Osteosarcoma MG-63 Cells Research Articles

The Potential In Vitro Inhibitory Effects of Neurokinin-1 Receptor (NK-1R) Antagonist, Aprepitant, in Osteosarcoma Cell Migration and Metastasis.

Background Osteosarcoma, the most frequent osteogenic malignancy, has become a serious public health challenge due to its high morbidity rates and metastatic potential. Recently, the neurokinin-1 receptor (NK-1R) is proved to be a promising target in cancer therapy. This study is aimed at determining the effect of aprepitant, a safe and Food and Drug Administration (FDA) approved NK-1R antagonist, on osteosarcoma cell migration and metastasis, and to explore its underlying mechanism of action. Methods Colorimetric MTT assay was employed to assess cell viability and cytotoxicity. A wound-healing assay was used to examine migration ability. The desired genes' protein and mRNA expression levels were measured by western blot assay and quantitative real-time PCR (qRT-PCR), respectively. Gelatinase activity was also measured by zymography. Results We found that aprepitant inhibited MG-63 osteosarcoma cell viability in a dose-dependent manner. We also observed that aprepitant inhibited the migrative phenotype of osteosarcoma cells and reduced the expression levels and activities of matrix metalloproteinases (MMP-2 and MMP-9). Aprepitant also reduced the expression of an angiogenic factor, VEGF protein, and NF-κB as an important transcriptional regulator of metastasis-related genes. Conclusion Collectively, our observations indicate that aprepitant modulates the metastatic behavior of human osteosarcoma cells, which may be applied to an effective therapeutic approach for patients with metastatic osteosarcoma.

Strontium Substituted β-Tricalcium Phosphate Ceramics: Physiochemical Properties and Cytocompatibility.

Sr2+-substituted β-tricalcium phosphate (β-TCP) powders were synthesized using the mechano-chemical activation method with subsequent pressing and sintering to obtain ceramics. The concentration of Sr2+ in the samples was 0 (non-substituted TCP, as a reference), 3.33 (0.1SrTCP), and 16.67 (0.5SrTCP) mol.% with the expected Ca3(PO4)2, Ca2.9Sr0.1(PO4)2, and Ca2.5Sr0.5(PO4)2 formulas, respectively. The chemical compositions were confirmed by the energy-dispersive X-ray spectrometry (EDX) and the inductively coupled plasma optical emission spectroscopy (ICP-OES) methods. The study of the phase composition of the synthesized powders and ceramics by the powder X-ray diffraction (PXRD) method revealed that β-TCP is the main phase in all compounds except 0.1SrTCP, in which the apatite (Ap)-type phase was predominant. TCP and 0.5SrTCP ceramics were soaked in the standard saline solution for 21 days, and the phase analysis revealed the partial dissolution of the initial β-TCP phase with the formation of the Ap-type phase and changes in the microstructure of the ceramics. The Sr2+ ion release from the ceramic was measured by the ICP-OES. The human osteosarcoma MG-63 cell line was used for viability, adhesion, spreading, and cytocompatibility studies. The results show that the introduction of Sr2+ ions into the β-TCP improved cell adhesion, proliferation, and cytocompatibility of the prepared samples. The obtained results provide a base for the application of the Sr2+-substituted ceramics in model experiments in vivo.

Lipoyl-Based Antagonists of Transient Receptor Potential Cation A (TRPA1) Downregulate Osteosarcoma Cell Migration and Expression of Pro-Inflammatory Cytokines.

Osteosarcoma is a heterogeneous tumor intimately linked to its microenvironment, which promotes its growth and spread. It is generally accompanied by cancer-induced bone pain (CIBP), whose main component is neuropathic pain. The TRPA1 ion channel plays a key role in metastasis and is increasingly expressed in bone cancer. Here, a novel TRPA1 inhibitor is described and tested together with two other known TRPA1 antagonists. The novel lipoyl derivative has been successfully assessed for its ability to reduce human osteosarcoma MG-63 cell viability, motility, and gene expression of the CIBP pro-inflammatory cytokines interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α). A putative three-dimensional (3D) model of the inhibitor covalently bound to TRPA1 is also proposed. The in vitro data suggest that the novel inhibitor described here may be highly interesting and stimulating for new strategies to treat osteosarcomas.

Caffeic Acid Induces Intrinsic Apoptotic Pathway in MG-63 Osteosarcoma Cells Through Bid Truncation and Cytochrome c Release

BACKGROUND: Caffeic acid has been reported to induce apoptosis in MG-63 osteosarcoma cells via caspases activation. However, apoptotic pathway that is involved in the caffeic acid-induced apoptosis is still unclear. Present study aimed to investigate the role of cytochrome c (Cyt c) release and BH3-interacting death (Bid) activation in caffeic acid-induced apoptosis in MG-63 osteosarcoma cells.METHODS: MG-63 cells were cultured, pretreated with/without Z-VAD FMK and treated with/without 10 μg/mL caffeic acid. Treated MG-63 cells were then lysed, homogenized, and processed further to prepare cell lysate and mitochondrial fraction. Immunoblotting method was used to measure the amount of Bid and truncated Bid (t-Bid) as well as mitochondrial and cytosolic Cyt c.RESULTS: The amount of Bid and mitochondrial Cyt c in MG-63 cells decreased in a time-dependent manner, while the amount of t-Bid and cytosolic Cyt c increased in a time-dependent manner. By pretreatment of 100 μM Z-VAD-FMK for 2 h, the amount of Bid and mitochondrial Cyt c was significantly higher, while the amount of t-Bid and cytosolic Cyt c was significantly lower after caffeic acid treatment for 6 and 12 h compared to MG-63 cells that were not pretreated.CONCLUSION: Caffeic acid could induce Cyt c release through the activation of Bid in MG-63 osteosarcoma cells.KEYWORDS: caffeic acid, osteosarcoma, MG-63 cells, Bid, t-Bid, cytochrome c, Z-VAD-FMK

Cisplatin and Exogenous Hydrogen Sulfide Inhibit the Progression of Osteosarcoma via EGFR/P13K/Akt Signaling Pathway

We attempted to analyze the effects of cisplatin and exogenous hydrogen on the invasion and proliferation of human osteosarcoma cells through EGFR/P13K/Akt signaling pathway. The effects of exogenous hydrogen sulfide and cisplatin on the invasion, apoptosis and proliferation of MG-63 cells were detected by transwell chamber invasion experiment, flow cytometry and CCK-8 method. After 24 and 48 hours of drug action, the inhibition rate of cell proliferation was significantly higher than in the combined treatment. The number of cells decreased significantly, compared with the control group. Western blotting and qRT-PCR showed that the expression of Akt mRNA in MG-63 cells treated with cisplatin+hydrogen sulfide at different concentrations for 48 hours decreased. The expression of Akt, P13K and EGFR were decreased Flow cytometry analysis showed that after the drug treatment of MG-63 cells, the S phase of cisplatin and cisplatin+exogenous hydrogen concentrations with different prolonged concentrations was significantly shortened phase and the G0/G1 was significantly, and the apoptosis rate of each group was significantly increased. Cisplatin and exogenous hydrogen sulfide can synergistically inhibit the proliferation and invasion of human osteosarcoma MG-63 cells, and regulate the apoptosis mechanism of MG-63 cells through EGFR/P13K/Akt signaling pathway.

Compound Library Screening for Synergistic Drug Combinations: mTOR Inhibitor and Proteasome Inhibitor Effective Against Osteosarcoma Cells.

The development of new drugs is urgently needed for new treatment strategies that can improve the prognosis of osteosarcoma (OS). In this study, we attempted to identify combinations of new molecular-targeted agents for OS. A library containing 324 compounds was used. For the first screening, MG-63 OS cells were treated with each of the compounds and cell viability was measured. After the best candidate compound was decided, the compound was included in a second screening. The combination of most effective compounds was decided. The antiproliferative effect of the combination was examined and the cell signaling mechanism was evaluated by western blot analysis. 143B OS-bearing mice were used for in vivo antitumor testing. In the first screening, bortezomib was chosen as the effective drug. In the second screening with bortezomib, everolimus was chosen. This combination showed a synergistic inhibitory effect on cell proliferation when compared to monotherapy with each of these drugs alone. Compared to monotherapy, the combination therapy enhanced the levels of cleaved poly (ADP-ribose) polymerase, caspase-3, caspase-8 and caspase-9, phospho-c-Jun N-terminal kinase, and P38. In contrast, the levels of c-MYC proto-oncogene bHLH transcription factor, survivin, and phospho-cyclin D1 were reduced. The combination effectively induced apoptosis and interfered with cell cycle progression. In the in vivo analysis, the combination therapy significantly inhibited tumor growth. The combination of everolimus and bortezomib demonstrated a synergistic effect against OS both in vitro and in vivo. These results indicate that this combination may be useful as a novel therapeutic strategy for OS.

Mussel-inspired polydopamine decorated alginate dialdehyde-gelatin 3D printed scaffolds for bone tissue engineering application.

This study utilized extrusion-based 3D printing technology to fabricate calcium-cross-linked alginate dialdehyde-gelatin scaffolds for bone regeneration. The surface of polymeric constructs was modified with mussel-derived polydopamine (PDA) in order to induce biomineralization, increase hydrophilicity, and enhance cell interactions. Microscopic observations revealed that the PDA layer homogeneously coated the surface and did not appear to induce any distinct change in the microstructure of the scaffolds. The PDA-functionalized scaffolds were more mechanically stable (compression strength of 0.69 ± 0.02 MPa) and hydrophilic (contact angle of 26) than non-modified scaffolds. PDA-decorated ADA-GEL scaffolds demonstrated greater durability. As result of the 18-days immersion in simulated body fluid solution, the PDA-coated scaffolds showed satisfactory biomineralization. Based on theoretical energy analysis, it was shown that the scaffolds coated with PDA interact spontaneously with osteocalcin and osteomodulin (binding energy values of −35.95 kJ mol−1 and −46.39 kJ mol−1, respectively), resulting in the formation of a protein layer on the surface, suggesting applications in bone repair. PDA-coated ADA-GEL scaffolds are capable of supporting osteosarcoma MG-63 cell adhesion, viability (140.18% after 7 days), and proliferation. In addition to increased alkaline phosphatase secretion, osteoimage intensity also increased, indicating that the scaffolds could potentially induce bone regeneration. As a consequence, the present results confirm that 3D printed PDA-coated scaffolds constitute an intriguing novel approach for bone tissue engineering.

Zinc oxide decorated titania nanostructured layer over Ti metal as a biocompatible and antimicrobial surface for biomedical application

The present study is an attempt to develop a zinc oxide (ZnO) decorated titania (TiO2) nanostructured layer over Ti metal by a simple chemical and thermal treatment method. Ti metal upon sequential chemical treatment with sodium hydroxide and zinc nitrate solution generated a nanostructured network layer enfolded with Zn2+ ions. Subsequent heat treatment up to 500 °C favoured the formation of ZnO and anatase TiO2 while increasing the temperature to 600 °C resulted in the formation of ZnO and ZnTiO3 along with anatase TiO2. This nanostructured layer showed significant improvement in the hardness after heat treatment at 600 °C as measured by the nano-indentation technique. ZnO formed over the anatase TiO2 layer showed excellent antibacterial activity against both gram positive Staphylococcus aureus and gram negative Escherichia coli and was also found to be non-toxic towards MG-63 osteosarcoma cells. Hence, ZnO/ZnTiO3 modified anatase TiO2 over Ti metal is anticipated to reduce the risk of bacterial infection and enhance the proper functionalisation through its metabolic and cellular signalling and can be useful in dental and orthopaedic fields.

An 85-amino-acid polypeptide from Myrmeleon bore larvae (antlions) homologous to heat shock factor binding protein 1 with antiproliferative activity against MG-63 osteosarcoma cells in vitro.

Venomous arthropods have substances in their venom with antiproliferative potential for neoplastic cells. To identify a polypeptide from Myrmeleon bore (antlion) with antiproliferative activity against neoplastic cells, and to elucidate the molecular mechanism of the activity. We used gel filtration and ion exchange chromatography to purify a polypeptide with antiproliferative activity against MG-63 human osteosarcoma cells from a proteinaceous extract of antlion. The polypeptide was sequenced and the stability of its antiproliferative activity was tested under a range of conditions in vitro. An 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the antiproliferative activity of the polypeptide against the MG-63 osteosarcoma cells and MC3T3-E1 mouse calvarial osteoblasts, which were used as a non-neoplastic control. We used western blotting to compare the levels of expression of heat shock transcription factor 1 (HSF1), heat shock protein 90 (HSP90), cyclin-dependent kinase 4 (CDK4), and protein kinase B alpha (ATK1) in MG-63 osteosarcoma cells and their mouse homologs in MC3T3-E1 osteoblasts after their treatment with the antlion antiproliferative polypeptide (ALAPP). The 85-amino-acid ALAPP has a 56% sequence identity with the human heat shock factor binding protein 1 (HSBP1). The antiproliferative activity of the polypeptide is relatively insensitive to temperature, pH, and metal ions. ALAPP has a strong concentration-dependent antiproliferative activity against MG-63 osteosarcoma cells compared with its effect on MC3T3-E1 osteoblasts. ALAPP significantly upregulates the expression of HSF1 in MC3T3-EL osteoblasts, but not in MG-63 osteosarcoma. ALAPP significantly downregulated the expression of HSP90, CDK4, and AKT1 expression in MG-63 osteosarcoma, but not in the osteoblasts. ALAPP has significant antiproliferative activity against MG-63 osteosarcoma cells, but not nonneoplastic MC3T3-E1 osteoblasts. We speculate that non-neoplastic cells may evade the antiproliferative effect of ALAPP by upregulating HSF1 to maintain their HSP90, CDK4, and AKT1 expression at a relatively constant level.

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.

Mesenchymal Stem Cell Derived Exosomes as Nanodrug Carrier of Doxorubicin for Targeted Osteosarcoma Therapy via SDF1-CXCR4 Axis.

PurposeThe objective of this study was to investigate the antitumor activity, targeting capability, and mechanism of the developed nanodrug consisting of doxorubicin and exosome (Exo-Dox) derived from mesenchymal stem cells in vitro and in vivo.MethodsThe exosomes were isolated with Exosome Isolation Kit, and the Exo-Dox was prepared by mixing exosome with Dox-HCl, desalinizing with triethylamine and then dialyzing against PBS overnight. The exosome and Exo-Dox were examined by nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). The antitumor activity, targeting capability, and mechanism of the developed Exo-Dox were evaluated by cell viability assay, histological and immunofluorescence analysis and in vivo imaging system.ResultsNTA results showed the size of the exosomes had increased from 141.6 nm to 178.1 nm after loading with doxorubicin. Compared with free Dox, the Exo-Dox exhibited higher cytotoxicity against osteosarcoma MG63 cells, HOS cells, and 143B cells than free Dox, the half-maximal inhibitory concentrations (IC50) of Dox, Exo-Dox were calculated to be 0.178 and 0.078 μg mL−1 in MG63 cells, 0.294 and 0.109μg mL−1 in HOS cells, 0.315 and 0.123 μg mL−1 in 143B cells, respectively. The in vivo imaging showed that MSC derived Exo could serve as a highly efficient delivery vehicle for targeted drug delivery. The immunohistochemistry and histology analysis indicated that compared with the free Dox group, the Ki67-positive cells and cardiotoxicity in Exo-Dox group were significantly decreased.ConclusionOur results suggested that MSC-derived Exo could be excellent nanocarriers used to deliver chemotherapeutic drug Dox specifically and efficiently in osteosarcoma, resulting in enhanced toxicity against osteosarcoma and less toxicity in heart tissue. We further demonstrated the targeting capability of Exo was due to the chemotaxis of MSC-derived exosomes to osteosarcoma cells via SDF1-CXCR4 axis.

3D bioprinted poly(lactic acid)/mesoporous bioactive glass based biomimetic scaffold with rapid apatite crystallization and in-vitro Cytocompatability for bone tissue engineering

In the recent years, bone tissue engineering is regarded as the promising solution for treatment of bone defects which arises due to trauma, infection and surgical intervention. In view of this, several polymer or ceramic based constructs are envisaged for bone tissue engineering potential. However, scaffolds based on pure polymeric materials suffer from slow bioactivity characteristics. On the other hand, scaffolds based on ceramic materials do not offer sufficient strength for load bearing applications. In order to overcome these drawbacks, the current work aims to develop mixed matrix scaffolds based on poly (L-lactic acid)/mesoporous bioactive glass composite with the formulation of 30:70 weight ratio, which mimics the natural bone composition. In the current work, PLA/MBG (30:70) composite based bioink suitable for 3D bioprinting is indigenously developed and its rheological characteristics are evaluated. The 3D architecture for PLA/MBG composite scaffold is designed using Solidworks CAD 2015 and the scaffolds are fabricated using pneumatic based 3D bioprinting technology, which has not been documented earlier for this formulation in view of bone tissue engineering in the best of our knowledge. Followed by this, optimization of printing parameters in order to develop 3D PLA/MBG composite constructs with hierarchical pore architecture suitable for bone tissue engineering is performed. The SEM analysis confirmed that the pore size of the 3D printed PLA/MBG composite scaffolds falls in the range of 500–700 μm, which corresponds to the macroporous nature of the scaffolds useful for bone cell growth. The mechanical analysis confirmed the superior compressive modulus and yield strength for PLA/MBG composite scaffold in comparison with neat PLA. The in-vitro bioactivity assessment showed rapid apatite crystallization by attaining Ca/P ratio of 1.66 equivalent to natural bone mineral within 3rd day of SBF treatment for PLA/MBG composite scaffold, thus indicating the excellent bioactivity behaviour. The 3D bioprinted PLA/MBG composite scaffold showed promising response in terms of cell attachment and proliferation, mineralization as well as gene expression characteristics while assessed through of in-vitro biological assessment using MG-63 osteosarcoma cells. In this regard, the 3D bioprinted PLA/MBG scaffold could be applied as potential implant for bone tissue engineering application.

A 3D Printed Bone Tissue Engineering Scaffold Composed of Alginate Dialdehyde-Gelatine Reinforced by Lysozyme Loaded Cerium Doped Mesoporous Silica-Calcia Nanoparticles.

A novel biomaterial comprising alginate dialdehyde-gelatine (ADA-GEL) hydrogel augmented by lysozyme loaded mesoporous cerium doped silica-calcia nanoparticles (Lys-Ce-MSNs) is 3D printed to create bioactive scaffolds. Lys-Ce-MSNs raise the mechanical stiffness of the hydrogel composite scaffold and induce surface apatite mineralization, when the scaffold is immersed in simulated body fluid (SBF). Moreover, the scaffolds can co-deliver bone healing (Ca and Si) and antioxidant ions (Ce), and Lys to achieve antibacterial (and potentially anticancer) properties. The nanocomposite hydrogel scaffolds can hold and deliver Lys steadily. Based on the in vitro results, the hydrogel nanocomposite containing Lys assured improved pre-osteoblast cell (MC3T3-E1) proliferation, adhesion, and differentiation, thanks to the biocompatibility of ADA-GEL, bioactivity of Ce-MSNs, and the stabilizing effect of Lys on the scaffold structure. On the other hand, the proliferation level of MG63 osteosarcoma cells decreased, likely due to the effect of Lys. Last but not least, cooperatively, alongside gentamicin (GEN), Lys brought about a proper antibacterial efficiency to the hydrogel nanocomposite scaffold against gram-positive and gram-negative bacteria. Taken together, ADA-GEL/Lys-Ce-MSN nanocomposite holds great promise for 3D printing of multifunctional hydrogel bone tissue engineering (BTE) scaffolds, able to induce bone regeneration, address infection, and potentially inhibit tumor formation and growth.

Apoptotic and Antioxidant Activity of Gold Nanoparticles Synthesized Using Marine Brown Seaweed: An In Vitro Study.

A major paradigm shift in the field of nanobiotechnology is the invention of an eco-friendly, economical, and green approach for synthesis of metal nanoparticles. In the present study, we have synthesized gold nanoparticles (AuNPs) using aqueous extracts of marine brown seaweed Sargassum longifolium. The synthesized nanoparticle was subjected to characterization using different techniques such as UV-Vis spectroscopy, Fourier transform infrared spectroscopy, atomic force microscope, scanning electron microscope, transmission electron microscope, and elemental dispersive X-ray diffraction. Further, the seaweed extract and the synthesized AuNPs were evaluated for its anticancer effect using MG-63 human osteosarcoma cells besides in vitro antioxidant effect. The formation of S. longifolium-mediated synthesis of gold nanoparticles was demonstrated by UV-Vis spectroscopy. Presence of elemental gold was confirmed by EDX analysis. TEM analysis demonstrated spherical morphology of the synthesized AuNPs and SEM analysis revealed the particle size to be in the range of 10-60 nm. The FTIR showed the presence of hydroxyl functional groups. The toxicity of S. longifolium extract and the synthesized AuNPs was tested using brine shrimp lethality assay at different concentrations with results showing both seaweed extract and AuNPs to be nontoxic. Both S. longifolium and AuNPs exhibited significant antioxidant activity by scavenging DPPH free radicals and H2O2 radicals. Significant antiproliferative effect was observed against MG-63 osteosarcoma cells. It was also shown that the seaweed extract and the AuNPs induced cytotoxicity in cell lines by mechanism of apoptosis. In conclusion, this study provided insight on AuNPs synthesized from S. longifolium as a potent antioxidant and anticancer agent.

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.

No Effect on Tumorigenesis in MG63 Cells Induced by Co-Cultured Mesenchymal Stem Cells.

Osteosarcoma is a kind of bone tumor with an extremely high malignant degree and often occurs in adolescents. Mesenchymal stem cells are believed to play an important role in the microenvironment of osteosarcoma, but whether they promote or inhibit the development of osteosarcoma is controversial. In this study, the coexpression of mesenchymal stem cells (MSCs) with osteosarcoma cell MG63 was used to explore the effect of MSCs on MG63. We found that co-culture of MSCs with MG63 did not affect the proliferation, invasion, and migration of MG63 cells, nor did it significantly affect the epithelial- and glial-mesenchymal transformation of MG63 cells. Therefore, in this study, we obtained a new concept that MSCs neither promote nor inhibit the occurrence and development of osteosarcoma.

Development of Ultrafine–Grained and Nanostructured Bioinert Alloys Based on Titanium, Zirconium and Niobium and Their Microstructure, Mechanical and Biological Properties

For this paper, studies of the microstructure as well as the mechanical and biological properties of bioinert titanium, zirconium, and niobium alloys in their nanostructured (NS) and ultrafine-grained (UFG) states have been completed. The NS and UFG states were formed by a combined two-step method of severe plastic deformation (SPD), first with multidirectional forging (MDF) or pressing into a symmetrical channel (PSC) at a given temperature regime, and then subsequent multi-pass groove rolling (MPGR) at room temperature, with pre-recrystallization annealing. Annealing increased the plasticity of the alloys in the NS and UFG states without changing the grain size. The UFG structure, with an average size of structural elements of no more than 0.3 μm, was formed as a result of applying two-step SPD and annealing. This structure presented significant improvement in the mechanical characteristics of the alloys, in comparison with the alloys in the coarse-grained (CG) or small-grained (SG) states. At the same time, although the formation of the UFG structure leads to a significant increase in the yield strength and tensile strength of the alloys, their elastic modulus did not change. In terms of biocompatibility, the cultivation of MG-63 osteosarcoma cells on the polished and sandblasted substrates demonstrated high cell viability after 10 days and good cell adhesion to the surface.

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.