MTT Colorimetric Assay Research Articles

In vitro antimicrobial, anticancer evaluation, and in silico studies of mannopyranoside analogs against bacterial and fungal proteins: Acylation leads to improved antimicrobial activity

Carbohydrate analogs are an important, well-established class of clinically useful medicinal agents that exhibit potent antimicrobial activity. Thus, we explored the various therapeutic potential of methyl α-D-mannopyranoside (MαDM) analogs, including their ability to synthesize and assess their antibacterial, antifungal, and anticancer properties; additionally, molecular docking, molecular dynamics simulation, and ADMET analysis were performed. The structure of the synthesized MαDM analogs was ascertained by spectroscopic techniques and physicochemical and elemental analysis. In vitro antimicrobial activity was assessed and revealed significant inhibitory effects, particularly against gram-negative bacteria along with the prediction of activity spectra for substances (PASS). Concurrently, MαDM analogs showed good results against antifungal pathogens and exhibited promising anticancer effects in vitro, demonstrating dose-dependent cytotoxicity against Ehrlich ascites carcinoma (EAC) cancer cells while sparing normal cells from compound 5, with an IC50 of 4511.65 µg/mL according to the MTT colorimetric assay. A structure–activity relationship (SAR) study revealed that hexose combined with the acyl chains of decanoyl (C-10) and benzenesulfonyl (C6H5SO2-) had synergistic effects on the bacteria and fungi that were examined. Molecular docking was performed against the Escherichia coli (6KZV) and Candida albicans (1EAG) proteins to acquire insights into the molecular interactions underlying the observed biological activities. The docking results were further supported by 100 ns molecular dynamics simulations, which provided a dynamic view of the stability and flexibility of complexes involving MαDM and its targets. In addition, ADMET analysis was used to evaluate the toxicological and pharmacokinetic profiles. Owing to their promising drug-like properties, these MαDM analogs exhibit potential as prospective therapeutic candidates for future development.

The Effect of Chitosan Incorporation on Physico-Mechanical and Biological Characteristics of a Calcium Silicate Filling Material.

A tricalcium silicate-based cement, Biodentine™, has displayed antibiofilm activity when mixed with chitosan powder. This study aimed to assess the effect of chitosan incorporation on the physico-mechanical and biological properties of Biodentine™. In this study, medium molecular weight chitosan powder was incorporated into Biodentine™ in varying proportions (2.5 wt%, 5 wt%, 10 wt%, and 20 wt%). The setting time was determined using a Vicat apparatus, solubility was assessed by calculating weight variation after water immersion, radiopacity was evaluated and expressed in millimeters of aluminum, the compressive strength was evaluated using an Instron testing machine, and the microhardness was measured with a Vickers microhardness tester. In addition, surface topography of specimens was analyzed using scanning electron microscopy, and the effect of chitosan on the viability of human embryonic kidney (HEK 293) cells was measured by a colorimetric MTT assay. Incorporation of 2.5 wt% and 5 wt% chitosan powder delivered an advantage by speeding up the setting time of Biodentine material. However, the incorporation of chitosan compromised all other material properties and the crystalline structure in a dose-dependent manner. The chitosan-modified material also showed significant decreases in the proliferation of the HEK 293 cells, signifying decreased biocompatibility. Chitosan incorporation into calcium silicate materials adversely affects the physical and biological properties of the material. Despite the increased antimicrobial activity of the modified material, the diminution in these properties is likely to reduce its clinical value.

Comparison Between Moxifloxacin and Chloramphenicol for the Treatment of Bacterial Eye Infections

Background: Moxifloxacin is a bactericidal methoxyquinolone used for the treatment of conjunctivitis and prophylactic therapy in cataract and refractive surgeries. Chloramphenicol is a bacteriostatic organochlorine introduced into clinical practice in 1948 and used mainly in topical preparations because of its known toxicity. Objectives: The study aimed to evaluate the in vitro antibacterial effect and the ocular cytotoxicity of these broad-spectrum antibiotics. Methods: Antimicrobic activity was tested on four bacterial strains (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis), and determined through calculation of Minimum Inhibitory Concentration (MIC) and IC50 for each microorganism. Antibacterial activity was determined by microdilution method after 24 hours incubation with 2-fold serial dilutions (2.5 mg/ml to 4.883 µg/ml) of moxifloxacin and chloramphenicol. Disk diffusion test were performed according to EUCAST methodology. Biofilm formation inhibition (BIC) and biofilm Eradication Concentration (BEC) assay were conducted for P. aeruginosa and S. epidermidis using the microdilution method. Cytotoxicity of antibiotics was evaluated by MTT colorimetric assay on human corneal cell (HCE-2). Results: Cytotoxicity of antibiotics was evaluated on human epithelial corneal cells after 4 hours treatment by viability assay. Results showed that corneal cell viability was significantly higher after moxifloxacin treatment compared to Chloramphenicol (p<0.01). Moxifloxacin is characterized by a significantly lower MIC and IC50 values and a larger inhibition zone for all the strain tested, with high performance in controlling gram-negative growth, compared to Chloramphenicol. Moreover, Moxifloxacin showed higher activity compared to Chloramphenicol in the inhibition of biofilm formation and in the disruption of biofilm, especially against S. epidermidis biofilm. Conclusions: The lower corneal cell toxicity and the broader spectrum of antibacterial activity observed with Moxifloxacin suggests its use in ophthalmic solution for the treatment of bacterial eye infections.

Antiviral activity of sulphated specialized metabolites from sea urchin Clypeaster humilis: in vitro and in silico studies.

Chemical investigations of the sea urchin Clypeaster humilis has led to separation of twelve compounds including one new sulfonic acid derivative (7R) tridec-1-en-7-yl hydrogen sulphate (1), first isolated from natural source, pyridine-3-yl methane sulfonate (2), and first isolated from marine organisms, boldine (12), in addition to nine known compounds (3-11), which were isolated for the first time from the genus Clypeaster. Their structures were elucidated based on spectroscopic analyses (1D and 2D NMR), HR-ESI-MS as well as comparison with the previously reported data. The antiviral activity of the crude extract and sulphated compounds were evaluated using MTT colorimetric assay against Coxsackie B4 virus. The crude extract and compound 1 showed very potent antiviral activity with a percentage of inhibition equal to 89.7 ± 0.53% and 86.1 ± 0.92%, respectively. Results of the molecular docking analysis of the isolated compounds within Coxsackie Virus B4 (COX-B4) X-ray crystal structure and quantum chemical calculation for three sulphated compounds are in a consistent adaptation with the in vitro antiviral results. The pharmacokinetic properties (ADME) of isolated compounds were determined.

Investigating the therapeutic potential of monothiocarbohydrazones: A comprehensive in vitro evaluation of antioxidant, antimicrobial, and cytotoxic activities

Organic compounds, particularly those with nitrogen and sulfur heteroatoms, constitute over 99 % of clinically approved drugs. Among these, thiocarbohydrazones have been extensively studied, with a focus on symmetrical bis-substituted compounds. However, asymmetric and monosubstituted thiocarbohydrazones remain underexplored, despite their demonstrated high biological potential. This study presents an in-depth in vitro evaluation of the antioxidant, antimicrobial, and cytotoxic properties of eighteen previously synthesized and characterized monothiocarbohydrazones. The antioxidant potential was assessed using the DPPH assay, while the antimicrobial activity was determined against Gram-positive bacteria using a modified broth microdilution susceptibility method. The cytotoxic effect was evaluated on human hepatocellular carcinoma using the colorimetric MTT assay. The results reveal that the investigated monothiocarbohydrazones exhibit significant antioxidant and antimicrobial activities. Furthermore, their activity and cytotoxicity are influenced by the stereochemistry of the molecule and the nature and position of the substituents. These findings provide valuable insights for future in vivo examinations and underscore the potential of monothiocarbohydrazones in drug development.

Combination of Thymoquinone and Intermittent Fasting as a Treatment for Breast Cancer Implanted in Mice.

Breast cancer stands out as a particularly challenging form of cancer to treat among various types. Traditional treatment methods have been longstanding approaches, yet their efficacy has diminished over time owing to heightened toxicity, adverse effects, and the emergence of multi-drug resistance. Nevertheless, a viable solution has emerged through the adoption of a complementary treatment strategy utilizing natural substances and the incorporation of intermittent fasting to enhance therapeutic outcomes. This study aimed to assess the anticancer activity of thymoquinone (TQ), intermittent fasting, and their combination using in vivo and in vitro methods. The anti-proliferative activity of TQ and fasting (glucose/serum restriction) were evaluated against the T47D, MDA-MB-231, and EMT6 cell lines and compared to normal cell lines (Vero) using the MTT colorimetric assay method. Additionally, this study aimed to determine the half-maximal inhibitory concentration (IC50) of TQ. For the in vivo experiment, the antitumor activity of TQ and intermittent fasting (IF) was assessed by measuring the tumor sizes using a digital caliper to determine the change in the tumor size and survival rates. At the molecular level, the serum levels of glucose, β-hydroxybutyrate (β-HB), leptin, and insulin growth factor-1 (IGF-1) were measured using standard kits. Additionally, the aspartate transaminase (AST), alanine transaminase (ALT), and creatinine serum levels were measured. The inhibition of the breast cancer cell lines was achieved by TQ. TQ and intermittent fasting both had an additional anticancer effect against breast tumors inoculated in mice. The combination therapy was evaluated and found to significantly reduce the tumor size, with a change in tumor size of -57.7%. Additionally, the combination of TQ and IF led to a decrease in the serum levels of glucose, IGF-1 (24.49 ng/mL) and leptin (1.77 ng/mL) while increasing β-hydroxybutyrate in the mice given combination therapy (200.86 nM) with no toxicity on the liver or kidneys. In the mice receiving combination therapy, TQ and IF treated breast cancer in an additive way without causing liver or kidney toxicity due to decreased levels of glucose, IGF-1, and leptin and increased levels of β-hydroxybutyrate. Further investigation is required to optimize the doses and determine the other possible mechanisms exhibited by the novel combination.

Isolation and characterization of undescribed flavonoid from Abrus precatorius L. based on HPTLC-DPPH bioautography and its cytotoxicity evaluation

BackgroundNaturally derived compounds play a tremendous role as a drug as well as lead structure for the development of APIs. Therefore, isolation and characterization of compounds from nature are needed to alleviate life-threatening diseases. A. precatorius L. belongs to the family Leguminosae and is valued for its medicinal properties. Therefore, in this study, efforts are being made to isolate bioactive entity based on HPTLC-DPPH bioautography from APHA extract. Among all the separated compounds on TLC plate, the one (APSP-3) at Rf = 0.67 showed significant antioxidant activity, and hence, APSP-3 was further subjected to isolation, purification, and structural characterization using diverse analytical modus operandi such as 1D and 2D NMR, FTIR, HPLC–MS/MS, and elemental analysis. In addition, antioxidant and cytotoxicity evaluation of APHA extract and APSP-3 was pursued by standard DPPH and colorimetric MTT assays, respectively.ResultsAntioxidative isolated compound APSP-3 was scrutinized based on HPTLC-DPPH bioautography. The APSP-3 was found novel and spectroscopic data revealed the plausible structure; 7-hydroxy-3,5-dimethoxy-2-(4-((3,4,5-trihydroxy-6-(hydroxymethyl) tetrahydro-2H-pyran-2-yl)oxy) phenyl)-4H-chromen-4-one. Moreover, APSP-3 ascribed higher free radical scavenging activity with IC50 = 38.70 ± 3.5 µg/mL than standard ascorbic acid (75.19 ± 1.5 µg/mL). Cytotoxicity evaluation of APHA extract exhibited IC50 value 122.09 µg/mL for HepG2, 122.61 µg/mL for MCF-7, and 48.08 µg/mL for HCT116 cell lines, while APSP-3 displayed IC50 values 96.75 for HepG2, 61.67 for MCF-7, and 47.61 µg/mL for HCT116 cell lines.ConclusionsIn a nutshell, HPTLC-directed bioautography leads to the capturing of new flavonoid entity having antioxidant potency from APHA extract. The IC50 values obtained from cytotoxicity establish a dose–response relationship helping to determine the concentration at which a substance begins to exhibit toxic effects. This fundamental information is crucial for establishing safe dosage level in medical and pharmaceutical applications. Further, research engrossed in assessing other bioactivities involving in silico and in vivo studies obliged to offer a promising and secure portrayal for clinical implications.Graphical abstract

Effects of theasaponin E1 on the regulationglucose uptake of C2C12 myoblasts PI3K/Akt/mTOR signaling pathway

ABSTRACT Targeting disorders of glucose metabolism and the occurrence of hyperglycemia provides a potential therapeutic strategy for balancing energy to reduce insulin resistance such as type 2 diabetes. The present study investigated whether theasaponin E1 regulate energy metabolism in skeletal muscle cells. C2C12 mouse myoblasts were differentiated into myotubes . An MTT colorimetric assay verified cell viability. Theasaponin E1 (30, 45 or 60 mg/ ml), or metformin (2.5 mM), insulin (150 nM) for reference were used to treat the C2C12 myotubes. The expressions of energy metabolism were measured by western blotting in C2C12 myotubes. 2-NBDG kit to detect the glucose uptake capacity. Theasaponin E1 Treatment increases glucose uptake,GSK 3 β, as well as increases to the glycogen synthesis through the PI3K/AKT/mTOR signaling pathway, while reducing AMPK and ACC activities, and reducing the adipose synthesis pathway. These results suggest that theasaponin E1 may be increases glucose uptake by improving muscle energy metabolism.

Exploring the antiviral potential of justicidin B and four glycosylated lignans from Phyllanthus brasiliensis against Zika virus: A promising pharmacological approach

BackgroundZika virus (ZIKV) is an emerging arbovirus that in recent years has been associated with cases of severe neurological disorders, such as microcephaly in newborns and Guillain-Barré syndrome in adults. As there is no vaccine or treatment, the search for new therapeutic targets is of great relevance. In this sense, plants are extremely rich sources for the discovery of new bioactive compounds and the species Phyllanthus brasiliensis (native to the Amazon region) remains unexplored. PurposeTo investigate the potential antiviral activity of compounds isolated from P. brasiliensis leaves against ZIKV infection. MethodsIn vitro antiviral assays were performed with justicidin B (a lignan) and four glycosylated lignans (tuberculatin, phyllanthostatin A, 5-O-β-d-glucopyranosyljusticidin B, and cleistanthin B) against ZIKV in Vero cells. MTT colorimetric assay was used to assess cell viability and plaque forming unit assay to quantify viral load. In addition, for justicidin B, tests were performed to investigate the mechanism of action (virucidal, adsorption, internalization, post-infection). ResultsThe isolated compounds showed potent anti-ZIKV activities and high selectivity indexes. Moreover, justicidin B, tuberculatin, and phyllanthostatin A completely reduced the viral load in at least one of the concentrations evaluated. Among them, justicidin B stood out as the main active, and further investigation revealed that justicidin B exerts its antiviral effect during post-infection stages, resulting in a remarkable 99.9 % reduction in viral load when treatment was initiated 24 h after infection. ConclusionOur findings suggest that justicidin B inhibits endosomal internalization and acidification, effectively interrupting the viral multiplication cycle. Therefore, the findings shed light on the promising potential of isolated compounds isolated from P. brasiliensis, especially justicidin B, which could contribute to the drug development and treatments for Zika virus infections.

Andrographolide induced heme oxygenase-1 expression in MSC-like cells isolated from rat bone marrow exposed to environmental stress

Background and objectivesMesenchymal stem cells (MSC-like cells) are the most important stem cells that are used in transplantation clinically in various applications. The survival rate of MSC-like cells is strongly reduced due to adverse conditions in the microenvironment of transplantation, including environmental stress. Heme oxygenase-1 (HO-1) is a member of the heat shock protein, as well as a stress-induced enzyme, present throughout the body. The present study was conducted to investigate the effect of andrographolide, an active derivative from andrographolide paniculate, on HO-1 expression in mesenchymal stem cells derived from rat bone marrow. Materials and methodsThe rat bone marrow-derived mesenchymal stem cells (BMSC-like cells) were extracted and proliferated in several passages. The identity of MSC-like cells was confirmed by morphological observations and differential tests. The flow cytometry method was used to verify the MSC-specific markers. Isolated MSC-like cells were treated with different concentrations of andrographolide and then exposed to environmental stress. Cell viability was assessed using the MTT colorimetric assay. A real-time PCR technique was employed to evaluate the expression level of HO-1 in the treated MSC-like cells. ResultsIsolated MSC-like cells demonstrated fibroblast-like morphology. These cells in different culture mediums differentiated into osteocytes and adipocytes and were identified using alizarin red and oil red staining, respectively. As well, MSC-like cells were verified by the detection of CD105 surface antigen and the absence of CD14 and CD45 antigens. The results of the MTT assay showed that the pre-treatment of MSC-like cells with andrographolide concentration independently increased the viability and resistance of these cells to environmental stress caused by hydrogen peroxide and serum deprivation (SD). Real-time PCR findings indicated a significant increase in HO-1 gene expression in the andrographolide-receiving groups (p < 0.01). ConclusionOur results suggest that andrographolide creates a promising strategy for enhancing the quality of cell therapy by increasing the resistance of MSC-like cells to environmental stress and inducing the expression of HO-1.

Possibility of nanostructured lipid carriers encapsulating astaxanthin from Haematococcus pluvialis to alleviate skin injury in radiotherapy

Purpose The study aimed to protect patients’ skin against ionizing irradiation during radiotherapy by using astaxanthin-encapsulated nanostructured lipid carriers (NLC-ATX). Materials and methods NLC-ATX was prepared by a combined method of hot homogenization and sonication. Cytotoxicity of NLC-ATX was evaluated by MTT colorimetric assay. The in vitro radioprotection of NLC-ATX for human fibroblast (HF) cells was investigated based on the level of ROS (reactive oxygen species), DNA damage, and cell death caused by X-irradiation. In addition, the in vivo radioprotection was evaluated based on the appearance and histological structure of the irradiated skin. Results NLC-ATX was successfully prepared, with a mean particle size, zeta potential, and encapsulation efficiency of 114.4 nm, −34.1 mV, and 85.67%, respectively. Compared to the control, NLC-ATX, at an optimum ATX concentration under in vitro condition, reduced the amount of generated ROS and DNA damage of 81.6% and 41.6%, respectively, after X-radiation, resulting in a significant decrease in cell death by 62.69%. Under in vivo condition, after the 9th day of X-irradiation (equivalent to an accumulated dose of 14 Gy), the dorsal skin of five out of six NLC-ATX-untreated mice exhibited grade-1 skin damage, according to CTCAE v5.0, while treatment with NLC-ATX protected 6/6 mice from acute skin damage. Moreover, on the 28th day after the first X-irradiation, the histological images illustrated that NLC-ATX at an ATX concentration of 0.25 µg/mL exhibited good recovery of the skin, with barely any difference noted in the collagen fibers and sebaceous glands compared to normal skin. Conclusions NLC-ATX shows potential for application in skin protection against adverse effects of ionizing rays during radiotherapy.

Macromolecules: Synthesis, antimicrobial, POM analysis and computational approaches of some glucoside derivatives bearing acyl moieties

Macromolecules i.e., carbohydrate derivatives are crucial to biochemical and medical research. Herein, we designed and synthesized eight methyl α-D-glucopyranoside (MGP) derivatives (2–8) in good yields following the regioselective direct acylation method. The structural configurations of the synthesized MGP derivatives were analyzed and verified using multiple physicochemical and spectroscopic techniques. Antimicrobial experiments revealed that almost all derivatives demonstrated noticeable antifungal and antibacterial efficacy. The synthesized derivatives showed minimum inhibitory concentration (MIC) values ranging from 0.75 µg/mL to 1.50 µg/mL and minimum bactericidal concentrations (MBCs) ranging from 8.00 µg/mL to 16.00 µg/mL. Compound 6 inhibited Ehrlich ascites carcinoma (EAC) cell proliferation by 10.36% with an IC50 of 2602.23 μg/mL in the MTT colorimetric assay. The obtained results were further rationalized by docking analysis of the synthesized derivatives against 4URO and 4XE3 receptors to explore the binding affinities and nonbonding interactions of MGP derivatives with target proteins. Compound 6 demonstrated the potential to bind with the target with the highest binding energy. In a stimulating environment, a molecular dynamics study showed that MGP derivatives have a stable conformation and binding pattern. The MGP derivatives were examined using POM (Petra/Osiris/Molinspiration) bioinformatics, and as a result, these derivatives showed good toxicity, bioavailability, and pharmacokinetics. Various antifungal/antiviral pharmacophore (Oδ−, O′δ−) sites were identified by using POM investigations, and compound 6 was further tested against other pathogenic fungi and viruses, such as Micron and Delta mutants of SARS-CoV-2.

In-silico and in-vitro study reveals ziprasidone as a potential aromatase inhibitor against breast carcinoma

Aromatase enzyme plays a fundamental role in the development of estrogen receptors, and due to this functionality, the enzyme has gained significant attention as a therapeutic for reproductive disorders and cancer diseases. The currently employed aromatase inhibitors have severe side effects whereas our novel aromatase inhibitor is more selective and less toxic, therefore has greater potential to be developed as a drug. The research framework of this study is to identify a potent inhibitor for the aromatase target by profiling molecular descriptors of the ligand and to find a functional pocket in the target by docking and MD simulations. For assessing cellular and metabolic activities as indicators of cell viability and cytotoxicity, in-vitro studies were performed by using the colorimetric MTT assay. Aromatase activities were determined by a fluorometric method. Cell morphology was assessed by phase-contrast light microscopy. Flow cytometry and Annexin V-FITC/PI staining assay determined cell cycle distribution and apoptosis. This study reports that CHEMBL708 (Ziprasidone) is the most promising compound that showed excellent aromatase inhibitory activity. By using better drug design methods and experimental studies, our study identified a novel compound that could be effective as a high-potential drug candidate against aromatase enzyme. We conclude that the compound ziprasidone effectively blocks the cell cycle at the G1-S phase and induces cancer cell death. Further, in-vivo studies are vital for developing ziprasidone as an anticancer agent. Lastly, our research outcomes based on the results of the in-silico experiments may pave the way for identifying effective drug candidates for therapeutic use in breast cancer.

Improved Neural Differentiation of Human-induced Pluripotent Stem Cell [hiPSCs] on a Novel Polyurethane-based Scaffold Containing Iron Oxide Nanoparticles [Fe2O3 NPs

Repair of the nervous system in humans has always been complicated and faced difficulties. Cell transplantation approaches using biocompatible scaffolds might be an attractive therapeutic strategy for neuronal regeneration. We designed a cell delivery platform based on polyurethane [PU] and modified it with iron oxide nanoparticles [Fe2O3 NPs] for neural induction of human-induced pluripotent stem cells [hiPSC]. Forskolin, IBMX, and different ratios of FBS were employed to induce neurogenesis of hiPSCs. Neural differentiations were assessed at the level of genes and proteins. As was shown by MTT colorimetric assay, the proliferation and viability of SNL 76/7 on PU/ Fe2O3 were superior in comparison with pure PU and Fe2O3. hiPSCs cultured with PU/Fe2O3 exhibited an elevated expression of β3-tubulin, MAP2, NSE, OLIG2, as compared to controls. Furthermore, Acridine Orange staining assured the survival and viability of hiPSCs after 14 days of differentiation. All in all, our findings pointed out the biocompatibility and positive regulatory effect of PU/Fe2O3 on neural markers. We believe this scaffold could be a potential candidate for future nerve differentiation applications.

Cobalt (II), Nickel (II) and Palladium (II) complexes appended terpyridine‐based ligand: Synthesis, spectral characterization, anticancer activity and apoptosis investigation

Three new complexes containing 2‐thiophenylterpyridine (TP) ligand, namely, [Co(stpy)2]Cl2 (Co‐TP), [Ni(stpy)Cl2]2 (Ni‐TP) and [Pd(stpy)Cl]Cl (Pd‐TP) were synthesized and subsequently characterized employing various spectroscopic techniques including UV–Visible, IR, 1H NMR, EPR, and ESI‐MS analysis. ESI‐MS spectra suggested the complexes Co‐TP and Ni‐TP to be six coordinated with two ligand units and the complex Pd‐TP to be a four‐coordinated mononuclear square planar complex. The dimeric nature of the complex Ni‐TP (M2L2Cl4) was established by X‐ray diffraction analysis, which confirmed the bridging of two nickel centers by two chloride ions. The ligand and complexes were assessed for their cytotoxic behavior by using MTT colorimetric assay. The compounds TP, Co‐TP, Ni‐TP and Pd‐TP displayed significant antiproliferative activity on A549 (human lung carcinoma) and MM2 (human breast cancer) cell lines and had no toxic effect on human normal embryonic kidney cells (HEK 293). Further, the morphological changes associated with MM2 and A549 cells triggered by the tested compounds due to apoptosis have been validated with the help of AO‐EB and DAPI staining assays.

3D printed Styrax Liquidus (Liquidambar orientalis Miller)-loaded poly (L-lactic acid)/chitosan based wound dressing material: Fabrication, characterization, and biocompatibility results

The medicinal plant of Styrax liquidus (ST) (sweet gum balsam) which extracted from Liquidambar orientalis Mill tree, was loaded into the 3D printed polylactic acid (PLA)/chitosan (CS) based 3D printed scaffolds to investigate its wound healing and closure effect, in this study. The morphological and chemical properties of the ST loaded 3D printed scaffolds with different concentrations (1 %, 2 %, and 3 % wt) were investigated by Scanning Electron Microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FT-IR), respectively. In addition, the mechanical and thermal properties of the materials were investigated by Tensile test and Differential Scanning Calorimetry (DSC), respectively. The antimicrobial activities of the ST loaded 3D printed scaffolds and their incubation media in the PBS (pH 7.4, at 37 °C for 24 h) were investigated on two Gram-positive and two Gram-negative standard pathogenic bacteria with the agar disc diffusion method. The colorimetric MTT assay was used to determine the cell viability of human fibroblast cells (CCD-1072Sk) incubated with free ST, ST loaded, and unloaded 3D printed scaffolds. The 1 % and 2 % (wt) ST loaded PLA/CS/ST 3D printed scaffolds showed an increase in the cell number. Annexin V/PI double stain assay was performed to test whether early or late apoptosis was induced in the PLA/CS/1 % ST and PLA/CS/2 % ST loaded groups and the results were consistent with the MTT assay. Furthermore, a wound healing assay was carried out to investigate the effect of ST loaded 3D printed scaffolds on wound healing in CCD-1072Sk cells. The highest wound closure compared to the control group was observed on cells treated with PLA/CS/1 % ST for 72 h. According to the results, novel biocompatible ST loaded 3D printed scaffolds with antimicrobial effect can be used as wound healing material for potential tissue engineering applications.

Phytochemical Analysis, In Vitro Biological Activities, and Computer-Aided Analysis of Potentilla nepalensis Hook Compounds as Potential Melanoma Inhibitors Based on Molecular Docking, MD Simulations, and ADMET.

Potentilla nepalensis Hook is a perennial Himalayan medicinal herb of the Rosaceae family. The present study aimed to evaluate biological activities such as the antioxidant, antibacterial, and anticancer activities of roots and shoots of P. nepalensis and its synergistic antibacterial activity with antibacterial drugs. Folin-Ciocalteau and aluminium chloride methods were used for the calculation of total phenolic (TPC) and flavonoid content (TFC). A DPPH radical scavenging assay and broth dilution method were used for the determination of the antioxidant and antibacterial activity of the root and shoot extracts of P. nepalensis. Cytotoxic activity was determined using a colorimetric MTT assay. Further, phytochemical characterization of the root and shoot extracts was performed using the Gas chromatography-mass spectrophotometry (GC-MS) method. The TPC and TFC were found to be higher in the methanolic root extract of P. nepalensis. The methanolic shoot extract of P. nepalensis showed good antioxidant activity, while then-hexane root extract of P. nepalensis showed strong cytotoxic activity against tested SK-MEL-28 cells. Subsequently, in silico molecular docking studies of the identified bioactive compounds predicted potential anticancer properties. This study can lead to the production of new herbal medicines for various diseases employing P. nepalensis, leading to the creation of new medications.

Bioassay Techniques in Entomological Research

BIOASSAY (Bio - life, Assay - determination) is the measurement of the potency of any stimulus (physical, chemical, biological, and psychological) and the reactions that it produces in living matter. In Entomology, to know about the potency, Specificity of any compounds, standardization of doses, and identify modes of action bioassay is needed. In addition to the basic bioassay methods such as poison food, dry film techniques, etc., depending upon the researcher’s necessity, they are modified or newly developed. Diet covering and diet incorporation for lepidopterans and homopterans have been developed as advancements over the poison food assay method, by using insecticide-incorporated artificial diet. WHO cone, Wire ball test, Susceptibility test, CDC bottle assay and MCD assay developed for mosquito resistance monitoring and Larval Packet Technique (LPT) for cattle ticks, Sandwich method, TIBS, and Adult vial test are advancements over dry film technique. In Thrips Insecticide Bioassay System (TIBS), treated vials can be stored for up to 3 weeks at 4˚C for future efficacy assays at the field level. Adult Vial Tests have been developed for pentatomid and mirid adult bug bioassays. Larval Immersion Technique (LIT), Adult Immersion Technique (AIT) for cattle ticks, and MEBS have been developed as advancements over topical bioassay. In Mite Egg Bioassay System (MEBS), entomopathogenic fungi have been tested against Tetranychus cinnabarinus. MTT colorimetric assay and Trypan Blue Dye Exclusion assay helps to identify cell viability in insect cell line cultures. Spodoptera frugiperda cell line, Sf – 21 was tested against a scorpion, Leiurus quinquestriatus toxin β– BUTX – LQQ1a and the toxicity was determined through the viability of cell lines. Various basic and advanced bioassay techniques are discussed elaborately in this review.

Preparation of Novel Chitosan Oligosaccharide Quaternary Ammonium Derivatives Bearing Quinoline with Antioxidative and Antibacterial Activities

AbstractIn this study, five new chitosan oligosaccharide quaternary ammonium derivatives bearing quinoline are synthesized by reaction between 6‐O‐chloroacetyl‐2‐N, N, N‐trimethyl quaternary ammonium salt chitosan oligosaccharide (CTCOS, compound 1) and quinoline derivatives. The derivatives are characterized by analytical techniques including Fourier Transform Infrared (FTIR), 1H Nuclear Magnetic Resonance (1H NMR) spectroscopy, and elemental analysis. The obtained results confirm that quinoline groups are successfully introduced into the CTCOS molecule. Meanwhile, the antioxidant activities of the prepared chitosan oligosaccharide derivatives are evaluated in vitro. The experimental results show that the derivatives have excellent free radical scavenging ability. The free radical scavenging abilities are all over 75% at the concentration of 1.6 mg mL−1. Moreover, antibacterial tests show that derivative 2e has the best antibacterial activities. Its minimum inhibitory concentration (MIC) value and minimum bactericidal concentration (MBC) value against Staphylococcus aureus and Escherichia coli are 0.0625 and 0.03125 mg mL−1, respectively. In addition, the cytotoxicity of chitosan oligosaccharide and its derivatives is examined by MTT colorimetric assay in RAW 264.7 macrophages, observing that all derivatives are non‐cytotoxic. Consequently, the findings indicate that the enhanced antioxidant activity, antibacterial activity, and biocompatibility of these chitosan oligosaccharide derivatives can enlarge the scope of the application of chitosan oligosaccharides, particularly as antioxidant and antibacterial agents in food packaging, pharmaceutical, cosmetics industries, and other fields.

Synthesis, characterization and antiproliferative activity of platinum(II) complexes with 3-(2-pyridyl)-N1,2-methyl-1,2,4-triazoles

The present work describes the synthesis, investigation and in vitro cytotoxicity of Pt(II) complexes with 3-(2-pyridyl)-N1-methyl-1,2,4-triazole (L1) and 3-(2-pyridyl)–N2-methyl-1,2,4-triazole (L2). The crystal structures of PtL1Cl2 and PtL2Cl2 were established using X-Ray analysis, which revealed the square-planar PtN2Cl2 coordination geometry for the central metal ion. High-resolution mass spectrometry (HRMS), as well as one- and two-dimensional nuclear magnetic resonance (NMR) spectroscopy showed stability of the investigated complexes in DMF solution, in contrast to DMSO, where species were shown to partially dissociate. The cytotoxicity of the investigated compounds was determined by the colorimetric MTT assay against two human cancer cell lines, including cisplatin-sensitive human ovarian adenocarcinoma A2780 cells (IC50 = 26 ± 5 and 6.1 ± 0.6 for PtL1Cl2 and PtL2Cl2 correspondingly) and their cisplatin-resistant A2780cis analogue (IC50 = 24 ± 2 and 20 ± 3 for PtL1Cl2 and PtL2Cl2 correspondingly).