Proliferation Of MG-63 Cells Research Articles

In vitro and in vivo biocompatibility study of surface modified TiN deposited on Ti6Al4V using high-power impulse magnetron sputtering technique

Although Ti6Al4V alloy is commonly used as a surgical implant, the toxicity of vanadium is always a serious concern. As a stable and protective coating for Ti6Al4V, TiN deposited using a high-power impulse magnetron sputtering (HiPIMS) technique exhibits great polarization resistance in simulated body fluid and shows excellent cell viability of osteoblast-like MG63 cell. Herein, we report further study on the in vitro and in vivo biocompatibility of the HiPIMS-deposited TiN coatings with various surface modifications. For the in vitro study, the effect of O2 plasma treatment, acrylamide (AAm) graft polymerization, and bone morphogenetic protein-2 (BMP2) immobilization on the viability of osteoblast-like MG63 cell were examined using the MTT assay. For the in vivo study, the pro-inflammatory cytokines and skin pathology in mice using subcutaneous sensitivity test were conducted. In vitro investigation shows that cell viability is significantly improved for TiN having BMP2 on the surface. When BMP2 was coated on bare TiN, the MG63 cell proliferation decreases with time. However, with surface pre-treatment of TiN using O2 plasma and AAm grafting, BMP2 coated TiN exhibits increasing MG63 cell proliferation with time. Likewise, in vivo investigation shows such sample has enhanced wound healing capability, significant increase of the body weights, obvious decrease of pro-inflammatory cytokines, and improved skin section structure, as compared to the bare Ti6Al4V. The pathological result shows that the TiN treated with O2 plasma and AAm graft polymerization, and with a surface BMP2 layer gives long-term stable immobilization of BMP2 and the best in vivo biocompatibility among all the samples.

DHRS12 inhibits the proliferation and metastasis of osteosarcoma via Wnt3a/β-catenin pathway.

Aim: This experimental design was based on DHRS12 to explore its biological effects on osteosarcoma (OS). Materials & methods: The expression level of endogenous DHRS12 was analyzed by immunohistochemical analysis. DHRS12 was overexpressed in MG-63 and HOS cells by plasmid transfection. Cell proliferation, invasion, migration, apoptosis and western blot were used in the experiment. Results: The expression of DHRS12 was significantly reduced in OS. Overexpression of DHRS12 inhibited the proliferation, migration and invasion of MG-63 and HOS cells and induced apoptosis of OS cells. Overexpression of DHRS12 upregulated Bax, Caspase 9 and Caspase 3. Overexpression of DHRS12 resulted in inactivation of the Wnt3a/β-catenin signaling pathway. Conclusion: Overexpression of DHRS12 inhibited the progression of OS via the Wnt3a/β-catenin pathway.

Polydatin induces apoptosis and autophagy via STAT3 signaling in human osteosarcoma MG-63 cells

Polydatin, a natural product, is detected in many daily diets, such as grape juices and peanut. Autophagy regulation is recognized as a new potential strategy for cancer therapy, and previous studies demonstrated that polydatin showed remarkable anti-cancer ability. Nevertheless, the capability of polydatin to induce autophagy and its role in anti-osteosarcoma remains obscure. In this study, we investigated the anticancer effect of polydatin on human osteosarcoma cell line MG-63 and its underlying mechanism. Our results indicated that polydatin significantly inhibited proliferation of MG-63 cells in a dose- and time-dependent manner, and increased their apoptosis and autophagic flux. Further experiments showed that polydatin reduced the expression and phosphorylation (Y705) level of STAT3 (Signal transducer and activator of transcription 3), increased the expression of autophagy-related genes (Atg12, Atg14, BECN1, PIC3K3), and therewith triggered autophagic cell death in MG-63 cells. Of note, the cytotoxicity effect of polydatin was rescued by co-treatment with Colivelin (STAT3 activator), suggesting the dependency of MG-63 cells on STAT3 for survival in this process. Moreover, polydatin-triggered autophagy and apoptosis were remarkably reduced following exposure to autophagy inhibitor 3-methyladenine, while cell viability was increased. In conclusion, these data demonstrated that polydatin induced MG-63 cell death through inducing apoptosis, and autophagy which was mediated via the STAT3 signaling. Therefore, polydatin might be a potential clinical drug in the remedy of osteosarcoma.

A Mg/Zn-co-doped composite coating on a titanium surface enhances osteogenic activity through the Wnt/β-catenin pathway

Although Ti alloys have good biocompatibility and mechanical properties, they lack bioactivity and cannot integrate with surrounding bone tissue when used as implants; thus, their clinical application is limited. In this study, a Mg/Zn-co-doped micro/nanocomposite coating (TZM) was prepared on a Ti alloy surface by plasma electrolytic oxidation (PEO), and the effect of the coating on the biological behaviour of osteoblasts on its surface was observed in experiments conducted in vitro and in vivo. Based on these findings, the role of the Wnt/β-catenin pathway in the effect of TZM on osteogenic differentiation was studied. The results show that TZM had a porous micro/nanostructure and was uniformly co-doped with Mg and Zn. TZM promoted the adhesion, spreading, proliferation and differentiation of MG-63 cells. In vivo experiments showed that Ti implants with the TZM coating stimulated new bone formation and promoted early implant osseointegration. More importantly, the addition of Wnt3a cells cultured on pure Ti (PT) significantly upregulated the level of β-catenin, and the addition of Dkk1 to cells cultured on the TZM surface significantly inhibited β-catenin activity. In conclusion, the preparation of a Mg/Zn-co-doped micro/nanocomposite coating through PEO is a feasible method for enhancing the osteogenic activity of Ti implants, and the Wnt/β-catenin signalling pathway may be involved in the mechanism by which the coating promotes osteoblast differentiation.

In situ synthesis of fluorapatite‐ZnO nanocomposite powder via mechanical alloying for biomedical applications

AbstractThis research aimed to synthesize of fluorapatite(FA)–ZnO (FAZ) nanocomposite powder using a one‐step mechanical alloying process without any post‐calcification process. Furthermore, the role of various concentrations of ZnO (0, 4, 8, 11, and 15 wt.%) on the chemical and biological characteristics of FA was examined. Our research confirmed the successful synthesize of FAZ nanopowders with irregular‐shaped particles and particle size of 38‐52 nm. The crystallinity, crystallite size, and c‐axes parameter of FA decreased with increasing ZnO content. Moreover, while all FAZ powders revealed superior bioactivity than FA, FA‐8 wt.% ZnO nanopowder significantly enhanced MG63 cell proliferation. Besides, between various samples, FA‐11 wt.% ZnO nanopowder revealed drastically greater antibacterial activity. It could be concluded that the in‐situ synthesis of FAZ powder meaningfully improved the biological activity of FA making it a promising coating for orthopedic and dental implants.

Effect of Silane-Coupling Modification on the Performance of chitosan-poly vinyl Alcohol-Hybrid Scaffolds in Bone Tissue Engineering

Biocompatibility and biodegradability characteristics of some polymers make them an excellent candidate to fabricate porous scaffolds for tissue engineering applications. However, the scaffold mechanical properties and biodegradation rate are vital for bone tissue replacement applications, which can be improved using proper fabrication techniques and cross-linker. In this investigation, Chitosan-polyvinyl alcohol scaffolds were prepared by freeze-drying technique utilizing various weight ratios of 3-Glycidoxypropyl trimethoxysilane (GPTMS) as a bioactive inorganic crosslinker. SEM micrographs indicated interconnected porous structures of cross-linked scaffolds while the average diameter of pores increased as a function of cross-linker enhancement. FTIR analysis was performed to confirm interactions among organic and inorganic components. The mechanical strength test represented that increasing GPTMS content improves the compressive strength of samples. The absorption capacity of the scaffolds in the PBS solution exhibited a decrease in water uptake and biodegradation by increasing silane coupling agent concentration. The formation of needle-like apatite particles proved suitable bioactivity of cross-linked samples. Moreover, MTT assay and ALP expression showed an acceptable adhesion, spreading, proliferation, and differentiation of MG-63 cells on the silane-contained scaffolds. Obtained results warrant further preclinical and clinical evaluations.

A Ca-deficientca-deficient hydroxyapatite (CDHA)/MgF2 bi-layer coating with unique nano-scale topography on biodegradable high-purity Mg

Enhanced corrosion resistance and accelerated new bone formation are desired to make Mg and its alloys to be ideal candidate for bone biomaterial. For this purpose, a CDHA/MgF2 bi-layer coating was prepared on high purity Mg by a combination of fluoride treatment and hydrothermal treatment. The coating exhibited a nanoscale surface topography. Enhanced adhesion strength and corrosion resistance was obtained for the CDHA/MgF2 bi-layer coating. In vitro cell experiment showed that the adhesion, proliferation and differentiation of MG63 cells were significantly improved on Mg with CDHA/MgF2 coating compared to that on Mg with HA coating and MgF2 coating. In conclusion, this study provides a promising surface modification method for Mg metal with enhanced corrosion resistance and superior osteogenic bioactivity.

Physical, mechanical and biological performance of PHB-Chitosan/MWCNTs nanocomposite coating deposited on bioglass based scaffold: Potential application in bone tissue engineering

Nowadays, using the nanocomposite coatings on bioceramic scaffolds is a great interest for many researchers to improve the properties of these scaffolds. In this study, the effect of poly (3-hydroxybutyrate) PHB-Chitosan (Cs)/multi-walled carbon nanotubes (MWCNTs) nanocomposite coating deposited on nano-bioglass (nBG)-titania (nTiO2) scaffolds fabricated by foam replication method was investigated. Structural analyses such as XRD and FT-IR confirmed the presence of PHB, Cs and MWCNTs in the coated scaffolds. The results of SEM and porosity measurement showed that even with 1 wt% MWCNTs, scaffolds have a high percentage of interconnected porosity. The compressive strength of the scaffolds coated with PHB-Cs/MWCNTs (1 wt%) was increased up to 30 folds compared to nBG/nTiO2 scaffold. The surface roughness of the coated scaffolds, which was determined by AFM, was increased. The nanocomposite coating caused a decrease in contact angle and retaining the negative zeta potential of the coated scaffolds. The increase in pH and degradation rate was observed in the coated scaffolds. Increasing the apatite-like formation by the presence of PHB-Cs/MWCNTs was confirmed by SEM, EDAX and XRD tests. PHB-Chitosan/MWCNTs nanocomposite coating lead to more proliferation and viability of MG-63 cells and higher secretion of alkaline phosphatase.

Cold atmospheric plasma‑activated Ringer's solution inhibits the proliferation of osteosarcoma cells through the mitochondrial apoptosis pathway.

The present study aimed to investigate the effects of cold atmospheric plasma (CAP)‑activated Ringer's solution on osteosarcoma cell lines MG63 and U2OS, and to identify the molecular mechanism underlying these effects. CAP‑activated Ringer's solution was used to treat osteosarcoma cell lines MG63 and U2OS for 30min. Cell viability was measured using the MTT method. The apoptosis rate was detected using Annexin‑V and propidium iodide. The expression levels of cytochrome c, caspase‑3 and polyADP ribose polymerase (PARP) in MG63 cells were analyzed via western blotting. The change in mitochondrial membrane potential was detected via the JC‑1 dye method and verified by the level of reactive oxygen species (ROS). CAP‑activated Ringer's solution inhibited the proliferation of MG63 and U2OS cells in a dose‑ and time‑dependent manner. Furthermore, CAP‑activated Ringer's solution induced the apoptosis of MG63 cells, increased the intracellular ROS level, decreased the mitochondrial membrane potential level, and induced the release of cytochrome c. CAP‑activated Ringer's solution inhibits osteosarcoma cell proliferation through intracellular ROS‑mediated mitochondrial apoptosis.

Silencing METTL3 inhibits the proliferation and invasion of osteosarcoma by regulating ATAD2

BackgroundOsteosarcoma is the most common primary malignant bone tumor in children and young adults. RNA N6-methyladenosine (m6A) is the most abundant internal modification in mammalian mRNA, which is involved in tumorigenesis and tumor progression. It has been reported that methyltransferase-like 3 (METTL3), the first reported m6A “writer”, plays critical roles in cancer progression. However, its role and molecular mechanism in osteosarcoma is poor studied. In this study, we aimed to investigate the functional role and underlying mechanism of METTL3 in the progression of osteosarcoma. MethodsWe detected the mRNA expression of METTL3 in osteosarcoma cell lines, and immunofluorescence assay was performed to observe the location of METTL3. Cell lines with METTL3 gene overexpression or knockdown were established by pcDNA3.1-METTL3 or siRNA interferences in order to determine the function of METTL3 in osteosarcoma in vitro. Transcriptomic RNA sequencing (RNA-seq) were used to screen the target genes of METTL3 in osteosarcoma. ResultsWe found that METTL3 localized in cytoplasm and nucleus of osteosarcoma cells. Silencing METTL3 in SAOS-2 and MG63 cells significantly inhibited the m6A methylation level, proliferation, migration, and invasion abilities, as well as promoted cell apoptosis. However, up-regulation of METTL3 had no significant effect on the biological behaviors of U2OS cells. Further mechanism analysis suggested that METTL3 knockdown inhibited the expression of ATPase family AAA domain containing 2 (ATAD2). Moreover, ATAD2 knockdown inhibited the proliferation and invasion of SAOS-2 and MG63 cells, while its overexpression showed a significant increase in cell proliferation and invasion. Furthermore, METTL3 knockdown abrogated the promoting effects of ATAD2 overexpression on osteosarcoma cells proliferation and invasion. ConclusionOverall, our study revealed that METTL3 functions as an oncogene in the growth and invasion of osteosarcoma by regulating ATAD2, suggesting a potential therapeutic target for osteosarcoma treatment.

Upregulation of miRNA-154-5p prevents the tumorigenesis of osteosarcoma

BackgroundOsteosarcoma (OS) is a primary malignant bone sarcoma in human worldwide. It has been shown that the level of microRNA-154-5p (miR-154-5p) was downregulated in human OS tissues. However, the mechanisms by which miR-154-5p regulates the proliferation, apoptosis and invasion in OS remain unclear. Thus, the present study aimed to investigate the role of miR-154-5p during the tumorigenesis of OS. MethodsThe level of miR-154-5p in human OS tissues was detected by RT-qPCR. In addition, the effects of miR-154-5p on apoptosis and invasion of OS cells were assessed by flow cytometry and transwell assays, respectively. Meanwhile, the dual luciferase reporter system assay was performed to explore the interaction of miR-154-5p and E2F5. ResultsThe level of miR-154-5p was downregulated in OS tissues. Overexpression of miR-154-5p significantly inhibited the proliferation, migration and invasion of MG63 cells. In addition, upregulation of miR-154-5p obviously induced apoptosis in MG63 cells via upregulation of Bax and cleaved caspase 3, and downregulation of Bcl-2. Moreover, luciferase reporter assay identified that E2F5 was the binding target of miR-154-5p. Meanwhile, overexpression of miR-154-5p induced cell cycle arrest in MG63 cells via inhibiting the expressions of E2F5, Cyclin E1 and CDK2. Furthermore, in vivo assays indicated that overexpression of miR-154-5p notably inhibited the tumor growth in an OS xenograft model. ConclusionThese results indicated that miR-154-5p may function as a potential tumor suppressor in OS. Therefore, miR-154-5p might be a novel therapeutic option for the treatment of OS.

Zn-HA/Bi-HA biphasic coatings on Titanium: Fabrication, characterization, antibacterial and biological activity

Hydroxyapatite (HA) coatings have been of important as biocompatible coatings for dental and bone tissue engineering application. However, the poor antibacterial performance and weak biological activity of HA coatings limited their clinical applications. As a strategy to improve the antibacterial performance and biological activity of HA, Zinc and bismuth ions were incorporated into HA lattice by substituting Ca2+ ions, respectively, and thus zinc substituted hydroxyapatite/bismuth substituted hydroxyapatite (Zn-HA/Bi-HA) biphasic coatings on titanium plates with various ratios were fabricated via sol-gel and dip-coating processes. The purity of the Zn-HA and Bi-HA phase was confirmed by X-ray diffraction (XRD) test. The biphasic coatings showed slower dissolution rate than pure HA coating. Furthermore, the Zn-HA/Bi-HA coatings reveal good biomineralization activity in simulated body fluid (SBF) by forming regular spherical apatite agglomerates. Moreover, the biphasic Zn-HA/Bi-HA coatings exhibited that improved antimicrobial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) as compared to pure HA coatings. The CCK-8 assays demonstrate Zn-HA/Bi-HA coatings showed no toxicity to MG63 cells, and the Zn-HA/Bi-HA2 (Zn-HA:Bi-HA=64:1) coating is more effective to enhance the proliferation of MG63 cells compared to other coatings. This finding suggests Zn-HA/Bi-HA biphasic coatings are promising candidates for biomedical applications.

Hydrolytic Expansion Induces Corrosion Propagation for Increased Fe Biodegradation.

Fe is regarded as a promising bone implant material due to inherent degradability and high mechanical strength, but its degradation rate is too slow to match the healing rate of bone. In this work, hydrolytic expansion was cleverly exploited to accelerate Fe degradation. Concretely, hydrolyzable Mg2Si was incorporated into Fe matrix through selective laser melting and readily hydrolyzed in a physiological environment, thereby exposing more surface area of Fe matrix to the solution. Moreover, the gaseous hydrolytic products of Mg2Si acted as an expanding agent and cracked the dense degradation product layers of Fe matrix, which offered rapid access for solution invasion and corrosion propagation toward the interior of Fe matrix. This resulted in the breakdown of protective degradation product layers and even the direct peeling off of Fe matrix. Consequently, the degradation rate for Fe/Mg2Si composites (0.33 mm/y) was significantly improved in comparison with that of Fe (0.12 mm/y). Meanwhile, Fe/Mg2Si composites were found to enable the growth and proliferation of MG-63 cells, showing good cytocompatibility. This study indicated that hydrolytic expansion may be an effective strategy to accelerate the degradation of Fe-based implants.

Dual functional modification of gellan gum hydrogel by introduction of methyl methacrylate and RGD contained polypeptide

For tissue fabrication and tissue regeneration, three-dimension (3D) hydrogel matrix with both cellular bioactivity and versatile fabrication ability for in-situ cell laden is always challenging and spotlight. In this study, we establish a facile approach to modify gellan gum (GG) for overcoming its disadvantages, including high gelling temperature which is incompatible for in-situ cell encapsulation and the native inactivity for cell adhesion. By esterification reaction, methacrylic anhydride modified GG (MAGG) is prepared, producing photo-reactivity for crosslinking and grafting with RGD-contained peptides. Therefore, RGD-grafted hydrogel (MAGG-RGD) is feasibly prepared. Detail characterizations show that MAGG hydrogel possesses improved physicochemical properties, with tunable strength varied with MA dose. Moreover, the introduction of RGD domains significantly improves the MG63 cells proliferation, adhesion and spreading both on two-dimension (2D) surface and in 3D matrix. Given the native photo-crosslinkable capacity, tunable physicochemical nature and good bioactivity, MAGG-RGD hydrogel is perspective in tissue fabrication and tissue engineering, and expands the potential material platform for cell carrier or tissue scaffold.

Down-regulation of miR-10b represses cell vitality in osteosarcoma and is inversely associated with prognosis via interacting with FAM46C: Running title: MiR-10b/FAM46C axis modulates OS progression

The purpose of this exploration was to detect the biological effects of miR-10b/FAM46C pair on osteosarcoma (OS) development. By accessing to the Gene Expression Omnibus (GEO) database, we achieved expressional profiles of miR-10b and FAM46C. Kaplan-Meier method was applied to determine the overall survival rates of OS patients. MiR-10b mimic/inhibitor were utilized to alter miR-10b expression. Overexpression of FAM46C was induced by pcDNA3.1-FAM46C. QRT-PCR and western blot were conducted to assess the expression levels. Cell counting kit-8 (CCK-8) and transwell assays were employed to evaluate the proliferative, invasive and migratory properties of OS cells. Pearson correlation analysis was performed to confirm the association between miR-10b and FAM46C. Dual-luciferase reporter assay was conducted to determine the target of miR-10b. The overall survival of OS patients was inversely correlated with miR-10b expression. MiR-10b was increased in OS compared with normal controls. Depletion of miR-10b attenuated the proliferation, invasion and migration of MG-63 cells. FAM46C was considered as a target gene of miR-10b and inversely related with miR-10b. Overexpression of FAM46C could inhibit cell growth, invasion and migration in OS; furthermore, it also can enforced the miR-10b inhibitor-induced effects on cell behaviors of OS cells. Down-regulation of miR-10b played a suppressive effect on the cell activity in OS cells, which provides a novel insight into the advance of OS therapeutic therapies.

Effects of long-chain non-coding HIF2PUT on migration and invasion of osteosarcoma cells

Objective To observe the effects of long-chain non-coding RNA (lncRNA) HIF2PUT on the proliferation, migration and invasion of osteosarcoma cells. Methods Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the expression of lncRNA HIF2PUT in 47 cases of osteosarcoma tissues (28 males and 19 females, the oldest was 45 years old, the youngest was 8 years old with an average of 20.24 years old) and 2 osteosarcoma cell lines (MG63 and HOS). After screening for stable transfectants by lentivirus, the methyl thiazol tetrazolium (MTT) assay and Transwell assay were applied to detect the effects of lncRNA HIF2PUT on proliferation, migration and invasion of osteosarcoma cells. According to the cell transfection, the experimental group (overexpressed lncRNA HIF2PUT group), the blank transfection group and the blank control group were respectively set. Data were expressed as mean±standard deviations (SD), and SPSS 21.0 software was used for statistical analysis. Results The expression of lncRNA HIF2PUT in 47 cases of osteosarcoma (39.56±0.48) was significantly lower than that in normal tissues (79.56±1.16, t=3.516, P<0.05). The expression level of lncRNA HIF2PUT in MG63 cells was significantly higher than that in HOS cells. Cell function experiments showed that lncRNA HIF2PUT had a significant effect on the proliferation, migration and invasion of osteosarcoma cells. Proliferation of MG63 and HOS cells was significantly inhibited by lncRNA HIF2PUT, and the proliferation index was significantly lower in lncRNA HIF2PUT group than in blank transfection group and blank control group (t=4.785, P<0.05). For the migration experiment, the migrating number of MG63 cells was (154.36±13.85), (196.32±13.52) and (258.63±12.35), respectively (t=3.968, P<0.01), and that of HOS cells was (131.24±10.58), (247.36±17.52) and (224.52±14.52) respectively (t=3.052, P<0.01). For the invasion experiment, the penetrating number of MG63 cells was (73.50±13.89), (98.63±12.57) and (112.30±15.29), respectively (t=23.178, P<0.01), and that of HOS cells was (70.74±13.49) (98.40±16.46) and 89.74±12.47, respectively (t=14.275, P<0.01). The migration and invasion ability of MG63 and HOS cells in lncRNA HIF2PUT group was lower than that in blank transfection group and blank control group. Conclusion The expression of lncRNA HIF2PUT was significantly low in osteosarcoma tissues and MG63 and HOS cell lines. The expression of lncRNA HIF2PUT significantly inhibited the proliferation, migration and invasion of osteosarcoma cells. Key words: Osteosarcoma; Long-chain non-coding RNA; Migration; Invasion

Investigation of the effect of hydroalcoholic mumie extract on expression of osteoprotegerin (OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL)

Background and Aim: Osteoporosis is one of the most common bone diseases that is caused by an imbalance between the activity of osteoblasts and osteoclasts. Osteoprotegerin (OPG) plays an active role in inhibiting osteoclasts by binding to the receptor activator of nuclear factor kappa-B ligand (RANKL). The balance in the OPG/RANKL ratio is important in bone remodeling. The current study investigated and compared the effects of mumie extract and chemical drugs on cell proliferation, expression of OPG and RANKL in MG63 cells. Materials and Methods: The effects of 100, 200 and 300 μg/ml of mumie extract on cell proliferation, expression of OPG and RANKL in MG63 cells were investigated in experimental groups, positive and negative control groups. Data were analyzed by ANOVA test. Results: Significant increases were observed in the proliferation of MG63 cells after exposure to 100 and 200 μg/ml concentrations of mumie extract. 200 μg/ml concentration of the extract significantly decreased the expression of RANKL and increased the expression of OPG. It also increased OPG/RANKL ratio significantly in the experimental groups, compared to those in the negative control groups. Although the effects of 100 μg/ml and 200 μg/ml concentrations were similar to each other, they showed no significant effects on the variables. 300 μg/ml concentration of the extract showed a reverse and insignificant effect. Conclusion: 200 μg/ml concentration of mumie extract had a significant effect on the expression of two important factors in bone remodeling. Therefore, individuals susceptible to osteoporosis can use mumie extract as an alternative to chemical drugs. © 2018 the Author (s).

LncRNA MEG3 inhibits proliferation and promotes apoptosis of osteosarcoma cells through regulating Notch signaling pathway.

To explore the effect of long non-coding ribonucleic acid (lncRNA)-maternally expressed gene 3 (MEG3) on the Notch signaling pathway, and its influences on the proliferation and apoptosis of osteosarcoma MG-63 cells. LncRNA MEG3 was overexpressed in osteosarcoma MG-63 cells, and the cells were divided into Blank group, Len-con group, and Len-MEG3 group. The expression level of MEG3 in each group was detected via quantitative Polymerase Chain Reaction (qPCR), the cell proliferation level in each group was detected via Cell Counting Kit-8 (CCK-8) assay, and the apoptosis in each group was detected via Hoechst 33258 staining. Moreover, the content of the inflammatory factors in each group was determined using the Enzyme-Linked Immunosorbent Assay (ELISA), and the expression levels of apoptosis-related proteins and Notch signaling pathway-related proteins were determined through Western blotting. The expression level of lncRNA MEG3 in Len-MEG3 group was significantly higher than that in the Blank group and Len-con group (p<0.01). The overexpression of lncRNA MEG3 could significantly weaken the proliferation (p<0.01) and enhance the apoptosis of osteosarcoma cells (p<0.01). The overexpression of lncRNA MEG3 could significantly increase the content of the inflammatory factor interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) (p<0.01), and remarkably decrease the content of the anti-inflammatory factor IL-10 in osteosarcoma cells (p<0.01). Besides, the overexpression of lncRNA MEG3 could evidently raise the expression of Caspase3 (p<0.01) and reduce the Bcl-2/Bax expression in osteosarcoma cells (p<0.01). Finally, the overexpression of lncRNA MEG3 could remarkably reduce the protein expressions of Jagged1, Notch1, and NICD1 in osteosarcoma cells (p<0.01). The overexpression of lncRNA MEG3 can inhibit the proliferation and promote the apoptosis of osteosarcoma MG-63 cells by suppressing the Notch signaling pathway.

Phytoconstituents of the rhizome of Drynaria bonii and the effect of phytoconstituents on the cell proliferation of osteoblast MG-63 cells

In folk medicine of Vietnam, the plant Drynaria bonii Heinrich Christ was used for the treatment of bone disease, osteoporosis, tinnitus and stimulate hair growth, instead of Drynaria fortunei. Up to now, there are very little studies on its phytochemical as well as bioactivities of Drynaria bonii. Treatments for osteoporosis through stimulating bone formation via osteoblasts is currently concerned In this paper, we report the isolation and chemical structural determination six compounds including of 24-methylen-cycloartan-3ß-ol (1), triphyllol (2), 5-hydroxymethylfurfural (3), protocatechuic acid (4), rutin (5) and nicotiflorin (6) from the methanol extract of the rhizomes of Drynaria bonii. The cell proliferative effects of isolated compounds on MG63 osteoblast cell were also investigated. The result indicated that compounds 1, 2 and 4 stimulated the proliferation of MG63 osteoblast cells by 8.49%; 11.18 % and 7.68 % at the concentrations of 100 μg/ml, respectively. This is the first notification of six isolated compounds from Drynaria bonii and their cell proliferation activity in osteoblast-like cells.

Effects of rapamycin on osteosarcoma cell proliferation and apoptosis by inducing autophagy.

To investigate the influences of rapamycin on proliferation and apoptosis of human osteosarcoma MG-63 cells and the mechanisms of action. The human osteosarcoma MG-63 cells were randomly divided into Control group, Rapamycin group, and Rapamycin + Beclin-1 plasmid transfection group. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was adopted to detect the viability of MG-63 cells in each group, and the 5-Ethynyl-2'-deoxyuridine (EdU) staining and Hoechst staining were applied to determine the proliferation and apoptosis, respectively, of MG-63 cells in each group. The levels of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) were measured using enzyme-linked immunosorbent assay (ELISA) kits, and the protein expression levels of Beclin-1 and Vps34 in each group of MG-63 cells were tested using the Western blotting. Compared with the Control group, Rapamycin group, and Rapamycin + Beclin-1 plasmid transfection group had markedly weakened the viability of MG-63 cells, inhibited cell proliferation, remarkably increased cell apoptosis rate, elevated Bax level, notably declined Bcl-2 level, and significantly raised the levels of Beclin-1 and Vps34 proteins in MG-63 cells. Besides, the effects in Beclin-1 plasmid transfection group were stronger. Rapamycin may decrease the viability, inhibit the proliferation, and promote the apoptosis of MG-63 cells by activating autophagy.