigate the antibacterial effects on two pathogenic bacteria and evaluate the product preservative ability. Ethanol extraction was prepared and subjected to characterize their triterpenoid contents. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and minimum biofilm eradication concentration (MBEC) were determined for Pseudomonas aeruginosa and Staphylococcus aureus. The product's stability and preservative qualities, along with its ability to scavenge free radicals through antioxidant activity, were also assessed. The gram-positive S. aureus showed greater susceptibility to the treatment. In additional, CTSD possessed significant free radical scavenging activity invitro and cultured normal human skin fibroblast CCD-966SK cells under nontoxic concentration. The challenge test and accelerated storage test confirmed the CTSD containing formulated emulsion is eligible for commercialization. CTSD has the potential to be developed as an alternative agent to control microbial biofilm formation, or can be used as an adjuvant compound for infectious disease control.

Evaluation of cytotoxic and genotoxic potential of avobenzone and octocrylene on human skin fibroblast cells

We investigated the cytotoxic and genotoxic effects of the organic UVA filter avobenzone (AVB), and the UVB filter octocrylene (OCT), found alone or in combination in different sunscreen formulations, on normal human skin fibroblast cells (CCD-1118Sk). To achieve this purpose, we used XTT viability assay, single cell gel electrophoresis (comet) assay and reactive oxygen species (ROS) assay. For cytotoxicity, CCD-1118Sk cells were exposed to various concentrations of AVB (32–800 µM), and OCT (130–6500 μM). The IC50 values were 100.2 and 1390.95 µM for AVB and OCT, respectively. IC12.5, IC25 and IC50 concentrations were chosen for genotoxicity testing. The comet assay revealed DNA damage at the two highest concentrations of AVB and all OCT concentrations. The rate of DNA damage was significantly higher than in the control groups. Co-treatment with AVB and OCT increased the level of DNA damage and intracellular oxidative stress compared with AVB and OCT treatment alone. Finally, our study showed that the UVA filter AVB, with the UVB filter OCT, may induce cytotoxic and genotoxic effects on human skin fibroblast cells. Highlights Avobenzone and octocrylene are UV filters used in sunscreen products. Healthy human skin fibroblast cell line CCD-1118Sk was used for cytotoxic and genotoxic assays. Cotreatment with avobenzone and octocrylene caused more severe DNA damage than their treatment alone. Our findings will contribute to the limited number of cytotoxicity and genotoxicity studies and lead to more detailed studies on the use of these two chemicals.

Nanoemulsion of Zanthoxylum rhetsa (Roxb.) DC: Formulation, release characteristics and its potential use for anti-inflammation applications

Nanoemulsions of Zanthoxylum rhetsa (Roxb.) DC fruit extract combined with eucalyptus oil (ZRNEs) were formulated using mixed surfactants at various Tween 80/Span 80 mass ratios of 20/80 (ZRNE-6), 50/50 (ZRNE-10), and 80/20 (ZRNE-13). These formulations had corresponding hydrophilic-lipophilic balance (HLB) values of 6, 10, and 13, respectively. Effect of the HLB on properties of the nanoemulsions were evaluated. Among these formulations, ZRNE-10, was found to be optimal for loading ZR extract with eucalyptus. It exhibited the smallest particle size of 402 ± 12 nm with a PDI of 0.41 ± 0.01, as analyzed by dynamic light scattering technique. ZRNE-10 showed the highest encapsulation efficiency for terpinen-4-ol at 83.42 ± 0.51%. It demonstrated a gradual release profile, releasing 30% of terpinen-4-ol within 48 h in a pH 7.4 PBS buffer at 37 °C. Interestingly, higher HLB values accelerated the release rate of terpinen-4-ol. The release kinetics of ZRNE-10 were best described by the Higuchi model (R2 = 0.903). ZRNE-10 displayed robust antioxidant activity (59.5 ± 1.6%) and effective inhibition of nitric oxide (76.9 ± 0.8%). It also exhibited low cytotoxicity on normal human skin fibroblasts (IC50 ≈ 3 mg/mL). Stability assessments revealed that all ZRNE formulations maintained their physical attributes and bioactivities even after one month of storage at 25 °C, and even at an elevated temperature of 45 °C. Notably, the antioxidant activities of ZRNEs surpassed those of unencapsulated ZR extract. Given their favorable anti-inflammatory properties, low cytotoxicity, and maintained efficacy, ZRNEs hold promise as potential ingredients for topical use.

Optimization and characterization of antileukemic l-asparaginase produced by Fusarium solani endophyte

l-asparaginase is an antileukemic enzyme that hydrolyzes l-asparagine into l-aspartic acid and ammonia, causing cell starvation and apoptosis in susceptible leukemic cell populations. Currently, l-asparaginase obtained from bacterial sources is constrained by several issues, including lesser productivity, stability, selectivity, and higher toxicity. The goal of this study is to provide fungal l-asparaginase with in-vitro effectiveness towards different human carcinomas. l-asparaginase from endophytic Fusarium solani (Gene Bank accession number MW209717) isolated from the roots of the medicinal plant Hedera helix L. was characterized and optimized experimentally for maximum l-asparaginase production in addition to evaluating its subsequent cytotoxicity towards acute monocytic leukemia and human skin fibroblast cell lines. The enzyme production was maximized using potato dextrose media (15.44 IU/ml/hr) at the 5th and 6th days of fermentation with incubation temperature 30 °C, 3% asparagine, 150–180 rpm agitation rate and a 250 ml flask. Enzyme characterization studies revealed that the enzyme maintained its thermal stability with temperatures up to 60 °C. However, its optimal activity was achieved at 35 °C. On measuring the enzymatic activity at various temperatures and different pH, maximum enzyme activity was recorded at 40 °C and pH 8 using 0.1 M asparagine concentration. Results also revealed promising cytotoxic activity against acute monocytic leukemia with IC50 = 3.66 µg/ml and low cytotoxicity against tested normal human skin fibroblast cell line which suggested that it might have selective toxicity, and consequently it could be used as a less toxic alternative to the current formulations.

Utilization of biosynthesized silver nanoparticles from Agaricus bisporus extract for food safety application: synthesis, characterization, antimicrobial efficacy, and toxicological assessment

The emergence of antimicrobial resistance in foodborne bacterial pathogens has raised significant concerns in the food industry. This study explores the antimicrobial potential of biosynthesized silver nanoparticles (AgNPs) derived from Agaricus bisporus (Mushroom) against foodborne bacterial pathogens. The biosynthesized AgNPs were characterized using various techniques, including UV–visible spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, high-resolution scanning electron microscopy with energy dispersive X-ray spectroscopy, dynamic light scattering, and zeta potential analysis. The antibacterial activity of the AgNPs was tested against a panel of foodborne bacterial strains, and their cytotoxicity was evaluated on normal human skin fibroblasts. Among the tested strains, Pseudomonas aeruginosa ATCC 27853 showed the highest sensitivity with an inhibition zone diameter (IZD) of 48 mm, while Klebsiella quasipneumoniae ATTC 700603 and Bacillus cereus ATCC 11778 displayed the highest resistance with IZDs of 20 mm. The silver cations released by AgNPs demonstrated strong bactericidal effects against both Gram-positive (G + ve) and Gram-negative (G − ve) bacteria, as evidenced by the minimum inhibitory concentration/minimum bactericidal concentration (MBC/MIC) ratio. Moreover, cytotoxicity testing on normal human skin fibroblasts (HSF) indicated that AgNPs derived from the mushroom extract were safe, with a cell viability of 98.2%. Therefore, AgNPs hold promise as an alternative means to inhibit biofilm formation in the food industry sector.

Green Synthesis of Silver and Titanium Oxide Nanoparticles Using Tea and Eggshell Wastes, Their Characterization, and Biocompatibility Evaluation

Using biodegradable wastes represents a viable alternative to creating a sustainable economy that benefits all humans. The present study aimed to use daily used waste products, tea (TE) and eggshell (ES) wastes, to synthesize silver (AgNPs) and titanium oxide (TiO2NPs) nanoparticles, respectively. Firstly, the green-synthesized nanoparticles were characterized using an ultraviolet-visible spectrophotometer (UV-VIS), Scanning (SEM), transmission electron microscope (TEM), Dynamic light scattering (DLS), zeta potential analysis, X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. Then, followed by their cytotoxic assessment against normal human skin fibroblast (HSF) cells using sulforhodamine B (SRB) assay, AgNPs_TE (300 and 470 nm) and TiO2NPs_ESE (320 nm) formation was confirmed using UV-vis spectra. SEM and XRD showed their crystalline shape. TEM images determined the nano-size of AgNPs_TE (25 nm) and TiO2NPs_ESE (120 nm), which appeared smaller in comparison with DLS analysis (299.8 and 742.9 nm), with zeta potentials of −20.5 mV and −12.6 mV, respectively. There was a great difference in both NPs’ sizes using TEM and DLS measurements because DLS is known to be more sensitive to larger particles due to their light scattering. FTIR detected the functional groups found in TE and ESE that were responsible for the synthesis, capping, and stabilization of the synthesized AgNPs and TiO2NPs. The SRB assay reveals the safety of TiO2NPs on normal HSF cells with an IC50 > 100, while AgNPs have a high cytotoxic effect with an IC50 = 54.99 μg/mL.

Design and Study of Nanoceria Modified by 5-Fluorouracil for Gel and Polymer Dermal Film Preparation.

In this work we studied nanoceria (CeO2NPs) and nanoceria modified by 5-fluorouracil (5FU) as potential APIs. Nanoceria were synthesized by precipitation in a matrix of hydroxyethyl cellulose or hydroxypropylmethyl cellulose, using cerium (III) nitrate and meglumine. Nanoceria properties were estimated by UV, FTIR and X-ray photoelectron spectra; scanning electron and atomic force microscopy; powder X-ray diffraction patterns and energy dispersive X-ray microanalysis. The cytotoxicity of nanoceria and polymer-protected nanoparticles was evaluated using the established cell line NCTC clone 929 (C3H/An mouse, subcutaneous connective tissue, clone of L. line). The morphology and metabolic activity of nanoparticles at 10 μg∙mL-1 of cells was not significant. In addition, the cytotoxic effects of nanoceria were assessed on two human colorectal cancer cell lines (HT29 and HCT116), murine melanoma B16 cells and normal human skin fibroblasts. An inhibitory effect was shown for HCT116 human colorectal cancer cells. The IC50 values for pure CeO2NPs and CeO2NPs-5FU were 219.0 ± 45.6 μg∙mL-1 and 89.2 ± 14.0 μg∙mL-1, respectively. On the other hand, the IC50 of 5FU in the combination of CeO2NPs-5FU was 2-fold higher than that of pure 5FU, amounting to 5.0 nmol∙mL-1. New compositions of nanoceria modified by 5-fluorouracil in a polymer matrix were designed as a dermal polymer film and gel. The permeability of the components was studied using a Franz cell.

Characteristics of Serum Exosomes after Burn Injury and Dermal Fibroblast Regulation by Exosomes In Vitro.

(1) Background: Exosomes (EXOs) have been considered a new target thought to be involved in and treat wound healing. More research is needed to fully understand EXO characteristics and the mechanisms of EXO-mediated wound healing, especially wound healing after burn injury. (2) Methods: All EXOs were isolated from 85 serum samples of 29 burn patients and 13 healthy individuals. We characterized the EXOs for morphology and density, serum concentration, protein level, marker expression, size distribution, and cytokine content. After a confirmation of EXO uptake by dermal fibroblasts, we also explored the functional regulation of primary human normal skin and hypertrophic scar fibroblast cell lines by the EXOs in vitro, including cell proliferation and apoptosis. (3) Results: EXOs dynamically changed their morphology, density, size, and cytokine level during wound healing in burn patients, which were correlated with burn severity and the stages of wound healing. EXOs both from burn patients and healthy individuals stimulated dermal fibroblast proliferation and apoptosis. (4) Conclusions: EXO features may be important signals that influence wound healing after burn injury; however, to understand the mechanisms by which EXOs regulates the fibroblasts in healing wounds, further studies will be required.

Sustainable Eco-Friendly Synthesis of Zinc Oxide Nanoparticles Using Banana Peel and Date Seed Extracts, Characterization, and Cytotoxicity Evaluation

This study describes the use of banana peel (BPEs) and date seed extracts (DSEs) as waste products in the sustainable and eco-friendly biological synthesis of zinc oxide nanoparticles (ZnONPs). ZnONPs_BPE and ZnONPs_DSE were characterized using an ultraviolet-visible spectrophotometer (UV-VIS), Scanning (SEM), transmission electron microscope (TEM), X-ray diffraction (XRD), dynamic light scattering (DLS), zeta potential analysis, and Fourier transform infrared (FTIR) spectroscopy. Moreover, the biocompatibility of ZnONPs was analyzed against the normal human skin fibroblast (HSF) cell line. Peaks of UV spectra were 300 nm and 400 nm for ZnONPs-BPE and for ZnONP _DSE, respectively, confirming the ZnONPs’ formation. XRD revealed their hexagonal structure. SEM showed the nanocrystals of ZnONPs_BPE, which are interlinked to one another in a uniform shape, while ZnONPs_DSE appear as large and small chunky crystals. The mean size of ZnONPs_BPE and ZnONPs_DSE was 50 nm and 62 nm using TEM, respectively. On the contrary, their mean size was bigger using DLS with the zeta potential of ZnONPs_BPE = −12.7 mV and ZnONPs_DSE = −5.69 mV. The FTIR analysis demonstrated the presence of carboxyl, hydroxyl, and C–H of cellulose, hemicelluloses, and lignin polymers on ZnONPs surfaces that act as reducing, capping, and stabilizing agents. ZnONPs_BPE (IC50 > 100) have lower cytotoxic effects on HSF cells than ZnONPs_DSE (IC50 = 29.34 μg/mL). The present study indicates the successful synthesis of ZnONPs using agro-wastes that could help in waste management and recycling. Furthermore, ZnONPs_BPE is safe to use for further applications.

Herniarin-loaded solid lipid nanoparticles: promising molecular mechanism and therapeutic potential against pancreatic cancer line.

The notion of cancer therapy is intrinsically subjected to multiple challenges due to the drug resistance and drug toxicity for normal tissues. Herniarin (7-methoxycoumarin) belongs to the naturally occurring aromatic phytochemicals and coumarins. Considering the boosting effect of nanocarriers in drug delivery, we investigated the proapoptotic, anti-metastatic properties, and molecular mechanism of herniarin-loaded solid lipid nanoparticles on human gastric adenocarcinoma (AGS), human colon adenocarcinoma (HT-29), human pancreatic carcinoma (Panc-1), and normal human skin fibroblast (HFF) cell lines. The cytotoxicity of synthesized nanoparticle have been tested using MTT assay. The obtained results manifested that concentration of herniarin that exerts 50% cell growth inhibition (IC50) against HT-29, AGS, and Panc-1 was calculated 138.34, 123.46, and 83.744 µL, respectively. Given that nanoparticles showed lowest IC50 values on Panc-1 cell line, these cells were selected for further analysis. The apoptosis induction and cell cycle arrest were examined performing real-time PCR, flow cytometry, and DAPI/acridine orange-propidium iodide staining. The expression of apoptosis-related genes, including BCL-2, was decreased, while the expression of CASP9, CASP8, and CASP3 was increased in response to the treatment. Moreover, the expression of metastasis-related gene (MMP2) was significantly suppressed under Her-SLN-NPs treatment. According to the flow cytometry findings, we observed no cell cycle arrest at any stage. Our funding manifested herniarin encapsulated solid lipid nanoparticles has potent therapeutic target against Panc-1 cell line.

Design of novel water-soluble isoxazole-based antimicrobial agents and evaluation of their cytotoxicity and acute toxicity

Based on the readily available 3-organyl-5-(chloromethyl)isoxazoles, a number of previously unknown water-soluble conjugates of isoxazoles with thiourea, amino acids, some secondary and tertiary amines, and thioglycolic acid were synthesized. The bacteriostatic activity of aforementioned compounds has been studied against Enterococcus durans B-603, Bacillus subtilis B-407, Rhodococcus qingshengii Ac-2784D, and Escherichia coli B-1238 microorganisms (provided by All-Russian Collection of Microorganisms, VKM). The influence of the nature of the substituents in positions 3 and 5 of the isoxazole ring on the antimicrobial activity of the obtained compounds has been determined. It is found that the highest bacteriostatic effect is observed for compounds containing 4-methoxyphenyl or 5-nitrofuran-2-yl substituents in position 3 of the isoxazole ring as well as methylene group in position 5 bearing residues of l-proline or N-Ac-l-cysteine (5a–d, MIC 0.06–2.5 µg/ml). The leading compounds showed low cytotoxicity on normal human skin fibroblast cells (NAF1nor) and low acute toxicity on mice in comparison with the well-known isoxazole-containing antibiotic oxacillin.

Core-Shell Nanostructured Drug Delivery Platform Based on Biocompatible Metal-Organic Framework-Ligated Polyethyleneimine for Targeted Hepatocellular Carcinoma Therapy.

Multifunctional nanosized metal-organic frameworks (NMOFs) have advanced rapidly over the past decade to develop drug delivery systems (DDSs). These material systems still lack precise and selective cellular targeting, as well as the fast release of the quantity of drugs that are simply adsorbed within and on the external surface of nanocarriers, which hinders their application in the drug delivery. Herein, we designed a biocompatible Zr-based NMOF with an engineered core and the hepatic tumor-targeting ligand, glycyrrhetinic acid grafted to polyethyleneimine (PEI) as the shell. The improved core-shell serves as a superior nanoplatform for efficient controlled and active delivery of the anticancer drug doxorubicin (DOX) against hepatic cancer cells (HepG2 cells). In addition to their high loading capacity of 23%, the developed nanostructure DOX@NMOF-PEI-GA showed an acidic pH-stimulated response and extended the drug release time to 9 days as well as enhanced the selectivity toward the tumor cells. Interestingly, the DOX-free nanostructures showed a minimal toxic effect on both normal human skin fibroblast (HSF) and hepatic cancer cell line (HepG2), but the DOX-loaded nanostructures exhibited a superior killing effect toward the hepatic tumor, thus opening the way for the active drug delivery and achieving efficient cancer therapy applications.