Non-cytotoxic Effect Research Articles

Abstract C008: Phase I biomarker analysis of farletuzumab ecteribulin (FZEC), formerly MORAb-202, effects on cancer responses in patients with platinum resistant ovarian cancer

Abstract Background: FZEC (formerly MORAb-202) is an antibody-drug conjugate consisting of farletuzumab and eribulin. FZEC is cytotoxic to FRα+ cancer cells (via direct binding) and to adjacent tumor cells (via bystander effects); it also exerts noncytotoxic bystander effects in the tumor microenvironment (Zhang et al, EORTC 2022). Study 101 is a first-in-human dose escalation followed by dose expansion study in Japanese patients (pts) with selected solid tumors to determine safety and preliminary efficacy of FZEC; antitumor activity across several doses was demonstrated in pts with platinum-resistant ovarian cancer (PROC) and FRα+ tumors (Nishio ASCO 2022). This exploratory biomarker study evaluated association of FZEC-induced pharmacological effects and clinical responses in pts with PROC to examine the mechanism of action (MoA) of FZEC. Methods: Serum samples from up to 58 pts with PROC from a FZEC 0.9 mg/kg cohort (equivalent to 33 mg/m2) or from a combined dose cohort (FZEC 0.3, 0.45, 0.68, 0.9, 1.2 mg/kg) were collected predose on day 1 of treatment cycles 1–6 and analyzed using 139 exploratory Luminex, Simoa, or OLINK protein assays. While these doses are no longer being evaluated, as a result of a change to body-surface-area-based dosing, the biomarkers observed to be preliminarily associated with response in this analysis should be further explored in ongoing studies. Markers with >30% unevaluable data points were excluded. The 1-sample Wilcoxon signed-rank test and 2-sample Wilcoxon rank-sum test were used to assess changes from baseline and differences between 2 best overall response groups (responders [CR/PR] and nonresponders [SD/PD]) at each visit. Fisher’s method was used to summarize statistical difference across visits (meta P<0.05) between the 2 groups. Results: Changes from baseline to at least 1 visit postdose (P<0.05) in either the responder or nonresponder groups, and differences between responders and nonresponders (meta P<0.05), were seen in 29 markers in the combined-dose cohort, and in 14 markers in the 0.9 mg/kg cohort. There was an overlap of 11 markers in both the FZEC 0.9 mg/kg and combined-dose cohorts that showed a significant change from baseline and significant differences in changes between responders and nonresponders. Among these 11 markers, SP-D and IGFBP-2 showed persistent changes (≥3 visits) in responders. Decreases in markers with reported protumor functions (eg, ANG-2, KLK-5, KLK-7, MCP-1, FRTN, and MMP-12), and increases in SP-D, were persistent (≥4 of 5 visits) in responders. Some biomarkers (eg, IGFBP-1, FLT3LG, and VEGF-D) showed significant increases in nonresponders. Conclusions: The persistent responder-associated biomarker changes suggest that FZEC may modulate functions associated with these markers (eg, immune regulation, vascular remodeling, extracellular matrix protease activities, iron metabolism) and this may contribute to the antitumor effect of FZEC. In the limited number of pts analyzed, preliminary pharmacodynamic effects trended with the antitumor effects observed with FZEC 0.9 mg/kg. Citation Format: Yan Zhang, Bob Zimmermann, Shuyu Li, Toshimitsu Uenaka, Keiji Furuuchi, Kan Yonemori, Toshio Shimizu, Shin Nishio, Mayu Yunokawa, Koji Matsumoto, Kazuhiro Takehara, Kosei Hasegawa, Yasuyuki Hirashima, Hidenori Kato, Yohei Otake, Takuma Miura, Junji Matsui, Meijuan Li. Phase I biomarker analysis of farletuzumab ecteribulin (FZEC), formerly MORAb-202, effects on cancer responses in patients with platinum resistant ovarian cancer [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr C008.

Silver Nanoparticle-Incorporated Hydroxyapatite-Chitosan-PVP-Based Scaffold for Bone Tissue Engineering: Physicochemical Characterization, Antibacterial Activity, and In Vitro Ceftriaxone Release Study

The development of biocompatible scaffolds is rapidly growing in several fields of human health, particularly for the treatment of bone defects. In this study, silver nanoparticle (AgNPs)-incorporated hydroxyapatite (HA), chitosan (Chi), and polyvinylpyrrolidone (PVP)-based scaffolds were fabricated using the freeze-drying method for bone tissue engineering. Physicochemical properties such as porosity, density, swelling ratio, and biodegradability, as well as blood biocompatibility, brine shrimp lethality, antibacterial activities, FTIR-ATR, tensile strength, cytotoxicity, and SEM, were conducted. A study on the in vitro release of ceftriaxone (Cef), a routinely prescribed medication for treating osteomyelitis, was done. The selected scaffolds had desirable porosity, density, swelling capability, and biodegradation rate, which would meet the basic prerequisites for osteogenesis. The scaffolds showed qualitative and quantitative antibacterial activities against Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Scaffolds were minimally cytotoxic to brine shrimp and RBC biocompatible. FTIR analysis confirmed intermolecular interaction among components of scaffolds. Scaffold had a tensile strength of 2.9 ± 0.22 N/mm2 and a non-cytotoxic effect on Vero cell lines. Surface morphology, pore shape, and sizes (98–382 μm) were observed through SEM. The percentage of drug (Cef) loading was found to be 63.16 ± 4.85%. An initial burst release of Cef was 40.21 ± 1.50% in the first 24 h. A sustained release of Cef was observed after 4 days, and about 60.36 ± 2.38% was released within 14 days. Therefore, selected AgNPs-incorporated HA-Chi-PVP scaffold could be a potential aid for bone tissue engineering.

Production and structural characterization of eco-friendly bioemulsifier SC04 from Saccharomyces cerevisiae strain MYN04 with potential applications

BackgroundBioemulsifiers are natural or microbial-based products with the ability to emulsify hydrophobic compounds in water. These compounds are biodegradable, eco-friendly, and find applications in various industries.ResultsThirteen yeasts were isolated from different sources in Alexandria, Egypt, and evaluated for their potential to produce intracellular bioemulsifiers. One yeast, isolated from a local market in Egypt, showed the highest emulsification index (EI24) value. Through 26S rRNA sequencing, this yeast was identified as Saccharomyces cerevisiae strain MYN04. The growth kinetics of the isolate were studied, and after 36 h of incubation, the highest yield of cell dry weight (CDW) was obtained at 3.17 g/L, with an EI24 of 55.6%. Experimental designs were used to investigate the effects of culture parameters on maximizing bioemulsifier SC04 production and CDW. The study achieved a maximum EI24 of 79.0 ± 2.0%. Furthermore, the crude bioemulsifier was precipitated with 50% ethanol and purified using Sephadex G-75 gel filtration chromatography. Bioemulsifier SC04 was found to consist of 27.1% carbohydrates and 72.9% proteins. Structural determination of purified bioemulsifier SC04 was carried out using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDX), high-performance liquid chromatography (HPLC), and nuclear magnetic resonance spectroscopy (NMR). FTIR spectroscopy revealed characteristic bands associated with carboxyl and hydroxyl groups of carbohydrates, as well as amine groups of proteins. HPLC analysis of monosaccharide composition detected the presence of mannose, galactose, and glucose. Physicochemical characterization of the fraction after gel filtration indicated that bioemulsifier SC04 is a high molecular weight protein-oligosaccharide complex. This bioemulsifier demonstrated stability at different pH values, temperatures, and salinities. At a concentration of 0.5 mg/mL, it exhibited 51.8% scavenging of DPPH radicals. Furthermore, in vitro cytotoxicity evaluation using the MTT assay revealed a noncytotoxic effect of SC04 against normal epithelial kidney cell lines.ConclusionsThis study presents a new eco-friendly bioemulsifier, named SC04, which exhibits significant emulsifying ability, antioxidant and anticancer properties, and stabilizing properties. These findings suggest that SC04 is a promising candidate for applications in the food, pharmaceutical, and industrial sectors.

Structure and properties of biogenic hydroxyapatite bioceramics modified by graphene-like structures

Today bone tissue engineering is one of the most used technologies for treat bones injure. Materials containing hydroxyapatite and graphene have received much attention recently. The aim of this study was preparation of biogenic hydroxyapatite bioceramics modified by graphene-like structures investigation effect of graphene on the structure and properties of material. Biogenic hydroxyapatite bioceramics modified by graphene-like structures were successfully prepared by chemical vapor deposition (CVD) method. Subsequently, microstructure, composition, specific surface area, skeleton density, resorption rate in physiological solution and cytotoxicity were evaluated. XRD, IR spectroscopy, micro-Raman spectroscopy and SEM proved graphene oxide’s formation on biogenic hydroxyapatite as well as on silica single crystal for comparison. Although the coating of graphene-like structures on biogenic hydroxyapatite bioceramics reduces the specific surface area, it allows to 4 times increase resorption rate of biogenic hydroxyapatite bioceramics in physiological solution and does not affect the overall assessment of the cytotoxicity. MTT assay established non-cytotoxic effect and indicated a high potential of biogenic hydroxyapatite bioceramics modified by graphene-like structures using CVD method for medical application.

Evaluation of physicochemical properties, release kinetics, and in vitro/in vivo wound healing activity of the electrospun nanofibres loaded with the natural antioxidant oil from Pistacia atlantica

Herbal medicine is an extraordinary source of bioactive compounds, and developing them can aid in the wound healing process. This study focuses on the optimal production of nanofibres containing an effective herbal extract. The nanofibres were produced by electrospinning a polymer solution containing polyvinyl alcohol (PVA) and sodium alginate (ALG) with Pistacia atlantica oil (PAO) (0, 1.5%, and 2.5% (w/v)). The PAO is rich in different biological properties including antioxidant, anti-inflammatory and antibacterial activities. The addition of 1.5% (w/v) PAO to PVA/ALG solutions increased surface tension and conductivity by 3.5% and 3.86%, respectively. Besides, the scanning electron microscopy (SEM) images revealed that the diameter of the nanofibres increased from 191 nm ± 15 nm–259 nm ± 10 nm. According to differential scanning calorimetry (DSC) results, the nanofibres loaded with 1.5% (w/v) PAO exhibited higher thermal stability compared to PVA/ALG nanofibres without PAO. Based on the in vitro drug release experiments, the PAO release from nanofibres followed the Korsmeyer-Peppas model and the pseudo-Fickian mechanism. Cell culture and cytotoxicity assay results illustrated that the PAO-loaded nanofibres had non-cytotoxic effects against L-929 cells with suitable fibroblast cell proliferation and cell viability. Animal test results showed that the PAO-loaded nanofibres significantly enhanced wound closure by 92.07% compared to PAO-unloaded nanofibres (p < 0.02). The wound healing ability of PAO was also confirmed by the histopathological findings, with PAO increasing re-epithelization and successfully attenuating tissue inflammation. Overall, this study introduces a new type of wound dressing containing PAO, a wound healing stimulant with excellent properties such as uniform structure, antibacterial activity, and breathability. Consequently, the results suggest that the investigated nanofibres may merit being considered as an ideal wound dressing for future clinical investigations into wound therapy and regeneration.

A first-in-human, open label, multiple dose, dose escalation, and cohort expansion phase I study to investigate the safety, tolerability, pharmacokinetics and antitumor activity of BB-1701 in patients with locally advanced/metastatic HER2-expressing solid tumors.

3029 Background: BB-1701 is an antibody-drug conjugate consisting of a humanized IgG1κ monoclonal antibody and eribulin. When BB-1701 targets HER2-expressing cancer cells and is internalized, free eribulin is cleaved from the ADC by cathepsin b and causes cytotoxicity to cancer cells. The neighboring cells were affected with bystander effects, like cytotoxicity on tumor cells and non-cytotoxic effects on microenvironment by free eribulin. Here, we report preliminary results from this phase 1 study (NCT04257110). Methods: This dose escalation study followed an accelerated titration and traditional "3 + 3" design with 6 dose levels from 0.4 to 2.6 mg/kg Q3W. Patients with confirmed advanced/metastatic HER2 overexpressing solid tumors, who had progressed on, or were intolerant to prior standard therapies, aged ≥ 18 years, with ECOG PS ≤ 2, and measurable disease, were enrolled. HER2 expressions were determined by IHC or next generation sequencing. Tumor imaging was conducted every 9 weeks. The primary objective is the assessment of safety and tolerability, and the secondary objective is the assessment of PK and preliminary anti-tumor activity. Results: A total of 29 patients were enrolled in this study. Tumor types were breast cancer, gastric/gastroesophageal junction cancer, colorectal cancer, gallbladder cancer, ovarian cancer and urothelial cancer. The medium prior lines of systemic therapy were 3 (range 1 – 6). During this study, no DLT was observed and the MTD was not reached. The most commonly reported (≥20%) treatment-related adverse events (TRAEs) were peripheral neuropathy, aspartate aminotransferase increased, neutrophil count decreased, white blood cell count decreased, alanine aminotransferase increased, hypertriglyceridemia, anemia, platelet count decreased. The most commonly reported ≥ grade 3 TRAEs (≥10 %) was peripheral neuropathy. There were two grade 4 adverse events: neurological symptom and sepsis. There was no interstitial lung disease observed during the study. Nine patients discontinued the study treatment due to peripheral neuropathy. Thirteen patients out of 26 patients evaluable for antitumor activity achieved partial response (PR), and 11 patients had stable diseases (SD). The best overall response rate (BOR) was 50.0% (13/26) and the disease control rate (DCR) was 92.3% (24/26). For 17 breast cancer patients in the study, 12 patients achieved PR, and 5 patients had SD. The BOR was 70.5% (12/17) and the DCR was 100% (17/17). For 5 gastric cancer patients in the study, all patients had SD. The duration of treatment ranged from 2 weeks to 66 weeks. Conclusions: BB-1701 was generally well tolerated up to 2.6 mg/kg dose levels and has shown promising antitumor activity in patients with HER2-overexpression solid tumors. Clinical trial information: NCT04257110 .

Evaluation of biocomposite putty with strontium and zinc co-doped 45S5 bioactive glass and sodium hyaluronate

The application of powder or granule formed bioactive glasses in the defect area with the help of a liquid carrier to fill the defects is a subject of interest and is still open to development. In this study, it was aimed to prepare biocomposites of bioactive glasses incorporating different co-dopants with a carrier biopolymer and to create a fluidic material (Sr and Zn co-doped 45S5 bioactive glassessodium hyaluronate). All biocomposite samples were pseudoplastic fluid type, which may be suitable for defect filling and had excellent bioactivity behaviors confirmed by FTIR, SEM-EDS and XRD. Biocomposites with Sr and Zn co-doped bioactive glass had higher bioactivity considering the crystallinity of hydroxyapatite formations compared to biocomposite with undoped bioactive glasses. Biocomposites with high bioactive glass content had hydroxyapatite formations with higher crystallinity compared to biocomposites with low bioactive glass. Furthermore, all biocomposite samples showed non-cytotoxic effect on the L929 cells up to a certain concentration. However, biocomposites with undoped bioactive glass showed cytotoxic effects at lower concentrations compared to biocomposites with co-doped bioactive glass. Thus, biocomposite putties utilizing Sr and Zn co-doped bioactive glasses may be advantageous for orthopedic applications due to their specified rheological, bioactivity, and biocompatibility properties.

Comparative Evaluation of Alginate-Gelatin Hydrogel, Cryogel, and Aerogel Beads as a Tissue Scaffold

Hydrogels are frequently used in tissue engineering and regenerative medicine, drug delivery, and environmental remediation. Alginate and gelatin, which are frequently used natural polymers to form hydrogels, were chosen in this study to form a core-shell structured hydrogel. Cryogels and aerogels were obtained by drying hydrogels with different methods, freeze-drying, and the continuous flow of supercritical CO2, respectively. The potential use of hydrogels, aerogels, and cryogels as a tissue scaffold was evaluated comparatively. Characterizations of materials were determined morphologically by scanning electron microscope and computed-micro tomography, chemically by energy dispersive spectroscopy, and mechanically by the dynamic mechanical analyzer. In addition, the cytotoxic effect of all structures was analyzed by the WST-1 method and the localization of the cells in these structures was determined by microscopic methods. All scaffolds show non-cytotoxic effects. Cryogels have the highest porosity (85.21 %) and mean pore size values (62.3±26.8 µm). Additionally, cryogels show high water retention capacity (782±53.5%) than aerogels (389±2.5%) for 24 h. The elastic modulus values were

Genotoxicity evaluation of two derived products from Allium extracts: s-propylmercaptocysteine and s-propyl mercaptoglutathione

Propyl-propane-thiosulfonate (PTSO) is one of the main organosulfur compounds present in Allium essential oils with a widely documented biological activity. For this reason, it could be used as a food and feed additive in the agri-food industry. A genotoxicity evaluation of substances and their metabolites present in food is necessary to guarantee the consumer's health following the recommendations of the European Food Safety Authority (EFSA). To evaluate the toxicological profile of derivatives of PTSO, the cytotoxicity, an Ames test, a micronucleus test and the comet assay were performed. Results showed that non-cytotoxic effects were observed in Caco-2 exposed to s-propyl mercaptocysteine (CSSP) and s-propyl mercaptoglutathione (GSSP) (0–450 μM). The mutagenicity index remained in the range of 0.6–1.4 for both compounds, showing no mutagenic effects for the concentrations of 5000–312.5 μg GSSP/plate and 250–15.63 μg CSSP/plate. Moreover, the % binucleated cells with micronuclei were 1.3–2.2 and 1.6–2.7 for GSSP and GSSP, respectively. For comet assays there was no DNA-genotoxic or oxidative damage in a concentration range of 112.5–450 μM. Therefore, we can conclude that these compounds are not genotoxic at the conditions tested. These results support that the presence of CSSP and GSSP in the food/feed is not of concern, although further studies are needed to complete their safety profile.

Poly (β-amino Ester) Nanoparticles Modified with a Rabies Virus-derived peptide for the Delivery of ASCL1 Across a 3D In Vitro Model of the Blood Brain Barrier.

Gene editing has emerged as a therapeutic approach to manipulate the genome for killing cancer cells, protecting healthy tissues, and improving immune response to a tumor. The gene editing tool achaete-scute family bHLH transcription factor 1 CRISPR guide RNA (ASCL1-gRNA) is known to restore neuronal lineage potential, promote terminal differentiation, and attenuate tumorigenicity in glioblastoma tumors. Here, we fabricated a polymeric nonviral carrier to encapsulate ASCL1-gRNA by electrostatic interactions and deliver it into glioblastoma cells across a 3D in vitro model of the blood-brain barrier (BBB). To mimic rabies virus (RV) neurotropism, gene-loaded poly (β-amino ester) nanoparticles are surface functionalized with a peptide derivative of rabies virus glycoprotein (RVG29). The capability of the obtained NPs, hereinafter referred to as RV-like NPs, to travel across the BBB, internalize into glioblastoma cells and deliver ASCL1-gRNA are investigated in a 3D BBB in vitro model through flow cytometry and CLSM microscopy. The formation of nicotinic acetylcholine receptors in the 3D BBB in vitro model is confirmed by immunochemistry. These receptors are known to bind to RVG29. Unlike Lipofectamine that primarily internalizes and transfects endothelial cells, RV-like NPs are capable to travel across the BBB, preferentially internalize glioblastoma cells and deliver ASCL1-gRNA at an efficiency of 10 % causing non-cytotoxic effects.

Influence of the Biological Medium on the Properties of Magnesium Doped Hydroxyapatite Composite Coatings

In this paper, the stability of magnesium-doped hydroxyapatite/chitosan (MHC) suspension obtained with the sol-gel approach was evaluated using nondestructive ultrasound measurements. The MHC coatings obtained by the spin-coating technique were characterized before and after immersion for 7 and 14 days, respectively, in Dulbecco’s modified eagle medium (DMEM) by scanning electron microscopy, equipped with an EDAX detector. Also, the functional groups present on the MHC coatings surface were analyzed with the aid of attenuated total reflection (ATR) Fourier transform infrared (FTIR) spectroscopy. The surface microstructure was evaluated using two commentary techniques, namely atomic force microscopy (AFM) and metallographic microscopy (MM). The influence of immersion in DMEM on the biological properties was studied with in vitro studies using primary osteoblast and HCT-8 cell lines. Our results revealed that both surface morphology and chemical composition of the MHC coatings allow rapid development of a new apatite layer on their surface after immersion in DMEM. Preliminary in vitro biological studies underlined the noncytotoxic effect of the studied samples on the proliferation of primary osteoblast and HCT-8 cell lines, which makes them a promising candidate for applications in fields such as orthopedics or dentistry. The antifungal assay of the MHC coatings was assessed using Candida albicans ATCC 10231 and their results showed a good inhibitory effect. The coatings made on the basis of the MHC composite could contribute to increasing the degree of success of implants by decreasing the risk of infections and postoperative inflammation.

In silico study of novel niclosamide derivatives, SARS-CoV-2 nonstructural proteins catalytic residue-targeting small molecules drug candidates

The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-mediated coronavirus disease 2019 (COVID-19) infection remains a global pandemic and health emergency with overwhelming social and economic impacts throughout the world. Therapeutics for COVID-19 are limited to only remdesivir; therefore, there is a need for combined, multidisciplinary efforts to develop new therapeutic molecules and explore the effectiveness of existing drugs against SARS-CoV-2. In the present study, we reported eight (SCOV-L-02, SCOV-L-09, SCOV-L-10, SCOV-L-11, SCOV-L-15, SCOV-L-18, SCOV-L-22, and SCOV-L-23) novel structurally related small-molecule derivatives of niclosamide (SCOV-L series) for their targeting potential against angiotensin-converting enzyme-2 (ACE2), type II transmembrane serine protease (TMPRSS2), and SARS-COV-2 nonstructural proteins (NSPs) including NSP5 (3CLpro), NSP3 (PLpro), and RdRp. Our correlation analysis suggested that ACE2 and TMPRSS2 modulate host immune response via regulation of immune-infiltrating cells at the site of tissue/organs entries. In addition, we identified someTMPRSS2 and ACE2 microRNAs target regulatory networks in SARS-CoV-2 infection and thus open up a new window for microRNAs-based therapy for the treatment of SARS-CoV-2 infection. Our in vitro study revealed that with the exception of SCOV-L-11 and SCOV-L-23 which were non-active, the SCOV-L series exhibited strict antiproliferative activities and non-cytotoxic effects against ACE2- and TMPRSS2-expressing cells. Our molecular docking for the analysis of receptor-ligand interactions revealed that SCOV-L series demonstrated high ligand binding efficacies (at higher levels than clinical drugs) against the ACE2, TMPRSS2, and SARS-COV-2 NSPs. SCOV-L-18, SCOV-L-15, and SCOV-L-09 were particularly found to exhibit strong binding affinities with three key SARS-CoV-2’s proteins: 3CLpro, PLpro, and RdRp. These compounds bind to the several catalytic residues of the proteins, and satisfied the criteria of drug-like candidates, having good adsorption, distribution, metabolism, excretion, and toxicity (ADMET) pharmacokinetic profile. Altogether, the present study suggests the therapeutic potential of SCOV-L series for preventing and managing SARs-COV-2 infection and are currently under detailed investigation in our lab.

In Vivo Skin Hydrating Efficacy of Fish Collagen from Greenland Halibut as a High-Value Active Ingredient for Cosmetic Applications.

The industrial processing of fish for food purposes also generates a considerable number of by-products such as viscera, bones, scales, and skin. From a value-added perspective, fish by-products can act also as raw materials, especially because of their collagen content (particularly in fish skin). Interestingly, the potential of marine collagen for cosmetic applications is enormous and, remarkably, the extraction of this protein from fish skins has been established for different species. Using this approach, we investigated the integration of marine collagen (COLRp_I) extracted from the skin of the Greenland halibut as an active ingredient in a cosmetic hydrogel formulation. In this study, extracts of marine collagen at concentrations up to 10 mg/mL showed a non-cytotoxic effect when cultured with fibroblast cells for 3 days. In addition, marine collagen extract, when incorporated into a cosmetic hydrogel formulation, met criterion A of ISO 11930:2019 regarding the efficacy of the preservative system (challenge test). In addition, the cosmetic formulations based on marine collagen at dosages of 0.1, 0.25 and 0.5% were tested in a clinical study on the skin of the forearms of 23 healthy volunteers, showing a sightly hydration effect, suggesting its potential for beauty applications. Moreover, this work illustrates that the circular economy concept applied to the fish processing industry can represent important benefits, at innovation, environmental and economic levels.

Dopamine and epigallocatechin-3-gallate cross-linked coating demonstrates improved osteointegration of polyetheretherketone in rabbits

The increasing demand for enhanced osteointegration calls for next-generation polyetheretherketone (PEEK) that can offer fast bone integration. Whether polydopamine (PDA) and epigallocatechin-3-gallate (EGCG) cross-linked coatings could improve osteointegration of PEEK remains unclear. Here, we fabricated a series of PDA and EGCG cross-linked coatings (PE coatings: P2E0.1, P2E0.5, P2E1, and P2E2 coatings) to enhance osteointegration of PEEK. The material characteristics of coatings were studied using scanning electron microscopy, X-ray photoelectron spectroscopy, micro-BCA tests, and static water contact angle tests. We evaluated the cytotoxicity, cell adhesion, and osteogenic effects of the coatings in vitro. Furthermore, the P2E0.5 coating was selected to coat porous PEEK scaffold to observe osteointegration using micro-computed tomography and histology in a rabbit. The results show that the PE coatings provide phenolic hydroxyl and amino groups and enhanced hydrophilicity of PEEK, especially in the P2E0.5 coating. Several lines of cellular evidence suggest that P2E0.5 coatings exhibit a noncytotoxic and excellent osteogenic effect. Compared with PEEK, the P2E0.5-coated PEEK shows higher values of bone volume, trabecular number, and trabecular thickness. This study suggests that the PE coating is a promising candidate for surface modification of PEEK and may lay the foundation for further application of PE coatings in clinical practice.

Farrerol Inhibits Vascular Smooth Muscle Cell Proliferation and Protects Them From Oxidative Injury via Bidirectional Modulation of the PI3K/Akt/mTOR Signaling Pathway

The inhibition of intimal hyperplasia (IH) is an effective strategy to improve the long-term outcome of endovascular therapy and prevent restenosis. Farrerol, a naturally occurring dihydroflavone with a variety of bioactivities, exerts inhibitory effects against balloon injury-induced IH in rats. In the present study, bioinformatics analysis, in combination with in vitro experimental validation, was performed to elucidate the underlying inhibitory mechanisms. The protein–protein interaction (PPI) network was assessed to identify farrerol-related protein targets in the context of IH, based on which biological functions and pathway enrichment were analyzed. The proliferation and cell cycle distribution of vascular smooth muscle cells (VSMCs) were investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2 H-tetrazolium bromide and 5-ethynyl-2-deoxyuridine incorporation assays and flow cytometric analysis, respectively. The level of pro-inflammatory cytokines in the cell culture medium was estimated using an enzyme-linked immunosorbent assay (ELISA). Protein expression in A7r5 cells was determined by western blotting. Forty-six IH-related targets of farrerol were identified, and the PI3K/Akt/mTOR pathway was highly enriched among the 43 predicted pathways ( P &lt; .05). In serum (10% fetal bovine serum)-induced A7r5 cells, farrerol inhibited proliferation through non-cytotoxic effects, induced cell cycle arrest in the G0/G1 phase , and suppressed the activation of the PI3K/Akt/mTOR pathway. In H2O2 (300 µM)-induced A7r5 cells, farrerol reduced the release of IL-1 β and TNF- α and reversed the suppressive effect on the PI3K/Akt/mTOR pathway in response to H2O2 stimulation. In conclusion, farrerol inhibits the proliferation of VSMCs and protects VSMCs from oxidative injury via the bidirectional modulation of the PI3K/Akt/mTOR signaling pathway, which might contribute to the suppression of neointima formation.

Colistin sulfate-sodium deoxycholate sulfate micelle formulations; molecular interactions, cell nephrotoxicity and bioactivity

This study aimed to develop a micelle formulation of colistin with several mole ratios of sodium deoxycholate sulfate (SDCS) carrier to reduce its nephrotoxicity. The molar ratio and surface properties of colistin and SDCS were determined. Colistin was incorporated into SDCS micelles followed by lyophilisation. The formulated micelles were evaluated in terms of particle size, zeta potential, morphology, thermal properties, encapsulation efficiency (EE) and release profile. The interactions were analysed with FTIR, NMR and also in-silico studies using molecular docking. The MIC and MBC were calculated, while time-kill kinetics were observed. Furthermore, a cytotoxicity study of the formulations against kidney cells was carried out and data were analysed. Particle sizes were in the range of 140.9–162.6 nm with spherical shapes, and the zeta potential values were between −35.3 and −22.8 mV. The EE of colistin was between 17.73 and 40.15%. FTIR and DSC data indicated an interaction between the colistin and SDCS, and the NMR analysis revealed hydrogen bonding and electrostatic interaction as the main interaction. Molecular docking simulation showed that multiple hydrogen bonds were present between the hydrophilic ring of colistin and SDCS. The formulations showed good safety profiles with non-cytotoxic effects against the kidney cell while maintaining antibacterial activity. The overall results indicated that micelle formulations with SDCS can be a good option for the delivery of colistin to reduce its nephrotoxicity with better antibacterial activity, while further studies in the animal model are important.

New Butyroside D from Argan Press Cake Possess Anti-Melanogenesis Effect via MITF Downregulation in B16F10 and HEM Cells.

Hyperpigmentation is a skin condition where patches of skin become darker in color due to excess melanin production upon UV exposure leading to melasma, which are lentigines or post inflammatory hyperpigmentation that psychologically affecting a great number of people. The present study investigates the anti-melanogenic effect of Butyroside D and the underling mechanism. After the confirmation of the non-cytotoxic effect of Butyroside D on B16F10 cells, we proceeded with analyzing the impact of the treatment at low and high concentration (i.e., 0.2 μM and 2 μM) using gene profiling analysis and examined the differentiation in gene expression. Our results identify cyclic adenosine monophosphate (cAMP), Wnt/β-catenin and Mitogen-Activated Protein Kinase (MAPK) signaling pathways to be downregulated upon treatment with Butyroside D. These pathways were targeted to further validate the effect of Butyroside D on membrane receptors melanocortin 1 receptor (MC1R) and receptor tyrosine kinase (c-Kit), related microphthalmia-associated transcription factor (MITF) and consequently tyrosinase (TYR), and tyrosine-related protein-1 (TYRP-1) that were all shown to be downregulated and, therefore, leading to the repression of melanin biosynthesis. Finally, the anti-melanogenic effect of Butyroside D was confirmed on human epidermal melanocytes (HEM) cells by inhibiting the activation of cAMP pathway generally mediated through α-melanocyte-stimulating hormone (α-MSH) and MC1R. Overall, this study suggests the potential applicability of this purified compound for the prevention of hyperpigmentation conditions.

Unravelling the sanitization potential of slightly acidic electrolyzed water combined Thymus vulgaris based nanoemulsion against foodborne pathogens and its safety assessment

Ecofriendly disinfectants are being extensively investigated for their potential sanitiser efficacy and food industry application.This study aimed to explore the inhibitory effect of slightly acidic electrolyzed water (SAEW) combined with essential oil (EO) based on surfactant monolaurin (ML) on the growth of Escherichia coli O157:H7 43894, Salmonella enteritidis ATCC 13076, Listeria monocytogenes Scott ATCC 43251, Staphylococcus aureus ATCC 13565, Candida albicans KCCM 11282. The SAEW's (ACC 5, 10 and 20 mg/L) antimicrobial activity was significantly enhanced for nanoemulsion (NE). The stability of the NE was enhanced based on sonication and the particle size, and zeta potential analysis was determined. The results revealed that the addition of ML (nonionic surfactant) and high-intensity ultrasound sonication influences the formulation of oil-in-water nanoemulsion. Higher antimicrobial activity of the essential oil and electrolyzed water in both individual and combined (nanoemulsion) effect were confirmed. The nanoemulsion appeared to amplify the antimicrobial effect of SAEW with particle size (117.3 nm) suggesting the formation of SAEW–TV complexes probably through hydrophobic interaction. The zeta potential showed the enhanced stability (−37.4 mV) of the emulsion was confirmed based on sonication. The mechanism of antimicrobial ability to disrupt cell membrane integrity was investigated based on electron microscopy (TEM) analysis and based on the molecular binding, the mode of action of epinastine, oseltamivir and 2-isopropyl -5-methylphenol (−12.47, −7.98 and −9.456 kcal/mol) binding affinity was confirmed respectively. In addition, the MTT assay was assessed to evaluate the non-cytotoxic effect of the combination of (SAEW + TR + ML) as a disinfectant.

Cytotoxicity, Phytochemical Screening and Genetic analysis of Ginger (Zingiber officinale Rosc.) Callus and Rhizome

• Ginger rhizome extracts displayed strong cytotoxic effect against HT29, HT116 and MCF-7 cancer cells lines with highest activity against HT29. • Ginger callus extracts recorded non-cytotoxic effect against all examined cancer cells with IC50 >100 µg/mL. • Ginger rhizome extracts contained more phenols and terpenes than the callus ones. • The principal components of ginger; 6-gingerol and 6-shogaol were not detected in callus extracts. • Incidence of genetic changes in DNA of ginger callus during in vitro culture in 2,4-D containing medium. • Most polymorphic bands (11 out of 12) were generated in callus during in vitro culture with total polymorphism 23.07% between callus and plant. Ginger ( Zingiber officinale Rosc.) is well known as distinctive spice with considered therapeutic values. In the current work we aimed to investigate cytotoxic effect, phytochemical screening and genetic analysis of ginger callus and their rhizome. Cytotoxicity of ginger rhizome and callus extracts was examined against some human cancer cell lines HT29, HT116 and MCF-7 by MTT method. Phytochemicals were screened by thin layer chromatography (TLC) and genetic analysis was performed by ISSR markers. Ginger rhizome principle components, 6-gingerol and 6-shogoal were used as standards to detect their presence in callus. Results showed that, rhizome extracts displayed a strong cytotoxicity towards all tested cancer cells with the highest activity (IC 50 20.4 ± 3.0 µg/mL; P< 0.05) against HT29. IC 50 of ginger callus extracts were >100 µg/mL in all examined cancer cell lines. TLC chromatograms of rhizome and callus extracts revealed the presence of phenols and terpenes, whereas 6-gingerol and 6-shogaol were only detected in rhizome extracts. Genetic analysis by ISSR markers revealed occurrence some genetic changes in DNA of callus compared to that of rhizome with total polymorphism 23.07%. In conclusion, ginger cytotoxicity to HT29, HT116 and MCF-7 cancer cells was mainly associated with the presence of 6-shogaol and 6-gingerol in extracts, which appeared as strong cytotoxic effect in rhizome compared to non-cytotoxic effect was evaluated in the callus extracts. Therefore, further studies on the callus elicitation and precursors feeding to induce biosynthesis of ginger bioactive molecules are recommended.

Antiviral Effect of Stenocline ericoides DC. and Stenocline inuloides DC., Two Flavonoid-Rich Endemic Plants from Madagascar, against Dengue and Zika Viruses.

Dengue and Zika viruses are identified as the most medically important arthropod-borne viral pathogens. Over the past 20 years, the global dengue incidence has dramatically increased with epidemics of severe dengue where the case fatality rate can reach up to 20% in untreated patients. The association between Zika virus infection and severe congenital anomalies was first reported in 2015. Today no specific antiviral therapies are available for dengue and Zika virus infections, accentuating the need of adapted antiviral strategies based on medicinal plant drug discovery. Plants are a potential source of antiviral phytocompounds which act primarily by blocking virus entry in the host-cell. In the present study, we evaluated whether crude extracts from Stenocline ericoides DC. and Stenocline inuloides DC., two endemic plants from Madagascar, may have antiviral effects against dengue and Zika viruses. We showed that S. ericoides has virucidal action whereas S. inuloides inhibits the early steps of virus infection with a non-cytotoxic effect in human cells. The administration of S. ericoides and S. inuloides extracts in zebrafish had no effect on the behavior of animals at the active doses against dengue and Zika viruses, suggesting the absence of adverse effects at these doses. LC-HRMS2 and molecular networking analyses revealed the richness of these two plants in polyphenols and flavonoid with the presence of clusters of phytocompounds specific to each Stenocline species. Consequently, S. ericoides and S. inuloides represent potential sources for natural and safe antiviral phytocompounds against flaviviruses of medical concern.