Microculture Tetrazolium Assay Research Articles

Anticancer effect of combined cinnamon–saffron versus its nanoparticles on oral squamous cell carcinoma cell line

Background Oral squamous cell carcinoma (OSCC), the eighth most common malignant neoplasia, often requires surgery, radiation therapy, and chemotherapy. However, cancer cells can become resistant, leading to the use of natural components in anticancer drugs. Objectives Evaluate the anticancer effect of combined cinnamon–saffron extract as compared to nanoform of this compound and doxorubicin on the cells of OSCC cell lines in relation to cell viability, migration and apoptotic efficiency. Materials and methods (SCC-25) OSCC cell lines were used. Scanning electron microscope was used to prepare and characterize the negative control group, cinnamon–saffron, nano cinnamon–saffron, and doxorubicin. The microculture tetrazolium assay viability assay was used to evaluate each study group that was examined, and the IC50 value was then determined. Cell cycle and apoptosis analysis in varied research groups were evaluated using flow cytometry and the IC50 dosages. Additionally, a wound-healing assay was used to assess the invasion and migration ability of the cells. Results When treated with cinnamon–saffron, nano cinnamon–saffron, and doxorubicin, the various experimental groups showed dose-dependent reductions in their % viability and IC50 values, which have potential effects against OSCC cell lines, according to our findings. It was discovered that treated cells exhibited cytotoxic, cell cycle arrest, apoptotic, and antimigratory effects as compared to untreated cells. Interestingly, the results of the present study when compared to the untreated group pointed out that, cinnamon–saffron, nano cinnamon–saffron, and doxorubicin reduced the viability of OSCC cells and increased the percentage of total apoptotic cells and necrotic cell death. Conclusion Nano cinnamon–saffron exerts potent cell cycle arrest, cytotoxic, and apoptotic effects on oral squamous carcinoma cell lines.

Evaluation of bioactivity and antibacterial properties of Ti6Al4V-based green biocomposite implant encompassing TiO2 nanotube arrays and garlic extract

This study involved the incorporation of an antibacterial garlic extract into titanium oxide nanotubes (TNTs) formed via the anodization of Ti6Al4V implants. The garlic extract, obtained through low-temperature extraction aided by ultrasound waves, was loaded into the nanotubes. The presence of the nanotubes was confirmed through X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDS), and scanning electron microscopy (SEM). Fourier-transform infrared spectroscopy (FT-IR) and gas chromatography-mass spectrometry (GC-MS) were used to investigate the presence of bioactive compounds, particularly sulfur compounds responsible for garlic's antibacterial effects. The impact of loading two concentrations (0.1 and 0.2 g per milliliter) of garlic extract on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) bacteria was examined. Results indicated a decrease in the growth range of S. aureus from 109 to 106 (CFU/ml) and E. coli from 1011 to 109 (CFU/ml) upon treatment. Additionally, cell adhesion and viability tests conducted on MG63 cells revealed an 8% increase in cell viability with the 0.1 g per milliliter concentration and a 35% decrease with the 0.2 g per milliliter concentration of garlic extract after 72 h of incubation (They have been evaluated by Microculture tetrazolium (MTT) assay). GC-MS analysis identified the presence of diethyl phthalate compounds in the garlic extract, suggesting a potential correlation with cellular toxicity observed in the sample with the higher concentration (0.2 g per milliliter) of garlic extract. Overall, the TNTs loaded with 0.1 g per milliliter of garlic extract simultaneously demonstrated antibacterial activity, cell viability, adhesion, and growth enhancement.

Design, synthesis and molecular docking study of coumarin pyrazoline derivatives against MCF-7 breast cancer cell line

A new eight series of 3-(2-oxo-2H-chromen-3-yl)-5-(substituted phenyl)-1H-pyrazole-1-carbaldehyde derivatives (9–16) were designed and created from coumarin-chalcone derivatives (1–8). The structures of the derivatives were established by using melting point, mass spectrum, IR, 1HNMR, and 13C NMR spectroscopic methods. In vitro antiproliferative activities were evaluated against MCF-7 breast cancer cell line using Microculture Tetrazolium (MTT) assay. The results showed that the compounds 9, 12- 14 has a moderate activity against MCF-7 breast cancer cell line with IC50 61.44, 70.11, 22.6 and 25.99 µg/mL respectively, while the compounds 10,11, 15 and 16 were found to be inactive against studied cell line within IC50 > 100 µg/mL. The possible binding interaction between studied compounds (9–16) and human ER-α (PDB ID: 1ERR) were studied by molecular docking. The results revealed that only the compounds 11 and 16 form π -H interaction with ER-α (PDB ID: 1ERR) within the highest negative values of binding affinity -7.04260 and -7.17308 kcal.mol-1 respectively than the other compounds, while Raloxifene used here as a positive control form a strong ionic bonding with Asp 351 within the binding affinity -9.61928 kcal/mol which is more negative value than the studied compounds.

Screening The Anticancer Activity for New Schiff Bases of Natural Steroids

The significance of cholesterol and ergosterol (provitamin d2) in biological systems has prompted interest in these compounds as pharmaceuticals. It has been reported on the effective straightforward synthesis of Schiff base modifications of cholesterol aldehyde and ergosterol aldehyde and their assessment as possible agents against cancer (esophagus natural cell lines and human cancer cells). Through a process called Steglish esterification, the initial cholesterol and ergosterol are converted into their respective aldehyde derivatives, by reacting with amines such as 2-amino 6-fluoro benzothiazole, carbohydrazide, and thiosemicarbazide, these aldehyde derivatives are transformed into the appropriate Schiff base derivatives, and then their anticancer activity is checked using the MTT (Microculture Tetrazolium Assay) assay method. For the purpose of analyzing their cytotoxic effects, these compounds were tested on cancerous (SK-GT-4, human esophageal adenocarcinoma) and normal (normal cell line, Rat Embryonic fibroblast (REF)) cell lines. The MTT assay results revealed that the compounds (a1, a2, c1, c2, compound 1) were more harmful to human esophageal cancer cell line than to healthy cell line other than compounds (t1, t2, compound 2, ergosterol and cholesterol alone). Overall, our research indicated that provitamin D2 (ergosterol), a1, a2, c2, t2, compound 1, had a growth-inhibiting effect on both cell lines compared to c1, t1, compound 2, and cholesterol alone.

Comprehensive Metabolite Profiling of Chemlali Olive Tree Root Extracts Using LC-ESI-QTOF-MS/MS, Their Cytotoxicity, and Antiviral Assessment.

The large quantity of olive roots resulting from a large number of old and unfruitful trees encouraged us to look for ways of adding value to these roots. For this reason, the current research work is devoted to the valorization of olive roots by identifying active phytochemicals and assessing their biological activities, including the cytotoxicity and antiviral potential of different extracts from the Olea europaea Chemlali cultivar. The extract, obtained by ultrasonic extraction, was analyzed using the liquid chromatography-mass spectrometry technique (LC-MS). The cytotoxicity was evaluated through the use of the microculture tetrazolium assay (MTT) against VERO cells. Subsequently, the antiviral activity was determined for HHV-1 (Human Herpesvirus type 1) and CVB3 (Coxsackievirus B3) replication in the infected VERO cells. LC-MS analysis allowed the identification of 40 compounds, classified as secoiridoids (53%), organic acids (13%), iridoids (10%), lignans (8%), caffeoylphenylethanoid (5%), phenylethanoids (5%),sugars and derivatives (2%), phenolic acids (2%), and flavonoids (2%). It was found that extracts were not toxic to the VERO cells. Moreover, the extracts did not influence the appearance of HHV-1 or CVB3 cytopathic effects in the infected VERO cells and failed to decrease the viral infectious titer.

Fabrication and Characterization of Piezoelectric PEO/SF/BaTiO3 Scaffolds for Cardiac Tissue Engineering

The existence of an intrinsic electrical platform responsible for the formation and transmission of impulses is essential, especially in cardiac tissue. However, most cardiac tissue made from biodegradable polymeric materials lacks conductive characteristics; this delays regional conduction, potentially causing arrhythmias. This study proposes a conductive polyethylene oxide (PEO)/silk fibroin (SF)-based material conjugated with conductive nanoparticles as a cardiac patch to fix any infarcted heart part. A new composite of PEO/15 wt%SF/0.2 wt%BaTiO3 was prepared and characterized in vitro. The obtained patches were characterized by conventional Bragg-platinum-conductive action (XRD), FTIR spectroscopy, Raman spectra, and thermogravimetric analysis. A PiezoTester device was used to evaluate the piezoelectric properties. The produced samples of 500 μm thickness were assessed in tapping mode. The applied load was selected to be as low as possible, and the frequencies were adjusted to simulate the heartbeats, ranging from 10 to 100 Hz. The results showed that a maximum of around 1100 mV was obtained at a load of 20 N. A maximum of about 80 mV was received at an applied force of 1 N and a frequency of 100 Hz, which matches the electricity generated by the human heart. The cytotoxicity effect of prepared films was tested against AC16 cells using microculture tetrazolium assay (MTT). The pristine PEO cell viability either was not affected by adding SF or slightly decreased. However, the cell viability dramatically increased by adding BaTiO3 to the PEO/SF composites. The confocal microscope images proved that the cells showed a spread morphology. The cells adhered to the PEO membranes and demonstrated a well-spread morphology. Overall, our study suggests that the PEO/SF/BaTiO3 composite can be a promising cardiac patch material for repairing infarcted heart tissue, as it is conductive, has good mechanical properties, and is biocompatible.

Gentamycin-loaded halloysite-based hydrogel nanocomposites for bone tissue regeneration: fabrication, evaluation of the antibacterial activity and cell response

Biocompatible hydrogels are promising approaches for bone repair and engineering. A novel therapeutic nanocomposite hydrogel was designed based on triblock copolymer poly e-caprolactone (PCL)-polyethylene glycol-PCL and natural gelatin (PCEC/GEL) and reinforced with halloysite nanotube (HNT). Gentamicin (GM) loaded HNT was immobilized in polymeric hydrogel matrix to fabricate scaffolds using the freeze-drying method. Scaffolds were characterized via Fourier transform infrared (FT-IR), x-ray powder diffraction, and scanning electron microscope (SEM) methods. The swelling ratio, density, porosity, degradation, and mechanical behavior were evaluated to investigate the effects of HNT on the physicochemical properties of the composite. Cell viability and cell attachment were investigated by microculture tetrazolium (MTT) assay and SEM. Cell proliferation was observed without any cytotoxicity effect on human dental pulp-derived mesenchymal stem cells (h-DPSCs). Alizarin red staining and real-time reverse transcription polymerase chain reaction (QRT-PCR) assay were carried out to monitor the osteoconductivity of scaffolds on h-DPSCs which were seeded drop wise onto the top of scaffolds. The quantification of the messenger RNA (mRNA) expression of osteogenic marker genes, bone morphogenetic protein 2, SPARK, bone gamma-carboxyglutamate protein and runt-related transcription factor 2 over a period of 21 d of cell seeding, demonstrated that cell-encapsulating PCEC/GEL/HNT-GM hydrogel scaffolds supported osteoblast differentiation of h-DPSCs into osteogenic cells through the up-regulation of related genes along with moderate effects on cell viability. Moreover, the antibiotics loading reduced bacterial growth while maintaining the osteogenic properties of the scaffold. Therefore, the bactericidal PCEC/GEL/HNT-GM hydrogel nanocomposite, with enhanced durability, maintenance the functionality of seeded cells in vitro that can be a remarkable dual-functional candidate for hard tissue reconstruction and customized bone implants fabrication via the direct incorporation of bactericidal drug to prevent infection.

Bioinductive and anti-inflammatory properties of Propolis and Biodentine on SHED

ObjectivesThis study aimed to evaluate and compare the cell viability, differentiation potential and anti-inflammatory potential of propolis and Biodentine™ on stem cells isolated from human exfoliated deciduous teeth (SHED). Materials and methodsSHED were segregated and cultured from the dental pulp of children after therapeutic extraction. Microculture Tetrazolium Assay (MTT) assay was carried out for assessing cell proliferation potential of propolis and Biodentine at different concentrations. As per the results from cell proliferation assay, cell differentiation potential of SHED was evaluated at concentration of 12.5 μg/ml using Alizarin Red staining. The anti-inflammatory potential of test materials was evaluated using gelatin zymography by detecting MMP-2 and MMP-9. ResultsThe maximum cell proliferation percentage of SHED treated with propolis and Biodentine was observed at a concentration of 12.5 μg/ml, on day 7, 14 and 21 with Biodentine having maximum cell proliferation potential followed by propolis. SHED treated with Biodentine showed maximum cell differentiation on day 7 (107.16), 14 (106.29) and 21 (107.72). However, anti-inflammatory activity against MMP-2 was 95 % with propolis and 85 % with Biodentine and whereas, against MMP-9 it was 65 % for propolis and 47 % for Biodentine. ConclusionPropolis shows comparable cell viability, cell proliferation and differentiation potential on SHED when compared to Biodentine. It also exhibits better invitro anti-inflammatory activity on SHED compared to Biodentine. Further studies are warranted to validate the application of propolis as an effective and economical alternative biocompatible agent to Biodentine for vital pulp therapies.

Cytotoxic cardiac glycosides from the root of Streblus asper

Bioassay-guided separation of the root of Streblus asper led to the identification of six undescribed cardiac glycosides, including a rare cardiac glycoside dimer, along with twelve previously reported analogues. Their structures were determined on the basis of analyses of spectroscopic methods (1D and 2D-NMR spectroscopy), high-resolution electrospray ionization mass spectrometry (HRESIMS), circular dichroism (CD), and comparison of their spectroscopic data with previously reported data. Regarding their cytotoxic activities, microculture tetrazolium assays showed that all isolated cardiac glycosides strongly inhibited MCC-803, T24, SKOV-3, HepG2, Wi-38, and A549 cancer cell lines, with IC50 values ranging from 0.075 μM to 0.752 μM. One cardiac glycoside, a rare cardiac glycoside dimer, exhibited the strongest activity against the six cancer cell lines, with IC50 values ranging from 0.075 μM to 0.214 μM. In addition, the structure-activity relationships (SARs) of cardiac glycosides were investigated. In summary, S. asper showed marked cytotoxicity to several cancer cell lines, which could be meaningful for discovering new anticancer agents.

Determination of Adsorption of Methylene Blue Dye by Incense Stick Ash Waste and Its Toxicity on RTG-2 Cells

Incense stick ash (ISA) has traces of toxic heavy metals which have an adverse effect on the environment. Every year, tonnes of ISA are disposed of in rivers and other water bodies which leads to water pollution and affects the natural water resources. ISA has several value-added minerals which could be modified or functionalized for environmental cleanup. Here, in the current research work, ISA was transformed into a flower-like noble porous material by mixing ISA and NaOH in a 1 : 1 ratio followed by calcination at 600°C for six hours in a muffle furnace. The developed material was analyzed by sophisticated instruments for the identification of the properties. The microscopic techniques revealed the micron-sized flower-like structure, while the XRD showed peaks at 30–33° which indicates the transformation of the calcite and silicate phases into new-phase mineral. FTIR also revealed bands in regions of 500–1200 cm−1 and new bands near 450 cm−1. EDS confirmed the presence of Na in the sintered product and the transformation of the ISA. Finally, the sintered product potential was assayed for the removal of methylene blue dye from wastewater using an adsorption mechanism. The removal efficiency of dye reached up to 70% within one hour only. It was found that the ISA sintered product has the potential to remove MB dye efficiently from wastewater and also reduce solid waste pollution. Microculture tetrazolium assay (MTT) and lactate dehydrogenase (LDH) assays were performed to evaluate the cytotoxicity of the sintered incense stick ash product on RTG-2 cells. The sintered incense stick ash product induced cytotoxicity on RTG-2 cells in a dose-dependent manner. Sintered ISA products have the potential to remove methylene blue dye efficiently from wastewater and reduce solid waste pollution.

Characterization and Anticancer Activity from Gaharu (Aquilaria malaccensis) Stem Bark Extract

Aquilaria malaccensis is a plant species from the Thymeleaceae family. A. malaccensis contains secondary metabolites of phenolic groups, namely benzophenone, chromone, coumarin, xanthone, and flavonoids with various biological activities such as antioxidants and anticancer. This study aims to determine the potential of A. malaccensis stem bark extract as an anticancer and to determine the characteristics of secondary metabolite compounds contained in A. malaccensis stem bark extract. Extraction of the stem bark of A. malaccensis by maceration method in stages using n -hexane, ethyl acetate and methanol as solvents. The anticancer activity test used the microculture tetrazolium (MTT) assay against Murine Leukemia P-388 cells. Compound separation using thin layer chromatograph (TLC) and gravity column chromatography (CC) methods. Determination of chemical compounds using liquid chromatography mass spectroscopy/mass spectroscopy (LCMS/MS). The extracts of n -hexane, ethyl acetate, and methanol each had anticancer activity which was classified as active with an IC 50 value of 20.64, 26.61, and 14.49 µg/mL. Based on the results of separation and identification of compounds in methanol extract with TLC, CC, and LCMS/MS, methanol extract contains acetaminophen, aminosamptothesin, isophorone, diethyl phthalate, 3-hydroxy-6H-benzochromen-6-one, isolongifolenic, 2-(2-phenethyl) chromone, 4′,7-dihydroxy-2′,5-dimethoxyisoflavone, 4-hydroxy-3-(1-phenylpropyl)-2H-chromone-2-one; 6-methylchromone-2-carboxylic acid; 3,4-dimethyl-2- phenylmorpholine; and β-calacoren compounds.

Synthesis and Biological Profiles of Some Benzimidazolyl-chalcones as Anti-leishmanial and Trypanocidal Agents

Background: Benzimidazole constitutes a starting point for the development of new antiprotozoal agents since this nucleus exists in several pharmacologically significant molecules, in particular possessing antifungal, antiviral, antibacterial, and antiparasitic properties.
 Objective: The present study aimed to identify a molecular hit likely to be developed as an anti-leishmanial and antitrypanosomal drug candidate.
 Methods: Thus, 12 hybrids of chalcone or benzimidazolyl-arylpropenones were synthesized and screened in vitro for anti-leishmanial activity against promastigotes of Leishmania donovani. The microculture tetrazolium assay was used to determine their potential to inhibit 50% of a parasite growth (IC50).
 Results: Two compounds among 5-chlorobenzimidazole-chalcones (4a and 4c), which exhibited potent activity (IC50<1 μM) against L. donovani and one derivative (4d) poorly effective against L. donovani (IC50>50 μM) were selected to check their trypanocidal activity. Lethal concentration (LC100) values of these compounds were estimated by using observations on the viability of trypomastigotes of Trypanosoma brucei brucei in MEM medium with Earle’s salts and L-glutamine. Seven of the tested compounds (4a, 4b, 4c, 4e, 4g, 4h, and 4j) showed particularly higher inhibitory activity than pentamidine (IC50= 7.6 μM) against L. donovani promastigotes with IC50 values in a range from 0.5 to 1.8 μM. In addition, the 2’-chlorine derivative (4c), displayed potent anti-trypanosomal activity comparable to those of melarsoprol, used as reference drug.
 Conclusion: These results support this series of benzimidazole holding arylpropenone group in position 2 as a starting point for future optimization to get novel agents active against leishmaniasis and trypanosomiasis.

Anticancer Effect of Cardamom Extract on Hep-2 Cell Line

To investigate the anticancer effect of Cardamom extract on Hep-2 cell line which may aid in development of a novel treatment modality to Head and Neck Squamous Cell Carcinoma. Hep-2 cell line was divided into five groups: one control group, two groups treated with Cardamom extract, and another two treated with Doxorubicin, each of the two treatments was applied for 24 and 48 hours, respectively. Then, cellular viability was measured using microculture tetrazolium assay, cell cycle analysis was done using Flow Cytometry and eventually, apoptotic activity was evaluated using enzyme-linked immunosorbent assay to measure the concentration of BAX protein and real time polymerase chain reaction to measure the fold change for caspase-3 enzyme. Cardamom extract succeeded to decrease the percentage of viable and proliferating cells with increasing dose. On the other hand, it increased the percentage of apoptotic cells and levels of caspase-3 and BAX protein. Cardamom extract has a potential cytotoxic effect on Head and Neck Squamous Cell Carcinoma cell line in a dose and time dependent manner, and exerts this action through induction of apoptosis, and its action is comparable to Doxorubicin action.

Circular RNA Phosphatidylinositol-4-Phosphate 5-Kinase Type 1 Alpha Affects Prostate Cancer Lymph Node Carcinoma of Prostate-Cell Multiplication and Apoptosis via Regulating MicroRNA-299-3p Expression

Prostate cancer is a common cancer in aging men. This research explores the its molecular mechanisms of the circular RNA (circRNA) circPIP5K1A and its role in the multiplication and apoptosis of prostate cancer LNCaP cells. Cancerous tissue and adjacent non-cancerous tissue samples were selected from 37 prostate cancer patients hospitalized from January 2016 to January 2020. Quantitative reverse transcription PCR (RT-qPCR) was used to amplify circPIP5K1A and detect the miR-299-3p levels. Cells from the LNCaP prostate cancer cell line were divided into the si-circPIP5K1A group, si-negative control (NC) group, miR-299-3p group, miR-NC group, si-circPIP5K1A + anti-miR-299-3p group, and si-circPIP5K1A + anti-miR-NC group. Microculture tetrazolium (MTT) assay was used to assess the cell viability; flow cytometry was used to detect and analyze cell apoptosis; western blot was used to study the protein expression; the dual-luciferase reporter assay was used to determine the targeted regulation of miR-299-3p by circPIP5K1A. CircPIP5K1A expression in prostate cancer tissue increased when miR-299-3p expression was reduced (P < 0.05) compared to those in the adjacent tissue. After inhibition of circPIP5K1A or overexpression of miR-299-3p, the activity of LNCaP cells decreased, the apoptosis rate of LNCaP cells increased, Ki-67 and Bcl-2 expressions in LNCaP decreased, and Bax expression increased (P < 0.05). Due to CircPIP5K1A regulation of miR-299-3p expression, mir-299-3p interference reversed the effect of circPIP5K1A inhibition of LNCaP-cell multiplication and apoptosis. Inhibiting the expression of circPIP5K1A may inhibit LNCaP-cell multiplication and expedite cell apoptosis by up-regulating miR-299-3p.

Evaluation of Anti-cancer Effect of Different Concentrations of Black Raspberries Extract on Oral Squamous Cell Carcinoma Cell Line (In Vitro Study)

Background: Black Raspberry has a wide range of pharmacological and therapeutic aspects including anti-inflammatory and antioxidant activities. Moreover, the effect of Black Raspberry has been studied in many cancers such as human cervical, esophageal and colorectal cancer and showed cytotoxic and anti-proliferative effects against these types of cancer. Aim: To identify the possible anti-cancer effect of Black Raspberry on Oral Squamous Cell Carcinoma cell line. Material and methods: Human tongue squamous cell carcinoma cell line was divided into 3 groups: one control group, second group treated with Black Raspberry, and the third group treated with Doxorubicin, each of the two treatments was applied for 24, 48, 72 hours, respectively. Then, cellular viability was measured using Microculture Tetrazolium assay, cell cycle analysis was done using Flow Cytometry and eventually, apoptotic activity was evaluated using Enzyme-linked Immunosorbent assay. Results: Black Raspberry succeeded to decrease the percentage of viable and proliferating cells with increasing time. On the other hand, it increased the percentage of apoptotic cells and levels of caspase-3. Conclusion: Black Raspberry has a potential cytotoxic effect on oral squamous cell carcinoma cell line in a dose and time dependent manner, and exerts this action through induction of apoptosis, and its action is comparable to Doxorubicin action.

Bupivacaine and Lidocaine Induce Apoptosis in Osteosarcoma Tumor Cells.

Osteosarcoma is the most common type of bone cancer in adolescents. There have been no significant improvements in outcomes since chemotherapy was first introduced. Bupivacaine and lidocaine have been shown to be toxic to certain malignancies. This study evaluates the effect of these medications on two osteosarcoma cell lines. (1) Does incubation of osteosarcoma cells with bupivacaine or lidocaine result in cell death? (2) Does this result from an apoptotic mechanism? (3) Is a specific apoptotic pathway implicated? Two cell lines were chosen to account for the inherent heterogeneity of osteosarcoma. UMR-108 is a transplantable cell line that has been used in multiple studies as a primary tumor. MNNG/HOS has a high metastatic rate in vivo. Both cell lines were exposed bupivacaine (0.27, 0.54, 1.08, 2.16, 4.33 and 8.66 mM) and lidocaine (0.66, 1.33, 5.33, 10.66, 21.32 and 42.64 mM) for 24 hours, 48 hours, and 72 hours. These concentrations were determined by preliminary experiments that found the median effective dose was 1.4 mM for bupivacaine and 7.0 mM for lidocaine in both cell lines. Microculture tetrazolium and colony formation assay determined whether cell death occurred. Apoptosis induction was evaluated by phase-contrast micrographs, flow cytometry, DNA fragmentation and reactive oxygen species (ROS). The underlying pathways were analyzed by protein electrophoresis and Western blot. All testing was performed in triplicate and compared with pH-adjusted controls. Quantitative results were analyzed without blinding. Both medications caused cell death in a dose- and time-dependent manner. Exposure to bupivacaine for 24 hours reduced viability of UMR-108 cells by 6 ± 0.75% (95% CI 2.9 to 9.11; p = 0.01) at 1.08 mM and 89.67 ± 1.5% (95% CI 82.2 to 95.5; p < 0.001) at 2.16 mM. Under the same conditions, MNNG/HOS viability was decreased in a similar fashion. After 24 hours, the viability of UMR-108 and MNNG/HOS cells exposed to 5.33 mM of lidocaine decreased by 25.33 ± 8.3% (95% CI 2.1 to 48.49; p = 0.03) and 39.33 ± 3.19% (95% CI 30.46 to 48.21; p < 0.001), respectively, and by 90.67 ± 0.66% (95% CI 88.82 to 92.52; p < 0.001) and 81.6 ± 0.47% (95% CI 79.69 to 82.31; p < 0.001) at 10.66 mM, respectively. After 72 hours, the viability of both cell lines was further reduced. Cell death was consistent with apoptosis based on cell morphology, total number of apoptotic cells and DNA fragmentation. The percentage increase of apoptotic UMR-108 and MNNG/HOS cells confirmed by Annexin-V positivity compared with controls was 21.3 ± 2.82 (95% CI 16.25 to 26.48; p < 0.001) and 21.23 ± 3.23% (95% CI 12.2 to 30.2; p = 0.003) for bupivacaine at 1.08 mM and 25.15 ± 4.38 (95% CI 12.9 to 37.3; p = 0.004) and 9.11 ± 1.74 (95% CI 4.35 to 13.87; p = 0.006) for lidocaine at 5.33 mM. The intrinsic apoptotic pathway was involved as the expression of Bcl-2 and survivin were down-regulated, and Bax, cleaved caspase-3 and cleaved poly (ADP-ribose) polymerase-1 were increased. ROS production increased in the UMR-108 cells but was decreased in the MNNG/HOS cells. These findings provide a basis for evaluating these medications in the in vivo setting. Studies should be performed in small animals to determine if clinically relevant doses have a similar effect in vivo. In humans, biopsies could be performed with standard doses of these medications to see if there is a difference in biopsy tract contamination on definitive resection. Bupivacaine and lidocaine could potentially be used for their ability to induce and enhance apoptosis in local osteosarcoma treatment. Outcome data when these medications are used routinely during osteosarcoma treatment can be evaluated compared with controls. Further small animal studies should be performed to determine if injection into the tumor, isolated limb perfusion, or other modalities of treatment are viable.

SECONDARY METABOLITES FROM RICE CULTURE OF ASPERGILLUS SP. ISOLATED FROM MELALEUCA SUBULATA (CHEEL) CRAVEN LEAVES AND THEIR ANTICANCER ACTIVITY

Objective: Aspergillus fungus is a rich source of natural products with broad biological activities. This study was conducted to identify secondary metabolites from the rice culture of Aspergillus species isolated from Melaleuca subulata leaves and evaluated their anticancer activity.&#x0D; Methods: Ethyl acetate extract was fractionated on silica gel and Sephadex columns. Structures of the compounds were established using physical and chemical methods. Cytotoxic activities of the extract and pure compounds against two human cancer cell lines (Mcf-7and Hep G2) were evaluated using microculture tetrazolium assay as well as the mode of the cytotoxicity was evaluated. Molecular docking studies have been performed using the Hsp 90 enzyme as an anticancer target.&#x0D; Results: Methyl linoleate (1), arugosin C (2), ergosterol (3), sterigmatocystin (4), diorcinol (5), alternariol-5-O-methyl ether (6), averufin (7), averufanin (8), and alternariol (9) were identified from ethyl acetate extract. All tested compounds exhibit week activity against MCF-7 and Hep G2 cell lines but a mixture of compounds 7 and 8 is considered to be more active towards both MCF-7 and Hep G 2 in comparison to other compounds. Compound 4 exhibits moderate activity against Hep G2 only as well as the ethyl acetate extract exerts moderate activity against MCF-7 cell line Moreover, compound 4 and a mixture of 7 and 8 caused a decrease in the number of Hep G2 cancer cells due to apoptotic and necrotic processes. Most active anticancer candidates 7 and 8 showed binding to the active site similar to geldanamycin reference ligand.&#x0D; Conclusion: Secondary metabolites identified from Aspergillus sp. and their anticancer activity were evaluated. Molecular docking suggested active candidates as Hsp 90 inhibitors.

The effects of HBXIP on the biological functions of tongue squamous cell carcinoma cells and correlation with PI3K/Akt.

BackgroundTongue squamous cell carcinoma (TSCC) is the most malignant oral cancer, having a high mortality rate.MethodsThe effects of hepatitis B X-interacting protein (HBXIP) overexpression on the proliferation, migration, and invasion of TSCC cells were measured by micro-culture tetrazolium assay (MTT) assay, transwell assay and scratch test, respectively, and the effects of HBXIP mRNA overexpression on the protein expression levels of AKT, p-AKT, PI3K, p-PI3K and S100A4 were detected by western blotting.ResultsMTT assay showed that there were significantly more proliferating cells than in the experimental group. In the scratch test and transwell assay, the migration rate and the number of invading cells were remarkably greater in the experimental group. The expression levels of p-AKT, p-PI3K and S100A4 were increased in the experimental group after HBXIP overexpression.ConclusionsHBXIP mRNA overexpression can influence the proliferation, invasion, and migration of TSCC cells and promote their proliferation and migration by increasing the protein expression levels of p-AKT, p-PI3K and S100A4.

Inhibition of Cervical Cancer Cell Line Hela by Human Wharton’s Jelly Stem Cells Through Induction of Apoptosis

Background: Cervical cancer is one of the most common cancers of women in the world, which causes high mortality. The human umbilical cord Wharton's jelly stem cells (hWJSCs) can inhibit various cancer cells. Objectives: This study aimed to investigate the effects of conditioned medium and cellular extract of human umbilical cord hWJSCs on cervical cancer cell line, Hela. Methods: After isolation and primary culture of hWJSCs, conditioned medium and cellular extracts of hWJSCs were prepared, and its anti-proliferative effects were evaluated on cervical cancer cells, Hela using micro-culture tetrazolium (MTT) assay. After total RNA extraction and cDNA synthesis, expression of apoptosis-related BCL-2 and BAX genes were evaluated using real-time PCR. Results: The results showed that conditioned medium (55% concentration in 72 hours) and cellular extraction (10% concentration in 24 hours) caused death of 50% cancer cells (IC50). The anti-cancer effects of conditioned medium and cellular extraction were concentration- and time-dependent. The conditioned medium and cellular extract of hWJSCs significantly down-regulated and up-regulated mRNA expression of apoptosis-related BCL-2 and BAX genes, respectively. Conclusions: Our study showed that conditioned medium and cellular extract of human umbilical cord hWJSCs inhibit viability and proliferation of cervical cancer cells. However, further studies on animal models are necessary for more accurate results.

Proximate Analysis and Safety Profile of Farmed Edible Bird's Nest in Malaysia and Its Effect on Cancer Cells.

Edible bird's nest (EBN) which is solidified swiftlet's saliva contains high nutritional value. It is widely consumed in countries like Malaysia, Indonesia, and Thailand. However, previous proximate analysis of Malaysia EBN was not representative of all the regions in Malaysia. In recent years, safety issues such as high nitrate and nitrite contents, presence of heavy metal, adulteration, fungal infection, and cancer cell stimulation were associated with EBN. Hence, this study aimed to determine the proximate analysis, safety profile during normal weather and hazy periods, and its effect on cancer cells stimulation in Malaysia-farmed EBN. Seven raw cleaned EBN samples were sourced from 6 different regions in Malaysia. Proximate analysis and safety profile were performed using official AOCA methods and Malaysian Standard. High protein (53.03–56.37%) and carbohydrate content (27.97–31.68%) with an acceptable level of moisture (10.8–14.04%) and ash (2.22–3.38%) were reported. A good safety profile was obtained with low nitrite and nitrate levels, with undetectable heavy metals and no significant growth of pathogenic microorganism except mould. Epidermal growth factor was detected but below the quantification level with the chicken EGF ELISA kit. The microculture tetrazolium (MTT) assay was performed for growth stimulation assessment comparing human EGF and EBN. There was no significant cell growth observed in cancer cells after EBN treatment. In conclusion, EBN Malaysia has a good nutritional profile, free of heavy metals, and an acceptable level of nitrate, nitrite, and microorganism profile except for mould contents. Furthermore, the in vitro study indicated that EBN was not associated with cancer cell growth.