Fibroblast Cell Line L929 Research Articles

In vitro evaluation of cytotoxic potential of essential oil extracted from leaves of Croton heliotropiifolius Kunth in human tumor cells

ABSTRACT Croton heliotropiifolius Kunth, popularly known as “velame,” is a shrub that resides in northeastern Brazil. The essential oil of C. heliotropiifolius contains high concentrations of volatile compounds in the leaves and is widely used in folk medicine for many purposes as an antiseptic, analgesic, sedative, and anti-inflammatory agent. Due to the apparent limited amount of information, the aim of this study was to determine the cytotoxic potential of essential oil extracted from leaves of C. heliotropiifolius, utilizing different human cancer cell lines (HL-60, leukemia; HCT-116, colon; MDA-MB435, melanoma; SF295, glioblastoma) and comparison to murine fibroblast L929 cell line. The chemical characterization of the essential oil revealed the presence of large amounts of monoterpenes and sesquiterpenes, the majority of which were aristolene (22.43%), germacrene D (11.38%), ɣ-terpinene (10.85%), and limonene (10.21%). The essential oil exerted significant cytotoxicity on all cancer cells, with low activity on murine L929 fibroblasts, independent of disruption of cell membranes evidenced by absence of hemolytic activity. The cytotoxicity identified was associated with oxidative stress, which culminated in mitochondrial respiration dysfunction and direct or indirect DNA damage (strand breaks and oxidative damage), triggering cell death via apoptosis. Our findings suggest that extracts of essential oil of C. Heliotropiifolius may be considered as agents to be used therapeutically in treatment of certain cancers.

Gold nanoparticle conjugation enhances berberine's antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA)

Nanoparticle (NP) conjugation with various biomolecules is one of the most promising approaches for targeting Methicillin-resistant Staphylococcus aureus (MRSA). In this study, berberine (BER) was conjugated with gold nanoparticles (AuNPs) to enhance its antibacterial activity against MRSA. Chemically synthesized AuNPs were characterized by UV–vis spectroscopy, size distribution and Field Emission-Scanning Electron Microscope (FE-SEM) analysis. Berberine was conjugated with AuNPs and the conjugants were characterized using UV–vis spectroscopy and Fourier Transform Infrared (FTIR). The cytotoxicity of free and conjugated BER was also investigated. Comparative studies were conducted based on the Minimum Inhibitory Concentration (MIC) and anti-biofilm activities of conjugants and free BER against MRSA isolates. To verify cell membrane disruption and intracellular imbalance following treatment exposure, reactive oxygen species (ROS) and live-dead staining experiments were performed. In vivo antibacterial efficacy of treated groups was also assessed in a BALB/c mouse-infected skin model. DLS measurement, FE-SEM, and UV–vis spectroscopy confirmed the synthesis of AuNPs with a narrow size distribution of 49.38 nm and a zeta potential of −31.9 mV. The results from UV–vis spectroscopy and FTIR provided support for the functionalization of AuNPs by BER functional groups. The In vitro antibacterial results demonstrated that the conjugated BER exhibited a lower MIC value against MRSA (109.5 μg/ml) compared to free BER (165 μg/ml). Free and conjugated BER, at their MIC concentrations, demonstrated anti-biofilm activity, resulting in biofilm eradication of 13.9 and 22.33 %, respectively. The highest level of ROS production (93 %) was associated with the conjugated BER at a concentration of 27.37 μg/ml. This finding indicates a disruption in cell membrane integrity and a reduction in bacterial viability, as demonstrated by ROS and live/dead staining assays. The cytotoxicity study on the mouse L929 fibroblast cell line revealed approximately 100 % cell viability when exposed to free or conjugated BER at their MIC concentration. This result indicates the biosafety of both of the compounds. The in vivo study in the infected skin model groups treated with conjugated and free BER revealed MRSA survival rate of 2.7 % and 26 %, respectively. These findings suggest that conjugated BER could be an effective nanoformulation candidate with a potential role in managing MRSA associated infections.

Preparation of Self-Assembled, Curcumin-Loaded Nano-Micelles Using Quarternized Chitosan-Vanillin Imine (QCS-Vani Imine) Conjugate and Evaluation of Synergistic Anticancer Effect with Cisplatin.

Nano-micelles are self-assembling colloidal dispersions applied to enhance the anticancer efficacy of chemotherapeutic agents. In this study, the conjugate of quarternized chitosan and vanillin imine (QCS-Vani imine) was synthesized using the reaction of a Schiff base characterized by proton-NMR (1HNMR), UV-Vis spectroscopy, and FT-IR. The critical micelle concentration (CMC), particle size, and zeta potential of the resulting product were determined. The QCS-Vani imine conjugate was used as a carrier for the development of curcumin-loaded nano-micelles, and their entrapment efficiency (%EE), drug-loading capacity (%LC) and in vitro release were investigated using HPLC analysis. Moreover, the nano-micelles containing curcumin were combined with various concentrations of cisplatin and evaluated for a possible anticancer synergistic effect. The anticancer activity was evaluated against lung cancer A549 and mouse fibroblast L929 cell lines. The percent yield (%) of the QCS-Vani imine conjugate was 93.18%. The curcumin-loaded QCS-Vani imine nano-micelles were characterized and found to have a spherical shape (by TEM) with size < 200 nm (by DLS) with high %EE up to 67.61% and %LC up to 6.15 ± 0.41%. The loaded lyophilized powder of the nano-micelles was more stable at 4 °C than at room temperature during 120 days of storage. pH-sensitive release properties were observed to have a higher curcumin release at pH 5.5 (cancer environment) than at pH 7.4 (systemic environment). Curcumin-loaded QCS-Vani imine nano-micelles showed higher cytotoxicity and selectivity toward lung cancer A549 cell lines and exhibited lower toxicity toward the normal cell (H9C2) than pure curcumin. Moreover, the curcumin-loaded QCS-Vani imine nano-micelles exhibited an enhanced property of inducing cell cycle arrest during the S-phase against A549 cells and showed prominently induced apoptosis in lung cancer cells compared to that with curcumin. The co-treatment of cisplatin with curcumin-loaded QCS-Vani imine nano-micelles presented an enhanced anticancer effect, showing 8.66 ± 0.88 μM as the IC50 value, in comparison to the treatment with cisplatin alone (14.22 ± 1.01 μM). These findings suggest that the developed QCS-Vani imine nano-micelle is a potential drug delivery system and could be a promising approach for treating lung cancer in combination with cisplatin.

Synergistic effects of ozone with doxorubicin on the proliferation, apoptosis and metastatic profile of luminal-B type human breast cancer cell line

ObjectiveLuminal-B type human breast cancer cell line (BT-474) to assess the synergistic effects of ozone applied after chemotherapeutic treatment with various dosages of doxorubicin, and compare the results with the effects on L929 fibroblast cell line. MethodsDoxorubicin (1–50 M) was added to each cell lines and left to sit for 24 h at 37 °C. Then, as combination groups, half of the groups were incubated with 30 g/mL ozone for 25 min. Tumor necrosis factor alpha (TNF-α), transforming growth factor beta (TGF-β), and matrix metalloproteinase-2 and − 9 (MMP-2 and MMP-9) levels were measured using the MTT test, flow cytometry, and immunocytochemistry, respectively. ResultsWhen compared to simply doxorubicin-applied cells without ozone treatment, each dose of doxorubicin + ozone treatment considerably boosted L929 viability but significantly decreased BT-474 viability. Additionally, the combination increased the apoptotic impact of doxorubicin on BT-474 but not L929 (P < 0.001). TGF-, MMP-2, and MMP-9 levels of L929 after combination were substantially higher than those of the other groups (P < 0.01). Doxorubicin's effect on BT-474's protein levels, which had significantly decreased in comparison to those of the other groups, was reversed by the combination treatment (P < 0.05). ConclusionDoxorubicin's anti-proliferative and apoptotic effects were enhanced by ozone treatment in BT-474 cells, but it also repaired and healed healthy fibroblast cells that had been harmed by the cytotoxicity of the chemotherapy drug. If doxorubicin and ozone treatment are coupled, BT-474 cells may develop resistance to it through expressions of TNF-α, TGF-β, MMP-2, and MMP-9.

A Novel Brevinin2 HYba5 Peptide against Polymicrobial Biofilm of Staphylococcus aureus and Enterococcus faecalis.

Brevinin2 HYba5 (Peptide 29) is a novel cationic peptide identified from an endemic frog, Hydrophylax bahuvistara. Staphylococcus aureus and Enterococcus faecalis are troublesome biofilm-forming pathogens associated with nosocomial and community-acquired infections and contribute to the severity of infections associated with implanted devices and chronic wounds. Co-existence of both pathogens in biofilm mode contributes to an increased antibiotic resistance, treatment failure and hence persistent disease burden. Identifying a novel and stable, less toxic compound targeting multispecies biofilm with a lower probability of acquiring resistance in comparison to antibiotics is highly warranted. Evaluate the activity of Brevinin2 HYba5 against S. aureus and E. faecalis mixed biofilm. The anti-biofilm activity of peptide 29 was tested by Crystal violet assay, Confocal laser scanning Microscopy (CLSM) and MTT Assay. Cytotoxicity of the peptide was tested in RBC and L929 fibroblast cell line. Biofilm inhibitory activity of the peptide was evaluated at different temperatures, pH, serum and plasma concentrations. The antibiofilm potential of the peptide was tested against polymicrobial biofilm by Fluorescent in situ hybridisation (FISH) and plate counting on HiCromeTM UTI Agar media. The peptide 29 could inhibit biofilm formation of S. aureus and E. faecalis individually as well as in polymicrobial biofilm at 75 µM concentration. The peptide maintained its antibiofilm potential at different temperatures, serum and plasma concentrations. Activity of the peptide was high at acidic and neutral pH but found to get reduced towards alkaline pH. The peptide is nonhemolytic and does not exhibit significant cytotoxicity against the L929 fibroblast cell line (92.80% cell viability). The biofilm inhibition property makes peptide 29 a promising candidate for the management of S. aureus and E. faecalis biofilm, especially in catheter-associated devices to prevent the initial colonization and thus can ease the burden of pathogenic biofilm-associated infections.

Synthesis and screening for anticancer activity of two novel telluro-amino acids: 1,3-Tellurazolidine-4-carboxylic acid and tellurohomocystine.

Tellurium (Te) containing amino acids and their derivatives have the potential to participate in biological processes, which are currently being studied extensively to understand the function of Te in biological and pharmacological activities. Here, we are reporting the synthesis of two novel Te-containing unnatural amino acids; 1,3-Tellurazolidine-4-carboxylic acid [Te{CH2CH(COOH)NHCH2}] 5, and 4,4'-(1,2-Ditellurdiyl)bis(2-aminobutanoic acid), i.e., tellurohomocystine [TeCH2CH2CH(NH2)COOH]2 7, synthesized from tellurocystine, and L-methionine as precursors, respectively. These telluro-amino acids were thoroughly characterized by multinuclear (1H, 13C, 125Te) NMR spectroscopy, high-resolution ESI-mass spectrometry (ESI-MS), and elemental analysis. The telluro-amino acids 5 and 7 demonstrated good biocompatibility when in vitro cytotoxicity was analyzed on two fibroblast cell lines L929 and NIH/3T3. The treatment of telluro-amino acids 1,3-Tellurazolidine-4-carboxylic acid 5 and tellurohomocystine 7 on breast cancer cell line MCF-7 showed anticancer activity with IC50 values of 7.29 ± 0.27µg/mL and 25.36 ± 0.12µg/mL, respectively. The cell cycle distribution studies also revealed arrest at the sub-G1 phase suggesting telluro-amino acids to be apoptotic.

5′-O-methyl-14-hydroxyarmillane, a new armillane-type sesquiterpene from cultures of Guyanagaster necrorhiza

Protoilludene-type sesquiterpene aryl esters are a unique and very diverse compound class that were exclusively isolated from members of the genus Armillaria (Agaricomycetes, Physalacriaceae) up to this point. Herein, we describe the isolation and structural characterization of 5′-O-methyl-14-hydroxyarmillane (1), a new armillane-type derivative, that was obtained from submerged cultures of Guyanagaster necrorhiza (CBS 138623) together with the known congeners melleolide G (2), melleolide B (3), and 10-dehydroxy-melleolide B (4). ROESY data and coupling constants assigned the relative configurations of 1, while common absolute configurations were confirmed from comparison of its ECD spectrum to the one of 10-hydroxy-5′-methoxy-6′-chloroarmillane (5). Additionally, the configuration of melleolide G (2) was revised based on observed ROESY correlations. It is the first time that protoilludene-type sesquiterpene aryl esters were isolated from another genus, namely, Guyanagaster, that is closely related to Armillaria. 1–4 were evaluated for their biological activities in a serial dilution assay against several yeast, fungi, and bacteria, as well as in a cytotoxicity assay against different cell lines. Compound 4 was moderately active against Bacillus subtilis, Staphylococcus aureus, and Mucor hiemalis. Furthermore, 1, 3, and 4 showed weak cytotoxic effects against the mouse fibroblast cell line L929 and the cervix carcinoma cell line KB3.1.

Vitex Agnus‐Castus L. Nanoparticles: Preparation, Characterization and Assessment of Antimicrobial and Anticancer Activity

AbstractThis study was aimed to synthesise antimicrobial and therapeutic silver nanoparticles (AgNPs) by green synthesis method using Vitex agnus‐castus L. (VAC) seed extract. The synthesised VAC seed extract coated AgNPs were characterised by UV‐Vis spectrometry, Zeta analysis and FT‐IR spectrometry. The antifungal, antimicrobial, cytotoxicity by MTT assay, and anticarcinogenic activities on HeLa cells line were investigated. VAC‐AgNPs exhibited high antimicrobial activity against S. aureus with a zone diameter of 11.3±0.6 mm. In addition, the cytotoxicity of VAC‐AgNPs was evaluated using the L929 fibroblast cell line and the HeLa cells line, with AgNPs applied at six different concentrations. At a concentration of 5.0 mM, viability was found to be 47.6 %±4.2 % in L929 fibroblast cells and 39.6 %±7.9 % in HeLa cells, suggesting a potential antiproliferative effect of VAC‐AgNPs on HeLa cancer cells. The IC50 values of VAC‐AgNPs against the HeLa cell line were calculated as 2.94 μg/ml.

Activation of receptor-interacting protein 3-mediated necroptosis accelerates periodontitis in mice.

To investigate the involvement and role of receptor-interacting protein 3 (RIP3)-mediated necroptosis in periodontitis. A periodontitis murine model was established by oral infection with Porphyromonas gingivalis, and activation of necroptosis pathway was identified by immunohistochemistry. Adeno-associated virus was used to knock down Rip3 and the effect of Rip3 knockdown on periodontal inflammation was examined by Micro-CT, qRT-PCR and histological staining. In vitro, P. gingivalis-LPS was used to infect fibroblast cell line L929 and siRNA was used to knock down Rip3. Necroptosis pathway signalling and inflammation in cells were detected by cell viability and death assay, Western Blot, qRT-PCR and immunofluorescence analysis. Phosphorylation of RIP3 and mixed lineage kinase domain-like protein (MLKL) was increased in the periodontal ligament of mice infected with P. gingivalis. RIP3 knockdown reduced osteoclastogenesis and inflammatory cytokines in the periodontal area, and alleviated alveolar bone loss in vivo. In vitro, P. gingivalis-LPS-induced RIP3-mediated necroptosis in L929 cells, and knockdown of RIP3 by siRNA decreased the expression of inflammatory cytokines. RIP3-mediated necroptosis is activated in periodontitis and blocking necroptosis alleviates disease progression, indicating that RIP3 may be a potential target for periodontitis treatment.

2D Collagen Membranes from Marine Demosponge Chondrosia reniformis (Nardo, 1847) for Skin-Regenerative Medicine Applications: An In Vitro Evaluation.

Research in tissue engineering and regenerative medicine has an ever-increasing need for innovative biomaterials suitable for the production of wound-dressing devices and artificial skin-like substitutes. Marine collagen is one of the most promising biomaterials for the production of such devices. In this study, for the first time, 2D collagen membranes (2D-CMs) created from the extracellular matrix extract of the marine demosponge Chondrosia reniformis have been evaluated in vitro as possible tools for wound healing. Fibrillar collagen was extracted from a pool of fresh animals and used for the creation of 2D-CMs, in which permeability to water, proteins, and bacteria, and cellular response in the L929 fibroblast cell line were evaluated. The biodegradability of the 2D-CMs was also assessed by following their degradation in PBS and collagenase solutions for up to 21 days. Results showed that C. reniformis-derived membranes avoided liquid and protein loss in the regeneration region and also functioned as a strong barrier against bacteria infiltration into a wound. Gene expression analyses on fibroblasts stated that their interaction with 2D-CMs is able to improve fibronectin production without interfering with the regular extracellular matrix remodeling processes. These findings, combined with the high extraction yield of fibrillar collagen obtained from C. reniformis with a solvent-free approach, underline how important further studies on the aquaculture of this sponge could be for the sustainable production and biotechnological exploitation of this potentially promising and peculiar biopolymer of marine origin.

Green synthesis of copper oxide nanoparticles using okra (Abelmoschus esculentus) fruit extract and assessment of their cytotoxicity and photocatalytic applications

In this research, copper oxide nanoparticles (CuO-NPs) were biosynthesized through a simple, quick, and green manner by the exertion of okra (Abelmoschus esculentus) fruit extract as a stabilizer and reducing agent. TGA/DTA, UV–Vis, DRS, FTIR, XRD, BET, TEM, FESEM/EDX, and PSA analyses were used to characterize the CuO-NPs. The XRD analysis of CuO-NPs exhibited the monoclinic structure of crystallizes in a size of about 25 nm. The optical and surface characteristics of CuO-NPs were examined by the results of DRS and BET analyses, while their surface area was observed to be 5.6 m2/g. The absorption spectra of NPs revealed a peak at around 320 nm. In addition, as their FESEM/TEM/PSA images displayed their spherical shape in a size of about 20 nm. The effectiveness of CuO-NPs as a photocatalyst was examined by testing their ability to degrade various dyes under UV light. According to the outcomes after 210 min, the degradation percentages of Eriochrome black T (EBT), Rhodamine B (RhB), and Methylene blue (MB) dyes were approximately 88, 96, and 88%, respectively. The anticancer efficacy of CuO-NPs against the HeLa cancer cell line in 24, 48, and 72 h (IC50=1000, 500, and 250 μg/mL, respectively) was observed to be significant, whereas lower cytotoxic effects were induced on the normal fibroblast L929 cell line when compared to that of cancer cells. The viability of normal cells at a concentration of 1000 μg/mL was decreased to 74, 71, and 65% after 24, 48, and 72 h, respectively.

Chalcone-based dipolar cycloaddition of novel heteroaromatic compounds: Their anticancer examination

In this study, new chalcone-isoxazole-based hybrid derivatives (12, 13, 18, and 21) and chalcone-triazole hybrid molecules (26) were synthesized using the click chemistry approach. Among these hybrid molecules, 12, 13, 21, and 26 were obtained by combining azole pharmacophore groups such as thiazole, isoxazole, and 1,2,3-triazole. In addition, a new bioactive hybrid molecule (E)-3-(2-(1-((4- (3-(4-((3-(4-(benzyloxy) phenyl) isoxazol-5-yl) methoxy) phenyl)–3–oxoprop-1-en-1-yl) phenoxy)methyl)-1H-1,2,3-triazol-4-yl)thiazol-4-yl)-2H-chromen-2-one (18), which is a chalcone-isoxazole hybrid derivative with azido thiazole coumarin substitution, was synthesized. The chemical structures of synthesized compounds were characterized by spectroscopic techniques such as 1H NMR, 13C NMR, FT-IR, and MALDI-TOF MS (18, 21, and 26). These novel chalcone-azole hybrids (12, 13, 18, 21, and 26) were investigated for their in-vitro anticancer activity against A549 cells, Capan-1 cell lines, and a healthy L929 fibroblast cell line, as well as the apoptotic and necrotic effects of these compounds on Capan-1 cell lines. Among the compounds tested, hybrid 18 showed the best activity in the A549 cell line, while it showed moderate activity in the Capan-1 cell line.

Phytoextract-mediated synthesis of Cu doped NiO nanoparticle using cullon tomentosum plant extract with efficient antibacterial and anticancer property

In the present study, nickel oxide (NiO) and copper-doped nickel oxide (NiCuO) nanoparticles (NPs) were successfully synthesized using Cullen tomentosum plant extract with the co-precipitation method. This work focuses on the Phyto-mediated synthesis and characterization of NPs for their biological applications. Phytochemicals that exist in the plant extract acts as reducing and capping agent. The successful formation of the NPs was validated by various analysis as XRD, FESEM, EDAX, FT-IR, UV–Vis, and Photoluminescence. According to XRD studies, the average crystallite size of NiO and NiCuO NPs is 36 nm and 31 nm, respectively. The river stone and nanoflower like morphology for NiO and NiCuO NPs are confirmed by FESEM image. Furthermore, the synthesized NPs were tested against Gram-positive (Bacillus subtilis, Streptococcus pneumoniae) and Gram-negative (Klebsiella pneumoniae, Escherichia coli) bacteria, which showed enhanced antibacterial activity of NiCuO NPs. The cytotoxicity of NPs was investigated against human breast cancer cells (MDA-MB-231) and fibroblast L929 cell lines. Also, the IC50 value for human breast cancer cells is 11.8 μg/mL. According to these findings, NiCuO NPs are potential nanomaterials with advanced healthcare uses.

Formulation and Evaluation of Chitosan-PLGA Biocomposite Scaffolds Incorporated with Quercetin Liposomes Made by QbD Approach for Improved Healing of Oral Lesions.

The current research aims to develop and evaluate chitosan-PLGA biocomposite scaffolds in combination with quercetin liposomes to accomplish the desired impact in oral lesions where pharmacotherapeutic agent treatment through circulation could only reach the low content at the target. Optimization of quercetin-loaded liposomes was carried out using 32 factorial design. The preparation of porous scaffolds comprising produced quercetin-loaded liposomes by thin-film method was carried out in the current study using a unique strategy combining solvent casting and gas foaming procedures. The prepared scaffolds were tested for physicochemical properties, in vitro quercetin release study, ex vivo drug permeation and retention research using goat mucosa, antibacterial activity, and cell migration studies on fibroblast L929 cell lines. Improved cell growth and migration were seen in the order control < liposomes < proposed system. The proposed system has been examined for its biological and physicochemical features, and it has the potential to be utilized as an efficient therapy for oral lesions.

Fabrication of Zr-Ti-Si glassy metallic overlay on 3D printed Ti-6Al4V implant prototypes for enhanced biocompatibility

Implantable materials with enhanced corrosion resistance and bioactivity are highly desired for biomedical application. A 3D metal printed prototype surface was coated with amorphous alloy components of elements Zr, Ti and Si (Zr54Ti35 Si11) as a novel biomedical grade ternary system for the implants. A DC magnetron sputtering technique was used to sputter the films on Ti-6Al-4 V implant prototypes, exhibiting a smooth and denser coating with root means square surface roughness of 1.4 nm. Surface morphology and structural integrity of the thin film metallic glass (TFMG) was studied from electron microscopic techniques and X-ray diffraction analysis. The TFMG is thermally stable with a large supercooled liquid region (∆Tx) of 81.2 °C. Mechanical characteristics of the coatings were studied using the nanoindentation method to correlate the hardness and modulus of the implant materials. Under the simulated body fluid condition, Ti-6Al-4 V implant with 500 nm thickness coatings (Zr54Ti35Si11) exhibited the superior corrosion resistance rate 1 × 10−6 mpy with a charge transfer resistance of 34 kΩ promising for practical application. Electrochemical impedance spectra and Tafel analysis revealed that the corrosion resistance was prominent in the 500 nm thickness TFMG system than the 250 nm and uncoated bare counterparts. The in-vitro biocompatibility and cytotoxicity studies of the as-coated 3D printed implants were studied using the human osteoblast and L929 fibroblast cell lines, respectively. A hemocompatibility study on the implant materials revealed that erythrocytes remain biconcave without aggregation, platelet activation and thrombogenesis, supporting the biocompatibility of the implants. The integration of such unique properties makes these TFMG system ideal candidates for biomedical implants.

In vitroandin silicoevaluation of the design of nano-phyto-drug candidate for oral use againstStaphylococcus aureus

Onopordum acanthiumis a medicinal plant with many important properties, such as antibacterial, anticancer, and anti-hypotensive properties. Although various studies reported the biological activities ofO. acanthium, there is no study on its nano-phyto-drug formulation. The aim of this study is to develop a candidate nano-drug based on phytotherapeutic constituents and evaluate its efficiencyin vitroandin silico. In this context, poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) ofO. acanthiumextract (OAE) were synthesized and characterized. It was determined that the average particle size of OAE-PLGA-NPs was 214.9 ± 6.77 nm, and the zeta potential was −8.03 ± 0.85 mV, and PdI value was 0.064 ± 0.013. The encapsulation efficiency of OAE-PLGA-NPs was calculated as 91%, and the loading capacity as 75.83%. Thein vitrodrug release study showed that OAE was released from the PLGA NPs with 99.39% over the 6 days. Furthermore, the mutagenic and cytotoxic activity of free OAE and OAE-PLGA-NPs were evaluated by the Ames test and MTT test, respectively. Although 0.75 and 0.37 mg/mL free OAE concentrations caused both frameshift mutation and base pair substitution (p&lt; 0.05), the administered OAE–PLGA NP concentrations were not mutagenic. It was determined with the MTT analysis that the doses of 0.75 and 1.5 mg/mL of free OAE had a cytotoxic effect on the L929 fibroblast cell line (p&lt; 0.05), and OAE-PLGA-NPs had no cytotoxic effect. Moreover, the interaction between the OAE andS. aureuswas also investigated using the molecular docking analysis method. The molecular docking and molecular dynamics (MD) results were implemented to elucidate theS. aureusMurE inhibition potential of OAE. It was shown that quercetin in the OAE content interacted significantly with the substantial residues in the catalytic pocket of theS. aureusMurE enzyme, and quercetin performed four hydrogen bond interactions corresponding to a low binding energy of −6.77 kcal/mol with catalytic pocket binding residues, which are crucial for the inhibition mechanism ofS. aureusMurE. Finally, the bacterial inhibition values of free OAE and OAE–PLGA NPs were determined againstS. aureususing a microdilution method. The antibacterial results showed that the inhibition value of the OAE–PLGA NPs was 69%. In conclusion, from thein vitroandin silicoresults of the nano-sized OAE-PLGA NP formulation produced in this study, it was evaluated that the formulation may be recommended as a safe and effective nano-phyto-drug candidate againstS. aureus.

Mouse Fibroblast L929 Cell Line as a Useful Tool for Replication and Adaptation of Infectious Bursal Disease Virus.

Infectious bursal disease virus (IBDV) causes a highly contagious disease associated with immunosuppression in young chickens. Production of either egg-based or primary cell-based high-quality vaccines requires time-consuming and costly procedures. To determine a suitable cell line for IBDV replication, L929 cell line was a candidate for the growth kinetics processing of the virus. The L929 cells were proliferated in monolayer, and doubling time was calculated. Replication kinetics an IBDV isolate at the multiplicity of infection 0.1 PFU/cell were determined using virus titration. To adapt IBDV on L929 cells, seven consecutive passages were performed. Virus titer and levels of apoptosis were quantitatively analyzed at each passage. The viral VP2 gene was amplified and sequenced in three passages. An average doubling time of 21 h was estimated for monolayers of L929 cells. Although during early passages, virus growth did not produce a clear cytopathic effect (CPE), an increase in IBDV titers was observed. Serial passages led to the evidence of marked CPEs and an increase in the virus titer in the third passage. During the fourth to seventh passages, consistent CPEs characterized by the formation of granulated and round cells were evident within 24 to 48 hours post-inoculation. The titer of the virus was increased in the third passage onwards to peak in the fourth and constant at 5.9 TCID50 until the end passage. The IBDV replication in connection with DNA fragmentation and FITC, revealed the characteristic picture of apoptosis in a time-dependent manner. We found that the IBDV could easily be adapted to L929 cells, increasing virus yields by about two orders of magnitude. These results indicated that the cell line may be useful in the production of efficient virus particles.

Screening of AS101 analog, organotellurolate (IV) compound 2 for its in vitro biocompatibility, anticancer, and antibacterial activities.

Organotellurium compounds are being well researched as potential candidates for their functional roles in therapeutic and clinical biology. Here, we report the in vitro anticancer and antibacterial activities of an AS101 analog, cyclic zwitterionic organotellurolate (IV) compound 2 [Te-{CH2CH(NH3+)COO}(Cl)3]. Different concentrations of compound 2 were exposed to fibroblast L929 and breast cancer MCF-7 cell lines to study its effect on cell viability. The fibroblast cells with good viability confirmed the biocompatibility, and compound 2 also was less hemolytic on RBCs. A cytotoxic effect on MCF-7 breast cancer cell line investigated compound 2 to be anti-cancerous with IC50 value of 2.86 ± 0.02µg/mL. The apoptosis was confirmed through the cell cycle phase arrest of the organotellurolate (IV) compound 2. Examination of the antibacterial potency compound 2 was done based on the agar disk diffusion, minimum inhibitory concentration, and time-dependent assay for the Gram-positive Bacillus subtilis and Gram-negative Pseudomonas putida. For both bacterial strains, tests were performed with the concentration range of 3.9-500μg/mL, and the minimum inhibition concentration value was found to be 125μg/mL. The time-dependent assay suggested the bactericidal activity of organotellurolate (IV) compound, 2 against the bacterial strains.

Characterization of p(PmO), p(LO) and p(RO) organoparticles, their bioactivity properties and their effect on pancreatic cancer Capan-1 cell

For the first time in the literature, p (PmO), p (LO) and p (RO) organo-particles were synthesized from Peppermint oil, Lemon oil and Rose oil. Of the organo-particles L-929 cell line viability/cytotoxicity and anticancer effect against Capan-1 pancreatic cancer cell line were investigated. p (PmO), p (LO) and p (RO) organo-particles were featured by thermogravimetry (TGA), Fourier Transform Infrared Spectroscopy (FTIR), Scanning electron microscope (SEM), Particle size (DLS), and particle charge (zeta potential, Zeta) analyses. Antioxidant, biocompatible, and antimicrobial activities and in vitro cytotoxicity specialties were investigated. In studies on Capan-1 and L-929 cell lines, it was observed that p (PmO), p (LO) and p (RO) organo-particles were effective on L-929 fibroblast cell line on Capan-1 pancreatic cancer cell line. In addition, it was observed that p (PmO), p (LO) and p (RO) organo-particles were not toxic in L-929 cell lines at high doses. When the Capan-1 cell line MTT analysis results of p (PmO), p (LO) and p (RO) organo-particles were examined, a difference was observed between cell viability rates and apoptosis and necrosis values. The highest % apoptosis rate was observed in the p (RO) organo particle.

Green synthesis of multi-functional carbon dots from medicinal plant leaves for antimicrobial, antioxidant, and bioimaging applications

In this research work, carbon dots (CDs) were synthesized from the renewable leaves of an indigenous medicinal plant by the one-pot sand bath method, Azadirachta indica. The synthesized CDs were characterized for its optical properties using UV–Vis, Fluorescence and Fourier transform infrared (FT-IR) spectrophotometry and for structural properties using dynamic light scattering (DLS), X-ray Diffraction (XRD) and high resolution Transmission electron microscopy (HR-TEM). The synthesized CDs exhibited concentration dependent biocompatibility when tested in mouse fibroblast L929 cell line. The EC50 values of biomedical studies, free radical scavenging activity (13.87 μgmL−1), and total antioxidant capacity (38 μgmL−1) proved CDs were exceptionally good. These CDs showed an appreciable zone of inhibition when examined on four bacterial (two gram-positive and gram-negative) and two fungal strains at minimum concentrations. Cellular internalisation studies performed on human breast cancer cells (MCF 7- bioimaging) revealed the applicability of CDs in bioimaging, wherein the inherent fluorescence of CDs were utilised. Thus, the CDs developed are potential as bioimaging, antioxidants and antimicrobial agents.