Fibroblast Cell Line Research Articles

Chitosan films synthesized via thiol-ene click chemistry: Toward safe and versatile platforms for packaging, cosmetics and biomedical applications

The first synthesis of chitosan-based films using bio-based reticulating agents via Thiol-Michael addition click reaction was described, along with their successful application in packaging, cosmetics, and biomedical fields. The impact of chitosan (CS) concentration and the type of synthesized bio-sourced biacids (GD) and triacids (GT and TT) on the physicochemical properties of these films were assessed. Films prepared with dibasic and tribasic acids (4%-CS) demonstrated superior mechanical properties and lower solubility compared to those containing 2% CS. The incorporation of the bio-sourced GD, GT, and TT acids led to a reduction of 53%, 60% and 60.4%, respectively, in the water vapor permeability (WVP) of 4% CS-based films compared to the control film. Remarkably, CS-TT-4% films exhibited the highest mechanical strength (TS), elongation at break (%E), elasticity, water vapor, and light barrier properties. Cytotoxicity tests performed on the NIH 3T3 fibroblast cell line showed that CS-GD-4% films exhibited the highest compatibility. The efficiency of these films as drug carriers was tested with challenging anticancer and cosmetic molecules, particularly etoposide (ETP) caffeine (Caf), showing very high drug loading with a progressive release of less than 20% after 72h of incubation. Moreover, ex-vivo permeation studies in rat skin revealed that the percutaneous absorption of Caf is significantly increased by a factor of 5.5 compared to that of CS-GD-4%-Caf films with no significant histological alterations of skin layers. Altogether, this work evidences the most suitable casting solvent for developing safe chitosan films, promising their potential applications in the cosmetic and biomedical fields.

Genetic insights and therapeutic potential for colorectal cancer: mutation analysis of KRASgene and efficacy of Oleuropein-conjugated iron oxide nanoparticles.

This study aimed to address the challenges of treating advanced stages of colon cancer (CRC) by exploring potential therapeutic options. The research focused on the genetic aspects of CRC, specifically the mutation rate of the KRAS gene, along with other genes like TTN, APC, MUC16, and TP53, using the TCGA dataset. Additionally, the study investigated the efficacy of Oleuropein, a polyphenolic compound found in olives, in combating CRC by using iron oxide nanoparticles coated with glucose and conjugated with Oleuropein. The study characterized the physicochemical properties of the nanoparticles, and the cytotoxic effects of the nanoparticles were evaluated on CRC and normal fibroblast cell lines, demonstrating significantly higher cytotoxicity against CRC cells compared to normal cells. Furthermore, the study analyzed gene expression changes using the GSE124627 dataset to understand the influence of KRAS alterations. It identified numerous upregulated and downregulated genes in KRAS-overexpressing samples, suggesting their involvement in critical cancer-related pathways. These findings suggest that KRAS-influenced genes could serve as potential therapeutic targets for CRC treatment. The study also examined the expression levels of identified genes in CRC samples compared to normal samples. Among the upregulated genes, 22 showed significant increases in cancer samples, while 14 downregulated genes exhibited decreased expression in both KRAS-influenced and cancer samples. Cox regression analysis identified specific upregulated genes, including ANKZF1, SNAI1, PPFIA4, SIX4, and NOTUM, associated with poor prognosis. Kaplan-Meier analysis further confirmed the correlation between increased expression of these genes and higher patient mortality rates. In conclusion, this study provided valuable insights into the genetic aspects of CRC and potential therapeutic strategies. The use of Oleuropein-conjugated iron oxide nanoparticles showed promising cytotoxic effects on colon cancer cells. These findings contribute to advancing our understanding of CRC and offer potential targets for further investigation and the development of novel therapeutic approaches.

Evaluation of the Genotoxic Effects of Grape Seed Extract and Marine Collagen Peptide on the Fibroblast Cell Line: An In Vitro Study.

Introduction Collagen plays a vital role in maintaining the structural integrity of dentin, and its modification with bioactive compounds can enhance its mechanical properties and bonding capabilities. Aim This study aimed to evaluate the genotoxic effects of grape seed extract (GSE) and marine collagen peptide(MCP) on dental pulp-derived primary cells. Methodology Human dental pulp stem cells were isolated,cultivated, and then treated with GSE and marine collagen peptides. DNA fragmentation was assessed using DAPI (4',6-diamidino-2-phenylindole) staining. Statistical analysis was performed using SPSS version 20 (IBM Corp., Armonk, NY, USA). Results The results showed that GSE exhibited a minimum level of cell death compared to marine collagen peptides. The viable cell count increased steadily over three days in all groups, with the control group showing the highest number of viable cells. The differences in viable cell count among the groups were statistically significant. Conclusion This study suggests that GSE and marine collagen peptides are highly biocompatible with dental pulp cells and could be considered for further clinical studies.

Exploring CK2 transcriptional regulation and signaling pathways in cancer.

e15199 Background: CK2 genes have been implicated in oncogenic processes in different cancer types, and their expression is known to be increased in many cancers relative to their normal tissue types. However the regulation of the three CK2 alpha transcripts and beta transcript is not well understood. Studies investigating the alpha gene ( CSNK2A1) and beta gene ( CSNK2B) found NFκB, ETS1, and SP1 as likely transcription factor regulators in lymphoid and fibroblast cell lines. Our study seeks to explore correlations between CK2 gene expression and the expression of these transcription factors in cancer. Since CK2 also regulates intracellular signaling through pathways such as PI3K-Akt, Jak-Stat, NFkB, Wnt, IL-6, TGF-β, Ras and Notch, this study also seeks to explore the correlation between CK2 expression and gene expression of known downstream signaling pathways across a variety of cancers. Methods: 32 non-overlapping cancer datasets were obtained from the The Cancer Genome Atlas (TCGA) and Broad GDAC Firehose projects which were obtained via cBioPortal.org and analyzed using a combination of cBioportal and RStudio statistical analysis packages. For each cancer type, mRNA expression of each of the four CK2 genes in cancer was compared to expression of transcription factors both previously identified in the literature and predicted generated by TFBIND analysis (via the transcription factor database TRANSFAC R3.4) of the CK2 minimal promoter regions. CK2 gene expression was also compared to expression of genes in known CK2 signaling pathways. Results: Our analysis found CSNK2A1 and CSNK2A3 expression to have a significant, strong positive correlation in nearly all cancer types studied, except pancreatic adenocarcinoma. CSNK2A1 demonstrated significant positive correlation with expression of the transcription factors NFKB1 and c-Rel in lymphoma, thymoma, and other cancers. CSNK2B demonstrated a significant, strong positive correlation with the transcription factors NELFE in every cancer examined. CK2 transcripts also showed significant correlations with other transcription factors and genes involved in IL-6, TGF-β, MAPK, and several other CK2 signaling pathways in many different cancer types. Conclusions: Overall this study demonstrates significant correlations between CK2 gene expression and previously proposed transcriptional regulation mechanisms as well as known CK2 signaling pathways in cancer.

MEK inhibitor PD0325901 upregulates CD34 expression in endothelial cells via inhibition of ERK phosphorylation

IntroductionCD34-positive endothelial progenitor cells (EPCs) promote angiogenesis and are a promising tool for regenerative cell therapy of ischemic diseases. However, the number and quality of CD34-positive cells decrease owing to various external and internal factors; thus, an efficient method is needed to establish CD34-positive EPCs. The generation of functional cells by reprogramming, that is, manipulating cell fate via gene transfer and/or treatment with chemical compounds, has recently been reported. Therefore, we aimed to generate CD34-positive cells by the reprogramming of endothelial cells (ECs). MethodsBased on previous reports, seven candidate chemical compounds were selected to reprogram human umbilical vein ECs (HUVECs) to CD34-positive cells. Following stimulation with the chemical compounds, the expression of CD34 was evaluated using quantitative PCR, flow cytometry, and immunocytochemistry. ResultsHUVECs treated with the compounds exhibited increased CD34 expression. We cultured cells in alternate media lacking one of the seven compounds and found no CD34 expression in cells treated with PD0325901-free media, suggesting that PD0325901—a MEK inhibitor—mainly contributed to the increase in CD34 expression. We found that 98% of cells were CD34-positive after PD0325901 treatment alone for 7 d. Western blotting revealed that the phosphorylation of ERK was suppressed in PD0325901-treated cells. No upregulation of CD34 was observed in fibroblast cell lines, even after PD0325901 treatment. These results suggested that PD0325901 induces CD34-positive cells by inhibiting ERK phosphorylation in ECs. ConclusionsCD34 expression was strongly induced in ECs by treatment with the MEK inhibitor PD0325901 in vitro. Our study provides a useful reference for the establishment of CD34-positive EPCs and will contribute to the development of regenerative therapies, especially for ischemic diseases.

A Novel Recombinant Modified Vaccinia Ankara Virus expressing Interleukin-13 Receptor α2 Antigen for Potential Cancer Immunotherapy.

Genetically altered recombinant poxviruses hold great therapeutic promise in animal models of cancer. Poxviruses can induce effective cellmediated immune responses against tumor-associated antigens. Preventive and therapeutic vaccination with a DNA vaccine expressing IL-13Rα2 can mediate partial regression of established tumors in vivo, indicating that host immune responses against IL-13Rα2 need further augmentation. The aim of the study is developing a recombinant modified vaccinia Ankara (MVA) expressing IL-13Rα2 (rMVA-IL13Rα2) virus and study in vitro infectivity and efficacy against IL-13Rα2 positive cell lines. We constructed a recombinant MVA expressing IL-13Rα2 and a green fluorescent protein (GFP) reporter gene. Purified virus titration by infection of target cells and immunostaining using anti-vaccinia and anti-IL-13Rα2 antibodies was used to confirm the identity and purity of the rMVA-IL13Rα2. Western Blot analysis confirmed the presence of IL-13Rα2 protein (~52 kDa). Flow cytometric analysis of IL-13Rα2 negative T98G glioma cells when infected with rMVA-IL13Rα2 virus demonstrated cell-surface expression of IL-13Rα2, indicating the infectivity of the recombinant virus. Incubation of T98G-IL13Rα2 cells with varying concentrations (0.1-100 ng/ml) of interleukin-13 fused to truncated Pseudomonas exotoxin (IL13-PE) resulted in depletion of GFP+ fluorescence in T98G-IL13Rα2 cells. IL13-PE (10-1000 ng/ml) at higher concentrations also inhibited the protein synthesis in T98G-IL13Rα2 cells compared to cells infected with the control pLW44-MVA virus. IL13- PE treatment of rMVA-IL13Rα2 infected chicken embryonic fibroblast and DF-1 cell line reduced virus titer compared to untreated cells. rMVA-IL13Rα2 virus can successfully infect mammalian cells to express IL-13Rα2 in a biologically active form on the surface of infected cells. To evaluate the efficacy of rMVA-IL13Rα2, immunization studies are planned in murine tumor models.

Fabrication of pH-stimuli hydrogel as bioactive materials for wound healing applications

Hydrogels exhibit exceptional suitability as wound dressing due to their remarkable three-dimensional (3D) characteristics. Here, we have reported the fabrication of hydrogels from biopolymers carboxymethyl cellulose (CMC), polyvinyl alcohol (PVA), and gelatin via a simple blending method to mimic the natural extracellular matrix. Scanning electron microscopy (SEM), water contact meters (WCM), and Fourier-transform infrared spectroscopy (FTIR) were used to evaluate the chemical structural, morphological, and wettability behavior. The wetting and degradation behavior were also found to be different for different formulations (Min. (51.60o) and Max. (113.60o)) and (Min. (38.82 mg) and Max. (3.72 mg)), respectively. Swelling was investigated in different media, including phosphate buffer saline solution (PBS) and aqueous media. It was observed that the hydrogel displayed the highest degree of swelling in an aqueous medium (Min. (597.32–1121.49 %) and Max. (1089.51–2139.73 %)) compared to PBS media (Min. (567.01–1021.85 %) and Max. (899.13–1639.17 %)). The release of Neomycin was studied in a PBS medium via the Franz diffusion method at 37 °C. The maximal release in various media demonstrated pH-responsive behavior. The viability and proliferation of fibroblast (3T3) cell lines were examined in vitro to evaluate cytocompatibility. Human Embryonic Kidney (HEK) 293 cells were used to evaluate the hydrogels' ability to promote vascularization and angiogenesis. Therefore, the data demonstrate that hydrogels that have been manufactured have qualities that make them promising for use as wound dressings in wound healing applications.

Barleria lupulina leaf extract-assisted biosynthesis of cobalt ferrite nanoparticles: Characterization and anticancer activity against MCF-7 cells

This study reports the synthesis of Cobalt ferrite (CoFe2O4) nanoparticles (NPs) by a green synthesis method using the leaf extract of Barleria Lupulina. The X-ray diffraction (XRD) analysis confirms the formation of a single-phase cubic (Fd3m) structure and from the Scherrer formula the average crystallite size (30.07 nm) calculated matches with that obtained from W-H plot(34.66 nm). A bandgap of 2.02 eV was observed from the UV–Vis-DRS analysis. Fourier transform infrared spectroscopy (FTIR) was used to confirm the characteristic functional groups present in the spinel ferrite. The investigation using scanning electron microscopy (SEM) shows aggregates of nanoparticles and EDX analysis gives the chemical composition of the green synthesized ferrite sample. The distribution of particle-sizes and colloidal-stability were estimated by DLS and zeta-potential measurements. The M−H plot confirmed the ferromagnetic nature of the prepared nanoparticles. Beyond the realm of material science, we ventured into the realm of biological activity. Our synthesized cobalt ferrite nanoparticles were assessed for their toxicity on human breast cancer cell lines (MCF-7) as well as normal L929 fibroblast cell lines by MTT assay. The results show potent anti-cancer activity of the ferrite sample (LC50 value 77.92 µg/ml) while sparing normal cells from excessive toxicity (LC50 value 252.34 µg/ml). The results of cell viability, changes in cell morphology, and cell cycle analysis revealed that cobalt ferrite nanoparticles triggered apoptosis, and hindered cell adhesion and migration of MCF-7 cells, potentially through elevated reactive oxygen species generation. This dual action, where the nanoparticles combat cancer effectively while minimizing harm to healthy cells, holds significant promise in the field of oncology.

Insights into cellular interactions of characterized Mg–Al Layered Double Hydroxide on L929 cells

Layered double hydroxides are members of an anionic clay family, characterised by unique two-dimensional layered structures and lend versatility in various applications. These biocompatible compounds have the potential to get intercalated with biological compounds and physico-chemically adsorbed onto organic molecules. Thus, making them important candidates for pharmaceutical and biomedical purposes. This study aims to synthesise, characterise and investigate the cellular toxicity interactions of Mg–Al LDH towards the mouse fibroblast L929 cell line. The Mg–Al LDH was synthesized by a meticulous process of co-precipitation followed by the hydrothermal method to ensure a well-defined and stable structure for suitable biological application. Characterisation techniques like Dynamic Light Scattering, Zeta potential, Scanning Electron Microscopy, Fourier transform infrared, and X-ray diffraction analysis were employed to provide deeper insights into the physiochemical properties and structural integrity of the synthesized Mg–Al LDH. The investigation of cellular interactions with the L929 fibroblast cell line served to assess the biocompatibility and potential cytotoxic effects of Mg–Al LDH. This was observed by assessing the morphological changes and evaluating the cytotoxic effects of Mg–Al LDH by utilising various techniques like phase contrast microscopy, fluorescent staining, and Giemsa staining. The cellular metabolic activity was assessed by MTT assay, and the subcellular lysosomal alteration was examined using the fluorescent staining method by the acridine orange staining. The dose-dependent response observed in the cellular interaction underscores the importance of dosage considerations for potential biomedical applications. By elucidating the dose-response relationship, this study contributes valuable information for the safe and effective usage of LDH in biomedical contexts.

Effect of caffeine versus hydrogen peroxide on human skin fibroblast cell line cytotoxicity, cell cycle phases, and apoptosis.

Effect of caffeine versus hydrogen peroxide on human skin fibroblast cell line cytotoxicity, cell cycle phases, and apoptosis.

Sialidase activity in Glioblastoma cells treated with thyme oil

Background: Cancer is a genetic disease that causes aberrant cells to grow and spread uncontrollably throughout the body, where cerebral glioblastoma is one of the brain tumors. Thyme oil has many therapeutic properties such as antioxidants and anticancer.Aim of study: Estimate sialidase enzyme in human glioblastoma cancer cell line (AMGM5) and rat embryo fibroblasts cell line (REF) in supernatant and suspension cells treated with thyme oil extract and cells without it. Methods: Sialidase activity was measured by ELISA (Enzyme-linked immunosorbent assay) in supernatant and suspension cells of AMGM5 and REF cells treated and untreated with thyme oil extract. Results: Sialidase showed a high level in the Glioblastoma cell line ( AMGM5), while a low level in cells after exposure to diluted thyme oil (10μl/ml) after incubation for 24 hours, also showed a lower level in the Rat Embryo Fibroblast after being treated with thyme extract.Conclusions The sialidase enzyme showed increased in glioblastoma cells but decreased in its level in these cells after exposure to thyme oil extract, which means that thyme oil inhibited cancer cell growth.

Characterization and antioxidant effect of mucilage in leaves from Cocculus hirsutus.

Natural polysaccharides are attractive biodegradable polymers. Among the natural plant-based polysaccharides, mucilage is an interest for numerous biomedical applications. Hence, mucilage was isolated from the leaves of Cocculus hirsutus (Family; Menispermaceae) and tested for its phytochemicals, physio-chemical characteristics using standard procedure such as solubility, pH, swelling index etc., and structural characterization studies using FTIR, GC-MS and SEM followed by antioxidant and in vitro cytotoxic assays. The phytochemical results showed the presence of carbohydrates, amino acids, flavonoids, alkaloids, tannins, terpenes, saponin, glycosides and steroids. The yield percentage of mucilage was 26% and showed swelling index of 6.8-7.4. The FTIR spectra of mucilage showed characteristics strong peaks of major functional groups. The SEM image showed the porous and rough surface morphological characters of mucilage. The obtained mucilage showed antioxidant potential by DPPH, FRAP and Total reducing power assay and also exhibited non-cytotoxic character against fibroblast cell lines. Thus, the isolated mucilage showed promising characteristics which can be exploited for various biological applications from food to drug release studies.

Synthesis, structural investigations, in vitro cytotoxicity, apoptotic activity, cell cycle analysis, and molecular modeling studies of nano‐sized Zn(II) Schiff base complex

A new novel tetradentate ligand, [(1E,1′E)‐N,N′‐(1,2‐phenylene)bis(1‐(5‐(2‐fluorophenyl)furan‐2‐yl)methanimine) (PFPFMI), and its nano‐sized [Zn(PFPFMI)Cl2].3H2O complex were synthesized. The geometry of PFPFMI and its Zn(II) complex was described through CHN analyses, Fourier‐transform infrared spectroscopy (FTIR), UV–visible, X‐ray diffraction (XRD), thermal gravimetric analysis (TGA), and mass spectral data tests. The spectral data of the nano‐sized [Zn(PFPFMI)Cl2].3H2O complex indicates that the PFPFMI ligand functions as a tetradentate (N2O2) coordination through two azomethine nitrogen groups (N) and two furan ring oxygen (O) atoms. Density functional theory (DFT) calculations were performed using the molecular studio software to investigate the optimal geometry of PFPFMI and its [Zn(PFPFMI)Cl2].3H2O complex. The SEM, TEM, XRD, AFM, and energy dispersive X‐ray analysis (EDX) analyses of the studied complex unveil distinct and strong diffraction peaks, indicating its crystalline nature and providing evidence of its nano‐sized particle sizes. Additionally, the inhibition zone diameter was used to assess the in vitro antimicrobial action of PFPFMI and the particular complex against Gram‐positive bacteria Streptococcus pneumonia and Bacillus subtilis, Gram‐negative bacteria Pseudomonas aeruginosa, Salmonella typhi, and Escherichia coli, as well as fungi Aspergillus fumigatus, Geotricum candidum, Syncephalastrum racemosum, and Candida tropicalis. The [Zn(PFPFMI)Cl2].3H2O complex exhibits higher activity than the ligand PFPFMI. In vitro cytotoxicity of the synthesized [Zn(PFPFMI)Cl2].3H2O complex was explored using MCF7 (breast cancer), HCT116 (colon cancer), A549 (lung cancer), HePG‐2 (liver cancer), A375 (melanoma cancer), and normal lung fibroblast (WI38) cell lines. Based on IC50 and selective index (SI) values, it was shown that the complex exhibited higher potency against MCF7 cell lines. Moreover, the Zn(II) complex exhibited the ability to induce DNA damage in MCF7 cells, resulting in dose‐dependent cell apoptosis. Subsequent investigations revealed that complex 2 also induced cell cycle arrest in the G2 and S phases.

Examination of cytotoxic and antimicrobial effect of whitening toothpastes: an in vitro study.

Toothpastes are widely used to protect oral and teeth health. This study aims to examine the cytotoxic and antimicrobial effects of whitening toothpastes. In this study, extracts were prepared according to ISO 10993-12:2021 standard (0.2 g/mL) using whitening and conventional toothpastes. The prepared extracts were added to human gingival fibroblast cell lines (HGF-1) in different dilutions (1:1, 1:2, 1:4, 1:8, 1:16, and 1:32) and a cytotoxicity test was performed. Antimicrobial analysis of toothpastes was performed on Streptococcus mutans, Staphylococcus aureus, and Candida albicans using the hole-plate diffusion method. Cell viability and microbial analysis data were examined using two-way analysis of variance (ANOVA) and Tukey post-hoc test (p < 0.05). Toothpastes with sodium lauryl sulfate (SLS) in their composition showed statistically more toxic effects (p < 0.05). The activated carbon toothpastes without SLS showed over 90% cell viability after dilution. Although the dilution rate of toothpastes containing SLS increased, cell viability remained below 70%. All toothpastes used in the study showed antimicrobial effects on S. mutans, S. aureus, and C. albicans. Toothpaste containing hydrogen peroxide and SLS produced more antibacterial effects than activated carbon, blue covarine, microparticles, and conventional toothpaste. SLS-containing toothpastes showed more toxicity on HGF-1 cells. Toothpaste containing hydroxyapatite did not show toxic effects on HGF-1 cells. SLS, sodium lauryl sarcosinate and hydrogen peroxide in toothpastes increase antimicrobial effects.

Enhanced wound regeneration by PGS/PLA fiber dressing containing platelet-rich plasma: an in vitro study

Novel wound dressings with therapeutic effects are being continually designed to improve the wound healing process. In this study, the structural, chemical, physical, and biological properties of an electrospun poly glycerol sebacate/poly lactide acid/platelet-rich plasma (PGS/PLA-PRP) nanofibers were evaluated to determine its impacts on in vitro wound healing. Results revealed desirable cell viability in the Fibroblast (L929) and macrophage (RAW-264.7) cell lines as well as human umbilical vein endothelial cells (HUVEC). Cell migration was evident in the scratch assay (L929 cell line) so that it promoted scratch contraction to accelerate in vitro wound healing. Moreover, addition of PRP to the fiber structure led to enhanced collagen deposition (~ 2 times) in comparison with PGS/PLA scaffolds. While by addition PRP to PGS/PLA fibers not only decreased the expression levels of pro-inflammatory cytokines (IL-6 and TNF-α) in RAW-264.7 cells but also led to significantly increased levels of cytokine (IL-10) and the growth factor (TGF-β), which are related to the anti-inflammatory phase (M2 phenotype). Finally, PGS/PLA-PRP was found to induce a significant level of angiogenesis by forming branching points, loops, and tubes. Based on the results obtained, the PGS/PLA-PRP dressing developed might be a promising evolution in skin tissue engineering ensuring improved wound healing and tissue regeneration.

Polysilsesquioxane decorated ZIF-8 as a potential pH-responsive vehicle for topical delivery and release of acyclovir and tetracycline: Investigation of blood compatibility, cytotoxicity and antibacterial properties

In this research, poly-chloropropylmethyl-silsesquioxanen was prepared and decorated with ZIF-8 in order to investigate its loading capacity for acyclovir and tetracycline. Before and after drug loadings, the composites were characterized by FT-IR, SEM-EDS, XRD, and XPS analyses. Then, the in-vitro release of these drugs was investigated by UV–Vis spectroscopy in different buffers (pH = 5, 7.4, and 9.1). The results showed that the release of ACV reached a maximum amount of 41.3 mg at pH = 7.4 during 12 h. In comparison, the release of TC reached a maximum amount of 22.5 mg at pH = 5 during 6 h. The blood compatibility, in-vitro cytotoxicity on the L929 fibroblast cells line, and antibacterial assay against Staphylococcus aureus and Pseudomonas aeruginosa were also investigated for this composite as a drug carrier.

The influence of Leucidal, a natural cosmetic preservative, on fibroblast and keratinocytes. Studies on cells and on model membrane systems

The aim of this work was to investigate the influence of Leucidal® Liquid (abbr. Leucidal), which is recommended as a natural cosmetic ingredient of antimicrobial properties, on model membranes of keratinocytes and fibroblasts. The toxicity tests on cell lines were also performed to allow for a more detailed discussion of the results. As model membrane systems the lipid Langmuir monolayers were applied. During the investigations, the surface pressure/area measurements, penetration studies and Brewster Angle Microscopy (BAM) visualization were performed for one component and mixed lipid monolayers.It was evidenced that at the membrane – corresponding conditions, the components of Leucidal do not penetrate either model keratinocyte and fibroblast membranes or one component films composed of the major lipids of skin cell membranes. Leucidal makes these systems slightly more expanded and less stable, however this is not reflected in the changes in the film morphology. Only the ceramide systems were sensitive to the presence of Leucidal, i.e. the incorporation of Leucidal components manifested well in the decrease of the films’ condensation and alterations in their morphology. The tests on cells demonstrated that Leucidal is non toxic for these types of cells at the concentrations suggested by the producer. A thorough comparison of these results with those published for bacteria model membranes enabled us to discuss them in the context of the mechanism of action of Leucidal components. It was concluded that Leucidal components are of low affinity to the skin cellular model membranes of low content of Leucidal-sensitive ceramides and are not toxic for fibroblast and keratinocyte cell lines. Moreover, the lipid composition of the membrane and its molecular organization can be important targets for Leucidal components, decisive from the point of view of the activity and selectivity of the studied composition.

Insights into the antioxidant, anti-inflammatory and anti-microbial potential of Nigella sativa essential oil against oral pathogens

Oral disorders can exert systemic ramifications beyond their localized effects on dental tissues, implicating a wide array of physiological conditions. The utilization of essential oils (EOs) for protection of oral health represents a longstanding practice. Consequently, in this investigation, essential oil derived from Nigella sativa seeds (NSEO) underwent isolation via the hydro-distillation process, followed by a comprehensive evaluation of its antioxidant, anti-inflammatory, anti-fungal, antibacterial activities, and cytocompatibility. The isolated NSEO manifested as a pale-yellow substance and was found to harbor a diverse spectrum of bioactive constituents, including steroids, triterpenoids, flavonoids, phenols, proteins, alkaloids, tannin, sesquiterpenoid hydrocarbons, monoterpenoid alcohol, and monoterpenoid ketone (thymoquinone). Notably, the total phenolic content (TPC) and total flavonoid content (TFC) of NSEO were quantified at 641.23 μg GAE/gm and 442.25 μg QE/g, respectively. Furthermore, NSEO exhibited concentration-dependent inhibition of protein denaturation, HRBC membrane stabilization, and hemolysis inhibition. Comparative analysis revealed that NSEO and chlorhexidine (CHX) 0.2% displayed substantial inhibition of hemolysis compared to aspirin. While NSEO and CHX 0.2% demonstrated analogous antibacterial activity against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, NSEO showcased heightened efficacy against Lactobacillus acidophilus and Candida albicans. Additionally, NSEO exhibited pronounced effects against periodontal pathogens such as Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Prevotella intermedia. Importantly, no cytotoxicity was observed on human gingival fibroblast cell lines. These findings underscore the potential of NSEO as a potent antibacterial and antifungal agent in the management of oral microbial pathogens, thereby offering avenues for the development of innovative therapies targeting diverse oral inflammatory conditions. Nevertheless, further investigations are imperative to unlock its full therapeutic repertoire.

Phytoactive essential oils-composed water-free organogels: Development, characterization and proof of antibacterial activity

The objective of this study was to develop and analyze a gel-like formulation that does not contain water, made from natural compounds found in essential oils, which are known for their antimicrobial properties and ability to reduce inflammation. Lentisk tree (Pistacia lentiscus L.), yarrow (Achillea millefolium L.) and myrtle (Myrtus communis L.) grow in the mild climate of Eurasia and are receiving particular attention for their medicinal and aromatic value. Two distinct concentrations of selected essential oils (EOs) were tested, dispersed in organogels based on 3 % (m/V) Carbopol 974P NF in PEG 400. Prior to formulation development, selected EOs were characterized by mass spectroscopy to identify the dominating bioactives. A total of 32 compounds were quantified for lentisk leaf EO, 42 for yarrow EO, and 26 for myrtle leaf EO. The best formulations were found for the highest concentration of EOs (4 %, m/m) into Carbopol/PEG400 organogels. Organogels were produced by high-speed homogenization followed by microwave heating and showed acceptable pH between 5.8 and 6.0, being compatible with the skin, with an electrical conductivity highest for yarrow EO-based organogel (YIG4%, ∼64 mV), and the lowest for myrtle EO-based organogel (MIG4%, ∼48 mV). Texture analysis showed the highest adhesion properties for lentisk EO-based organogel (LIG4%), attributed to its higher viscosity recorded for the two tested temperatures (25 °C vs 37 °C). Rheological analysis confirmed that organogels have pseudoplastic behaviour. The antibacterial activity was compared between plain EOs and EOs-based organogels, with the yarrow EO and its organogel (YIG4%) depicting the highest activity against both tested bacterial strains (S. aureus and C. acnes). On the other hand, MIG4% contributed for the highest cell proliferation rate tested in a mouse embryo fibroblast cell line (3T3-L1), whereas with LIG4% the lowest viability value was recorded.

Evaluation of Antibiofilm, Antimicrobial, Cytotoxic and Antioxidant Effects of Some Wild Mushroom Species

In this study, ethanol extracts of some wild mushroom species, in the phylum Basidiomycota, [Armillaria mellea (Vahl) P. Kumm., Infundibulicybe geotropa (Bull.) Harmaja, Leucopaxillus gentianeus (Quél.) Kotl. and Trametes versicolor (L.) Lloyd] were tested for their antioxidant, antimicrobial, antibiofilm, and cytotoxic activities. Mushroom samples showed low antimicrobial activity on Enterococcus faecalis and Staphylococcus aureus, while biofilm inhibitory activity on test microorganisms ranging from 24.6% to 80.5%. At the end of the antioxidant activity studies A. mellea was the mushroom sample having the highest DPPH radical scavenging capacity (0.105±0.001 mg/mL) whereas T. versicolor was the one having the highest iron (III) ion reducing power (40.709±0.003 µg TE/100g). The highest polyphenol content was observed in T. versicolor (29.916±0.002 mg GAE/100g) samples, and the lowest in A. mellea (9.5±0.006 mg GAE/100g) samples. The cytotoxic effects of the samples were tested on MCF-7 and MDA-MB-231 (breast cancer) and L929 (mouse fibroblast) cell lines using the MTT method. As a result, it was observed that A. mellea and T. versicolor samples were more effective on MCF-7 cell line, A. mellea on MDA-MB-231 cell line.