Human Skin Fibroblast Cells Research Articles

Expression Strategy of Soluble Recombinant Human TGF-β3 in Escherichia coli: sfGFP -Fusion Tag

Transforming growth factor-beta 3 (TGF-β3) is an important cytokine involved in various biological processes. TGF-β3 is used as a scar-reducing antifibrotic agent for acute and chronic wounds and fibrosing disorders. TGF-β3, a valuable therapeutic protein, is produced recombinantly in different expression systems. TGF-β3, produced in the Escherichia coli (E. coli) expression system, widely used due to its various advantages in recombinant production, is commercially available. However, the main problem encountered in protein expression in E. coli cells is the formation of an inclusion body. Various approaches have been developed to solve this problem. The use of a fusion tag is one of the most powerful strategies used to obtain protein in the soluble active form in the E. coli expression system. Superfolder GFP (sfGFP) is one of the fusion tags used to increase the solubility of the fusion partner in E. coli. In this study, TGF-β3 with sfGFP fusion tag (sfGFP-TGFβ3) was successfully produced in soluble form in E. coli BL21 (DE3) in high yield and purity for the first time. Purified protein was identified by western blot and SDS-PAGE. 20 mg of protein with 98% purity was obtained from 1 L of bacterial culture. It was determined that the obtained high purity protein did not have a cytotoxic effect on BJ normal human skin fibroblast cells. The impact of sfGFP-TGFβ3 fusion protein on wound healing was evaluated with in vitro scratch wound healing assay. The results showed that the sfGFP-TGFβ3 fusion protein produced in soluble form in the E. coli expression system has the potential to support the wound healing process.

Biosynthesis of Novel Tellurium Nanorods by Gayadomonas sp. TNPM15 Isolated from Mangrove Sediments and Assessment of Their Impact on Spore Germination and Ultrastructure of Phytopathogenic Fungi

The biosynthesis of nanoparticles using green technology is emerging as a cost-efficient, eco-friendly and risk-free strategy in nanotechnology. Recently, tellurium nanoparticles (TeNPs) have attracted growing attention due to their unique properties in biomedicine, electronics, and other industrial applications. The current investigation addresses the green synthesis of TeNPs using a newly isolated mangrove-associated bacterium, Gayadomonas sp. TNPM15, and their impact on the phytopathogenic fungi Fusarium oxysporum and Alternaria alternata. The biogenic TeNPs were characterized using transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), Raman spectroscopy and Fourier transform infrared (FTIR). The results of TEM revealed the intracellular biosynthesis of rod-shaped nanostructures with a diameter range from 15 to 23 nm and different lengths reaching up to 243 nm. Furthermore, the successful formation of tellurium nanorods was verified by SEM-EDX, and the XRD pattern revealed their crystallinity. In addition, the FTIR spectrum provided evidence for the presence of proteinaceous capping agents. The bioinspired TeNPs exhibited obvious inhibitory effect on the spores of both investigated phytopathogens accomplished with prominent ultrastructure alternations, as evidenced by TEM observations. The biogenic TeNPs impeded spore germination of F. oxysporum and A. alternata completely at 48.1 and 27.6 µg/mL, respectively. Furthermore, an increase in DNA and protein leakage was observed upon exposure of fungal spores to the biogenic TeNPs, indicating the disruption of membrane permeability and integrity. Besides their potent influence on fungal spores, the biogenic TeNPs demonstrated remarkable inhibitory effects on the production of various plant cell wall-degrading enzymes. Moreover, the cytotoxicity investigations revealed the biocompatibility of the as-prepared biogenic TeNPs and their low toxicity against the human skin fibroblast (HSF) cell line. The biogenic TeNPs showed no significant cytotoxic effect towards HSF cells at concentrations up to 80 μg/mL, with a half-maximal inhibitory concentration (IC50) value of 125 μg/mL. The present work spotlights the antifungal potential of the biogenic TeNPs produced by marine bacterium against phytopathogenic fungi as a promising candidate to combat fungal infections.

One-pot microwave synthesis of chitosan-stabilized silver nanoparticles entrapped polyethylene oxide nanofibers, with their intrinsic antibacterial and antioxidant potency for wound healing

Different physical and chemical techniques could be used to prepare chitosan/Silver nanoparticle (CHS/AgNPs) nanocomposite. The microwave heating reactor was rationally adopted as a benign tool for preparing CHS/AgNPs owing to less energy consumption and shorter time required for completing the nucleation and growth particles. UV–Vis, FTIR, and XRD, provided conclusive evidence of the AgNPs creation, while TEM micrographs elucidated that the size was spherical (20 nm). CHS/AgNPs were embedded in polyethylene oxide (PEO) nanofiber via electrospinning, and their biological properties, cytotoxicity evaluation, antioxidant, and antibacterial activity assays were investigated. The generated nanofibers have mean diameters of 130.9 ± 9.5, 168.7 ± 18.8, and 186.8 ± 8.19 nm for PEO, PEO/ CHS, and PEO/ CHS (AgNPs), respectively. Because of the tiny AgNPs particle size loaded in PEO/CHS (AgNPs) fabricated nanofiber, good antibacterial activity with ZOI against E. coli was 51.2 ± 3.2, and S. aureus was 47.2 ± 2.1 for PEO/ CHS (AgNPs) nanofibers. Non-toxicity was observed against Human Skin Fibroblast and Keratinocytes cell lines (>93.5 %), which justifies its great antibacterial potential to remove or prevent infection in wounds with fewer adverse effects.

7S Globulin from Soybean Is Metabolized in Human Cell Cultures by a Specific Uptake and Degradation System

We examined the biological fate of 7S globulin from soybean in a hepatoma cell line (Hep G2) and in human skin fibroblasts (HSF) to gain new insights into the 7S globulin cell process, the final effect of which is an enhanced expression of the LDL-receptor. The ability of 7S globulin to bind and to be internalized and degraded by both cell types was investigated under different experimental conditions. In all cases, specific uptake (binding + internalization) and degradation of 125I-7S globulin were curvilinear functions of substrate concentration at 37°C. The two processes were saturated at around 80 mg/L, a concentration at which an up-regulation of LDL-receptor was previously reported. The specific uptake of 125I-7S globulin at 37°C was a curvilinear function of time, and achieved equilibrium after 6 and 12 h in HSF and Hep G2 cells, respectively. Binding experiments, conducted at 4°C in Hep G2 cells, showed a specific and saturable association of 7S globulin to the cell membrane. Linear Scatchard analysis demonstrated a single population of binding sites. The amount of 7S globulin bound at saturation (Bmax) was about 2.73 mg/L, with an apparent Kd of 21 µmol/L, assuming 175 kDa as the 7S globulin molecular weight. SDS-PAGE of Hep G2 membrane proteins incubated with 125I-7S globulin revealed a specific interaction of 7S globulin with a cell protein component with molecular weight between 14 and 21 kDa. Further studies are needed to ascertain whether this interaction is directly or indirectly related to the observed stimulation of the LDL-receptor.

Ultrasound-assisted synthesis of MIL-88(Fe) conjugated starch-Fe3O4 nanocomposite: A safe antibacterial carrier for controlled release of tetracycline

A constructing antibiotic carrier with a sustained release profile is a promising method to stop long-term bacterial infection, which is of ideal interest in different biomedical fields. To end this, the present study aims to design a novel carrier based on the modification of biopolymeric starch for the rising possible interaction between carrier and antibiotic agent. We established an in-situ ultrasound-assisted method was applied to grow and create MIL-88(Fe) framework in the structure of magnetic polysaccharide (i.e., St/Fe3O4) synthesized by precipitation method resulting in St/Fe3O4/MIL-88(Fe) nanocomposite. It was loaded with a high amount of Tetracycline (TC) through its immersion into the TC aqueous solution. The release profile of TC-loaded St/Fe3O4/MIL-88(Fe) displays a lower initial burst release (about 26 % after 12 h) and followed by a controlled and sustained release (about 73 % up to 168 h) in the simulated physiological environment at pH 7.4. The in vitro cytotoxicity showed good cytocompatibility against Human skin fibroblast (HFF-1) cells. TC-loaded St/Fe3O4/MIL-88(Fe) showed higher antibacterial activity against both S. aureus and E. coli with the MIC value of 64 and 128 μg·mL−1, respectively.

GLUT-Targeting Phototherapeutic Nanoparticles for Synergistic Triple Combination Cancer Therapy.

The combination of multimodal therapies into one nanocarrier system is promising for its potential to enhance treatment performance by overcoming the efficacy problems encountered in conventional monomodal therapy. In this study, targeted and multimodal therapeutic hybrid nanocarriers are fabricated for breast cancer treatments. In this context, the synthesized gold nanorods (AuNRd), photothermal therapy (PTT) agent, are coated with doxorubicin (DOX) conjugated, targeted, and biocompatible tetrablock glycopeptide (P(DMAEMA-b-HMBAMA-b-FrucMA)-b-P(Lys)/DOX, P-DOX) polymer. Here, fructose-based (Fruc) glycopeptide polymer enhances cellular uptake into breast cancer through GLUT5. A photosensitizer molecule, indocyanine green (ICG), was loaded into the particles to provide photodynamic therapy (PDT) upon NIR light at 808 nm. In the final step of the fabrication, the polymer-coated nanoparticles are integrated with antisense ISIS5132 oligonucleotides to prevent apoptotic resistance of cells against drug molecules. The biocompatibility and therapeutic efficacy of the nanoparticles are evaluated on both human normal skin fibroblast cell (CCD-1079Sk) and human breast cancer cell (MCF7) lines. These multimodal therapeutic AuNRd@P-DOX/ICG/ISIS5132 nanoparticles demonstrate an efficient triple synergistic effect of chemo-/PTT/PDT, which is desired for breast cancer treatment. We believe that this promising multimodal therapeutic nanoparticle system can promote the further clinical application in the treatment of breast cancer and can also be adapted to other types of cancer.

New Ionic Liquid Microemulsion-Mediated Synthesis of Silver Nanoparticles for Skin Bacterial Infection Treatments.

This work reports a new approach for the synthesis of extremely small monodispersed silver nanoparticles (AgNPs) (2.9-1.5) by reduction of silver nitrate in a new series of benzyl alkyl imidazolium ionic liquids (BAIILs)-based microemulsions (3a-f) as media and stabilizing agents. Interestingly, AgNPs isolated from the IILMEs bearing the bulkiest substituents (tert-butyl and n-butyl) (3f) displayed almost no nanoparticle agglomeration. In an in vitro antibacterial test against ESKAPE pathogens, all AgNPs-BAIILs had potent antibiotic activity, as reflected by antibacterial efficiency indices. Furthermore, when compared to other nanoparticles, these were the most effective in preventing biofilm formation by the tested bacterial strains. Moreover, the MTT assay was used to determine the cytotoxicity of novel AgNPs-BAIILs on healthy human skin fibroblast (HSF) cell lines. The MTT assay revealed that novel AgNPs-BAIILs showed no significant toxic effects on the healthy cells. Thus, the novel AgNPs-BAIILs microemulsions could be used as safe antibiotics for skin bacterial infection treatments. AgNPs isolated from BAIIL (3c) was found to be the most effective antibiotic of the nanoparticles examined.

Exploitation of wheat germ for the production of wheat germ oil and microbial PUFAs and their potential application as wound healing agents for Human Skin Fibroblast cell line.

Exploitation of wheat germ for the production of wheat germ oil and microbial PUFAs and their potential application as wound healing agents for Human Skin Fibroblast cell line.

Homogeneous preparation of water-soluble products from chitin under alkaline conditions and their cell proliferation in vitro

The purpose of this study was to prepare water-soluble products by homogeneous depolymerization of chitin with H2O2 under alkaline conditions and investigate their potential application in wound healing. For the first time, water-soluble products were successfully prepared using a chitin-NaOH/urea solution; the products were chitosans with molecular weights (Mw) of 3.48–33.5 kDa and degrees of deacetylation (DD) > 0.5. Their Mw, DD and yield were affected by the reaction temperature, reaction time, concentration of H2O2 and chitin DD. The deacetylation and depolymerization of chitin were achieved simultaneously. The depolymerization of chitin was caused by hydrogen abstraction of HO, whereas the deacetylation resulted from the cleavage of amide bonds by HO− and HO2−, although the latter played a more important role. All water-soluble chitosans markedly promoted the proliferation of human skin fibroblast (HSF) cells, but they inhibited the proliferation of human keratinocyte cells. For the proliferation of HSF, a low concentration of chitosans was important. In addition, water-soluble chitosans with an Mw of 3.48–16.4 kDa markedly stimulated the expression of growth factors such as PDGF and TGF-β by macrophages. Water-soluble chitosans could be used as a potential active component in wound dressings.

Green Synthesis of Highly Fluorescent Carbon Dots from Bovine Serum Albumin for Linezolid Drug Delivery as Potential Wound Healing Biomaterial: Bio-Synergistic Approach, Antibacterial Activity, and In Vitro and Ex Vivo Evaluation

A simple and green approach was developed to produce novel highly fluorescent bovine serum albumin carbon dots (BCDs) via facile one-step hydrothermal treatment, using bovine serum albumin as a precursor carbon source. Inherent blue photoluminescence of the synthesized BCDs provided a maximum photostability of 90.5 ± 1.2% and was characterized via TEM, FT-IR, XPS, XRD, UV-visible, and zeta potential analyses. By virtue of their extremely small size, intrinsic optical and photoluminescence properties, superior photostability, and useful non-covalent interactions with the synthetic oxazolidinone antibiotic linezolid (LNZ), BCDs were investigated as fluorescent nano-biocarriers for LNZ drug delivery. The release profile of LNZ from the drug delivery system (LNZ-BCDs) revealed a distinct biphasic release, which is beneficial for mollifying the lethal incidents associated with wound infection. The effective wound healing performance of the developed LNZ-BCDs were evaluated through various in vitro and ex vivo assays such as MTT, ex vivo hemolysis, in vitro antibacterial activity, in vitro skin-related enzyme inhibition, and scratch wound healing assays. The examination of LNZ-BCDs as an efficient wound healing biomaterial illustrated excellent biocompatibility and low cytotoxicity against normal human skin fibroblast (HSF) cell line, indicating distinct antibacterial activity against the most common wound infectious pathogens including Staphylococcus aureus (ATCC® 25922) and methicillin-resistant Staphylococcus aureus, robust anti-elastase, anti-collagenase, and anti-tyrosinase activities, and enhanced cell proliferation and migration effect. The obtained results confirmed the feasibility of using the newly designed fluorescent LNZ-BCDs nano-bioconjugate as a unique antibacterial biomaterial for effective wound healing and tissue regeneration. Besides, the greenly synthesized BCDs could be considered as a great potential substitute for toxic nanoparticles in biomedical applications due to their biocompatibility and intense fluorescence characteristics and in pharmaceutical industries as promising drug delivery nano-biocarriers for effective wound healing applications.

Comparative assessment of Cucurbita moschata seed polypeptides toward the protection of human skin cells against oxidative stress-induced aging.

Skin aging has attracted much attention among the current aging population of society. The seeds of Cucurbita moschata possess a variety of potential biological activities as a healthy diet. However, limited information is available on the skin-antiaging properties of C. moschata seed protein and its hydrolysate. Herein, we developed a novel strategy for protecting human skin cells against oxidative stress-induced aging by C. moschata seed polypeptides. C. moschata seed polypeptides (CSPs) with different molecular weight distributions were successfully prepared by controlling the protease hydrolysis time. The proportions of < 1,000 Da polypeptides of P-1, P-2, and P-3 were 0.11, 20.26, and 92.72%, respectively. P-3 contained the highest proportion of polypeptides of size < 1,000 Da, which was observed to promote human skin fibroblast (HSF) growth by MTT assay, cell cycle, and morphology. P-3 has an efficient repair effect on the H2O2-induced aging of HSF cells. To explain this phenomenon, cell lifespan, intracellular ROS level, superoxide dismutase (SOD) activity, and glutathione (GSH) content were investigated to reveal the interactions between P-3 and antiaging. With the increase in P-3 concentration, the ROS level significantly decreased, and the SOD activity and GSH content significantly increased in H2O2-induced HSF cells. These findings indicated that CSPs have the potential to inhibit skin aging, which could be advantageous in the health industry for providing personal care.

In vitro investigation of genoprotective and antioxidant effects of santalol in CCD-1079Sk human skin fibroblast cells.

In vitro investigation of genoprotective and antioxidant effects of santalol in CCD-1079Sk human skin fibroblast cells.

Comparative Metabolic Study of Tamarindus indica L.’s Various Organs Based on GC/MS Analysis, In Silico and In Vitro Anti-Inflammatory and Wound Healing Activities

The chemical composition of the n-hexane extract of Tamarindus indica's various organs-bark, leaves, seeds, and fruits (TIB, TIL, TIS, TIF)-was investigated using gas chromatography-mass spectrometry (GC/MS) analysis. A total of 113 metabolites were identified, accounting for 93.07, 83.17, 84.05, and 85.08 % of the total identified components in TIB, TIL, TIS, and TIF, respectively. Lupeol was the most predominant component in TIB and TIL, accounting for 23.61 and 22.78%, respectively. However, n-Docosanoic acid (10.49%) and methyl tricosanoate (7.09%) were present in a high percentage in TIS. However, α-terpinyl acetate (7.36%) and α-muurolene (7.52%) were the major components of TIF n-hexane extract. By applying a principal component analysis (PCA) and hierarchal cluster analysis (HCA) to GC/MS-based metabolites, a clear differentiation of Tamarindus indica organs was achieved. The anti-inflammatory activity was evaluated in vitro on lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. In addition, the wound healing potential for the n-hexane extract of various plant organs was assessed using the in-vitro wound scratch assay using Human Skin Fibroblast cells. The tested extracts showed considerable anti-inflammatory and wound-healing activities. At a concentration of 10 µg/mL, TIL showed the highest nitric oxide (NO) inhibition by 53.97 ± 5.89%. Regarding the wound healing potential, after 24 h, TIB, TIL, TIS, and TIF n-hexane extracts at 10 g/mL reduced the wound width to 1.09 ± 0.04, 1.12 ± 0.18, 1.09 ± 0.28, and 1.41 ± 0.35 mm, respectively, as compared to the control cells (1.37 ± 0.15 mm). These findings showed that the n-hexane extract of T. indica enhanced wound healing by promoting fibroblast migration. Additionally, a docking study was conducted to assess the major identified phytoconstituents' affinity for binding to glycogen synthase kinase 3-β (GSK3-β), matrix metalloproteinases-8 (MMP-8), and nitric oxide synthase (iNOS). Lupeol showed the most favourable binding affinity to GSK3-β and iNOS, equal to -12.5 and -13.7 Kcal/mol, respectively, while methyl tricosanoate showed the highest binding affinity with MMP-8 equal to -13.1 Kcal/mol. Accordingly, the n-hexane extract of T. indica's various organs can be considered a good candidate for the management of wound healing and inflammatory conditions.

Gentamicin-loaded chitosan/folic acid-based carbon quantum dots nanocomposite hydrogel films as potential antimicrobial wound dressing

BackgroundTo provide effective healing in the wound, various carbohydrate polymers are commonly utilized that are highly potent platforms as wound dressing films. In this work, novel antibacterial flexible polymeric hydrogel films were designed via crosslinking polymeric chitosan (CS) with folic acid-based carbon quantum dots (CQDs). To end this, folic acid as a bio-precursor is used to synthesize CQDs through the hydrothermal technique. The synthesized CQDs as a crosslinking agent was performed at different concentrations to construct nanocomposite hydrogel films via the casting technique. Also, gentamicin (GM), L-Arginine and glycerol were supplemented in the formulation of nanocomposite since their antibiotic, bioactivity and plasticizing ability, respectively.ResultsThe successful construction of films were verified with different methods (FT-IR, UV-Vis, PL, SEM, and AFM analyses). The GM release profile displayed a controlled release manner over 48 h with a low initial burst release in the simulated wound media (PBS, pH 7.4). Antibacterial and in vitro cytotoxicity results showed a significant activity toward different gram-positive and negative bacterial strains (about 2.5 ± 0.1 cm inhibition zones) and a desired cytocompatibility against Human skin fibroblast (HFF-1) cells (over 80% cell viability), respectively.ConclusionThe obtained results recommend CQDs-crosslinked CS (CS/CQD) nanocomposite as a potent antimicrobial wound dressing.Graphical

Bioderived deep eutectic solvent-based topical chemotherapy for squamous cell carcinoma of the skin

The properties of deep eutectic solvents (DESs) are tunable, providing a route to new functions. Choline and geranic acid (CAGE) class of DESs shows antimicrobial properties, acts as drug delivery systems, and functions as anti-obesity agents. We hypothesize that new properties are attainable by rational variation of the molar ratios of choline and geranic acid. This study assessed the anticancer properties of a series of CAGE DESs, aiming to develop novel topical chemotherapy for squamous cell carcinoma of the skin. Cell viability assay demonstrated that varying the molar ratio of choline and geranic acid provided a strategy to significantly reduce the viability of human squamous carcinoma of the skin cell line (A431) without compromising the viability of human normal skin fibroblast cell line (CRL-2091). This molar ratio did not induce CRL-2091 to release interleukin–1β (IL–1β), a biomarker of acute and chronic inflammation, suggesting that the therapeutic dose of CAGE is unlikely to trigger skin inflammation. The anticancer property of CAGE is attributable to geranic acid, which also reduced the viability of A431 and CRL-2091 cell lines. With regards to mechanism, the therapeutic dose of CAGE did not rupture the cell membrane but induced oxidative stress on the A431 cell line. Concluding, rationally varying choline and geranic acid molar ratios provides a strategy to endow new properties on CAGE as demonstrated by selective activity against squamous cell carcinoma of the skin.

Synthesis of polymeric composite grafted with mineral particles/graphene oxide-based biomaterial: A promising robust hemostatic bandage

The loss of blood is one of the great challenges nowadays, leading to death in many acute circumstances like disasters, militant war, traumatic injury, and surgical procedures requiring newer hemostatic materials. In this aspect, the present study describes the synthesis of inexpensive polymer (polyvinyl alcohol and silk fibroin)-mineral-metal (hydroxyapatite)/carbon (graphene oxide) (PSHG) composite as a hemostatic dressing material for controlling the loss of blood. Initially, silk fibroin (SF), hydroxyapatite (HA), and graphene oxide (GO) were synthesized separately. The esterification of polyvinyl acetate (PVAc) to produce PVA. The mixture of exfoliated GO and HA was mixed at the time of gelation of PVA to produce PVA-HA-GO homogenous solution. The SF solution was mixed into the PVA-HA-GO homogenous solution to produce grey colour gel and cast on Petri dishes to produce PSHG biomaterial. The prepared PSHG biomaterials were subjected to biochemical test assays like hemolysis, protein adsorption, platelet aggregation, and blood clotting ability. The PSHG-based biomaterials biocompatibility was ascertained using Human skin fibroblast (HSK) cell lines. The results suggested that the prepared PSHG biomaterial has high absorption ability (~1040%), hemolysis (~2.38%), platelets aggregation (~35%), protein loading (~30 mg/cm 2 ), and ~ 89% blood clot within 15 s. Moreover, PSHG-based biomaterials significantly improved the number of fibroblast cells with high biocompatibility and controlled blood loss. The method synthesizing PSHG-based hemostatic material in the present study is novel, facile, and economically viable.

EGR1/NOX4 pathway regulates oxidative stress and further facilitates fibrosis progression in keloids responses to TGF-β1

BackgroundAs classic benign fibroproliferative tumors, keloids remain a major therapeutic challenge due to their complex pathological mechanisms. ObjectiveTo determine the functional role of transforming growth factor β1 (TGF-β1)/early growth response factor-1 (EGR1)/NADPH oxidases 4 (NOX4) axis in the pathogenesis of keloid fibrosis. MethodsDifferentially expressed genes in keloid tissues and normal skins were analyzed by RNA sequencing. Then, the human skin fibroblast cell line was treated with TGF-β1 at a dose of 10 ng/mL in order to stimulate the TGF-β1/SMAD pathway and the pathway was blocked using the SB431542. Furthermore, EGR1/NOX4 was over-expressed and inhibited by transfecting overexpression plasmids and small interfering RNAs, respectively. The levels of intracellular reactive oxygen species were measured using the DCFH-DA assay, and the expression levels of fibrosis‐related genes were assessed by Western blot analysis. Alternately, dual-luciferase reporter analysis verified the targeting relationship between EGR1 and NOX4. ResultsThe TGF-β1/SMAD signaling pathway was significantly activated in keloid tissues to promote dermal fibrosis. The level of ROS was increased in keloid fibroblasts. Moreover, TGF-β1 could facilitate the expression of EGR1 through regulating the SMAD pathway in keloids and promoting the fibrotic phenotype of keloid fibroblasts. EGR1 could regulate the production of ROS by targeting NOX4. Furthermore, NOX4-derived ROS could promote fibrotic-like phenotype of keloid fibroblasts and play an important role in keloid fibrosis. ConclusionOur findings provide new insights into the mechanisms of the TGF-β1/EGR1/NOX4 pathway in keloid fibrosis, and the TGF-β1/EGR1/NOX4 axis may serve as a potential therapeutic target for keloids.

The Effect of Plasma on Bacteria and Normal Cells in Infected Wound.

Infected wound is one of the most common and serious problem in wound management. Cold atmospheric plasma (CAP) is considered to have a good effect in wound healing as a new type medicine. However, there is a key issue that has not been addressed in the treatment of infected wounds by plasma. Bacteria are always found in the deep region of the wound. When plasma is used to treat wounds, it also acts on normal tissue cells while decontaminating. What is the difference between the same dose of plasma acting on bacteria and normal cells? In this study, the most common bacteria (S. aureus, P. aeruginosa, and E. coli) in infected wound and two kinds of normal skin cells (human keratinocyte and human skin fibroblasts (HSF)) were selected to study the difference of the effects of the same dose of plasma on bacteria and cells. The results reveal that three kinds of 106 CFU mL bacteria could be effectively inactivated by 5 order after plasma treatment 3 min, and P. aeruginosa was more sensitive to plasma (could be inactivated 5 order after 2 min treatment). The 104 mL keratinocyte and HSF were treated with the same dose of plasma; keratinocyte can maintain over 90% of the activity and HSF cells can maintain over 70% of the activity. Moreover, the level of collagen I secreted by HSF increased. Therefore, cells can remain a high activity when a plasma dose capable of inactivating bacteria is applied to them.

Full factorial design, physicochemical characterization, ex vivo investigation, and biological assessment of glutathione-loaded solid lipid nanoparticles for topical application

l-Glutathione (GSH) has exceptional antioxidant activities against UVA irradiation-induced oxidative stress and is used widely for combatting skin ageing. However, topical administration of GSH is challenging due to its inability to penetrate the stratum corneum (SC). This study aims to evaluate the solid lipid nanoparticles (SLNs) carrier system for improving the skin penetration and stability of GSH. The GSH-loaded SLNs (GSH-SLNs) were prepared by the double emulsion technique and were optimized by a full factorial design. The optimized GSH-SLNs formulation had a mean particle size of 305 ± 0.6 nm and a zeta potential of + 20.1 ± 9.5 mV, suitable for topical delivery. The ex-vivo penetration study using human skin demonstrated a 3.7-fold improvement of GSH penetration across SC with GSH-SLNs when compared with aqueous GSH. GSH-SLNs prolonged antioxidant activity on UVA irradiated fibroblast cells when compared to GSH solution, preventing UVA-induced cell death and promoting cell growth for times over 48 h. This research has illustrated that as a carrier system, SLNs were able to enhance the physicochemical stability, skin penetration, and drug deposition in the viable epidermis and dermis layers of the skin for GSH, while also maintaining the ability to protect human skin fibroblast cells against oxidative stress caused by UVA irradiation. This delivery system shows future promise as a topical delivery platform for the topical delivery of GSH and other chemically similar bioactive compounds for improving skin health.

Safety and Efficacy of Pure and a Nanosuspension of D-limonene for Controlling Pigeon Lice.

This study investigated the safety and efficacy of two forms of D-limonene (DL) against Columbicola columbae (pigeon feather lice); pure and a nanoemulsion formulation (DLN). The cell cytotoxicity of the prepared forms of DL/DLN was investigated using skin cell lines. In vitro and ex vivo bioassays were applied on lice. The ex vivo bioassay was done on cut feathers containing lice eggs. The in vivo experiment was conducted on pigeons naturally infested by lice. The infested pigeons were treated with DL, DLN, or deltamethrin (D) as a positive control. Both forms of D-limonene were found to be safe when applied to the normal human skin fibroblast cell line, but DLN was toxic to skin cell carcinoma. The in vitro and ex vivo results of both DL and DLN forms were similar. All eggs treated with DL, DLN, and D failed to hatch (100%). The in vivo results showed complete elimination of lice 24 h post-treatment (PT), and biochemical analysis showed that the treated birds retained normal kidney and liver functions. Treated groups also showed improved productivity in the 4 months PT. In conclusion, DL and DLN are safe and effective in controlling feather lice infestation in pigeons and successful treatment encourages bird productivity.