Osteosarcoma Cell Line Saos-2 Research Articles

Polyacrylonitrile‒Chitosan IPN composite scaffolds that closely mimic the human trabecular bone structure for tissue engineering

Successful bone tissue engineering involves managing several important parameters, such as the design of intercommunicating porous structures, pore sizes, and the material and mechanical suitability of the material. In our work, we focused on the preparation of a synthetic scaffold that morphologically mimics the structure of human trabecular bone. The scaffolds were fabricated through the thermal modification (TM) of polyacrylonitrile. The scaffold strength was supported by a crosslinked chitosan supporting network. The prepared scaffold has imprinted pores of the appropriate size to facilitate the ingrowth and proliferation of human osteoblasts throughout the entire pore volume created by a primary porogen, sodium chloride. The resulting material has a dual porous morphological structure, in which adjustable larger pores support cellular ingrowth into the scaffold, whereas smaller pores, created using succinonitrile (SCN) as a secondary porogen, increase the diffusion of oxygen and nutrients to developing cells. The mechanical properties of the scaffold were promoted by the use of a secondary interpenetrating network (IPN) based on chitosan. The incorporation of secondary IPNs led to a significant improvement in the mechanical characteristics of the scaffold. Two crosslinking agents were used: the widely utilized glutaraldehyde (GA) and its green, nontoxic alternative, genipin (GEN).The current study introduces a synthetic scaffold that effectively mimics the structure of human trabecular bone, providing a conducive environment for osteoblast proliferation and ingrowth. We incorporated a dual porous morphology and a secondary IPN based on chitosan and found that the scaffold exhibited improved mechanical properties and nutrient diffusion capabilities. This research highlights the potential of these scaffolds for major advancements in bone tissue engineering, demonstrating their noncytotoxic nature and ability to support the adhesion and proliferation of the human osteosarcoma cell line SAOS-2. The described scaffold preparation technique may offer distinct advantages over current high-end methods such as 3D printing. The primary benefit lies in its very simple instrumentation, which enables the creation of a complex trabecular bone morphology with both macro- and microporosity in the bulk, which is still challenging for common 3D printers. Additionally, IPN with chitosan can be postmodified, substantially expanding the potential applications of these scaffolds.

Characterization of Iron Oxide Nanotubes Obtained by Anodic Oxidation for Biomedical Applications-In Vitro Studies.

To improve the biocompatibility and bioactivity of biodegradable iron-based materials, nanostructured surfaces formed by metal oxides offer a promising strategy for surface functionalization. To explore this potential, iron oxide nanotubes were synthesized on pure iron (Fe) using an anodic oxidation process (50 V-30 min, using an ethylene glycol solution containing 0.3% NH4F and 3% H2O, at a speed of 100 rpm). A nanotube layer composed mainly of α-Fe2O3 with diameters between 60 and 70 nm was obtained. The effect of the Fe-oxide nanotube layer on cell viability and morphology was evaluated by in vitro studies using a human osteosarcoma cell line (SaOs-2 cells). The results showed that the presence of this layer did not harm the viability or morphology of the cells. Furthermore, cells cultured on anodized surfaces showed higher metabolic activity than those on non-anodized surfaces. This research suggests that growing a layer of Fe oxide nanotubes on pure Fe is a promising method for functionalizing and improving the cytocompatibility of iron substrates. This opens up new opportunities for biomedical applications, including the development of cardiovascular stents or osteosynthesis implants.

Investigating the Anticancer Potential of Zinc and Magnesium Alloys: From Base Materials to Nanocoated Titanium Implants.

In this work, we show the in vitro anticancer potential of surgical wires, obtained from zinc (ZnMg0.004) or magnesium (MgCa0.7) alloys by spatial technology comprising casting, extrusion, and final drawing processes. We also present the selective anticancer effects of applied soluble multilayer nanocoatings of zinc and magnesium onto titanium surfaces using the pulse laser deposition method. In the latter, the titanium samples were produced via 3D printing using the selective laser melting method and coated with various combinations of zinc and magnesium layers. For cytotoxicity studies, human dental pulp-derived stem cells (hDPSCs) and human osteosarcoma SaOS-2 cell line were used as representatives of healthy and cancer cells. Cells were examined against the 0.3-3.0 cm2/mL material extract ratios obtained from experimental and steel surgical wires, the latter being the current clinical industry standard. The MgCa0.7 alloy wires were approx. 1.5 times more toxic to cancer cells at all examined extract ratios vs. the extracts from steel surgical wires that exhibited comparable toxicity towards healthy and cancer cells. The ZnMg0.004 alloy wires displayed increased toxicity towards cancer cells with decreasing extract ratios. This was also reflected in the increased anticancer effectiveness, calculated based on the viability ratio of healthy cells to cancer cells, from 1.1 to 4.0 times. Healthy cell viability remained at 80-100%, whereas cancer cell survival fluctuated at 20-75%, depending on the extract ratio. Furthermore, the culture of normal or cancer cells on the surface of Zn/Mg-coated titanium allowed us to select combinations of specific coating layers that yielded a comparable anticancer effectiveness to that observed with the experimental wires that ranged between 2 and 3. Overall, this work not only demonstrates the substantial anticancer properties of the studied wires but also indicates that similar anticancer effects can be replicated with appropriate nanocoatings on titanium samples. We believe that this work lays the groundwork for the future potential development of the category of new implants endowed with anticancer properties.

Allicin affects immunoreactivity of osteosarcoma cells through lncRNA CBR3-AS1

ObjectiveTo analyze the effect of allicin on the immunoreactivity of osteosarcoma (OS) cells and further explore whether its mechanism is related to the long non-coding Ribonucleic Acid (lncRNA) CBR3-AS1/miR-145-5p/GRP78 axis, so as to provide clinical evidence. MethodsThe human OS cell line Saos-2 was treated with allicin at 25, 50, and 100 μmol/L, respectively, to observe changes in cell biological behaviors. Subsequently, CBR3-AS1 abnormal expression vectors were constructed and transfected into Saos-2 to discuss their influence on OS. Furthermore, the regulatory relationship between allicin and the CBR3-AS1/miR-145-5p/GRP78 axis was validated by rescue experiments. Finally, a nude mice tumorigenesis experiment was carried out to analyze the effects of allicin and CBR3-AS1/miR-145-5p/GRP78 axis on the growth of living tumors. Alterations in T-lymphocyte subsets were also detected to assess the effect of allicin on OS immunoreactivity. ResultsWith the increase of allicin concentration, Saos-2 activity decreased and apoptosis increased (P < 0.05). In addition, the expression of CBR3-AS1 and GRP78 decreased after allicin intervention, while miR-145-5p increased (P < 0.05). Silencing CBR3-AS1 led to reduced Saos-2 activity, enhanced apoptosis, and activated mitophagy and endoplasmic reticulum stress (P < 0.05). In the rescue experiment, the effect of CBR3-AS1 on OS cells was reversed by silencing miR-145-5p, while the impact of miR-145-5p was reversed by GRP78. Finally, the tumorigenesis experiment in nude mice confirmed the regulatory effects of allicin and CBR3-AS1/miR-145-5p/GRP78 on tumor growth in vivo. Meanwhile, it was seen that allicin activated CD4+CD8+ in OS mice, confirming that allicin has the effect of activating OS immunoreactivity. ConclusionsAllicin activates OS immunoreactivity and induces apoptosis through the CBR3-AS1/miR-145-5p/GRP78 molecular axis.

Self-Assembled Hydrogel Membranes with Structurally Tunable Mechanical and Biological Properties.

Using supramolecular self-assembled nanocomposite materials made from protein and polysaccharide components is becoming more popular because of their unique properties, such as biodegradability, hierarchical structures, and tunable multifunctionality. However, the fabrication of these materials in a reproducible way remains a challenge. This study presents a new evaporation-induced self-assembly method producing layered hydrogel membranes (LHMs) using tropocollagen grafted by partially deacetylated chitin nanocrystals (CO-g-ChNCs). ChNCs help stabilize tropocollagen's helical conformation and fibrillar structure by forming a hierarchical microstructure through chemical and physical interactions. The LHMs show improved mechanical properties, cytocompatibility, and the ability to control drug release using octenidine dihydrochloride (OCT) as a drug model. Because of the high synergetic performance between CO and ChNCs, the modulus, strength, and toughness increased significantly compared to native CO. The biocompatibility of LHM was tested using the normal human dermal fibroblast (NHDF) and the human osteosarcoma cell line (Saos-2). Cytocompatibility and cell adhesion improved with the introduction of ChNCs. The extracted ChNCs are used as a reinforcing nanofiller to enhance the performance properties of tropocollagen hydrogel membranes and provide new insights into the design of novel LHMs that could be used for various medical applications, such as control of drug release in the skin and bone tissue regeneration.

Natural melanin nanoparticles (MNPs) extracted from Sepia officinalis: A cost-effective, chemo-photothermal, synergistic nanoplatform for osteosarcoma treatment

Osteosarcoma conventional chemotherapeutics are known for their side effects, limited options, and induction of drug resistance. This creates the need to develop new therapeutics capable of effectively destroying cancer cells with low toxicity, improving patient survival rate and their life quality. This work reports a novel drug delivery nanoplataform made of Natural Melanin Nanoparticles (MNPs), obtained from Sepia officinalis ink, with 99% incorporation efficiency of doxorubicin (Dox) without the use of non-toxic solvents. A significant photothermal effect was shown by a 36ºC increment after 10 min of laser irradiation, surpassing reported values for synthetic melanin. A sustained drug release of ca. 23% with photothermal stimuli was observed, compared to 15% without stimuli, after 48 h. This nanoplatform is obtained as a food industry side product, which makes it a natural cost-effective biomedical material. Natural MPs were applied in an osteosarcoma cell line (SaOs-2), and internalized by the cells in less than 2 h, showing cytocompatibility up to 1000 µg/mL after 72 h of contact with cells. On the contrary, when natural MNPs loaded with Dox (Dox-MNPs) were placed in contact with the SaOs-2 cells and were simultaneously receiving NIR light it was observed a 93% reduction in cancer cells in 48 h, revealing a synergistic effect between chemotherapy and phototherapy. To our knowledge this is the first time that natural MNPs extracted from Sepia officinalis were tested on an osteosarcoma cell line as chemo-photothermal agent, showing these NPs are an effective, cost-effective, reproducible, non-toxic nanoplatform for osteosarcoma treatment using combined effects.

The Oncolytic Effect of Aerva lanata on Osteosarcoma Cell Lines via the Apoptotic Signaling Pathway.

Introduction Osteosarcoma, a malignant bone tumor, poses significant treatment challenges, necessitating the development of alternative therapeutic strategies. Aerva lanata (A. lanata), a medicinal plant with traditional use in various healthcare systems, has anti-cancer properties. This study looks at the oncolytic effect of A. lanata extract on osteosarcoma cell lines (sarcoma osteogenic-Saos2). Aim The aim of this study was to investigate the oncolytic effect of Aerva lanata on Saos2 cell lines through the apoptotic signaling pathway. Materials and methods A. lanata extract was prepared using Soxhlet extraction, and its cytotoxic effects on Saos2 cells were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Real-time polymerase chain reaction (RT-PCR) analysis of gene activity was used to assess the extract's effect on apoptotic signaling pathways. Results The MTT assay demonstrated a dose-dependent decrease in Saos2 proliferation following treatment with A. lanata extract at concentrations ranging from 50 μg to 200 μg. The standard deviations observed ranged from 1.414 to 7.071. Gene expression analysis revealed that the extract led to a reduction in the messenger ribonucleic acid (mRNA) levels of the anti-apoptotic marker B-cell lymphoma 2 (Bcl2), with standard deviations ranging from 1 to 0.535. Conversely, it induced an increase in the mRNA levels of the tumor suppressor protein p53, with standard deviations ranging from 1 to 1.835. These findings suggest that the extract modulates the apoptotic pathways of the Bcl2 and p53 genes. Conclusion A. lanata extract exhibits promising anti-cancer activity against Saos2 osteosarcoma cell lines, inducing apoptosis by downregulating Bcl2 and increasing p53. The study's findings suggest that A. lanata may be useful as a natural treatment for osteosarcoma.

Osteosarcoma cell death induced by innovative scaffolds doped with chemotherapeutics.

Osteosarcoma (OS) cancer treatments include systemic chemotherapy and surgical resection. In the last years, novel treatment approaches have been proposed, which employ a drug-delivery system to prevent offside effects and improves treatment efficacy. Locally delivering anticancer compounds improves on high local concentrations with more efficient tumour-killing effect, reduced drugs resistance and confined systemic effects. Here, the synthesis of injectable strontium-doped calcium phosphate (SrCPC) scaffold was proposed as drug delivery system to combine bone tissue regeneration and anticancer treatment by controlled release of methotrexate (MTX) and doxorubicin (DOX), coded as SrCPC-MTX and SrCPC-DOX, respectively. The drug-loaded cements were tested in an in vitro model of human OS cell line SAOS-2, engineered OS cell line (SAOS-2-eGFP) and U2-OS. The ability of doped scaffolds to induce OS cell death and apoptosis was assessed analysing cell proliferation and Caspase-3/7 activities, respectively. To determine if OS cells grown on doped-scaffolds change their migratory ability and invasiveness, a wound-healing assay was performed. In addition, the osteogenic potential of SrCPC material was evaluated using human adipose derived-mesenchymal stem cells. Osteogenic markers such as (i) the mineral matrix deposition was analysed by alizarin red staining; (ii) the osteocalcin (OCN) protein expression was investigated by enzyme-linked immunosorbent assay test, and (iii) the osteogenic process was studied by real-time polymerase chain reaction array. The delivery system induced cell-killing cytotoxic effects and apoptosis in OS cell lines up to Day 7. SrCPC demonstrates a good cytocompatibility and it induced upregulation of osteogenic genes involved in the skeletal development pathway, together with OCN protein expression and mineral matrix deposition. The proposed approach, based on the local, sustained release of anticancer drugs from nanostructured biomimetic drug-loaded cements is promising for future therapies aiming to combine bone regeneration and anticancer local therapy.

Surface modification of Ti40Cu40Zr11Fe3Sn3Ag3 amorphous alloy for enhanced biocompatibility in implant applications

This study focuses on the design of a new Ti based multicomponent amorphous alloy for the development of biocompatible implant materials with enhanced hemocompatibility and cytocompatibility. While this class of amorphous alloys has shown its potential for biomedical implant applications, there are major concerns due to the presence of elements such as copper which can lead to cytotoxicity in the human body during long term implantation. Nevertheless, copper is indispensable in the development of an amorphous alloy. Thus, the objective of this work is to selectively remove copper from the surface of the Ti40Cu40Zr11Fe3Sn3Ag3 (at%) amorphous alloy using the dealloying technique and produce a patterned protective passivated surface rich in Ti and Zr oxides. Nitric acid (HNO3) has been found to be effective in depleting copper from the sample surface. Optimization of treatment parameters such as temperature (70 °C and room temperature) yielded drastic differences in the morphology of the samples studied using Field-Emission Scanning Electron Microscopy. The treated sample surface demonstrated good hemocompatibility and cytocompatibility with primary human osteoblast cells (HOb) and human osteosarcoma cell line (Saos-2). Additionally, the treated samples showed higher ability to produce reactive oxygen species with respect to pristine samples, which could be convenient for preserving the implant from bacterial contamination. These findings contribute to the advancement of producing copper-depleted nanostructured Ti based amorphous alloys for biomedical implant applications.

Abstract 3912: ATM and PARP combined inhibition demonstrate synergistic antitumor efficacy in osteosarcoma models

Abstract Signatures of BRCAness are found in many osteosarcoma (OS) tumors, driving an interest in PARP inhibition (PARPi) for OS treatment. Although PARPi have shown limited efficacy as OS monotherapy, rational combination strategies have not yet been explored for this disease. We performed a genome-scale loss-of-function CRISPR-Cas9 screen in two OS cell lines in the presence and absence of olaparib, a small molecule PARPi. ATM knockout (KO) scored highly as a potent synergizer of PARPi across both cell lines. Here we report this screen finding as well as in vitro validation of PARPi and ATM inhibition (ATMi) synergy in OS models. OS cell lines SAOS2 and CAL72 were infected with the Avana genome-scale lentiviral CRISPR library, targeting 20,000 genes with 4 uniquely barcoded guides (sgRNAs) per gene. After infection and antibiotic selection, cells were grown in an IC40 dose of olaparib or matched control for 18 days. Following DNA sequencing of barcodes and rigorous quality control, sgRNA dropout hits were calculated against the control arm. ATM KO sensitizing to PARPi was first validated utilizing a CRISPR-based genetic approach. ATM KO in CAL72 and SAOS2 was performed via lentiviral infection. After KO confirmation by western immunoblotting (WB), ATM KO and control KO cells were treated with PARPi in a dose range of 19.5 nM-10 µM. ATP-based and Incucyte assays were used to quantify differential cell viability. Secondly, the combination of a small molecule ATMi and a PARPi was studied in a panel of OS cell lines. Synergistic cytotoxicity was quantified using the Zero Interaction Potency (ZIP) model. DNA damage levels after PARPi and ATMi (by chemical inhibition and genetic KO) were assessed by WB for γH2AX, a marker of DNA double-stranded breaks (DSBs). In our genome-scale CRISPR screens, ATM arose as a top druggable hit across both lines. Reassuringly, PARP1 KO scored as a top resistance mediator to PARPi, confirming biologic relevance of the screen and supporting the known PARP-trapping mechanism of these drugs in OS. At low-throughput, ATM KO lines trended toward increased sensitivity to PARPi in short-term viability assays. Combined chemical inhibition yielded highly synergistic ZIP scores across OS cell lines; synergy was achieved with both drugs in the low nanomolar range. γH2AX levels were substantially increased in the setting of combined PARPi and ATMi (via chemical inhibition or genetic KO) as compared to inactivation of either PARP or ATM alone, indicating increased presence of DNA DSBs. In conclusion, these data demonstrate synergy of ATM and PARP inhibition, providing promise for their potential combined use in the treatment of OS. Accumulation of DNA DSBs in response to PARP and ATM inactivation suggests a shared role in DNA damage repair as the molecular basis of synergism. Ongoing studies are investigating anti-tumor potential in in vivo OS models and exploring the mechanistic impact of the combination in OS tumors. Citation Format: Sona N. Kocinsky, Janeala J. Morsby, William C. Wright, Monika Wierdl, Caroline S. Wechsler, Gabriela Alexe, Paul Geeleher, Kimberly Stegmaier, Lillian M. Guenther. ATM and PARP combined inhibition demonstrate synergistic antitumor efficacy in osteosarcoma models [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 3912.

Innovative hydroxyapatite/submicron mesoporous SiO2/HA particles composite coatings for enhanced osteogenic activity of NiTi bone implants: A comprehensive investigation of materials and biological interactions

In this study, hydroxyapatite (HA) incorporated in mesoporous SiO2 particles (SiO2/HA) was initially synthesized using a soft template method. The resulting structure owned a high surface area (796.62 m2 g-1) and a substantial pore volume (0.757 cm3 g-1). The TEM and FESEM images indicated that the synthesized mesoporous particles possessed a spherical morphology, with a diameter of approximately 200 nm, and pores of slit-shape. The mission was to improve the surface characteristics of NiTi implants by composite coatings. This investigation emphasized specifically the impact of different concentrations of mesoporous SiO2/HA particles within the electrolyte on the properties of HA/mesoporous composite coatings. The surface characteristics of the coatings were examined using ATR-FTIR, FESEM, EDS, AFM, and hydrophilicity measurements. Furthermore, the osteogenic activity of the coatings was evaluated by culturing the osteosarcoma cell line SAOS-2 on the coated samples. The introduction of mesoporous particles resulted in a decrease in the coatings’ surface roughness along with an improvement in their wettability. Moreover, there was an enhancement in the cell viability (99% after 7 days), proliferation, adhesion, and growth of SAOS-2 cells at the optimized concentration of mesoporous SiO2/HA particles (1000 mg/L). DAPI images illustrated that the mesoporous particles did not cause any adverse impact on the cellular nuclei. Finally, in the apoptosis test, the composite coating displayed a reduced number of necrotic cells compared to the pure HA coating.

Analysis of Osteosarcoma Cell Lines and Patient Tissue Using a 3D In Vivo Tumor Model—Possible Effects of Punicalagin

Osteosarcomas are the most common primary malignant bone tumors and mostly affect children, adolescents, and young adults. Despite current treatment options such as surgery and polychemotherapy, the survival of patients with metastatic disease remains poor. In recent studies, punicalagin has reduced the cell viability, angiogenesis, and invasion in cell culture trials. The aim of this study was to examine the effects of punicalagin on osteosarcomas in a 3D in vivo tumor model. Human osteosarcoma biopsies and SaOs-2 and MG-63 cells, were grown in a 3D in vivo chorioallantoic membrane (CAM) model. After a cultivation period of up to 72 h, the tumors received daily treatment with punicalagin for 4 days. Weight measurements of the CAM tumors were performed, and laser speckle contrast imaging (LSCI) and a deep learning-based image analysis software (CAM Assay Application v.3.1.0) were used to measure angiogenesis. HE, Ki-67, and Caspase-3 staining was performed after explantation. The osteosarcoma cell lines SaOs-2 and MG-63 and osteosarcoma patient tissue displayed satisfactory growth patterns on the CAM. Treatment with punicalagin decreased tumor weight, proliferation, and tumor-induced angiogenesis, and the tumor tissue showed pro-apoptotic characteristics. These results provide a robust foundation for the implementation of further studies and show that punicalagin offers a promising supplementary treatment option for osteosarcoma patients. The 3D in vivo tumor model represents a beneficial model for the testing of anti-cancer therapies.

BRD4-targeting PROTACs Synergize With Chemotherapeutics Against Osteosarcoma Cell Lines.

Osteosarcoma at an advanced stage has a poor outcome, and novel targeted therapies are needed, especially for metastatic disease. Bromodomain inhibitors (BETi) are epigenetic modulators that broadly impair the expression of oncogenic proteins and exert antitumor effects. BETi can be combined with chemotherapeutics to increase therapeutic responses with superior effects in the form of proteolysis targeting chimeras (PROTACs) that degrade proteins of interest (POI) in multiple cycles. This work aimed to investigate the efficacy of BETi, such as JQ1, dBET57, and MZ1 PROTACs in combination with cytotoxic drugs against osteosarcoma cell lines. Chemosensitivity of the osteosarcoma cell lines HOS, Saos-2, MG-63, and G292 were tested with BET-directed agents alone or in combination with cytotoxic drugs comprising cisplatin, doxorubicin, topotecan, and gemcitabine using cell viability assays. The BET degraders exhibited highest toxicity to HOS cells and showed synergistic activity in combination with the chemotherapeutics, except for the degrader - topotecan/gemcitabine combinations. Highest synergy between BET agents and chemotherapeutics were found for the more chemoresistant Saos-2 cells and potentiation of toxicity in MG-63 cells for the BET agents - doxorubicin combinations and the MZ1-topotecan pair. HOS and Saos-2 cell lines had reduced protein expression of AXL, BCL-X, e-cadherin, CAIX, EpCAM, ErbB2, and vimentin in response to JQ1, MZ1, and BET57. The study suggests that the application of novel BET PROTACs in combination with chemotherapeutics could represent a new therapeutic option to improve the therapy of osteosarcomas. First orally available PROTACs have reached clinical trials.

Effect of Staphylococcal Nuclease and Tudor Domain Containing 1/SLC7A11 on the Occurrence and Development of Osteosarcoma by Inhibiting Ferroptosis

Objective To investigate the effect of staphylococcal nuclease and tudor domain containing 1(SND1) on the biological function of osteosarcoma cells and decipher the mechanism of SND1 in regulating ferroptosis in osteosarcoma cells via SLC7A11. Methods Human osteoblasts hFOB1.19 and osteosarcoma cell lines Saos-2,U2OS,HOS,and 143B were cultured,in which the expression level of SND1 was determined.Small interfering RNA was employed to knock down the expression of SND1(si-SND1) in the osteosarcoma cell line HOS and 143B.The CCK8 assay kit,colony formation assay,and Transwell assay were employed to examine the effect of SND1 expression on the biological function of osteosarcoma cells.Furthermore,we altered the expression of SND1 and SLC7A11 in osteosarcoma cells to investigate the effect of SND1 on osteosarcoma ferroptosis via SLC7A11. Results The mRNA and protein levels of SND1 in Saos-2,U2OS,HOS,and 143B cells were higher than those in hFOB1.19 cells(all P<0.01).Compared with the control group,transfection with si-SND1 down-regulated the expression level of SND1 in HOS and 143B cells(all P<0.01),decreased the viability of HOS and 143B cells,reduced the number of colony formation,and inhibited cell invasion and migration(all P<0.001).The ferroptosis inducer Erastin promoted the apoptosis of HOS and 143B cells,while the ferroptosis inhibitor Ferrostatin-1 improved the viability of HOS and 143B cells(all P<0.001).After SND-1 knockdown,Erastin reduced the viability of HOS and 143B cells,while Ferrostatin-1 restored the cell viability(all P<0.001).After treatment with Erastin in the si-SND1 group,the levels of iron and malondialdehyde were elevated,and the level of glutathione was lowered(all P<0.001).The results of in vivo experiments showed that SND1 knockdown inhibited the mass of the transplanted tumor in 143B tumor-bearing nude mice(P<0.001).Knocking down the expression of SND1 resulted in down-regulated SLC7A11 expression(all P<0.001) and increased ferroptosis in HOS and 143B cells(P<0.001,P=0.020). Conclusions SND1 presents up-regulated expression in osteosarcoma cells.It may inhibit ferroptosis by up-regulating the expression of SLC7A11,thereby improving the viability of osteosarcoma cells.

Three-dimensional (3D) polylactic acid gradient scaffold to study the behavior of osteosarcoma cells under dynamic conditions.

This study adopts an in vitro method to recapitulate the behavior of Saos-2 cells, using a system composed of a perfusion bioreactor and poly-L-lactic acid (PLLA) scaffold fabricated using the low-cost thermally-induced phase separation (TIPS) technique. Four distinct scaffold morphologies with different pore sizes were fabricated, characterized by Scanning electron microscopy and micro-CT analysis and tested with osteosarcoma cells under static and dynamic environments to identify the best morphology for cellular growth. In order to accomplish this purpose, cell growth and matrix deposition of the Saos-2 osteosarcoma cell line were assessed using Picogreen and OsteoImage assays. The obtained data allowed us to identify the morphology that better promotes Saos-2 cellular activity in static and dynamic conditions. These findings provided valuable insights into scaffold design and fabrication strategies, emphasizing the importance of the dynamic culture to recreate an appropriate 3D osteosarcoma model. Remarkably, the gradient scaffold exhibits promise for osteosarcoma applications, offering the potential for targeted tissue engineering approaches.

Genistein Induces Antiproliferative Activity and Apoptosis in Human Osteosarcoma Saos-2 Cells.

Genistein (4', 5, 7-trihydroxyisoflavone) and daidzein (4', 7-dihydroxyisoflavone) are isoflavones derived from soybean and have anti-cancer effects in various cells. However, the effects of genistein and daidzein on the human osteosarcoma cell line Saos-2 has not been investigated before. Human osteosarcoma Saos-2 cells were treated with genistein for 24 and 48 hours. Cytotoxicity and apoptosis were measured. Genistein significantly inhibited proliferation of Saos-2 cells stronger than daidzein in a dose-dependent manner (0 to 80 μM). Genistein also significantly suppressed Saos-2 cell viability in a dose-dependent manner (0 to 100 μM). In contrast, daidzein did not affect Saos-2 cell viability. Real-time PCR revealed that genistein caused G1-arrest by increasing the expression of p21 and p27 mRNAs in Saos-2 cells. In addition, genistein induced apoptosis through the up-regulation of effector caspase-3/7 activity in Saos-2 cells. Genistein also enhanced initiator caspase-9 and TNF-α mRNA expression in cells. Genistein may inhibit proliferation through the up-regulation of p21 and p27 and viability by inducing apoptosis in Saos-2 cells.

Combinatorial anticancer effects of multi metal ion and drug substitute with hydroxyapatite coatings on surgical grade 316LSS stainless steel alloys towards biomedical applications

The present study highlights the metal ions and existing drug ampicillin-loaded nano-hydroxyapatite (nHAP) coated on 316 stainless steel alloy (316LSS) using the electrodeposition method. In addition, the sample phase purity and crystallinity of the HAp composite coatings on 316LSS were analyzed by X-ray diffraction analysis (XRD) and Fourier transform infrared (FT-IR) spectroscopy using the KBr pellet method. The coating microstructure/morphology was characterized using a scanning electron microscope, which showed a flower-like morphology with the attachment of energy-dispersive X-ray spectroscopy (EDX) to confirm the major element groups in the synthesized sample. The TGA results showed that weight loss occurred in the temperature range of up to 800 °C for oxidation and reduction reactions. Additionally, the beat-optimized nanomaterial showed promising in vitro bioactivity against disease-causing microorganisms, displaying a significant zone of inhibition. Furthermore, cytotoxicity assessment using the MTT assay demonstrated that the synthesized samples effectively inhibited cell proliferation in human osteosarcoma cell lines (Saos-2). AO/EB double staining performed under a fluorescence microscope indicated changes in the morphology of the apoptotic cell nuclei. In addition, the relationship between ROS and nanomaterial-induced apoptosis in Saos-2 cells was analyzed by flow cytometry. MMP (ΔѰmt) profiling showed that the HAp composite increased the effective mitochondrial membrane damage or depolarization in the human osteosarcoma cell line. Moreover, the TUNEL assay suggested that treated nanomaterial with Saos-2 cells showed enhanced green fluorescence intensity, indicating terminal DNA damage. The obtained results meet the recommended physiological standard for synthesized 316LSS HAp composites to guide bone tissue regeneration.

Boldine Treatment Induces Cytotoxicity in Human Colorectal Carcinoma and Osteosarcoma Cells.

Introduction Cancer continues to be a significant health issue worldwide, with colorectal cancer (CRC) standing out as one of the most prevalent forms of cancer on a global scale. The lifetime risk of developing CRC is about one in 23 (4.3%) for men and one in 25 (4.0%) for women. Moreover, children and adolescents are frequently reported with osteosarcoma with a low five-year survival rate (69% and 67%, respectively). Aim The aim of the study was to analyze the cytotoxic effects of boldine against human CRC (HCT-116) and osteosarcoma cell lines (Saos-2). Materials and methods HCT-116 and Saos-2 cell lines were subjected to different concentrations of boldine treatment (5, 10, 20, 30, 40, and 50 μg/mL) and (10, 20, 40, 60, and 80 µg/mL), respectively, for 24 hours. The cytotoxicity was analyzed by MTT assay, AO/EBstaining, DCFH-DA assay, and scratch assay. Results The MTT assay, microscopic analysis, and staining showed that boldine had dose-dependent cytotoxic effects against HCT-116 and Saos-2 cell lines by inhibiting their proliferation, viability, and migration, and inducing ROS-mediated apoptosis. Conclusion The study concluded that boldine had a concentration-dependent cytotoxic effect on human CRC and osteosarcoma cell lines.

Menthol causes mitochondrial Ca2+-influx, affects structure-function relationship and cools mitochondria

Menthol is a small bioactive compound able to cause several physiological changes and has multiple molecular targets. Therefore, cellular response against menthol is complex, and still poorly understood. In this work, we used a human osteosarcoma cell line (Saos-2) and analysed the effect of menthol, especially in terms of cellular, subcellular and molecular aspects. We demonstrate that menthol causes increased mitochondrial Ca2+ in a complex manner, which is mainly contributed by intracellular sources, including ER. Menthol also changes the Ca2+-load of individual mitochondrial particles in different conditions. Menthol increases ER-mito contact points, causes mitochondrial morphological changes, and increases mitochondrial ATP, cardiolipin, mitochondrial ROS and reduces mitochondrial membrane potential (ΔΨm). Menthol also prevents the mitochondrial quality damaged by sub-lethal and lethal doses of CCCP. In addition, menthol lowers the mitochondrial temperature within cell and also serves as a cooling agent for the isolated mitochondria in a cell free system too. Notably, menthol-induced reduction of mitochondrial temperature is observed in diverse types of cells, including neuronal, immune and cancer cells. As the higher mitochondrial temperature is a hallmark of several inflammatory, metabolic, disease and age-related disorders, we propose that menthol can serve as an active anti-aging compound against all these disorders. These findings may have relevance in case of several pharmacological and clinical applications of menthol. Significance statementMenthol is a plant-derived bioactive compound that is widely used for several physiological, behavioural, addictive, and medicinal purposes. It is a well-established “cooling and analgesic agent”. However, the exact cellular and sub-cellular responses of menthol is poorly understood. In this work, we have characterized the effects of menthol on mitochondrial metabolism. Menthol regulates mitochondrial Ca2+, ATP, superoxides, cardiolipin, membrane-potential, and ER-mito contact sites. Moreover, the cooling agent menthol also cools down mitochondria and protects mitochondrial damage by certain toxins. These findings may promote use of menthol as a useful supplementary agent for anti-aging, anti-cancer, anti-inflammatory purposes where higher mitochondrial temperature is prevalent.

Yeni PDI tabanlı antiproliteratif kromoforlar

The primary objective of the work was to monitor the antiproliferative activity of two newly synthesized and described perylene diimides. The MTT test was used to investigate the antiproliferative effect against PC3 the prostat cancer cell line, Panc1 pancreatic cancer cell line, Saos2 osteosarcoma cell line, Hep3B hepatoma cell line and HUVEC non-cancerous human umbilical vein endothelial cells. To PC3, Panc1, and Saos 2 cells, both compounds were found to be considerably cytotoxic; however, a comparable impact was not seen in Hep3B cells. Compound 2 had the lowest IC50 value of 40,88 µg/ml and significantly inhibited PC3 cell proliferation when compared to other cell lines.