Human Osteosarcoma Saos-2 Cells Research Articles

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.

Fish scale-derived hydroxyapatite for alveolar ridge preservation.

Alveolar ridge resorption following tooth extraction poses significant challenges for future dental restorations. This study investigated the efficacy of fish scale-derived hydroxyapatite (FSHA) as a socket preservation graft material to maintain alveolar bone volume and architecture. FSHA was extracted from *Labeo rohita* fish scales and characterized using Fourier transform infrared (FTIR) analysis. In vitro, biocompatibility and osteogenic potential were assessed using Saos-2 human osteosarcoma cells. Cell viability, migration, and proliferation were evaluated using MTT and scratch assays. In vivo performance was assessed in a rat model, and FSHA was compared to a commercial xenograft (Osseograft) and ungrafted controls. Histological analysis was performed at 8-week post-implantation to quantify new bone formation. FTIR confirmed the purity and homogeneity of FSHA. In vitro, FSHA enhanced Saos-2 viability, migration, and proliferation compared to controls. In vivo, FSHA demonstrated superior bone regeneration compared to Osseograft and ungrafted sites, with balanced graft resorption and new bone formation. Histological analysis revealed an active incorporation of FSHA into new bone, with minimal gaps and ongoing remodeling. Approximately 50%-60% of FSHA was resorbed by 8 weeks, closely matching the rate of new bone deposition. FSHA stimulated more bone formation in the apical socket region than in coronal areas. In conclusion, FSHA is a promising biomaterial for alveolar ridge preservation, exhibiting excellent biocompatibility, osteogenic potential, and balanced resorption. Its ability to promote robust bone regeneration highlights its potential as an effective alternative to currently used graft materials in socket preservation procedures.

Exploring the Anti-Cancer Potential of Hispidin: A Comprehensive in Silico and in Vitro Study on Human Osteosarcoma Saos2 Cells.

Hispidin was initially discovered in basidiomycete Inonotus hispidus (Bull.) P. Karst and this extraordinary compound possesses immense potency and can be extracted from the wild mushroom through specialized bioreactor cultivation techniques. In our study, we isolated it from Inonotus hispidus (Bull.) P. Karst., with a yield of 3.6 %. We identified and characterized hispidin through the implementation of spectroscopic techniques such as FTIR, NMR, and MS. Additionally, we utilized Thermogravimetric Analysis for thermal characterization of the compound. Computational studies based on DFT were performed to investigate the molecular structure, electronic properties, and chemical reactivity of hispidin. PASS analysis for hispidin demonstrated that 19 of them are anti-neoplastic activities. The Pharmacology prediction of hispidin confirm that it is not toxic, non-carcinogenesis with a good human intestinal absorption. The effect of hispidin on the viability of bone cancer cells was evaluated by MTT assay. The results showed that hispidin significantly reduced SaoS2 cell viability in a dose-dependent manner. Molecular docking was carried out using five targets related to bone cancer to determine the interactions between hispidin and the studied proteins. The results demonstrate that hispidin is a good inhibitor for the five targets. Dynamic simulation shows a good stability of the complex hispidin-protein.

Preparation of trivalent rare-earth element-substituted bioactive glass robocast scaffolds for osteosarcoma treatment: structural, morphological, mechanical, drug delivery, and biological properties

In this study, trivalent rare-earth ion (Eu3+, Gd3+, and Yb3+)-substituted silicate-based bioactive glass scaffolds were prepared by robocasting method using sol–gel-derived bioactive glass powders for tissue engineering applications and cancer therapy. The structural, morphological, and mechanical properties of the prepared scaffolds as well as their in vitro bioactivity in simulated body fluid (SBF) were investigated in detail. In addition, an anticancer drug (5-FU) adsorption and release behavior of the scaffolds was studied as a function of time. In vitro, cytotoxicity and alkaline phosphatase activity were investigated using human skin fibroblast BJ and osteosarcoma SaOS-2 cells. Results showed that using lanthanide ion-containing (0.5, 1, 3, and 5 wt%) sol–gel-derived bioactive glass powders it was possible to successfully fabricate periodic, mesh-like patterned robocast glass scaffolds. All of the scaffolds prepared in the study sintered at 675 °C showed an amorphous structure. The compressive strength of scaffolds was in the range of 8.8 MPa to 13.6 MPa and the highest strength values were obtained in the Yb3+-containing scaffolds. Hydroxyapatite formation was obtained for the scaffolds immersed in SBF for 28 days. The fluorouracil adsorption amount was calculated to be ~ 25% for all types of scaffolds and the cumulative drug release was in the range of 20–25% depending on the dopant concentration. Results of the in vitro cell culture experiments revealed that all of the scaffolds fabricated in the study were not cytotoxic to fibroblast and osteosarcoma cells for up to 7 days under in vitro conditions. An increase was obtained for the ALP activities for both types of cells as the incubation time was increased.

Combination Treatment with Liposomal Doxorubicin and Inductive Moderate Hyperthermia for Sarcoma Saos-2 Cells.

Despite efforts in osteosarcoma (OS) research, the role of inductive moderate hyperthermia (IMH) in delivering and enhancing the antitumor effect of liposomal doxorubicin formulations (LDOX) remains unresolved. This study investigated the effect of a combination treatment with LDOX and IMH on Saos-2 human OS cells. We compared cell viability using a trypan blue assay, apoptosis and reactive oxygen species (ROS) measured by flow cytometry and pro-apoptotic Bax protein expression examined by immunocytochemistry in response to IMH (42 MHz frequency, 15 W power for 30 min), LDOX (0.4 μg/mL), and LDOX plus IMH. The lower IC50 value of LDOX at 72 h indicated increased accumulation of the drug in the OS cells. LDOX plus IMH resulted in a 61% lower cell viability compared to no treatment. Moreover, IMH potentiated the LDOX action on the Saos-2 cells by promoting ROS production at temperatures of <42 °C. There was a 12% increase in cell populations undergoing early apoptosis with a less heterogeneous distribution of Bax after combination treatment compared to those treated with LDOX (p < 0.05). Therefore, we determined that IMH could enhance LDOX delivery and its antitumor effect via altered membrane permeabilization, ROS generation, and a lower level of visualized Bax heterogeneity in the Saos-2 cells, suggesting the potential translation of these findings into in vivo studies.

Bortezomib-encapsulated metal-phenolic nanoparticles for intracellular drug delivery.

Bortezomib (BTZ) is an important boronate proteasome inhibitor that is widely used in cancer therapy. However, the clinical application of BTZ suffers from poor stability and serious adverse effects. Herein, we fabricated metal-polyphenol nanoparticles for the covalent encapsulation of BTZ. BTZ-encapsulated tannic acid (TA)-Fe3+ nanoparticles can be prepared by mixing BTZ, TA, and ferric chloride owing to the formation of metal-polyphenol coordination interaction and dynamic boronate ester bonds. The BTZ-encapsulated TA-Fe3+ nanoparticles (BTZ NPs) are stable in physiological environment (pH 7.4) with minimal drug leakage. However, BTZ NPs can be disassembled in an acidic environment. Therefore, BTZ can be rapidly released from BTZ NPs in an acidic environment (pH 5.0). More than 50% BTZ can be released from BTZ NPs after 8 h incubation at pH 5.0. BTZ NPs exhibited high cytotoxicity against human osteosarcoma Saos-2 cells and human multiple myeloma OPM-2 cells. The metal-polyphenol nanoparticles can be a promising nanoplatform for the delivery of BTZ with simultaneously enhanced therapeutic efficacy and reduced side effects.

Bioactivity, Cytotoxicity, and Tribological Studies of Nickel-Free Austenitic Stainless Steel Obtained via Powder Metallurgy Route.

In the present study, the bioactivity, cytotoxicity, and tribological properties of a nickel-free austenitic stainless steel produced via the mechanical alloying of elemental iron, chromium, and manganese nitride powders following by hot isostatic pressing was investigated. Powders after 90 h of mechanical alloying were consolidated via hot isostatic pressing at 1150 °C (1425 K) and heat treated at 1175 °C (1448 K) for 1 h in a vacuum with furnace cooling. Tribological tests were performed to determine the resistance of the as-received nickel-free steel. It was noticed that applying heat treatment after hot isostatic pressing decreases the average friction coefficient and wear rate of the austenitic steel. An immersion test in a simulated body fluid for 28 days at 37 ± 1 °C has been used to determine the biocompatibility of the tested material. The SEM-EDS analysis allowed us to characterise the morphology of the films and the elements of the steel on the thin-film layer. Elements typical of apatite (calcium and phosphorus) were detected on the surface of the sample. Cellular toxicity tests showed no significant toxic side effects for Saos-2 human osteosarcoma cells and the number of Saos-2 human osteosarcoma cells on the nickel-free steel was greater than on the 316LV grade steel.

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.

Anticancer Potential of Farnesol Against Human Osteosarcoma Saos-2 Cells and Human Colorectal Carcinoma HCT-116 Cells.

Increased colorectal carcinoma (CRC) and osteosarcomaprevalence, low survival rate, poor prognosis, and the limitations of existing anticancer therapies like side effects of drugs, non-specificity, short half-life, etc., pose a need for novel anticancer drugs.Farnesol, an organic sesquiterpene compound, found in the essential oils of various plants has been shown to possess antioxidant, anti-inflammatory, and anticancer properties. However, the anticancer effect of farnesol againstCRCand osteosarcoma has not yet been adequately elucidated. The aim of the study was to analyze the anticancer effects of farnesol against human osteosarcoma and CRC cell lines. Human osteosarcoma (Saos-2) and colorectal carcinoma (HCT-116) cell lines were procured and cultured at37oC and 5% CO2. The cells were treated with 10, 20, 40, 60, 80, and 100 µM/ml and 20, 40, 60, 80, 100, and 120 µM/ml of farnesol for 24 hours, respectively.3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromideassay was performed to assess the cytotoxicity of farnesol on Saos-2 and HCT-116 cells. Acridine orange/ethidium bromidestaining was carried out to analyze apoptosis.4',6-diamidino-2-phenylindolestaining was done to observe the nuclear changes.Dichloro-dihydro-fluorescein diacetatestaining was performed to assess the farnesol-induced reactive oxygen species (ROS)-mediated cell death. Farnesol reduced the viability and proliferation of Saos-2 and HCT-116 cells in a dose-dependent manner. Farnesol was able to alter the cellular and nuclear morphology of Saos-2 and HCT-116 cells, promoting cell death. Farnesol-induced apoptosis inhuman osteosarcoma and colorectal carcinoma cell lines. Early apoptosis was observed in farnesol-treated HCT-116 cells. Additionally, ROS-mediated apoptotic cell death was reported in Saos-2 cells. Farnesol has the potential to induce cytotoxicity against human osteosarcoma and CRC cell lines.

In Vitro Characterization of Doped Bioglass 45S5/HAp Coatings Obtained by CoBlastTM Deposition

Bone replacement is one of the major medical procedures in the oral surgery field due to the progressive ageing population and to illness or trauma in younger age groups. The use of implants without biological activity and effective osseointegration increases the chances of implant failure. This work aims to improve the interaction between implants and bone by using Bioglass 45S5 (BG)/hydroxyapatite (HAp) mixtures, including copper-, zinc-, and cerium-doped BG, as well as co-doping by the mentioned metals, as coatings produced by the CoBlastTM technique. All coatings present a uniform coverage of the Ti-6Al-4V substrate. Furthermore, in vitro testing using human osteosarcoma Saos-2 cells indicated that BG/HAp coatings have no cytotoxic effect, and the used of doping agents did not alter cell adhesion, proliferation, or alkaline phosphatase (ALP) expression when compared to undoped coating. These results demonstrate that BG/HAp by CoBlastTM can be a solution to improve implants’ osseointegration.

Molecular Analysis of the Superior Efficacy of a Dual Epidermal Growth Factor Receptor (EGFR)-DNA-Targeting Combi-Molecule in Comparison with Its Putative Prodrugs 6-Mono-Alkylamino- and 6,6-Dialkylaminoquinazoline in a Human Osteosarcoma Xenograft Model.

Background: ZR2002 is a dual EGFR-DNA-targeting combi-molecule that carries a chloroethyl group at the six-position of the quinazoline ring designed to alkylate DNA. Despite its good pharmacokinetics, ZR2002 is metabolized in vivo into dechlorinated metabolites, losing the DNA-alkylating function required to damage DNA. To increase the DNA damage activity in tumor cells in vivo, we compared ZR2002 with two of its 6-N,N-disubstituted analogs: "JS61", with a nitrogen mustard function at the six-position of the quinazoline ring, and "JS84", with an N-methyl group. Methods: Tumor xenografts were performed with the human Saos-2 osteosarcoma cell line expressing EGFR. Mice were treated with ZR2002, JS84 or JS61, and the tumor burden was measured with a caliper and CT/PET imaging. Drug metabolism was analyzed with LC-MS. EGFR and ɣ-H2AX phosphorylation were quantified via Western blot analysis and immunohistochemistry. Results: In vivo analysis showed that significant tumor growth inhibition was only achieved when ZR2002 was administered in its naked form. The metabolic dealkylation of JS61 and JS84 did not release sufficient concentrations of ZR2002 for the intratumoral inhibition of P-EGFR or enhanced levels of P-H2AX. Conclusions: The results in toto suggest that intratumoral concentrations of intact ZR2002 are correlated with the highest inhibition of P-EGFR and induction of DNA damage in vivo. ZR2002 may well represent a good drug candidate for the treatment of EGFR-expressing osteosarcoma.

Influences of surface topography of porous titanium scaffolds manufactured by powder bed fusion on osteogenesis

Powder bed fusion (PBF) has emerged as a useful metal-three-dimensional (3D) printing technology for manufacturing porous structures; it has been widely employed for the production of medical implants. Multiple previous reports have suggested that the bone ingrowth characteristics of porous structures, among the most important factors for medical implants, are affected by 3D printing parameters (e.g., pore shape). The topography of 3D-printed metal surfaces can also influence cellular responses, including osteoblast differentiation. The surface topography may change according to the shape of the printed pores. Nevertheless, it remains unclear how the surface topography changes. In addition, it is still unknown how the pore shape and the resulting surface topography can further impact the bone ingrowth efficiency of the porous structure. Therefore, to better understand the relationship, we prepared Ti–6Al–4V specimens with different pore shapes (i.e., circular, triangular, and rectangular) using the PBF method, then analyzed the surface topographies of these 3D scaffolds based on curvature and roughness profiles. Through subsequent in vitro studies, we investigated how changes in pore shape and surface topography affect the activity of Saos-2 human osteosarcoma cells. The results of this study confirmed that the curvature radii and local surface roughness change according to pore shape because of the characteristics of PBF. Furthermore, differences in local surface topography and pore shape led to differences in cell proliferation and differentiation. Through our findings, we anticipate that the meticulous design of pore shape and sophisticated control of the processing parameters, such as laser speed and paths, in the fabrication of orthopedic and dental implants will result in significantly enhanced bone ingrowth efficiency.

Preparation and In Vitro Characterization of Magnetic CS/PVA/HA/pSPIONs Scaffolds for Magnetic Hyperthermia and Bone Regeneration.

Conventional bone cancer treatment often results in unwanted side effects, critical-sized bone defects, and inefficient cancer-cell targeting. Therefore, new approaches are necessary to better address bone cancer treatment and patient's recovery. One solution may reside in the combination of bone regeneration scaffolds with magnetic hyperthermia. By incorporating pristine superparamagnetic iron oxide nanoparticles (pSPIONs) into additively manufactured scaffolds we created magnetic structures for magnetic hyperthermia and bone regeneration. For this, hydroxyapatite (HA) particles were integrated in a polymeric matrix composed of chitosan (CS) and poly (vinyl alcohol) (PVA). Once optimized, pSPIONs were added to the CS/PVA/HA paste at three different concentrations (1.92, 3.77, and 5.54 wt.%), and subsequently additively manufactured to form a scaffold. Results indicate that scaffolds containing 3.77 and 5.54 wt.% of pSPIONs, attained temperature increases of 6.6 and 7.5 °C in magnetic hyperthermia testing, respectively. In vitro studies using human osteosarcoma Saos-2 cells indicated that pSPIONs incorporation significantly stimulated cell adhesion, proliferation and alkaline phosphatase (ALP) expression when compared to CS/PVA/HA scaffolds. Thus, these results support that CS/PVA/HA/pSPIONs scaffolds with pSPIONs concentrations above or equal to 3.77 wt.% have the potential to be used for magnetic hyperthermia and bone regeneration.

Development of anti-cancer bionanoparticles isolated from corn for bone cancer treatment

Nanoparticles (NPs) are particles ranging from 1 to 100 nm in size. NPs can be derived from various sources such as copper, gold, and even plants such as kiwis and corn. Previous studies have shown that nanoparticles derived from corn (cNPs) displayed anticancer properties, but their effects were not tested on bone cancer samples. Current bone cancer treatments are limited as anticancer drugs often either cannot reach the tumor or have little effect on it. Therefore, in this study, we hypothesize that cNPs may have anti-proliferative effects on bone cancer and metastasized bone cancer. First, we found that SAOS-2 (human osteosarcoma) cell viability decreased to 0% when treated with a 20% cNP concentration, and NCI-H526 (human lung carcinoma metastasized to bone marrow) cell viability decreased to 0% when treated with only a 15% cNP concentration. These results indicate that NCI-H526 is more sensitive to cNPs than SAOS-2. Secondly, we found that cNP treatment reduced the expression of BCL2, a gene that prevents apoptosis, suggesting that the cNP treatment induced BCL2-dependent apoptosis. Overall, these results indicate that cNPs have anti-proliferative effects on bone cancer cells and cancer cells that metastasize to the bone. Therefore, our study suggests that cNPs may be applied to the development of bone cancer treatment.

SIRT2 promotes the viability, invasion and metastasis of osteosarcoma cells by inhibiting the degradation of Snail

Osteosarcomas (OS) are highly metastatic and usually lead to poor outcomes. Epithelial-mesenchymal transition (EMT) is reported to be a critical event in metastasis. SIRT2 exerts dual functions in many different tumors. However, the underlying molecular mechanisms of SIRT2 in osteosarcoma cell metastasis and the question of whether SIRT2 regulates EMT have not been fully explored. In this study, we confirmed that SIRT2 was highly-expressed in human osteosarcoma MG63 and Saos-2 cell lines. The viability, migration and invasion of osteosarcoma cells were inhibited by knockdown of SIRT2 and were enhanced by overexpression of SIRT2. Moreover, SIRT2 positively regulated EMT and upregulated the protein levels of the mesenchymal markers N-cadherin and Vimentin and the levels of MMP2 and MMP9. A xenograft mouse model showed that SIRT2 knockdown in osteosarcoma cells led to reduced tumor growth, decreased expression of mesenchymal markers and impaired lung and liver metastasis in vivo. Furthermore, we showed that SIRT2 interacted with and upregulated the protein level of the EMT-associated transcription factor Snail. SIRT2 inhibited Snail degradation via its deacetylase activity. Knockdown of Snail abrogated the promoting effects of SIRT2 on migration and invasion of osteosarcoma cells. In conclusion, SIRT2 plays a crucial role in osteosarcoma metastasis by inhibiting Snail degradation and may serve as a novel therapeutic target to manage osteosarcoma.

Nox4 promotes osteoblast differentiation through TGF-beta signal pathway.

NADPH oxidase 4 (Nox4) is the main source of reactive oxygen species, which promote osteoclast formation and lead to bone loss, thereby causing osteoporosis. However, the role of Nox4 in osteoblasts during early development remains unclear. We used zebrafish to study the effect of Nox4 deletion on bone mineralization in early development. nox4-/- zebrafish showed decreased bone mineralization during early development and significantly reduced numbers of osteoblasts, osteoclasts, and chondrocytes. Transcriptome sequencing showed that the TGF-β signaling pathway was significantly disrupted in nox4-/- zebrafish. Inhibiting TGF-β signaling rescued the abnormal bone development caused by nox4 deletion and increased the number of osteoblasts. We used Saos-2 human osteosarcoma cells to confirm our results, which clarified the role of Nox4 in human osteoblasts. Our results demonstrate the mechanism of reduced bone mineralization in early development and provide a basis for the clinical treatment of osteoporosis.

Production and bioactivities of nanoparticulated and ultrasonic-degraded levan generated by Erwinia tasmaniensis levansucrase in human osteosarcoma cells

Levan is a bioactive polysaccharide that can be synthesized by various microorganisms. In this study, the physicochemical properties and bioactivity of levan synthesized by recombinant levansucrase from Erwinia tasmaniensis were investigated. The synthesis conditions, including the enzyme concentration, substrate concentration, and temperature, were optimized. The obtained levan generally appeared as a cloudy suspension. However, it could transform into a hydrogel at concentrations exceeding 10 % (w/v). Then, ultrasonication was utilized to reduce the molecular weight and increase the bioavailability of levan. Dynamic light scattering (DLS) and gel permeation chromatography (GPC) indicated that the size of levan was significantly decreased by ultrasonication, whereas Fourier transform infrared spectroscopy, 1H-nuclear magnetic resonance, and X-ray powder diffraction revealed that the chemical structure of levan was not changed. Finally, the bioactivities of both levan forms were examined using human osteosarcoma (Saos-2) cells. The result clearly illustrated that sonicated levan had higher antiproliferative activity in Saos-2 cells than original levan. Sonicated levan also activated Toll-like receptor expression at the mRNA level. These findings suggested the important beneficial applications of sonicated levan for the development of cancer therapies.

Moringa oleifera leaf extract induces osteogenic-like differentiation of human osteosarcoma SaOS2 cells

IntroductionMoringa oleifera is known as a ‘natural nutrition of the tropics’ because it provides vital nutritional supplements and a variety of pharmacological benefits. The focus of this study was to elucidate the dose dependent effects of Moringa oleifera leaf (MOL) extract on the growth of the human osteoblast-like osteosarcoma SaOS-2 cell line and primary osteoblast cells. MethodsTrypan blue & tetrazolium assay, intracellular ROS generation, chromatin condensation, cell cycle analysis, alkaline phosphatase (ALP), mineralization, and osteogenic gene expression were tested on both treated and untreated osteosarcoma SaOS-2 cells. ResultsAs revealed by cell viability assay, growth activity was observed at concentrations 25 and 50 μg/mL of MOL extract, whereas 100 and 200 μg/mL doses decreased the proliferation activity, resulting in ROS production and chromatin condensation. Cell cycle study revealed that MOL extract at 50 and 100 μg/mL concentrations arrested the cells in the G2/M phase. Low doses increased the ALP levels, mineralization, and expression of the bone morphogenetic protein 2 (BMP2) and runt-related transcription factor 2 (Runx2) genes in osteoblast-like SaOS-2 cells, however, high doses inhibited the proliferation properties of MOL extract. Through AutoDock Vina and iGEMDOCK 2.1, the interaction of active components of MOL, such as β-sitosterol, quercetin and kaempferol, with BMP2 and Runx2 proteins revealed a reasonable binding affinity. Moreover, these components did not show any Lipinski's rule of five violation and showed predictable pharmacokinetic properties. ConclusionThe results of the biphasic dose-response of MOL extract on the growth activity of osteoblast-like SaOS-2 cells and in silico binding interface, may provide a therapeutic and/or preventive implication in prospective drug development.

Cyclophosphamide exerts an anti-metastatic effect by reducing the expression of MMP-2 and -9 in Saos2 osteosarcoma cells

Purpose: Although osteosarcoma is the most common primary malignant bone tumor among adolescents and young adults, the molecular mechanisms underlying the development of cancer are not fully elucidated. Cyclophosphamide (CYC) is an alkylating antineoplastic agent widely used in the treatment regimens of various cancers and chronic inflammatory diseases. The aim of this study is to reveal the effect of CYC on Saos-2 cells at the molecular level by determining the expression levels of some matrix metalloprotease proteins (MMP-2 and MMP-9) of cancer progression related genes.&#x0D; Materials and Methods: Cytotoxicity analyzes were performed by applying CYC to the developed human osteosarcoma (Saos-2) cell cultures. The determined dose of CYC was applied to the 2D cell lines for 12, 24 and 48 hours. As a result of agent application, the expression levels of MMPs, which are metastatic markers in these cultures, were determined by immunocytochemical method.&#x0D; Results: While MMP-2 and MMP-9 protein expression levels increased in the control (PBS) groups in correlation with the extend in duration in the cell cultures created; decreased with CYC administration.&#x0D; Conclusion: It was determined that the expression levels of genes that play a role in metastasis such as MMP-2 and MMP-9 were decreased by CYC application to Saos-2 cells. These results shed light on molecular studies for the treatment of osteosarcoma cancer.

Fabrication and characterization of strontium substituted chitosan modify hydroxyapatite for biomedical applications

The present study aimed to preparation of hydroxyapatite using strontium (Sr2+) and chitosan (CS) for the preparation of porous scaffold material like SrHA-CS. These synthesized nano-materials were alsofurther characterized by Fourier transform infrared spectroscopy and X-ray diffraction analysis to confirm the phase purity and crystallinity. The crystal-like morphology of the SrHA-CS was observed using a high-resolution transmission electron microscope (HRTEM) which revealed rod-like morphology with the major elements confirmed by energy-dispersive X-ray spectroscopy (EDX). The size of the synthesized nanomaterial and their stability were analyzed through dynamic light scattering (DLS)and zeta potential. The hemolytic study indicates that SrHA-CS is non-toxic red blood cells of the humans. Besides, SrHA-CS is also active on disease-causing microorganisms and exhibit a strong zone of inhibition. The cytotoxic (MTT) assay systemically investigatedthe synthesized samplesagainst SaOS-2 human osteosarcoma cell lines it resulting in effectively induce cell proliferation without generating any harmful effect. The acridine orange/Ethidium bromide dual staining (AO/EB) under the fluorescence microscope data suggests that apoptotic cell nuclei became irregularly changed. Our present study findings concluded that the strongly recommend the developed SrHA-CSscaffold excellent materialhas mechanical and functional properties including biological activities while compared to other synthetic nano-materials. Hence, the synthesizedSrHA-CSmaterial can be considered as a promising cost-effective nano-material for human bone regenerative applications.