Osteosarcoma Cell Line MG-63 Research Articles

Synthesis and preliminary cytotoxicity evaluation of water soluble pentacyclic triterpenoid phosphonates.

Synthesis, solubility and cytotoxicity evaluation of anionic phosphonates derived from betulin, betulinic acid, oleanolic acid and ursolic acid is reported. Phosphonate moieties were successfully installed at terpenoid C28 by carboxylic acid deprotonation/alkylation sequence using (dimethoxyphosphoryl)methyl trifluoromethanesulfonate as alkylation reagent. Also, betulin-derived and ether-linked bis-phosphonate is obtained and characterized. After demethylation in the presence of TMSI the resulting phosphonic acids were transformed into their disodium salts. All target compounds display excellent water solubility, which was determined by qNMR in D2O. Cytotoxicity tests were performed in different concentrations of each compound (10-50 µM) against human osteosarcoma cell line MG-63 and osteoblast precursor cell line MC3T3-E1. Improved aqueous solubility and low cytotoxicity profile of the newly designed triterpenoid phosphonates reveal high potential for various medicinal chemistry and pharmacological applications in the future.

Curcumin and Methotrexate: A Promising Combination for Osteosarcoma Treatment via Hedgehog Pathway Inhibition

Osteosarcoma (OS) is the most severe bone tumor in children. A chemotherapy regimen includes a combination of high-dose Methotrexate (MTX), doxorubicin, and cisplatin. These drugs cause acute and chronic side effects, such as infections, thrombocytopenia, neutropenia, DNA damage, and inflammation. Therefore, to identify new therapeutic strategies, effective and with a safety profile, is necessary. The Hedgehog (Hh) signaling pathway involved in tumorigenesis is active in OS. Hh components Patched receptor 1 (PTCH1), Smoothened (SMO), and glioma-associated oncogene homolog transcription factors (GLI1 and GLI2) are overexpressed in OS cell lines and patient samples. Curcumin (CUR)—with antioxidant and anti-cancer properties—downregulates Hh components in cancer, inhibiting progression. This study investigates CUR effects on the MG-63 OS cell line, alone and combined with MTX, to propose a novel therapeutic approach. Our study suggests CUR as a novel therapeutic agent in OS, particularly when combined with MTX. Targeting the Hh signaling pathway, CUR and MTX showed significant pro-apoptotic effects, increasing the BAX/Bcl-2 ratio and total apoptotic cell percentage. They reduced the expression of Hh pathway components (PTCH1, SMO, GLI1, and GLI2), inhibiting OS cell proliferation, survival, and invasion. CUR and MTX combined determined a β-Catenin decrease and a trend toward reducing NF-kB and matrix metalloproteinases (MMP-2 and MMP-9). Our findings suggest CUR as a support to OS treatment, improving outcomes and reducing the adverse effects of current therapies.

Eggshell derived scaffold of hydroxyapatite-ammonium bicarbonate nano-composite: Bioactivity and cytotoxicity studies

This work investigated the facile synthesis of porous scaffold eggshell derived hydroxylapatite (ESHAp) as a composite with ammonium bicarbonate (AMB) for potential biomaterial in tissue engineering application. The phase purity, composition, size, functional groups and morphology of the apatite were elucidated using high resolution transmission electron microscopy (HTEM), X-Ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FT-IR) and scanning electron microscopy (SEM). The results showed that hydroxylapatite (HAp) nanoparticles have round morphologies with average diameters between 20 nm and 80 nm, FT-IR analysis confirmed significant hydroxylapatite functional groups like carbonate, phosphate, and hydroxyl groups, while XRD analysis revealed a well crystalline monophasic HAp powder. The scaffold samples containing 10, 20, 25 and 30 % of AMB withstood a compressive stress up to 5, 20, 30 and 42 N/mm2 respectively which indicates that the compressive stress increased with the AMB content introduced as the pore forming agent. MTT assay performed using MG63 osteosarcoma cell lines showed that on comparing the sample of ESTHAp which contained 0 % AMB with other samples in the range of 0.01–1 mM, viability of above 85 % MG63 cells was achieved except for ESTHAp with 40 % AMB, which showed some level of toxicity. The cell adhesion studies of sintered ESTHAp porous scaffold with different weight percent of the pore forming agents using inverted microscopic images of MG 63 cells incubated with ESTHAp samples and treated with heat at 1000 °C appeared to be unstable in the media used with particle leaching observed, and no cells observed near to the samples.

Chemical Modification of Nanocrystalline Cellulose for Manufacturing of Osteoconductive Composite Materials.

Cellulose is one of the main renewable polymers whose properties are very attractive in many fields, including biomedical applications. The modification of nanocrystalline cellulose (NCC) opens up the possibility of creating nanomaterials with properties of interest as well as combining them with other biomedical polymers. In this work, we proposed the covalent modification of NCC with amphiphilic polyanions such as modified heparin (Hep) and poly(αL-glutamic acid) (PGlu). The modification of NCC should overcome two drawbacks in the production of composite materials based on poly(ε-caprolactone) (PCL), namely, (1) to improve the distribution of modified NCC in the PCL matrix, and (2) to provide the composite material with osteoconductive properties. The obtained specimens of modified NCC were characterized by Fourier-transform infrared spectroscopy and solid-state 13C nuclear magnetic resonance spectroscopy, dynamic and electrophoretic light scattering, as well as thermogravimetric analysis. The morphology of PCL-based composites containing neat or modified NCC as filler was studied by optical and scanning electron microscopy. The mechanical properties of the obtained composites were examined in tensile tests. The homogeneity of filler distribution as well as the mechanical properties of the composites depended on the method of NCC modification and the amount of attached polyanion. In vitro biological evaluation showed improved adhesion of human fetal mesenchymal stem cells (FetMSCs) and human osteoblast-like cells (MG-63 osteosarcoma cell line) to PCL-based composites filled with NCC bearing Hep or PGlu derivatives compared to pure PCL. Furthermore, these composites demonstrated the osteoconductive properties in the experiment on the osteogenic differentiation of FetMSCs.

Poly(styrene-co-methyl methacrylate)-silver/reduced graphene oxide-nano hydroxyapatite nanocomposites for bone cement applications

Herein, we report (polystyrene-co-polymethylmethacrylate/silver/reduced graphene oxide)-(nano-hydroxyapatite) ((PS-PMMA/Ag/RGO)-(nHA)) nanocomposites as potential bone cement. The polymer nanocomposites were characterized using Fourier Transform Infrared spectroscopy (FT-IR), X-ray diffraction (XRD), nano-indentation, thermal, and electron microscopic methods. The FT-IR and XRD results confirmed the successful formation of the nanocomposites. The elastic modulus (E) was found to be 6.85 GPa, and hardness (H) was 0.181 GPa for the nanocomposite bone cement samples, while the E and H values of commercial counterparts are 7.22 and 0.218 GPa, respectively. The thermogravimetric analysis (TGA) has revealed that the nanocomposite bone cement are stable up to 371 °C. The addition of nHA enhances the biocompatibility of bone cement. Cytotoxicity studies were conducted using human osteosarcoma MG63 cell lines. In-vitro studies showed that the material does not display any toxicity on the MG63 cell line, and increased concentration of nHA did not significantly impact the material’s toxicity, only impacting the curing kinetics of the cement.

LncRNA-PVT1 Inhibits Ferroptosis through Activating STAT3/GPX4 Axis to Promote Osteosarcoma Progression.

Osteosarcoma (OS) is a primary malignant bone tumor in the pediatric and adolescent populations. Long non-coding RNAs (LncRNAs), such as plasma-cytoma variant translocation 1 (PVT1), have emerged as significant regulators of OS metastasis. Recent studies have indicated that activation of signal transducer and activator of transcription 3 (STAT3) signaling, which might be controlled by PVT1, inhibits ferroptosis to promote the malignant progression of cancer. Therefore, the present study aimed to determine the role of PVT1 in OS pathogenesis and investigate whether PVT1 affects OS progression by regulating STAT3/GPX4 pathway-mediated ferroptosis. The human OS cell line MG63 were transfected with sh-PVT1 plasmid to inhibit PVT1 expression, with or without co-transfection with a STAT3 overexpression plasmid. The expression of PVT1 was determined by real-time quantitative polymerase chain reaction (RT-qPCR). The proliferation, migration, invasion, and apoptosis of MG63 cells were determined using the cell counting kit-8 (CCK8), Transwell assay, and flow cytometry. The levels of malondialdehyde (MDA), Fe2+, and glutathione (GSH) were determined by ELISA kits, whereas reactive oxygen species (ROS) level was determined by immunofluorescence. The protein expression levels of STAT3, p-STAT3, and glutathione peroxidase 4 (GPX4) were detected by western blot (WB). PVT1 expression was significantly increased in MG63 cells. When knocking down PVT1 with sh-PVT1 plasmid, the proliferation, migration, and invasion of MG63 cells were markedly inhibited, while the rate of apoptosis was upregulated. Further investigation revealed that MG63 cells with PVT1 knockdown exhibited elevated levels of MDA, Fe2+, and ROS. In addition, the inhibition of PVT1 expression resulted in decreased levels of GSH and inhibited expression of p-STAT3 and GPX4. When sh-PVT1 was co-transfected with STAT3 overexpression plasmid in MG63 cells, the increased levels of MDA, Fe2+, and ROS were downregulated, and the decreased expressions of GSH, p-STAT3, and GPX4 were upregulated. PVT1 promotes OS metastasis by activating the STAT3/GPX4 pathway to inhibit ferroptosis. Targeting PVT1 might be a novel therapeutic strategy for OS treatment.

Chitosan-copper microparticles as doxorubicin microcarriers for bone tumor therapy

Doxorubicin (DOX) is a chemotherapeutic drug used in osteosarcoma treatments, usually administrated in very high dosages. This study proposes novel DOX microcarriers based on chitosan (CHT) physically crosslinked with copper(II) ions that will act synergically to inhibit tumor growth at lower drug dosage without affecting the healthy cells. Spherical CHT-Cu microparticles with a smooth surface and an average size of 30.1 ± 9.1 µm were obtained by emulsion. The release of Cu2+ ions from the CHT-Cu microparticles showed that 99.4 % of added cupric ions were released in 72 h of incubation in a complete cell culture medium (CCM). DOX entrapment in microparticles was conducted in a phosphate buffer solution (pH 6), utilizing the pH sensitivity of the polymer. The successful drug-loading process was confirmed by DOX emitting red fluorescence from drug-loaded microcarriers (DOX@CHT-Cu). The drug release in CCM showed an initial burst release, followed by sustained release. Biological assays indicated mild toxicity of CHT-Cu microparticles on the MG-63 osteosarcoma cell line, without affecting the viability of human mesenchymal stem cells (hMSCs). The DOX@CHT-Cu microparticles at concentration of 0.5 mg mL‒1 showed selective toxicity toward MG-63 cells.

Anti-proliferative, antimicrobial, DFT calculations, and molecular docking 3D scaffold based on sodium alginate, chitosan, neomycin sulfate and hydroxyapatite

3D multifunctional scaffold has been designed based on Cs/SA/NS/NPHA. Nanoparticles hydroxyapatite (NPHA) was prepared via precipitation method of sodium dihydrogen phosphate in presence calcium chloride. Different ratios of Chitosan (CS)/Sodium Alginate (SA) were used to prepare Cs/SA scaffolds in presence of CaCl2 as a cross linker. NPHA was incorporated in CS/SA scaffold and neomycin sulfate (NS) was added as an antimicrobial agent. The structure and surface morphology of the scaffolds were investigated via infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscope (SEM) and thermal gravimetric analysis (TGA) techniques. Additionally, Antimicrobial activity of the scaffold has evaluated against Gram- negative and Gram- positive bacteria. The result showed promising antimicrobial activity against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans. Furthermore, cytotoxicity against MG63 osteosarcoma cell and fibroblast normal cell line has investigated. The result showed anti-proliferative against MG63. DFT calculations and molecular docking were used to study the reactivity of the compounds. The results exhibited that Cs/SA/NS/NPHA is potent expected to be used in bone tissue regeneration.

A comprehensive study on bioactivity, mechanical and tribological behavior of copper-doped borosilicate glass derived from natural waste for dental applications

The present study investigates in-vitro biomineralization, mechanical, and tribological properties of bioactive borosilicate glass composed of 31B2O3–20SiO2-(24.5-x)Na2O-24.5CaO and xCuO (mol%). Glass was manufactured using conventional melt-quench technique, utilizing chemicals partially derived from bio-waste, with varying copper oxide (CuO) content (x = 0, 1, 3, and 5). In-vitro study was conducted by immersing the specimens in simulated body fluid for 21 days. The formation of apatite layer was confirmed using FTIR, XRD and FESEM-EDS. Vicker's micro hardness tester and a universal testing machine measured hardness and compressive strength, while a pin-on-disc tribometer assessed wear. MTT assay was performed for all the specimens using osteosarcoma MG63 cell line. The study revealed that substituting copper in place of sodium increased the ion dissolution and improved the formation of the apatite layer hence, enhanced the bioactivity, however, accompanied by a gradual decrease in hardness and compressive strength. BSG-0 exhibited the highest hardness (6.64 GPa) and compressive strength (86.81 MPa), while wear resistance weakened with higher CuO content, notably in BSG-5 with the highest specific wear rate (0.072 mm³/N-m) under a 5 N load. Although MTT assay showed non-toxicity for all specimens, except BSG-5, the compounds demonstrated cell tolerance and growth stimulation. In conclusion, the study suggests that CuO-doped borosilicate glasses exhibit superior bioactivity, making them potentially effective for bone filling, wound healing, and dental applications, despite a compromise in mechanical strength and wear resistance.

Inhibition of proliferation of human osteosarcoma cell line (MG63) by ajwain (Trachyspermum ammi L.) plant extract

Trachyspermum ammi L. (TA), popularly called ajwain, is a well-known spice plant, with fruit pods that are used in Indian traditional medicine. Fruits of this plant were scientifically investigated for medicinal properties, but no studies are reported on the vegetative parts of the plant. Hence, the present investigation is aimed to evaluate the aerial parts of the plant for anti-proliferative and antioxidant effects. Methanolic extract (Me) of TA was subjected to sequential fractionation using the solvents in the increasing order of polarity viz. hexane (He), dichloromethane (Dcm), ethyl acetate (Ea), n-butanol (n-Bu), and water (Aq). Phytochemical analyses, DPPH radical scavenging assay, and in vitro antiproliferative activity on the MG63 osteosarcoma cell line (MTT assay) were performed. Preliminary phytochemical analyses revealed the presence of reducing and non-reducing sugars, amino acids, tannins, phenols, terpenoids, and saponins. Ethyl acetate fraction showed the highest amount of total phenolics (367.90±0.02), followed by n-Bu (287.50±0.04), Dcm (193.56±0.07), Me extract (190.14±0.07), Aq (142.20±0.02) and He (80.74±0.01) fractions. Ethyl acetate fraction exhibited higher DPPH radical scavenging activity (IC50 =116.74 μg/mL) followed by n-Bu (IC50=133.81 μg/mL) fraction with a correlation to the higher amount of phenolics and flavonoids. However, maximum antiproliferative activity was exhibited by hexane fraction on MG 63 human osteosarcoma cell line which can be attributed to the presence of thymol, a monoterpenoid phenol present in ajwain. The current study indicates that the aerial parts of T. ammi Linn. can be considered as potential antioxidants and therapeutic against osteosarcoma.

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.

Bioactive glass doped with zinc-based metal–organic frameworks (Zn/MOF) nanoparticles for antibiotic delivery applications

Zn-MOF/bioactive glass nanoparticles were successfully prepared using quick–alkali-modified sol–gel and tested for gentamicin drug delivery. Samples of 0 and 0.8 mol% Zn MOF contents (BG and BG/Zn MOF, respectively) were characterized using XRD, FTIR, TEM, and SEM/EDX. In addition, zeta characteristics and surface area parameters were also evaluated. Moreover, the cell viability of the prepared samples was tested against the osteosarcoma MG-63 cell line to evaluate the ability of prepared samples for bone regeneration. In addition, the antimicrobial activity of glass samples with and without antibiotics was determined against different microorganisms. Finally, the samples were loaded with gentamicin drug, and drug release profile and kinetics were studied too. XRD and FTIR results confirmed the physicochemical properties of the prepared samples. TEM results showed that particles were in the nano-sized range in both samples; however, the presence of Zn MOF increased the dispersity of particles. Both samples were bioactive, as indicated by hydroxyapatite formation on their surfaces. Cytotoxicity estimation of prepared materials exhibited their safe and nontoxic nature. BG/Zn MOF sample demonstrated prolonged drug release and excellent antimicrobial activity in comparison to BG alone. Therefore, the present study provides promising preliminary results regarding the usage of BG/Zn MOF nanoparticles loaded with gentamicin as an effective approach for bone regeneration.Graphical

Osteogenic assessment of leukocyte platelet-rich fibrin and injectable platelet-rich fibrin in the human osteosarcoma MG - 63 cell line in chronic periodontitis patients: An in vitro study.

The burgeoning interest in implant and regenerative dental care has led to a notable upsurge in the utilization of regenerative modalities. The intent of the present investigation was to evaluate the osteogenic ability of two different concentrated platelet groups at various concentrations in the human osteosarcoma MG-63 cell line. Blood samples from 21 volunteers with chronic periodontitis were collected which were then centrifuged in accordance with the protocols of Choukroun et al. and Miron et al. to produce leukocyte- and platelet-rich fibrin (L-PRF) and injectable platelet-rich fibrin (I-PRF), respectively. Following MG-63 cell culture, the osteogenic ability of 0, 4%, and 20% concentrations of both L-PRF and I-PRF were determined using the real-time polymerase chain reaction assay. The results showed that 20% I-PRF (1.52 ± 0.24) and 4% L-PRF (1.42 ± 0.37) had the highest amount of bone morphogenetic protein 2 and osteocalcin, respectively. I-PRF appears to promote the initial differentiation of cells.

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.

Doxorubicin loaded octacalcium phosphate particles as controlled release drug delivery systems: Physico-chemical characterization, in vitro drug release and evaluation of cell death pathway

Mastering new and efficient ways to obtain successful drug delivery systems (DDS) with controlled release became a paramount quest in the scientific community. Increase of malignant bone tumors and the necessity to optimize an approach of localized drug delivery require research to be even more intensified. Octacalcium phosphate (OCP), with a number of advantages over current counterparts is extensively used in bone engineering. The aim of the present research was to synthesize bioactive and biocompatible doxorubicin (DOX) containing OCP particles. DOX-OCP was successfully obtained in situ in an exhaustive range of added drug (1–20 wt%, theoretical loading). Based on XRD, above 10 wt% of DOX, OCP formation was inhibited and the obtained product was low crystalline α-TCP. In-vitro drug release was performed in pH 7.4 and 6.0. In both pH environments DOX had a continuous release over six weeks. However, the initial drug burst for pH 7.4, in the first 24 h, ranged from 15.9 ± 1.3 % to 33.5 ± 12 % and for pH 6.0 23.7 ± 1.5 % to 36.2 ± 12 %.The DOX-OCP exhibited an inhibitory effect on viability of osteosarcoma cell lines MG63, U2OS and HOS. In contrast, MC3T3-E1 cells (IC50 > 0.062 µM) displayed increased viability and proliferation from 3rd to 7th day. Testing of the DDS on ferroptotic markers (CHAC1, ACSL4 and PTGS2) showed that OCP-DOX does not induce ferroptotic cell death. Moreover, the evaluation of protein levels of cleaved PARP, by western blotting analysis, corroborated that apoptosis is the main pathway of programmed cell death in osteosarcoma cells induced by DOX-OCP.

Anticancer Property of Digera muricata Leaf Extract: An In Vitro Study.

Aim The aim was to evaluate the anticancer potential of Digera muricata ethanolicleaf extract on MG-63 osteosarcoma cell lines. Materials and methods The anti-cancer properties of Digera muricata ethanolic leaf extract were evaluated on osteosarcoma cell lines using 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the morphological changes in MG-63 cells were assessed after 24 hours using microscopic observation. Additionally, fluorescence microscopy was employed to evaluate the apoptotic changes after acridine orange/ethidium bromide (AO/EtBr) dual staining. Results The MTT assay revealed a dose-dependent cell death. The cell viability decreased with increase in concentrations of the extract, The cell viability was 89.98 ± 4.89 percentage at 25 μg/ml and 15.64 ± 3.64 percentage at 200 μg/ml concentrations. A concentartion of 116.95 μg/ml showed 50% inhibition (IC50). The morphological and dual staining studies also showed the extract's effectiveness in inducing apoptosis. Conclusion The ethanolic leaf extract of D. muricatacould impart good antiproliferative activity in MG-63 cell lines. The extract could also induce apoptosis and hence, it may be considered as a potential anticancer agent for the development of drug formulation for the treatment of osteosarcoma.

Biomineralized fluorocanasite-reinforced biocomposite scaffolds demonstrate expedited osteointegration of critical-sized bone defects.

The development of patient-specific bone scaffolds that can expedite bone regeneration has been gaining increased attention, especially for critical-sized bone defects or fractures. Precise adaptation of the scaffold to the region of implantation and reduced surgery times are also crucial at clinical scales. To this end, bioactive fluorcanasite glass-ceramic microparticulates were incorporated within a biocompatible photocurable resin matrix following which the biocomposite resin precursor was 3D-printed with digital light processing method to develop the bone scaffold. The printing parameters were optimized based on spot curing investigation, particle size data, and UV-visible spectrophotometry. In vitro cell culture with MG-63 osteosarcoma cell lines and pH study within simulated body fluid demonstrated a noncytotoxic response of the scaffold samples. Further, the in vivo bone regeneration ability of the 3D-printed biocomposite bone scaffolds was investigated by implantation of the scaffold samples in the rabbit femur bone defect model. Enhanced angiogenesis, osteoblastic, and osteoclastic activities were observed at the bone-scaffold interface, while examining through fluorochrome labelling, histology, radiography, field emission scanning electron microscopy, and x-ray microcomputed tomography. Overall, the results demonstrated that the 3D-printed biocomposite bone scaffolds have promising potential for bone loss rehabilitation.

Anti-cancer targets and molecular mechanisms of formononetin in treating osteosarcoma based on network pharmacology.

Osteosarcoma (OS) is a multifactorial bone malignancy that accounts for most cancers in children and adolescents. Formononetin has been proven to exhibit various pharmacological effects including anti-tumor, anti-obesity, anti-inflammation, and neuroprotective effects. Few studies have examined the pharmacological activities of formononetin in OS treatment, but the mechanism has not yet been completely elucidated. Network pharmacology is a new method based on the theory of system biology for analyzing the network of biological systems and selecting specific signal nodes for multi-target drug molecular design. Here, we used network pharmacology to explore the possible mechanism of formononetin in OS treatment. Human OS cell line MG63 was processed with four concentrations (0, 2, 5, 8 μg/mL) of formononetin. Subsequently, an MTT assay was performed to test cell proliferation and a scratch test was used to evaluate the migration ability of cancer cells. Caspase-3, p53, p21, and bcl-2 expression levels incubated with different concentrations of formononetin in MG63 cells were determined using Western blotting. After treated with formononetin for 48 h, MG63 cells exhibited marked apoptosis. The results revealed that certain concentrations of formononetin significantly exerted inhibitory effects on MG63 cell proliferation. Furthermore, formononetin decreased the bcl-2 level in MG63 cells but increased caspase-3, p21, and p53 levels in a concentration-dependent manner. Additionally, formononetin suppressed the expression of SATB2. Therefore, formononetin could dose-dependently inhibit MG63 cell proliferation and induce apparent cell apoptosis, providing a candidate treatment for OS, whereas SATB2 could be a potential prognostic biomarker for screening OS and therapeutic target of formononetin.

Extrusion-based 3D printing of osteoinductive scaffolds with a spongiosa-inspired structure.

Critical-sized bone defects resulting from trauma, inflammation, and tumor resections are individual in their size and shape. Implants for the treatment of such defects have to consider biomechanical and biomedical factors, as well as the individual conditions within the implantation site. In this context, 3D printing technologies offer new possibilities to design and produce patient-specific implants reflecting the outer shape and internal structure of the replaced bone tissue. The selection or modification of materials used in 3D printing enables the adaption of the implant, by enhancing the osteoinductive or biomechanical properties. In this study, scaffolds with bone spongiosa-inspired structure for extrusion-based 3D printing were generated. The computer aided design process resulted in an up scaled and simplified version of the bone spongiosa. To enhance the osteoinductive properties of the 3D printed construct, polycaprolactone (PCL) was combined with 20% (wt) calcium phosphate nano powder (CaP). The implants were designed in form of a ring structure and revealed an irregular and interconnected porous structure with a calculated porosity of 35.2% and a compression strength within the range of the natural cancellous bone. The implants were assessed in terms of biocompatibility and osteoinductivity using the osteosarcoma cell line MG63 and patient-derived mesenchymal stem cells in selected experiments. Cell growth and differentiation over 14days were monitored using confocal laser scanning microscopy, scanning electron microscopy, deoxyribonucleic acid (DNA) quantification, gene expression analysis, and quantitative assessment of calcification. MG63 cells and human mesenchymal stem cells (hMSC) adhered to the printed implants and revealed a typical elongated morphology as indicated by microscopy. Using DNA quantification, no differences for PCL or PCL-CaP in the initial adhesion of MG63 cells were observed, while the PCL-based scaffolds favored cell proliferation in the early phases of culture up to 7days. In contrast, on PCL-CaP, cell proliferation for MG63 cells was not evident, while data from PCR and the levels of calcification, or alkaline phosphatase activity, indicated osteogenic differentiation within the PCL-CaP constructs over time. For hMSC, the highest levels in the total calcium content were observed for the PCL-CaP constructs, thus underlining the osteoinductive properties.

Thermal necrosis in orthopedic bone tumors: experimental research.

The extent of surgical resection in orthopedic oncology differs according to tumor biology. While malignant bone tumors are operatively managed with wide resection, benign bone tumors and metastatic carcinomas are often treated through intralesional excision and adjuvant modalities, including the elimination of residual neoplastic cells through thermal necrosis. This study investigates in vitro temperature thresholds for thermal necrosis in common orthopedic bone tumors. Eleven cell lines, including metastatic carcinomas to bone (A549, A498, FU-UR-1, PC3, MDA-MB-231, TT, MCF7, and K1), giant cell tumor of bone, osteosarcoma (HG-63), and control non-neoplastic cells (HEK293) were cultured. Cells were exposed to thermal stress at varying times and temperatures and evaluated for survival and viability with crystal violet and MTT assays. Both the MTT and crystal violet assay demonstrated statistically superior rates of viability and survival for A549 (lung carcinoma), FU-UR-1 (renal carcinoma), K1 (thyroid carcinoma), and MG-63 (osteosarcoma) cell lines compared to control (HEK293 cells) at 60°C. Additionally, the MTT assay demonstrated superior viability for PC3 (prostate carcinoma), MCF7 (breast carcinoma), and A498 (renal carcinoma) compared to control. All cell lines demonstrated significantly decreased survival and viability in temperatures more than 90°C. This study demonstrated in vitro thresholds for thermal necrosis for cell lines of common orthopedic tumors of bone. The A549 (lung carcinoma), K1 (thyroid carcinoma), and FU-UR-1 (renal carcinoma) cell lines demonstrated greater resistance to heat stress compared to non-neoplastic control cells. Temperatures in excess of 90°C are necessary to reliably reduce cell survival and viability to less than 10%.