Osteosarcoma MG-63 Research Articles

Application value of a selenium-hydroxyapatite nanodelivery system as osteosarcoma treatment

To investigate the anticancer and osteogenic properties and mechanism of action of a selenium (Se4+) hydroxyapatite (HA) nanodelivery system for osteosarcoma (OS) therapy. We loaded different concentrations of Na2SeO3 (3% and 6%) into HA nanoparticles (HANP) for OS therapy. Simultaneously, we tested the stability and pH responsiveness of Se-HANP in vitro experiments, and OS MG-63 cells and mice BMSCs were cultured to further evaluate its permeability, anticancer ability and mineralization ability [alkaline phosphatase (ALP), osteocalcin (OCN)]. In addition, We conducted in vivo experiments by constructing OS mouse models to further investigate the anticancer mechanism of Se-HANP. Se-HANP showed good stability in blood; an acidic environment (pH 5) was more conducive to Se4+ ion release, and could effectively penetrate into OS MG-63 cells. In cell experiments, compared with HANP, Se-HANP could more effectively improve BMSCs viability and enhanced their mineralization performance, such as improving ALP and OCN levels. In addition, Se-HANP could effectively inhibit the activity and invasiveness of MG-63 cells; 6% Se-HANP showed stronger anticancer effects. In OS in vivo mouse experiments, Se-HANP could effectively inhibit tumor tissue growth (volume and weight) in a concentration dependent manner. In addition, Se4+ activated endogenous and exogenous apoptotic pathways by increasing the level of reactive oxygen species (8-OHdG) in tumor cells, leading to tumor cell apoptosis. Se-HANP could effectively penetrate OS cells, and its good stability and pH responsiveness could improve the anticancer efficiency of Se4+. Moreover, its excellent mineralization ability could effectively repair the bone defects caused by OS. Accordingly, Se-HANP can have great potential in OS treatment.

Injectable, Anti-Cancer Drug-Eluted Chitosan Microspheres against Osteosarcoma.

The purpose of this study is to fabricate different anti-cancer drug-eluted chitosan microspheres for combination therapy of osteosarcoma. In this study, electrospray in combination with ground liquid nitrogen was utilized to manufacture the microspheres. The size of obtained chitosan microspheres was uniform, and the average diameter was 532 μm. The model drug release rate and biodegradation rate of chitosan microspheres could be controlled by the glutaraldehyde vapor crosslinking time. Then the 5-fluorouracil (5-FU), paclitaxel (PTX), and Cis-dichlorodiammine-platinum (CDDP) eluted chitosan microspheres were prepared, and two osteosarcoma cell lines, namely, HOS and MG-63, were selected as cell models for in vitro demonstration. We found the 5-FU microspheres, PTX microspheres, and CDDP microspheres could significantly inhibit the growth and migration of both HOS and MG-63 cells. The apoptosis of both cells treated with 5-FU microspheres, PTX microspheres, and CDDP microspheres was significantly increased compared to the counterparts of control and blank groups. The anti-cancer drug-eluted chitosan microspheres show great potential for the treatment of osteosarcoma.

Synthesis and Characterization of Tween-20 Capped Biosynthesized Silver Nanoparticles for Anticancer and Antimicrobial Property.

The Vitrus vinifera fruit extract was used to make silver nanoparticles (AgNPs) utilizing a green chemical technique. The biosynthesized Tween-20/Vitrus vinifera-AgNPs were observed by UV-Vis spectrophotometry. Fourier transform infrared spectroscopy, scanning transmission electron microscopy, selected area electron diffraction, and energy-dispersive X-ray spectroscopy were used to characterize the physiochemical properties. The spherical form of AgNPs was confirmed by transmission electron microscopy. The peaks in the Tween-20/Vitrus vinifera-AgNPs have an average crystallite size that is found to be 46nm according to powder X-ray diffraction examination. Biosynthesized AgNPs had a significant effect on bone osteosarcoma MG63 cells with 55% inhibition, respectively, using MTT assay. The effective dangerous concentration of Tween-20/Vitrus vinifera with AgNP nanoparticles was less harmful to MG63 cells. The results of antibacterial activity showed that Tween-20/Vitrus vinifera-AgNPs effectively inhibited Eggerthella lenta and Staphylococcus epidermis bacteria.

α-Mangostin suppresses proliferation and invasion in osteosarcoma cells via inhibiting fatty acid synthase

• α-mangostin was able to induce human osteosarcoma cells apoptosis via downregulating fatty acid synthase (FASN) expression in dose-dependent manner. • α-mangostin, as a FASN inhibitor, plays a great role in regulated ER stress-related protein and signal protein expression. • α-mangostin induced FASN inhibition, which might influence osteosarcoma progression, invasion and migration. Fatty acid synthase (FASN) is highly expressed in multiple types of human cancers and is recognized as one of the therapeutic targets for treating cancer metastasis. α-Mangostin is a natural xanthone extracted from mangosteen fruit, and possesses various biological activities. In our previous studies, α-mangostin was found to inhibit FASN activity. The present study was designed to reveal the effects of α-mangostin on osteosarcoma and to reveal whether the mechanism of α-mangostin in anticancer activity is related to FASN inhibition. Cytotoxicity was assessed in osteosarcoma MG63, 143B and U2OS cells. Cell viability was detected by an MTT assay. The protein expression levels were detected by western blotting. Flow cytometry, Annexin V/propidium iodide dual staining and Hoechst 33258 staining were performed to assess the apoptotic effects. Wound healing and Transwell assays were used to detect the inhibitory effect of α-mangostin on osteosarcoma cells invasion and migration. We found that α-mangostin blocked FASN expression, inhibited osteosarcoma cell proliferation, invasion and migration in a dose-dependent manner. In addition, α-mangostin regulated endoplasmic reticulum transmembrane receptors and signal protein expression in osteosarcoma cells. These findings suggested that α-mangostin suppresses osteosarcoma cell proliferation and metastasis by inhibiting FASN expression, which provides a basis as a potential target for osteosarcoma treatment.

Efficacy of glycoprotein-based nanocurcumin/silk fibroin electrospun scaffolds: Perspective for bone apatite formation

In this research, a blend of poly (vinyl) alcohol (PVA), silk fibroin nanofibers (SF-NFs), hydroxyapatite nanoparticles (HA-NPs), and curcumin nanoparticles (Cu-NPs) were used to design a nanofibrous scaffold (NFS), by electrospun method, for their possible application in bone apatite formation. The prepared nanofibrous scaffold (NFS) was investigated using physicochemical, mechanical, cell viability, etc. The bioactivity test of NFS was evaluated after immersion in SBF (simulated body fluid). NFS from PVA, SF-NFs, HA-NPs, and Cu-NPs exhibited outstanding physicochemical, mechanical, and antibacterial properties. PVA: SF-NFs: HA-NPs: Cu-NPs scaffold excellent antimicrobial performance against E.coli (4.16 ± 0.16 mm) and S.aureus (4.88 ± 0.10 mm). Biocompatibility studies were assessed using human osteosarcoma MG63 cell line, which displayed 100% viable cell presence in PVA: SF-NFs: HA-NPs: Cu-NPs. Based on the characterization, results indicated that they are excellent scaffolds for bone tissue regeneration.

Highly active selenium nanotherapeutics combined with metformin to achieve synergistic sensitizing effect on NK cells for osteosarcoma therapy

Abstract NK cells-based cancer therapy combined with chemotherapeutic drugs for the treatment of tumors can enhance the immunosensitivity of NK cells, increase the expression of NK cell receptors, and eventually boost the killing effect of NK cells on cancer cells. Selenium (Se) with different chemical structures can be metabolized into selenoproteins to regulate tumor and immune cells’ fate and functions. Herein, we found that, functionalized Se nanoparticles (SeNPs) combining with metformin (met) could amply the immunotherapeutic effects of NK92 cells against osteosarcoma cancer. The results revealed that TW80-SeNPs combined with met had the optimum performance on NK92 cells for HepG2 cells, owing to the increased ROS in HepG2 cells and the augmented expression of cell surface receptor proteins ULBP-3/4, PD-L1, MICA, and NK92 cell surface receptor proteins PD-1 and FasL. Additionally, TW80-SeNPs were gradually metabolized into selenoproteins (Gpx4 and TR1) into human osteosarcoma MG63 cells to reinforce the anticancer effect of NK92 cells by regulating the redox balance in the tumor microenvironment. This study provides a therapeutic approach in treating cancer itself or diabetes coupled with cancer. Moreover, it provides a multidrug strategy to improve immune cell function in practical applications, especially for synergistic immunotherapy of osteosarcoma.

HO-3867 Induces Apoptosis via the JNK Signaling Pathway in Human Osteosarcoma Cells.

Metastatic osteosarcoma often results in poor prognosis despite the application of surgical en bloc excision along with chemotherapy. HO-3867 is a curcumin analog that induces cell apoptosis in several cancers, but the apoptotic effect and its mechanisms on osteosarcoma cells are still unknown. After observing the decrease in cellular viability of three human osteosarcoma U2OS, HOS, and MG-63 cell lines, and the induction of cellular apoptosis and arrest in sub-G1 phase in U2OS and HOS cells by HO-3867, the human apoptosis array showed that heme oxygenase (HO)-1 and cleaved caspase-3 expressions had significant increases after HO-3867 treatment in U2OS cells and vice versa for cellular inhibitors of apoptosis (cIAP)1 and X-chromosome-linked IAP (XIAP). Western blot analysis verified the results and showed that HO-3867 activated the initiators of both extrinsic caspase 8 and intrinsic caspase 9, and significantly increased cleaved PARP expression in U2OS and HOS cells. Moreover, with the addition of HO-3867, ERK1/2, and JNK1/2 phosphorylation were increased in U2OS and HOS cells. Using the inhibitor of JNK (JNK in 8), HO-3867’s increases in cleaved caspases 3, 8, and 9 could be expectedly suppressed, indicating that JNK signaling is responsible for both apoptotic pathways, including extrinsic and intrinsic, in U2OS and HOS cells caused by HO-3867. Through JNK signaling, HO-3867 has proven to be effective in causing both extrinsic and intrinsic apoptotic pathways of human osteosarcoma cells.

Expression of mTOR conduction pathway in human osteosarcoma MG-63 cells and their stem cells, and the inhibitory effect of different doses of rapamycin

Purpose: To investigate the expressions of rapamycin target protein (mTOR) conduction pathway in human osteosarcoma MG-63 cells and their stem cells, and to examine the inhibitory effect of different doses of rapamycin.Methods: mTOR mRNA in osteosarcoma stem-like cells and human osteosarcoma MG-63 cells were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The cells were treated with different doses of rapamycin and divided into low dose group (0.5 mg), medium dose group (1.0 mg), high dose group (2.0 mg) and blank (control) group. Apoptosis and cell cycle of MG-63 cells were determined by flow cytometry, while proliferation of MG-63 cells up was assessed by CCK-8 kit.Results: mTOR in human osteosarcoma MG-63 cells was significantly lower than that in osteosarcoma stem-like cells. Compared with the control group, mRNA expression levels of mTOR in MG-63 cells and osteosarcoma stem-like cells were significantly decreased after treatment with different concentrations of rapamycin (p < 0.05). MG-63 cells treated with various doses of rapamycin exhibited a significant decrease in their proliferation, compared with control group, while only the high rapamycin concentration group exhibited a significant decrease in osteosarcoma stem-like cell proliferation (p < 0.05). Treatment with rapamycin in MG-63 cells and osteosarcoma stem-like cells resulted in a significant increase in apoptosis, prolonged G0/G1 phase and shortened S phase (p < 0.05).Conclusion: Rapamycin inhibits the expression of mTOR mRNA in osteosarcoma stem-like and MG-63 cells. It also inhibits the proliferation and cell cycle formation of osteosarcoma stem-like cells and MG-63 cells via mTOR signal pathway. These findings may provide a new target for the treatment of osteosarcoma.

Targeting Akt/NF-κB/p53 Pathway and Apoptosis Inducing Potential of 1,2-Benzenedicarboxylic Acid, Bis (2-Methyl Propyl) Ester Isolated from Onosma bracteata Wall. against Human Osteosarcoma (MG-63) Cells.

Onosma bracteata Wall. is an important medicinal and immunity-enhancing herbs. This plant is commonly used in the preparation of traditional Ayurvedic drugs to treat numerous diseases. Inspired by the medicinal properties of this plant, the present study aimed to investigate the antiproliferative potential and the primary molecular mechanisms of the apoptotic induction against human osteosarcoma (MG-63) cells. Among all the fractions isolated from O. bracteata, ethyl acetate fraction (Obea) showed good antioxidant activity in superoxide radical scavenging assay and lipid peroxidation assay with an EC50 value of 95.12 and 80.67 µg/mL, respectively. Silica gel column chromatography of ethyl acetate (Obea) fraction of O. bracteata yielded a pure compound, which was characterized by NMR, FTIR, and HR-MS analysis and was identified as 1,2-benzene dicarboxylic acid, bis (2-methyl propyl) ester (BDCe fraction). BDCe fraction was evaluated for the antiproliferative potential against human osteosarcoma MG-63, human neuroblastoma IMR-32, and human lung carcinoma A549 cell lines by MTT assay and exhibited GI50 values of 37.53 μM, 56.05 μM, and 47.12 μM, respectively. In MG-63 cells, the BDCe fraction increased the level of ROS and simultaneously decreased the mitochondria membrane potential (MMP) potential by arresting cells at the G0/G1 phase, suggesting the initiation of apoptosis. Western blotting analysis revealed the upregulation of p53, caspase3, and caspase9 while the expressions of p-NF-κB, p-Akt and Bcl-xl were decreased. RT-qPCR studies also showed upregulation in the expression of p53 and caspase3 and downregulation in the expression of CDK2, Bcl-2 and Cyclin E genes. Molecular docking analysis displayed the interaction between BDCe fraction with p53 (−151.13 kcal/mol) and CDK1 (−133.96 kcal/mol). The results of the present work suggest that the BDCe fraction has chemopreventive properties against osteosarcoma (MG-63) cells through the induction of cell cycle arrest and apoptosis via Akt/NF-κB/p53 pathways. This study contributes to the understanding of the utilization of BDCe fraction in osteosarcoma treatment.

Green synthesized zinc oxide nanoparticles induce apoptosis by suppressing PI3K/Akt/mTOR signaling pathway in osteosarcoma MG63 cells

This study aimed to assess the apoptosis-inducing mechanism of zinc oxide nanoparticles (ZnO NPs) stabilized by Solanum xanthocarpum plant extract in human osteosarcoma MG63 cells. In the present study, we synthesized ZnO NPs from S. xanthocarpum extract and evaluated its anticancer mechanism on MG 63 cells. The synthesized ZnO NPs were characterized by ultraviolet spectroscopy, X-ray crystallography, transmission electron microscopy, energy dispersive X-ray, and Fourier-transform infrared spectroscopy analysis. The mean size of the synthesized ZnO NPs was 21.62 ± 7.45 nm and spherical in shape. The cytotoxicity of ZnO NPs on MG63 cells was determined by MTT assay. The Western blot analysis was carried out to examine the expression of apoptotic and autophagy-related proteins in MG63 cells. The findings of the study reveal that ZnO NPs treatment showed concentration-dependent cytotoxicity, increased lipid peroxidation, decreased antioxidant activity, increased reactive oxygen species generation, and increased DNA damage. In addition, ZnO NPs treatment increased the expression of apoptotic members such as p53, Bax, caspase-3, -8, and -9 while downregulating Bcl-2 expression in MG63 cells. Furthermore, ZnO NPs treatment suppressed the P13K/AKT/mTOR signaling pathway and increased the expression of LC3 and beclin-1 in MG63 cells. The present study demonstrated that ZnO NPs induced apoptosis and autophagy in MG63 cells through modifying apoptotic and autophagy-related proteins.

MG-63 and FetMSC Cell Response on Atomic Layer Deposited TiO2 Nanolayers Prepared Using Titanium Tetrachloride and Tetraisopropoxide

Titanium oxide nanocoatings were synthesized on the surface of monocrystalline silicon and ultra-fine-grained titanium by atomic layer deposition (ALD) using titanium tetrachloride (TiCl4) and titanium tetraisopropoxide (TTIP). The morphology of the samples was studied by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The structure and composition were studied by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), contact angle measurements, and energy-dispersive spectroscopy (EDS). The cytological response of osteosarcoma MG-63 and human fetal mesenchymal stem cells (FetMSCs) were studied by analyzing their morphology, viability, and alkaline phosphatase activity with and without the use of medium-induced differentiation in the osteogenic direction. A significant influence of the precursor type and ALD temperature on the crystal structure, morphology, composition, and surface free energy of TiO2 nanocoatings was found. The biocompatibility of amorphous non-stoichiometric and partially crystalline stoichiometric TiO2 coatings was compared. Both types of cells showed faster adhesion and improved spreading on the surface for the samples from TTIP compared to those from TiCl4 at the early stages of cultivation (2 h) due to the difference in composition and higher surface free energy. No cytotoxic effect was found on both types of coatings, nor was there a noticeable difference in cell differentiation. All ALD coatings provided excellent biocompatibility and osteoconductive properties.

α-Linolenic Acid Suppresses Proliferation and Invasion in Osteosarcoma Cells via Inhibiting Fatty Acid Synthase.

Fatty acid synthase (FASN) is highly expressed in multiple types of human cancers and is recognized as one of the targets for treating cancer metastasis. α-Linolenic acid is an omega-3 essential fatty acid and it possesses various biological activities. The present study was designed to reveal the effects of α-linolenic acid on osteosarcoma and to reveal whether the mechanism of α-linolenic acid in anticancer activity may be related to FASN inhibition. The cytotoxicity of α-linolenic acid was assessed in osteosarcoma MG63, 143B, and U2OS cells. Cell viability was detected by the MTT assay. The protein expression level was detected by western blotting. Flow cytometry, Annexin V/propidium iodide dual staining, and Hoechst 33258 staining were performed to assess the apoptotic effects. Wound healing assay was applied to detect the inhibitory effect of α-linolenic acid on osteosarcoma cells migration. The results showed that α-linolenic acid downregulated FASN expression. α-Linolenic acid inhibited osteosarcoma cell proliferation and migration in a dose-dependent manner. In addition, α-linolenic acid regulated endoplasmic reticulum transmembrane receptors and signal protein expression in osteosarcoma cells. The findings of the present study suggested that α-linolenic acid suppresses osteosarcoma cell proliferation and metastasis by inhibiting FASN expression, which provides a basis as a potential target for osteosarcoma treatment.

A Consistent Protocol Reveals a Large Heterogeneity in the Biological Effectiveness of Proton and Carbon-Ion Beams for Various Sarcoma and Normal-Tissue-Derived Cell Lines.

Simple SummaryUsing a consistent experimental protocol, we found a large heterogeneity in the relative biological effectiveness (RBE) values of both proton and carbon-ion beams in various sarcomas and normal-tissue-derived cell lines. Our data suggest that proton beam therapy may be more beneficial for some types of tumors. In carbon-ion therapy, for some types of tumors, large heterogeneity in RBE should prompt consideration of dose reduction or an increased dose per fraction. In particular, a higher RBE value in normal tissues requires caution. Specific dose evaluations for tumor and normal tissues are needed for both proton and carbon-ion therapies.This study investigated variations in the relative biological effectiveness (RBE) values among various sarcoma and normal-tissue-derived cell lines (normal cell line) in proton beam and carbon-ion irradiations. We used a consistent protocol that specified the timing of irradiation after plating cells and detailed the colony formation assay. We examined the cell type dependence of RBE for proton beam and carbon-ion irradiations using four human sarcoma cell lines (MG63 osteosarcoma, HT1080 fibrosarcoma, SW872 liposarcoma, and SW1353 chondrosarcoma) and three normal cell lines (HDF human dermal fibroblast, hTERT-HME1 mammary gland, and NuLi-1 bronchus epithelium). The cells were irradiated with gamma rays, proton beams at the center of the spread-out Bragg peak, or carbon-ion beams at 54.4 keV/μm linear energy transfer. In all sarcoma and normal cell lines, the average RBE values in proton beam and carbon-ion irradiations were 1.08 ± 0.11 and 2.08 ± 0.36, which were consistent with the values of 1.1 and 2.13 used in current treatment planning systems, respectively. Up to 34% difference in the RBE of the proton beam was observed between MG63 and HT1080. Similarly, a 32% difference in the RBE of the carbon-ion beam was observed between SW872 and the other sarcoma cell lines. In proton beam irradiation, normal cell lines had less variation in RBE values (within 10%), whereas in carbon-ion irradiation, RBE values differed by up to 48% between hTERT-HME1 and NuLi-1. Our results suggest that specific dose evaluations for tumor and normal tissues are necessary for treatment planning in both proton and carbon-ion therapies.

Nerolidol induced apoptosis via PI3K/JNK regulation through cell cycle arrest in MG-63 osteosarcoma cells.

The aim of the present study was to determine the cell proliferation, apoptotic pathway analysis through protein, mRNA and cell cycle arrest mechanism in nerolidol induced osteosarcoma MG-63 cells. The osteosarcoma MG-63 cells were treated with various doses of nerolidol (15 and 20μM/ml) for 24h. Cell proliferation was examined using assist method of MTT assay, fixed the IC50 value of nerolidol 15μM/ml. Reactive oxygen species (ROS) generation was analyzed by DCFH-DA dye, mitochondrial potential detected by Rh-123 dye, apoptotic morphological changes identified by AO/EtBr, PI, DAPI staining, and cell adhesion were detected by using fluorescence microscope. Cell proliferation, and apoptotic molecular protein and mRNA expressions such as ERK, P38, p-PI3K, p-JNK, Bcl-2, JNK, p-P38, cyclin-D1, and Bax were analyzed in osteosarcoma MG-63 cells. Nerolidol significantly suppressed the osteosarcoma cells progression in a dose dependent manner (p<.05) evident in the oxidative stress induction and apoptotic morphological changes. Nerolidol also regulated the protein PI3K/AKT mechanistically via induction of apoptosis Nerolidol suppresses osteosarcoma MG-63 cells by PI3K/AKT by cell cycle arrest at early phase of G0/G1. To sum up, nerolidol suppressed the growth of bone cancer cells and can be finally targeted as a potent drug for analyzing its chemotherapeutic effects in future.

Mechanical and biological properties of atmospheric plasma-sprayed carbon nanotube-reinforced tantalum pentoxide composite coatings on Ti6Al4V alloy

Ta2O5 coatings containing 0, 3, 5 and 7 vol% multi-walled carbon nanotubes (CNTs) were deposited on Ti6Al4V substrates by atmospheric plasma spraying (APS). The addition of CNTs had no obvious effect on the surface morphology and roughness of the coatings, but increased the coating porosity. The coatings consisted primarily of β-Ta2O5 phase, a small amount of α-Ta2O5, and minor C peaks corresponding to the CNTs. As the CNT content increased, the elastic modulus (E) and indentation fracture toughness also increased. However, the higher CNT content increased the coating porosity and reduced the microhardness. Following immersion in simulated body fluid (SBF) for 14 days, the surfaces of all the coatings were completely covered with hemispherical bone-like apatite. Furthermore, after incubation in osteoblast-like osteosarcoma MG-63 cell culture for 7 days, all of the coatings showed excellent cell attachment, growth and spreading. Overall, the present results show that the addition of CNTs to Ta2O5 raw powder improves the indentation fracture toughness of the resulting coatings without degrading their biological properties.

Effects of lysophosphatidic acid (LPA) signaling via LPA receptors on cellular functions associated with ATP reduction in osteosarcoma cells treated with ethidium bromide.

Lysophosphatidic acid (LPA) signaling via LPA receptors (LPA1 to LPA6) exhibits a variety of malignant properties in cancer cells. Intracellular ATP depletion leads to the development of necrosis and apoptosis. The present study aimed to evaluate the effects of LPA receptor-mediated signaling on the regulation of cancer cell functions associated with ATP reduction. Long-term ethidium bromide (EtBr) treated (MG63-EtBr) cells were established from osteosarcoma MG-63 cells. The intracellular ATP levels of MG63-EtBr cells were significantly lower than that of MG-63 cells. LPAR2, LPAR3, LPAR4 and LPAR6 gene expressions were elevated in MG63-EtBr cells. The cell motile and invasive activities of MG63-EtBr cells were markedly higher than those of MG-63 cells. The cell motile activity of MG-63 cells was increased by LPA4 and LPA6 knockdowns. In cell survival assay, cells were treated with cisplatin (CDDP) every 24h for 3 days. The cell survival to CDDP of MG63-EtBr cells was lower than that of MG-63 cells. LPA2 knockdown decreased the cell survival to CDDP of MG-63 cells. The cell survival to CDDP of MG-63 cells was inhibited by (2S)-OMPT (LPA3 agonist). Moreover, the cell survival to CDDP of MG-63 cells was enhanced by LPA4 and LPA6 knockdowns. These results indicate that LPA signaling via LPA receptors is involved in the regulation of cellular functions associated with ATP reduction in MG-63 cells treated with EtBr.

Development of Silver Doped Hydroxyapatite Thin Films for Biomedical Applications

Silver doped hydroxyapatite [AgHAp, Ca10−xAg(PO4)6(OH)2], due to its antimicrobial properties, is an advantageous material to be used for various coatings. The AgHAp thin films with xAg = 0.05 and xAg = 0.1 were achieved using the spin-coating method. The resulting samples were examined by X-ray diffraction (XRD), scanning electron microscopy (SEM), atomic force microscopy (AFM), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, and X-ray photoelectron spectroscopy (XPS). XRD analysis revealed that the particles of both samples are ellipsoidal. Also, in agreement with the results obtained by XRD measurements, the results of the SEM studies have shown that the particles shape is ellipsoidal. Optical properties of silver doped hydroxyapatite thin films deposited on Si substrate were investigated through Fourier transform infrared spectroscopy (FTIR) and Raman spectroscopy. The results obtained by the two complementary techniques highlighted that the molecular structure of the studied samples is not influenced by the increase of the silver concentration in the samples. Our studies revealed that the surface morphology of the obtained samples consist of uniform and continuous layers. The biocompatibility of the obtained thin films was also evaluated with the aid of human osteosarcoma MG63 (ATCC CRL 1427) cell line. Moreover, the in vitro antifungal activity against Candida albicans fungal strain of the AgHAp thin films was studied and the obtained results revealed their antifungal effect. The results of the biological assays showed that the AgHAp thin films are a very promising material for biomedical applications.

S-Adenosylmethionine: From the Discovery of Its Inhibition of Tumorigenesis to Its Use as a Therapeutic Agent.

Alterations of methionine cycle in steatohepatitis, cirrhosis, and hepatocellular carcinoma induce MAT1A decrease and MAT2A increase expressions with the consequent decrease of S-adenosyl-L-methionine (SAM). This causes non-alcoholic fatty liver disease (NAFLD). SAM administration antagonizes pathological conditions, including galactosamine, acetaminophen, and ethanol intoxications, characterized by decreased intracellular SAM. Positive therapeutic effects of SAM/vitamin E or SAM/ursodeoxycholic acid in animal models with NAFLD and intrahepatic cholestasis were not confirmed in humans. In in vitro experiments, SAM and betaine potentiate PegIFN-alpha-2a/2b plus ribavirin antiviral effects. SAM plus betaine improves early viral kinetics and increases interferon-stimulated gene expression in patients with viral hepatitis non-responders to pegIFNα/ribavirin. SAM prevents hepatic cirrhosis, induced by CCl4, inhibits experimental tumors growth and is proapoptotic for hepatocellular carcinoma and MCF-7 breast cancer cells. SAM plus Decitabine arrest cancer growth and potentiate doxorubicin effects on breast, head, and neck cancers. Furthermore, SAM enhances the antitumor effect of gemcitabine against pancreatic cancer cells, inhibits growth of human prostate cancer PC-3, colorectal cancer, and osteosarcoma LM-7 and MG-63 cell lines; increases genomic stability of SW480 cells. SAM reduces colorectal cancer progression and inhibits the proliferation of preneoplastic rat liver cells in vivo. The discrepancy between positive results of SAM treatment of experimental tumors and modest effects against human disease may depend on more advanced human disease stage at moment of diagnosis.

Potential Anti-Metastatic Role of the Novel miR-CT3 in Tumor Angiogenesis and Osteosarcoma Invasion.

Osteosarcoma (OS) is the most common primary bone tumor mainly occurring in young adults and derived from primitive bone-forming mesenchyme. OS develops in an intricate tumor microenvironment (TME) where cellular function regulated by microRNAs (miRNAs) may affect communication between OS cells and the surrounding TME. Therefore, miRNAs are considered potential therapeutic targets in cancer and one of the goals of research is to accurately define a specific signature of a miRNAs, which could reflect the phenotype of a particular tumor, such as OS. Through NGS approach, we previously found a specific molecular profile of miRNAs in OS and discovered 8 novel miRNAs. Among these, we deepen our knowledge on the fifth candidate renamed now miR-CT3. MiR-CT3 expression was low in OS cells when compared with human primary osteoblasts and healthy bone. Through TargetScan, VEGF-A was predicted as a potential biological target of miR-CT3 and luciferase assay confirmed it. We showed that enforced expression of miR-CT3 in two OS cell lines, SAOS-2 and MG-63, reduced expression of VEGF-A mRNA and protein, inhibiting tumor angiogenesis. Enforced expression of miR-CT3 also reduced OS cell migration and invasion as confirmed by soft agar colony formation assay. Interestingly, we found that miR-CT3 behaves inducing the activation of p38 MAP kinase pathway and modulating the epithelial-mesenchymal transition (EMT) proteins, in particular reducing Vimentin expression. Overall, our study highlights the novel role of miR-CT3 in regulating tumor angiogenesis and progression in OS cells, linking also to the modulation of EMT proteins.

Antihyperglycemic effect of Vernonia amygdalina and in vitro evaluation of its antiproliferative activity on human osteosarcoma MG-63.

the leaves of Vernonia amygdalina (V. amygdalina) are consumed as food in sub-Saharan Africa (SSA). In traditional medicine, this plant is widely used in the treatment of cancer and diabetes mellitus. In the present study, we evaluated the antihyperglycemic and the antiproliferative activities of the hydroalcoholic extract of V. amygdalina leaves (HAEVa). we conducted an experimental descriptive and analytical study with a prospective data collection from May 2019 to July 2020. For the in vivo study, the experiments were carried out on albino male rats of Wistar strain (Rattus norvegicus). Antihyperglycemic activity was performed in vivo in dexamethasone-induced insulin-resistant rats using the oral glucose tolerance test (OGTT). The biocompatibility and the antiproliferative activity of extract were performed in vitro respectively on rabbit primary dermal fibroblasts (RPDF) and human osteosarcoma MG-63 cells using the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The data were analyzed with the GraphPad Prism software version 5.0.3. The statistical analyses were obtained by the analysis of variance (ANOVA), followed by Bonferroni´s post-test. P<0.05 was considered as the minimal level of statistical significance. regarding to the antiproliferative investigation, extract at 125, 250 μg/mL exhibited a significant cytotoxic effect on human osteosarcoma MG-63 compared to the vehicle (p<0.001) in a dose-response manner after 24h, 48h of exposure to HAEVa. Interestingly, HAEVa in concentrations of 125 and 250μg/ml showed no cytotoxicity (p>0.05) on RPDF after the different times of exposure. However, HAEVa in a high concentration of 500 μg/mL wasn´t biocompatible with RPDF. HAEVa also prevented postprandial blood glucose level in dexamethasone-induced insulin-resistant rats at both doses tested (p>0.05 and p<0.01 at doses of 50 and 100 mg/kg respectively). the results of this study suggest that HAEVa has antiproliferative properties on MG-63 osteosarcoma in vitro and also inhibits in vivo the postprandial blood glucose level in dexamethasone-induced insulin-resistant rats.