Human Osteosarcoma MG-63 Cells Research Articles

Effect of TiO2 Nanotubes on Biological Activity of Osteoblasts and Focal Adhesion Kinase/Osteopontin Level

Osteoblasts are important cells for bone formation and play a major part in bone diseases and bone defects. Clinically, we usually adopt bone implants for related diseases. Also, nanotechnology is important in bones and joints. This study assessed the effects of TiO2 nanotubes of different diameters on osteoblast activity, FAK and OPN levels, aiming to provide an experimental foundation for selection of clinical bone implant materials. The morphology of MG-63 human osteosarcoma cells changed with expansion of TiO2 nanotubes’ diameter. From the biological activity, the cell proliferation and adhesion were enhanced as the diameter of the TiO2 nanotube was increased and its proliferation and adhesion were highest in the 100 nm TiO2 nanotube, which is related to increased ALP activity, FAK and OPN protein and mRNA expression. ELISA detected ALP activity and found that MG-63 cells cultured with 70 nm nanotube had strongest activity. Immune blotting and PCR results showed that, FAK and OPN activities were highest in 70 nm TiO2 nanotube cells. In summary, TiO2 nanotubes increased cell proliferation and adhesion by up-regulating the activities of FAK and OPN in a concentration-dependent relationship.

Isolation of 24-homoscalarane sesterterpenoids from a sponge of the genus Lendenfeldia

The chemical screening of an algae-containing marine sponge identified as Lendenfeldia species has isolated two 24-homoscalarane sesterterpenoids, including a known compound, felixin A (1) and a new analogue, lendenfeldarane V (2). The structure of 1, obtained in a previous study, was cited for the first time in this study via single-crystal X-ray diffraction analysis. The structure of 2 was ascertained via 2D NMR experiments and a literature review. The absolute configurations of 1 and 2 were delineated with DP4+ computation and specific optical rotation. Homoscalarane 1 exhibited cytotoxicity towards MG63 human osteosarcoma cells.

Laser texturing and oxidation of TiZrTa thin films to improve the biocompatible performance of titanium alloys

Titanium (Ti) and Ti alloys possess extensive applications in the medical field due to their non-toxic, biocompatible, and antibacterial properties. This study investigates the biocompatibility enhancement of a Ti-based alloy, Ti-6Al-4V, through a multi-step process involving cathodic arc ion plating of a TiZrTa thin film and subsequent laser-textured oxidation (LTO). LTO treatment produced a structured microtopography with regular groove patterns, including straight lines (L-s), circles (L-c), and checkboards (L-ch), resulting in surface oxidation and the formation of various oxides on the TiZrTa coating. Microstructure analyses revealed substantial alterations in surface topography induced by laser texturing. Antibacterial assessments against Staphylococcus aureus (S. aureus) showed that the films with LTO treatment did not exhibit significant advantages after 12 or 24 h of coculture because the laser-textured portions consisted of a combination of the film and the substrate, and the antibacterial effect was not obvious. Coatings with LTO treatment significantly improved L-929 mouse fibroblasts cell viability at 48 h, particularly for samples with a 50 μm spacing of straight and circular patterns (L-s and L-c). After two weeks of MG-63 human osteosarcoma cell culture, the viability of all samples reached similar levels. It suggested that the TiZrTa-coated Ti-6Al-4V exhibited good biocompatibility with both cells. This study contributed to the understanding of surface modification techniques for enhancing the biocompatibility of Ti-based alloys in biomedical applications.

Abstract 2878: Hyperthermia increases the efficacy of aminolevulinic acid-mediated photodynamic therapy in human osteosarcoma cells

Abstract Osteosarcoma is a malignant osteoid tumor that arises from mesenchymal cells exhibiting osteoblastic differentiation. Common treatments for this neoplasm include limb-salvage surgery and/or amputation of the affected limb, either of which can substantially decrease quality of life. Therefore, other, less invasive treatments should be explored. Photodynamic therapy (PDT) represents a more targeted, less invasive treatment possibility for osteosarcoma. Previous data from my laboratory demonstrated that aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) can be used to induce significant cell death in human osteosarcoma MG-63 cells in vitro. However, delivering high doses of drugs to deep osteosarcoma tumors could prove difficult, so we aim to explore the use of combination therapies to increase the efficacy of ALA-PDT in human osteosarcoma cells. Previous studies have shown that hyperthermia (increased temperature) can induce cell death in cancer cells. Furthermore, data suggest that combining PDT with hyperthermia can induce cell death in cancer cells synergistically by activating differential death-inducing pathways. However, hyperthermia in combination with ALA-PDT has not been explored in human osteosarcoma. Therefore, we aim to investigate the possibility that hyperthermia in combination with ALA-PDT can increase cell death in human osteosarcoma cells compared to ALA-PDT alone. Data from our lab show that compared to normal body temperature (37°C), cell death in MG-63 human osteosarcoma cells was not significantly increased by mild hyperthermia alone (40°C and 43°C respectively), while moderate hyperthermic temperatures alone (46°C and 49°C) did significantly increase cell death. We also found that at moderately hyperthermic temperatures, ALA-PDT does not increase cell death any further compared to moderate hyperthermia alone. Furthermore, we found that at the mildly hyperthermic temperature of 40°C, cell death was not significantly increased any further by ALA-PDT compared to ALA-PDT alone. However, our data do show that at a temperature of 43°C, hyperthermia plus ALA-PDT increased cell death compared to ALA-PDT alone. Therefore, our data suggest that inducing mild hyperthermia could increase the efficacy of ALA-PDT in human osteosarcoma cells. Citation Format: Paige J. Baugher. Hyperthermia increases the efficacy of aminolevulinic acid-mediated photodynamic therapy in human osteosarcoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2878.

Vitexicarpin Induces Apoptosis and Inhibits Metastatic Properties via the AKT-PRAS40 Pathway in Human Osteosarcoma.

Osteosarcoma, which has poor prognosis after metastasis, is the most common type of bone cancer in children and adolescents. Therefore, plant-derived bioactive compounds are being actively developed for cancer therapy. Artemisia apiacea Hance ex Walp. is a traditional medicinal plant native to Eastern Asia, including China, Japan, and Korea. Vitexicarpin (Vitex), derived from A. apiacea, has demonstrated analgesic, anti-inflammatory, antitumour, and immunoregulatory properties; however, there are no published studies on Vitex isolated from the aerial parts of A. apiacea. Thus, this study aimed to evaluate the antitumour activity of Vitex against human osteosarcoma cells. In the present study, Vitex (>99% purity) isolated from A. apiacea induced significant cell death in human osteosarcoma MG63 cells in a dose- and time-dependent manner; cell death was mediated by apoptosis, as evidenced by the appearance of cleaved-PARP, cleaved-caspase 3, anti-apoptotic proteins (Survivin and Bcl-2), pro-apoptotic proteins (Bax), and cell cycle-related proteins (Cyclin D1, Cdk4, and Cdk6). Additionally, a human phosphokinase array proteome profiler revealed that Vitex suppressed AKT-dependent downstream kinases. Further, Vitex reduced the phosphorylation of PRAS40, which is associated with autophagy and metastasis, induced autophagosome formation, and suppressed programmed cell death and necroptosis. Furthermore, Vitex induced antimetastatic activity by suppressing the migration and invasion of MMP13, which is the primary protease that degrades type I collagen for tumour-induced osteolysis in bone tissues and preferential metastasis sites. Taken together, our results suggest that Vitex is an attractive target for treating human osteosarcoma.

Vat photopolymerization 3D printing of yttria-stabilized ZrO2 ceramics: effects of a sintering additive (Na2O – 2SiO2), biocompatibility, and osteointegration

Effects of a liquid-phase sodium disilicate additive on the properties of ZrO2 ceramics containing 3 mol.% of Y2O3 and 2 wt% of Al2O3 are presented, including phase composition, microstructure, shrinkage, porosity, and bending strength. The distribution of elements along grain boundaries and the composition in a selected area of a lamella were investigated by transmission electron microscopy in pure and additive-containing samples. As a result of the introduction of the additive, sintering activity of the material increased, and the sintering temperature diminished down to 1150 °C. Thus, dense nanocrystalline materials with a crystallite size of 50–70 nm and a bending strength of up to 450 MPa were obtained. Optimized polymer-ceramic suspensions were used to produce objects with complex geometric shapes by 3D molding technology through layer-by-layer vat polymerization. According to in vitro assays, the obtained samples are nontoxic and cytocompatible with human osteosarcoma MG-63 cells, and according to in vivo experiments, are biocompatible and showing osseointegrative properties when implanted into the rat tibia.

Bio-functionalized carbon dots for signaling immuno-reaction of carcinoembryonic antigen in an electrochemical biosensor for cancer biomarker detection

Early diagnosis of cancer demands sensitive and accurate detection of cancer biomarkers in blood. Carbon dots (CDs) bio-functionalization with antibodies, peptides or aptamers have played significant role in cancer diagnosis and targeted cancer therapy. Herein, a biosensor for detection of cancer biomarker carcinoembryonic antigen (CEA) in blood serum has been designed using CDs bio-functionalized with HRP-conjugated CEA antibody (CUCDs@CEAAb2) as detection probe. CDs were synthesized by upscaling of cow urine, a nitrogen rich biomass waste, by hydrothermal method. Detection probe based on CDs resulted in 3.5 times higher sensitivity as compared to conventional electrochemical sandwich immunoassay. To further improve the sensor performance, hyper-branched polyethylenimine grafted poly amino aniline (PEI-g-PAANI) was used as the sensing interface, which enabled immobilization of higher amount of capture antibody. Detection of CEA in human blood serum coupled with wide linear range (0.5–50 ng/ml), good specificity, stability, reproducibility and low detection limit (10 pg/ml) signified the excellence of CUCDs based CEA immunosensor. CUCDs exhibited excitation wavelength dependent fluorescence property and showed strong blue emission under UV irradiation. MTT assay indicated that the material is not toxic towards human dental pulp stem cells (hDPSCs) and MG63 osteosarcoma cells (cell viability > 90%). The present study demonstrates a methodology for valorization of animal waste to a cost-effective carbon based functional nanomaterial for clinical detection of cancer biomarkers.

3D-Printed Collagen-Nanocellulose Hybrid Bioscaffolds with Tailored Properties for Tissue Engineering Applications.

Hybrid collagen (Coll) bioscaffolds have emerged as a promising solution for tissue engineering (TE) and regenerative medicine. These innovative bioscaffolds combine the beneficial properties of Coll, an important structural protein of the extracellular matrix, with various other biomaterials to create platforms for long-term cell growth and tissue formation. The integration or cross-linking of Coll with other biomaterials increases mechanical strength and stability and introduces tailored biochemical and physical factors that mimic the natural tissue microenvironment. This work reports on the fabrication of chemically cross-linked hybrid bioscaffolds with enhanced properties from the combination of Coll, nanofibrillated cellulose (NFC), carboxymethylcellulose (CMC), and citric acid (CA). The bioscaffolds were prepared by 3D printing ink containing Coll-NFC-CMC-CA followed by freeze-drying, dehydrothermal treatment, and neutralization. Cross-linking through the formation of ester bonds between the polymers and CA in the bioscaffolds was achieved by exposing the bioscaffolds to elevated temperatures in the dry state. The morphology, pores/porosity, chemical composition, structure, thermal behavior, swelling, degradation, and mechanical properties of the bioscaffolds in the dry and wet states were investigated as a function of Coll concentration. The bioscaffolds showed no cytotoxicity to MG-63 human bone osteosarcoma cells as tested by different assays measuring different end points. Overall, the presented hybrid Coll bioscaffolds offer a unique combination of biocompatibility, stability, and structural support, making them valuable tools for TE.

Blue light photobiomodulation induced apoptosis by increasing ROS level and regulating SOCS3 and PTEN/PI3K/AKT pathway in osteosarcoma cells

Blue light photobiomodulation (PBM) has attracted great attention in diminishing proliferation and inducing death of cancer cells recently. Osteosarcoma (OS) primarily occurring in children and adolescents, the limitations of drug resistance and limb salvage make it urgent to develop and identify new adjuvant therapeutic strategies. In this work, we attempted to research the anticancer effects and biological mechanisms of blue light PBM in human OS MG63 cells. The effects of various blue light parameters on MG63 cells indicated that suppressed cell proliferation and cell migration, induced cell apoptosis which are experimentally assessed using multiple assays including CCK, LDH, wound healing assay and Hoechst staining. Concurrently, the increases of ROS level and the inhibition of PI3K and AKT expression were identified under high-dose blue light PBM in MG63 cells. Meanwhile, SOCS3 is a major inducible anti-tumor molecule, we also found that blue light LED substantially promoted its expression. Thus, this study proposed that bule light PBM may be a hopeful therapeutic approach in OS clinical treatment in the future.

Pterostilbene-Isothiocyanate Inhibits Proliferation of Human MG-63 Osteosarcoma Cells via Abrogating β-Catenin/TCF-4 Interaction-A Mechanistic Insight.

Osteosarcoma, a highly metastasizing bone neoplasm, is a leading cause of death and disability in children and adolescents worldwide. Osteosarcoma is only suboptimally responsive to surgery and radio- and chemotherapy, that too with adverse side effects. Hence, there is a necessary need for safer alternative therapeutic approaches. This study evaluated the anticancer effects of the semi-synthetic compound, pterostilbene-isothiocyanate (PTER-ITC), on human osteosarcoma MG-63 cells through cytotoxicity, wound-healing, and transwell-migration assays. Results showed that PTER-ITC specifically inhibited the survival, proliferation, and migration of osteosarcoma cells. PTER-ITC induced apoptosis in MG-63 cells by disrupting mitochondrial membrane potential, as evident from the outcomes of different cytological staining. The antimetastatic potential of PTER-ITC was evaluated through immunostaining, RT-qPCR, and immunoblotting. In silico (molecular docking and dynamic simulation) and, subsequently, biochemical [co-immunoprecipitation (Co-IP) and luciferase reporter] assays deciphered the underlying mode-of-action of this compound. PTER-ITC increased E-cadherin and reduced N-cadherin levels, thereby facilitating the reversal of epithelial-mesenchymal transition (EMT). It also modulated the expressions of proliferative cell nuclear antigen (PCNA), caspase-3, poly [ADP-ribose] polymerase (PARP-1) and matrix metalloproteinase-2/9 (MMPs-2/9) at transcriptional and translational levels. PTER-ITC interfered with the β-catenin/transcription factor-4 (TCF-4) interaction in silico by occupying the β-catenin binding site on TCF-4, confirmed by their reduced physical interactions (Co-IP assay). This inhibited transcriptional activation of TCF-4 by β-catenin (as shown by luciferase reporter assay). In conclusion, PTER-ITC exhibited potent anticancer effects in vitro against human osteosarcoma cells by abrogating the β-catenin/TCF-4 interaction. Altogether, this study suggests that PTER-ITC may be regarded as a new approach for osteosarcoma treatment.

Upregulation of human GD3 synthase (hST8Sia I) gene expression during serum starvation-induced osteoblastic differentiation of MG-63 cells.

In this study, we have firstly elucidated that serum starvation augmented the levels of human GD3 synthase (hST8Sia I) gene and ganglioside GD3 expression as well as bone morphogenic protein-2 and osteocalcin expression during MG-63 cell differentiation using RT-PCR, qPCR, Western blot and immunofluorescence microscopy. To evaluate upregulation of hST8Sia I gene during MG-63 cell differentiation by serum starvation, promoter area of the hST8Sia I gene was functionally analyzed. Promoter analysis using luciferase reporter assay system harboring various constructs of the hST8Sia I gene proved that the cis-acting region at -1146/-646, which includes binding sites of the known transcription factors AP-1, CREB, c-Ets-1 and NF-κB, displays the highest level of promoter activity in response to serum starvation in MG-63 cells. The -731/-722 region, which contains the NF-κB binding site, was proved to be essential for expression of the hST8Sia I gene by serum starvation in MG-63 cells by site-directed mutagenesis, NF-κB inhibition, and chromatin immunoprecipitation (ChIP) assay. Knockdown of hST8Sia I using shRNA suggested that expressions of hST8Sia I and GD3 have no apparent effect on differentiation of MG-63 cells. Moreover, the transcriptional activation of hST8Sia I gene by serum starvation was strongly hindered by SB203580, a p38MAPK inhibitor in MG-63 cells. From these results, it has been suggested that transcription activity of hST8Sia I gene by serum starvation in human osteosarcoma MG-63 cells is regulated by p38MAPK/NF-κB signaling pathway.

Collagen-Coated Hyperelastic Bone Promotes Osteoblast Adhesion and Proliferation.

Successfully reconstructing bone and restoring its dynamic function represents a significant challenge for medicine. Critical size defects (CSDs), resulting from trauma, tumor removal, or degenerative conditions, do not naturally heal and often require complex bone grafting. However, these grafts carry risks, such as tissue rejection, infections, and surgical site damage, necessitating the development of alternative treatments. Three-dimensional and four-dimensional printed synthetic biomaterials represent a viable alternative, as they carry low production costs and are highly reproducible. Hyperelastic bone (HB), a biocompatible synthetic polymer consisting of 90% hydroxyapatite and 10% poly(lactic-co-glycolic acid, PLGA), was examined for its potential to support cell adhesion, migration, and proliferation. Specifically, we seeded collagen-coated HB with MG-63 human osteosarcoma cells. Our analysis revealed robust cell adhesion and proliferation over 7 days in vitro, with cells forming uniform monolayers on the external surface of the scaffold. However, no cells were present on the core of the fibers. The cells expressed bone differentiation markers on days 3 and 5. By day 7, the scaffold began to degrade, developing microscopic fissures and fragmentation. In summary, collagen-coated HB scaffolds support cell adhesion and proliferation but exhibit reduced structural support after 7 days in culture. Nevertheless, the intricate 3D architecture holds promise for cellular migration, vascularization, and early osteogenesis.

Down-Regulation of CYP3A4 by the KCa1.1 Inhibition Is Responsible for Overcoming Resistance to Doxorubicin in Cancer Spheroid Models

The large-conductance Ca2+-activated K+ channel, KCa1.1, plays a pivotal role in cancer progression, metastasis, and the acquisition of chemoresistance. Previous studies indicated that the pharmacological inhibition of KCa1.1 overcame resistance to doxorubicin (DOX) by down-regulating multidrug resistance-associated proteins in the three-dimensional spheroid models of human prostate cancer LNCaP, osteosarcoma MG-63, and chondrosarcoma SW-1353 cells. Investigations have recently focused on the critical roles of intratumoral, drug-metabolizing cytochrome P450 enzymes (CYPs) in chemoresistance. In the present study, we examined the involvement of CYPs in the acquisition of DOX resistance and its overcoming by inhibiting KCa1.1 in cancer spheroid models. Among the CYP isoforms involved in DOX metabolism, CYP3A4 was up-regulated by spheroid formation and significantly suppressed by the inhibition of KCa1.1 through the transcriptional repression of CCAAT/enhancer-binding protein, CEBPB, which is a downstream transcription factor of the Nrf2 signaling pathway. DOX resistance was overcome by the siRNA-mediated inhibition of CYP3A4 and treatment with the potent CYP3A4 inhibitor, ketoconazole, in cancer spheroid models. The phosphorylation levels of Akt were significantly reduced by inhibiting KCa1.1 in cancer spheroid models, and KCa1.1-induced down-regulation of CYP3A4 was reversed by the treatment with Akt and Nrf2 activators. Collectively, the present results indicate that the up-regulation of CYP3A4 is responsible for the acquisition of DOX resistance in cancer spheroid models, and the inhibition of KCa1.1 overcame DOX resistance by repressing CYP3A4 transcription mainly through the Akt-Nrf2-CEBPB axis.

Molecular mechanism of a novel root-end filling material containing zirconium oxide on the osteogenic/odontogenic differentiation of human osteosarcoma MG-63 cells.

Although the novel root-end filling material containing zirconium oxide (NRFM-Zr) which is hydroxyapatite-based may promote osteoblast differentiation, the molecular mechanism remains unclear. The aim of this study is to investigate it underlying the osteogenic/odontogenic differentiation of human osteosarcoma MG-63 cells induced by NRFM-Zr, compared with calcium silicate-based mineral trioxide aggregate (MTA), and glass ionomer cement (GIC). Firstly, three different types of root filling materials were co-cultured with MG-63 cells, and their cell toxicity, alkaline phosphatase (ALP) activity, and calcium ion concentration were evaluated. Next, gene expression profiling microarray was employed to analyze the impact of the materials on the gene expression profile of MG-63 cells. The results of cell viability revealed that NRFM-Zr group had no significant difference compared to the negative control group. After 5 and 7days of cultivation, both the NRFM-Zr and MTA groups exhibited significantly higher ALP activity compared to the negative control (p < 0.05). Moreover, the NRFM-Zr group had the highest calcium ion concentration, while the GIC group was the lowest (p < 0.05). Gene expression profiling microarray analysis identified 2915 (NRFM-Zr), 2254 (MTA) and 392 (GIC) differentially expressed genes, respectively. GO functional and KEGG pathway analysis revealed that differentially expressed genes of NRFM-Zr, MTA and GIC participated in 8, 6 and 0 differentiation-related pathways, respectively. Comparing the molecular mechanisms of osteogenic/odontogenic differentiation induced by hydroxyapatite-based NRFM-Zr and calcium silicate-based MTA, it was found that they shared similarities in their molecular mechanisms of promoting osteogenic differentiation. NRFM-Zr primarily promotes differentiation and inhibits cell apoptosis, thereby enhancing osteogenic/odontogenic differentiation of MG-63 cells. Furthermore, the inducing efficacy of NRFM-Zr was found to be superior to MTA.

Morphological and fractal features of cancer cells anchored on composite layers based on magnesium-doped hydroxyapatite loaded in chitosan matrix

In the present study, we report the development and characterization of composite layers (by spin coating) based on magnesium-doped hydroxyapatite in a chitosan matrix, containing human osteosarcoma MG63 cells anchored. Studies regarding the biocompatibility of the composite layers were performed with the aid of a MTT (3–4,5-Dimethylthiazol 2,5-diphenyltetrazolium bromide) assay. The data determined that the composite layers did not inhibit the growth and adhesion of MG63 cells to their surfaces exhibiting good biocompatibility properties. Furthermore, the attachment and development of MG63 cells on the surface of MgHApCh composite layers were investigated using atomic force microscopy (AFM). AFM topographical maps emphasized that the HApCh and 8MgHApCh composite layers surface promoted the attachment and proliferation of MG63 cells on their surface. Meanwhile, in the case of 30MgHApCh layers incubated for 48 h, a slight modification of the morphological features of the MG63 cells. In addition, the effects of the composite layers against Candida albicans ATCC 10231 were also evaluated. The data results from the in vitro antifungal assay depicted that the composite layers successfully inhibited the growth of the fungal cells onto their surface. Morphological and fractal analyses unveil cancer cell surfaces on Mg-containing composite layers with intricate 3D patterns, driven by high-frequency components. Their remarkable complexity and roughness arises from a strong multifractal nature, supporting more effective vertical growth compared to Si and HApCh surfaces. The cell viability reduced of uncoated Si surface is highlighted by its less intense 3D pattern growth. Our results show that the uncoated Si surface promotes lower viability of MG63 cancer cells, with less rough and complex 3D spatial patterns.

Prunetin Triggers ROS-mediated Apoptosis through the Suppression of MAPKs/STAT-3/NF-κB-p65 Signaling Pathway in Human Osteosarcoma Cells

Background Osteosarcoma (OS) is a highly metastatic primary bone malignancy, and the treatment options remain inadequate. Hence, exploring innovative natural medications is required. Prunetin (PRU) is an isoflavone that has been a proven anticancer agent in numerous cancer cell lines. However, the activity of PRU against OS remains uncertain. Materials and Methods Here, we studied the anticancer activity of PRU (20 and 25 µM) on human OS cells MG-63 and investigated its latent mechanism. The PRU activity of MG-63 cells cytotoxicity, intracellular ROS, metastasis, apoptosis, anti-apoptotic proteins, MAPK/STAT-3, and AKT signaling pathways was assessed by MTT assay, DCFH-DA, DAPI, PI, AO/EB, cell adhesion, and RT-PCR analysis. Findings unveiled that PRU could constrain MG-63 cell viability and adhesion through elevated intracellular ROS and elicited apoptosis. Results Likewise, PRU (20 and 25 µM) avert the MG-63 cell proliferation, which stimulates apoptosis by the enhancement of Bax and caspases, while it diminishes Bcl-2 in a dose-dependent way. Furthermore, PRU could reduce Pin-1, and anti-apoptotic elements, as well as trigger apoptotic signaling pathways. Our data established that PRU alleviates MG-63 cell proliferation and metastasis via ROS-mediated apoptosis, which triggers MAPKs/STAT3 and AKT pathways, suggesting that PRU is a promising natural remedy for OS. In order to comprehend the therapeutic target for cancer, we assessed the effect of PRU on the expression of Pin1, which is thought to be over-expressed in many human malignancies. According to our findings, PRU specifically suppressed Pin1 expression to reduce the expression of Akt, STAT3, P38, JNK, P65, and IL-6. We evaluated the impact of PRU on the expression of Pin1, which is allegedly over-expressed in many human malignancies, to better understand the therapeutic target for cancer. Researchers state that PRU inhibited the expression of Akt, STAT3, P38, JNK, P65, and IL-6 in particular, by suppressing Pin1 expression. Conclusion Together, these results suggest that PRU may be an effective treatment for bone cancer in people by preventing Pin1 expression.

Hyaluronic acid/cellulose acetate polymeric mixture containing binary metal oxide nano-hybrid as low biodegradable wound dressing

Wound dressing from a polymeric mixture of cellulose acetate (CA) and hyaluronic acid (HA) embedded with copper oxide (CuO) and magnesium oxide (MgO) nanoparticles as a scaffold was successfully fabricated. Several methods, including X-Ray diffraction analysis (XRD), contact angle, Fourier-transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM), were employed to characterize the prepared nanocomposite samples. The data obtained from XRD showed that pure HA with diffraction peaks at 2θ = 20.8°, 46.7°, and 63.2° that assigned to the semi-crystalline nature of HA. At the same time, the XRD pattern of the CuO@CA nanocomposites revealed the presence of a sharp crystalline peak at 2θ = 36°, representing the presence of the CuO nanomaterial. In addition, the typical cubic MgO peaks in the MgO@CA showed the pattern at 2θ = 43.1° and 62.5°. Furthermore, the diffraction pattern of CuO/MgO@HA/CA film shows strong crystallinity since its diffraction peaks are stated at 2θ = 8.8°, 13.01°, 29.6°, and 53.5° which represents CuO nanoparticles and the polycrystalline cubic structure of MgO nanoparticles exhibit several peaks at 2θ = 29.4°, 31°, 38.7°, 42.2°, and 63°. FTIR spectra of CuO/MgO@HA/CA displayed bands at 657 cm−1 and 843 cm−1 which correspond to the broadening vibration mode for the Mg-O-Mg and the existence of Mg-O vibrations, while the single band for CuO was shown at 1556 cm−1. SEM analysis revealed that the nanoparticles of copper oxide and magnesium oxide are evenly distributed throughout the polymer mass with the clean surface of the polymer with the appearance of nanoparticles with rough size and distribution in regions of varied densities by adding binary metal oxide nanoparticles (CuO/MgO@HA/CA). Furthermore, swelling rate percent, contact angle, and degradation behavior studies showed that the swelling rate percent of the polymer mixture film decreased when adding nanometal oxide. Contact angle was increased by incorporating the metal oxide nanoparticles in the polymeric mixture structure. Moreover, the degradation behavior of prepared samples showed lower rates of deterioration and the sample containing a mixture of metal oxide nanoparticles (CuO/MgO@HA/CA) exhibits a lower weight loss value when compared to HA, HA/CA, and CA samples. Human osteosarcoma MG-63 cells were used in in-vitro cytotoxicity tests. CuO/MgO@HA/CA sample treatment on cells suggests a non-cytotoxic impact, in contrast to the MTT test, which yields cell viability values of between 70% and 80% (72 h).

Hydrothermal Transformation of Eggshell Calcium Carbonate into Apatite Micro-Nanoparticles: Cytocompatibility and Osteoinductive Properties.

The eggshell is a biomineral consisting of CaCO3 in the form of calcite phase and a pervading organic matrix (1-3.5 wt.%). Transforming eggshell calcite particles into calcium phosphate (apatite) micro-nanoparticles opens the door to repurposing the eggshell waste as materials with potential biomedical applications, fulfilling the principles of the circular economy. Previous methods to obtain these particles consisted mainly of two steps, the first one involving the calcination of the eggshell. In this research, direct transformation by a one-pot hydrothermal method ranging from 100-200 °C was studied, using suspensions with a stoichiometric P/CaCO3 ratio, K2HPO4 as P reagent, and eggshells particles (Ø < 50 μm) both untreated and treated with NaClO to remove surface organic matter. In the untreated group, the complete conversion was achieved at 160 °C, and most particles displayed a hexagonal plate morphology, eventually with a central hole. In the treated group, this replacement occurred at 180 °C, yielding granular (spherulitic) apatite nanoparticles. The eggshell particles and apatite micro-nanoparticles were cytocompatible when incubated with MG-63 human osteosarcoma cells and m17.ASC murine mesenchymal stem cells and promoted the osteogenic differentiation of m17.ASC cells. The study results are useful for designing and fabricating biocompatible microstructured materials with osteoinductive properties for applications in bone tissue engineering and dentistry.

Expression of Concern: Adenovirus small interfering RNA targeting ezrin induces apoptosis and inhibits metastasis of human osteosarcoma MG-63 cells.

Expression of Concern: Adenovirus small interfering RNA targeting ezrin induces apoptosis and inhibits metastasis of human osteosarcoma MG-63 cells.

Fabrication of highly ordered willemite/PCL bone scaffolds by 3D printing: Nanostructure effects on compressive strength and in vitro behavior

In this study, first willemite (Zn2SiO4) micro and nano-powders were synthesized by the sol-gel method. X-ray diffraction (XRD), transmission electron microscopy (TEM), and dynamic light scattering (DLS) were applied to characterize the crystalline phases and particle size of powders. Then polycaprolactone (PCL) polymer scaffolds containing 20 wt% willemite were successfully fabricated by the DIW 3D printing (direct ink writing) method. The effects of willemite particle size on compressive strength, elastic modulus, degradation rate, and bioactivity of the composite scaffolds were investigated. The results showed that nanoparticle willemite/PCL (NW/PCL) scaffolds had 33.1% and 58.1% higher compressive strength and the elastic modulus of NW/PCL were 1.14 and 2.45 times better compared to micron size willemite/PCL (MW/PCL) and pure PCL scaffolds, respectively. Scanning electron microscopy (SEM) images and Energy-dispersive X-ray spectroscopy map (EDS map) results indicated that willemite nanoparticles, unlike microparticles, were smoothly embedded in the scaffold struts. In vitro tests also revealed an improvement in bone-like apatite formation ability and an increase in the degradation rate up to 2.17% by decreasing the willemite particle size to 50 nm. In addition, NW/PCL rendered significant enhancement in cell viability and cell attachment during the culture of MG-63 human osteosarcoma cell line. Nanostructure had also a positive effect on ALP activity and biomineralization in vitro.