Cell Line MG-63 Research Articles

Thiolated sodium alginate/polyethylene glycol/hydroxyapatite nanohybrid for bone tissue engineering

The present study aimed to develop a thiolated sodium alginate-based nanohybrid formulation reinforced with hydroxyapatite to improve bone architecture in bone degenerative disorders. The prepared thiolated sodium alginate was converted into a nanocomposite by combining it with polyethylene glycol and hydroxyapatite. The nanocomposite was characterized by scanning electron microscopy, atomic force microscopy, and dynamic light scattering. The developed nanocomposite was evaluated for cell toxicity by performing an MTT assay using the MG63 cell line. The bone targeting efficacy was investigated by analysis of trabecular architecture on ovariectomized rat model and the molecular mechanism was investigated by molecular docking studies. The results of particle size determination indicated that the nanocomposite was within the size range of 100–200 nm. FTIR and XRD studies confirmed the thiolation of sodium alginate and the development of a composite material. Cell line studies showed that the developed nanocomposite was not cytotoxic and well tolerated by the MG63 cell line. The scanning electron microscopy and histological studies of trabecular bone in substantiation with molecular docking study results confirmed the bone targeting and healing ability of the nanocomposite. The results of the study indicated that the developed nanocomposite can be a potential biomaterial for bone tissue regeneration.

Synthesis of nano hydroxyapatite from Hypopthalmichthys molitrix (silver carp) bone waste by two different methods: a comparative biophysical and in vitro evaluation on osteoblast MG63 cell lines.

More than a thousand tonnes of fish bone wastes can be transformed into biomedical products annually. Alkaline hydrolysis and thermal calcification were used to create nanosized hydroxyapatite (HAp) crystals from Silver carp bone wastes. Biophysical tests were used to determine the nano size and chemical composition of synthesised hydroxyapatite. Alkaline hydrolysis hydroxyapatite (AH-HAp) was 58.3nm, while Thermal calcination hydroxyapatite (TC-HAp) was 64.3nm in size, confirmed by Atomic Force Microscopy. Energy Dispersive X-ray Analysis studies showed Ca/P (Calcium phosphate) ratio of AH-HAp to be 1.65, whereas TC-HAp as 1.45, confirming AH-HAp to be organically rich along with a similar Ca/P ratio as natural HAp. Fourier Transform Infrared Spectroscopy spectra indicated HAp formation from both procedures, however AH-HAp had superior crystallinity than TC-HAp confirmed from X-Ray Diffraction spectra. MG63 osteoblast cell lines showed 91% cell viability in cytotoxicity studies and 70.1% proliferation efficiency in Alkaline Phosphatase assay, which was higher than TC-HAp. The present study shows that HAp produced via alkaline hydrolysis has better biocompatibility which enhances its applicability as a biomaterial, than HAp synthesized through thermal calcination, which tends to incinerate organic moieties.

The long non-coding RNA CRNDE promotes osteosarcoma proliferation and migration by sponging miR-136-5p/MRP9 axis.

BackgroundThe long-noncoding RNA colorectal neoplasia differentially expressed (CRNDE) gene has been found to be upregulated in several solid tumors. Whether CRNDE affects osteosarcoma (OS) and its underling mechanism remains unknown.MethodsTumor tissues and corresponding normal tissues were collected from 45 patients with OS. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was applied to determine lncRNA CRNDE level in the tissues. Participants were divided into a high CRNDE group and a low CRNDE group according to the median value of lncRNA CRNDE expression detected by in situ hybridization (ISH). The differences between high and low expression of lncRNA CRNDE in patients were compared clinically by chi-square test. Kaplan-Meier survival analysis was applied to analyze the relationship between lncRNA CRNDE expression and patient survival. Subsequently, silencing or overexpression of lncRNA CRNDE were performed in MG63 and 143B cell lines, qRT-PCR was applied to verify the expression of lncRNA CRNDE, miR-136-5p, and MRP9; dual-luciferase reporter assay was used to evaluate the targeting relationship between miR-136-5p, lncRNA CRNDE, and Cell Counting Kit-8 (CCK8), wound-healing, and Transwell assays were used to analyze for cell proliferation, migration, and invasion, respectively, and western blot was used to detect expression in cells.ResultsThe expression of CRNDE in OS tissues was higher than that in normal tissues. High lncRNA CRNDE expression was significantly associated with clinical stage, lung metastasis, and poor prognosis in OS patients. Additionally, overexpression of lncRNA CRNDE promoted proliferation and migration of OS cells. Bioinformatics analysis showed that lncRNA CRNDE competitively inhibited miR-136-5p through acting as a competitive endogenous RNA (ceRNA). It was also revealed that miR-136-5p is a binding target gene of lncRNA CRNDE and that MRP9 is involved in this process as a downstream target gene of miR-136-5p.ConclusionsThe lncRNA CRNDE promotes the proliferation and migration of OS cells by regulating the miR-136-5p/MRP9 pathway, and lncRNA CRNDE can be a significant marker of OS prognosis.

RNA 5-methylcytosine status is associated with DNMT2/TRDMT1 nuclear localization in osteosarcoma cell lines

Osteosarcoma (OS) is a pediatric malignant bone tumor with unsatisfying improvements in survival rates due to limited understanding of OS biology and potentially druggable targets. The present study aims to better characterize osteosarcoma U-2 OS, SaOS-2, and MG-63 cell lines that are commonly used as in vitro models of OS. We focused on evaluating the differences in cell death pathways, redox equilibrium, the activity of proliferation-related signaling pathways, DNA damage response, telomere maintenance, DNMT2/TRDMT1-based responses and RNA 5-methylcytosine status. SaOS-2 cells were characterized by higher levels of superoxide and nitric oxide that promoted AKT and ERK1/2 activation thus modulating cell death pathways. OS cell lines also differed in the levels and localization of DNA repair regulator DNMT2/TRDMT1. SaOS-2 cells possessed the lowest levels of total, cytoplasmic and nuclear DNMT2/TRDMT1, whereas in MG-63 cells, the highest levels of nuclear DNMT2/TRDMT1 were associated with the most pronounced status of RNA 5-methylcytosine. In silico analysis revealed potential phosphorylation sites at DNMT2/TRDMT1 that may be related to the regulation of DNMT2/TRDMT1 localization. We postulate that redox homeostasis, proliferation-related pathways and DNMT2/TRDMT1-based effects can be modulated as a part of anti-osteosarcoma strategy reflecting diverse phenotypic features of OS cells.

PDCD10 promotes proliferation, migration, and invasion of osteosarcoma by inhibiting apoptosis and activating EMT pathway.

Osteosarcoma, a common primary malignant tumor, occurs in children and adolescents with a poor prognosis. The current treatment methods are various, while the five-year survival rate of patients has not been significantly improved. As a member of the programmed death factor (PDCD) family, programmed death factor 10 (PDCD10) plays a role in regulating cell apoptosis. Several studies of PDCD10 in CCM and cancers have been reported before. However, there are no relevant research reports on the effects of PDCD10 on osteosarcoma. We used bioinformatics analysis, IHC, and clinical data to confirm the expression of PDCD10 and its correlation with prognosis in osteosarcoma. Then, we used shRNAs and cDNA to knock down or overexpress PDCD10 in U2OS and MG63 cell lines. A series of function assays such as CCK8, Wound healing test, Plate cloning formation assay, and Transwell were done to confirm how PDCD10 affects osteosarcoma. Animal assays were done to confirm the conclusions in cell lines. At last, WB was used to measure the protein expression levels of apoptosis and the EMT pathway. PDCD10 was highly expressed in patients with osteosarcoma and correlated with prognosis; PDCD10 knockdown inhibited osteosarcoma growth, proliferation, migration, and invasion; PDCD10 overexpression promoted osteosarcoma growth, proliferation, migration, and invasion. In vivo experiments confirmed the conclusions in cell lines; PDCD10 inhibited apoptosis and activated the EMT pathway. In this study, it was found that PDCD10 was highly expressed in patients with osteosarcoma, and it was closely related to patient prognosis. PDCD10 inhibited tumor cell apoptosis and promoted tumor progression by activating the EMT pathway. These findings may provide a potential target for gene therapy of osteosarcoma.

Physicochemical and biological studies of Ni(II), Cu(II) and Zn(II) ternary complexes of sulfaquinoxaline and 2,2’-bipyrimidine

Three monomeric ternary complexes of Ni(II) (1), Cu(II) (2), and Zn(II) (3) with sulfaquinoxaline (SQO), and 2,2’-bipyrimidine (bpm) as ligands have been prepared and their crystal structures have been determined. The complexes are formulated as M(SQO)2bpm·2H2O, completing MN6 coordination spheres. Hirshfeld surface analysis was employed to study the intermolecular interactions in the crystal lattices of the compounds. From these studies, it was found that hydrogen bonding and π-stacking interactions play an important role. Besides, the complexes have been characterized by FTIR and UV-Visible spectroscopies, and thermogravimetric techniques. The cytotoxicity of the complexes was screened against the tumoral A549 (human lung adenocarcinoma) and MG-63 (human osteosarcoma) cell lines. The complexes showed enhanced antibacterial activity against S. aureus and S. epidermidis as compared with the free SQO ligand.

Efficacy of glycoprotein-based nanocurcumin/silk fibroin electrospun scaffolds: Perspective for bone apatite formation

In this research, a blend of poly (vinyl) alcohol (PVA), silk fibroin nanofibers (SF-NFs), hydroxyapatite nanoparticles (HA-NPs), and curcumin nanoparticles (Cu-NPs) were used to design a nanofibrous scaffold (NFS), by electrospun method, for their possible application in bone apatite formation. The prepared nanofibrous scaffold (NFS) was investigated using physicochemical, mechanical, cell viability, etc. The bioactivity test of NFS was evaluated after immersion in SBF (simulated body fluid). NFS from PVA, SF-NFs, HA-NPs, and Cu-NPs exhibited outstanding physicochemical, mechanical, and antibacterial properties. PVA: SF-NFs: HA-NPs: Cu-NPs scaffold excellent antimicrobial performance against E.coli (4.16 ± 0.16 mm) and S.aureus (4.88 ± 0.10 mm). Biocompatibility studies were assessed using human osteosarcoma MG63 cell line, which displayed 100% viable cell presence in PVA: SF-NFs: HA-NPs: Cu-NPs. Based on the characterization, results indicated that they are excellent scaffolds for bone tissue regeneration.

Experimental study on effect of complement C3 on bone formation observed by cell culture in vitro

To explore effect of lentivirus-mediated complement C3 silencing vector on the osteogenic ability of human B lymphocyte Raji-osteoblast cell line MG63 co-culture system and its mechanism. The lentiviral complement C3 silencing vector was constructed and transfected into human B lymphocyte Raji to establish in vitro Raji-osteoblast cell line MG63 co-culture system. The cells were divided into blank control group (without special treatment), complement C3 silencing group (lentiviral complement C3 silencing vector transfection co-culture system) and model group(lentiviral vector transfection co-culture system). The expression of complement C3 in each group was detected by reverse transcription-polymerase chain reaction(RT-PCR) and Western-Blot at 24 h after culture, proliferation of MG63 cells was detected by CCK-8 at 0, 3, 6, 12, 24, 48 and 72 h, apoptosis of MG63 cells in each group was detected by flow cytometry at 24 h after culture, and alkaline phosphatase(AKP) activity of MG63 cells in each group was detected by AKP detection kit, and osteoprotegerin (OPG ) protein expression of MG63 cells in each group was detected by Western-Blot method. RT-PCR results showed that the expression level of C3 in complement C3 silencing group was lower than that in blank control group and model group, Western-Blot results showed that expression of C3 in complement C3 silencing group was lower than that in blank control group and model group, CCK-8 results showed that there was no difference in proliferation ability of MG63 among complement C3 silencing group and blank control group and model group at 3 and 6 h after culture;at 12 h after culture, proliferation ability of MG63 cells with C3 silencing was higher than that of blank control group and model group;at 24, 48 and 72 h after culture, proliferation ability of MG63 cell line with complement C3 silencing group were higher than that of blank control group and model group. Flow cytometry resluts showed that apoptosis of proliferation ability of MG63 cell line with complement C3 silencing group was lower than that of blank control group and model group;AKP detection kit suggested that AKP activity in complement C3 silencing group was higher than that in blank control group and model group, Western-Blot results showed that expression level of OPG protein in complement C3 silencing groupwas higher than that in blank control groupand model group. Silencing of complement C3 could enhance bone formation ability of osteoblast MG63, and it takes time to accumulate this ability. Complement C3 may affect osteogenesis by altering OPG / RANKL / RANK axis.

HO-3867 Induces Apoptosis via the JNK Signaling Pathway in Human Osteosarcoma Cells.

Metastatic osteosarcoma often results in poor prognosis despite the application of surgical en bloc excision along with chemotherapy. HO-3867 is a curcumin analog that induces cell apoptosis in several cancers, but the apoptotic effect and its mechanisms on osteosarcoma cells are still unknown. After observing the decrease in cellular viability of three human osteosarcoma U2OS, HOS, and MG-63 cell lines, and the induction of cellular apoptosis and arrest in sub-G1 phase in U2OS and HOS cells by HO-3867, the human apoptosis array showed that heme oxygenase (HO)-1 and cleaved caspase-3 expressions had significant increases after HO-3867 treatment in U2OS cells and vice versa for cellular inhibitors of apoptosis (cIAP)1 and X-chromosome-linked IAP (XIAP). Western blot analysis verified the results and showed that HO-3867 activated the initiators of both extrinsic caspase 8 and intrinsic caspase 9, and significantly increased cleaved PARP expression in U2OS and HOS cells. Moreover, with the addition of HO-3867, ERK1/2, and JNK1/2 phosphorylation were increased in U2OS and HOS cells. Using the inhibitor of JNK (JNK in 8), HO-3867’s increases in cleaved caspases 3, 8, and 9 could be expectedly suppressed, indicating that JNK signaling is responsible for both apoptotic pathways, including extrinsic and intrinsic, in U2OS and HOS cells caused by HO-3867. Through JNK signaling, HO-3867 has proven to be effective in causing both extrinsic and intrinsic apoptotic pathways of human osteosarcoma cells.

Exploration on in vitro bioactivity, antibacterial activity and corrosion behavior of Strontium doped Hydroxyapatite reinforced chitosan-polypyrrole/TNT for bone regeneration

This study aimed to investigate the effects of TNT/CS-PPy@Sr-HAP composite coatings on Ti alloys that were used to develop the coating by anodization via electrophoretic deposition method. The formation of TNT/CS-PPy@Sr-HAP composite coatings was characterized by FT-IR, XRD, and FESEM with EDAX analyses. The mechanical performance of the composite coating was investigated for its thickness, microhardness, and adhesion tests. Antibacterial tests showed that TNT/CS-PPy@Sr-HAP (II) composite coated sample recorded the maximum level of antimicrobial activity against E. coli and S. aureus bacteria was found to be 25 & 22 mm for 100 μL samples, respectively. The in vitro studies confirmed that TNT/CS-PPy@Sr-HAP (II) composite coating exhibited a greater number of viable cells found to be ∼99.7% after 7 days of incubation. Potentiodynamic polarization and electrochemical impedance studies were performed on TNT/CS-PPy@Sr-HAP (II) composite coatings resulting in its better corrosion behavior when compared to other coatings. All these results show that TNT/CS-PPy @ Sr-HAP (II) composite coated on Ti implant alloy will serve as a potential material for orthopedic applications.

Local Wisdom and Diversity of Medicinal Plants in Cha Miang Forest in Mae Kampong Village, Chiang Mai, Thailand, and Their Potential for Use as Osteoprotective Products.

“People-Forest-Miang” communities are villages located in the cultivated area of Camellia sinensis var. assamica, or Cha Miang, in northern Thailand. Cha Miang forests are a form of agriculture relying on forest-rich bioresources. This study focuses on a survey of the diversity of medicinal plants used by “People-Forest-Miang” communities in Mae Kampong Village, Chiang Mai, Thailand. The results demonstrated that 73 species of medicinal plants were used to prevent and treat various ailments. The highest number of species (30.14%) was used for musculoskeletal system disorders, followed by digestive system disorders (21.92%) and unspecified medicinal disorders (15.07%). The alkaline phosphatase (ALP) is the most widely recognized biochemical marker for osteoblast activity. The ALP activity of ethanol and deionized water extracts of the nine selected medicinal plants used for musculoskeletal system disorders were examined in the MG63 cell line. The results showed that the numerous water extracts, including MKP1, MKP2, MKP5, MKP6, MKP7, MKP8, and MKP9, and the ethanolic extracts—namely, MKP2, MKP3, MKP7, and MKP9—significantly increased ALP activity in the MG-63 cell line. The findings indicate that some medicinal plants may be further studied for active chemicals and developed as natural active pharmaceutical ingredients for osteoprotective products.

Licochalcone B Induced Apoptosis and Autophagy in Osteosarcoma Tumor Cells via the Inactivation of PI3K/AKT/mTOR Pathway.

Licochalcone B (LicB) is a flavonoid derived from the Chinese medicinal herb Glycyrrhiza uralensis Fisch. Several previous studies have demonstrated the wide range of pharmacological activities shown by LicB. In this study, we investigated the anticancer effects of LicB in osteosarcoma (OS) tumor cells and its underlying mechanisms. According to the Cell Counting Kit-8 (CCK8) analysis and 5-ethynil-2'-deoxyuridine (EdU) staining results, we found that LicB suppresses OS cells (MG-63 and U2OS) growth depending on its concentration. Furthermore, flow cytometry and Western blot revealed that LicB promoted autophagy and apoptosis in OS cells in a dose-dependent manner. LicB treatment not only decreased the levels of Bcl-2, p62, Caspase-3, and Ki67 protein in MG-63 and U2OS cell lines but also increased the levels of Cleaved Caspase-3, Beclin1, Bax, Atg7, and LC3B. Mechanistically, LicB induced cell apoptosis by promoting the apoptosis-related cleavage of Caspase-3, while suppressing the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway to induce autophagy. The present work is the first to illustrate that LicB can serve as a potential drug candidate for tumor treatment owing to its ability to enhance autophagy and apoptosis, and suppress OS proliferation by inactivating the PI3K/AKT/mTOR pathway.

Fabrication and evaluation of combined 3D printed/pamidronate-layered double hydroxides enriched electrospun scaffolds for bone tissue engineering applications

The combination of commonly used fabrication technologies, including electrospinning and additive manufacturing, is a new approach to produce tissue engineering scaffolds resembling the heterogeneous tissue structure. This study presents the fabrication and characterization of novel 3D-structured scaffolds of pamidronate (PAM)-loaded layered double hydroxides (LDH)/polycaprolactone (PCL) electrospun nanofibers combined with micro-strand PCL 3D printed grids. PAM-LDH was synthesized through the coprecipitation method, and the intercalation of PAM into LDH was confirmed via XRD and FTIR analysis. PCL and PAM-LDH/PCL nanofiber mats were fabricated by electrospinning. Adding PAM-LDH to PCL decreased fiber diameter from 527 ± 24 nm to 289 ± 43 nm and increased the surface roughness from 33.5 ± 17 nm to 65.6 ± 24 nm. Circular PCL grids, including strands of 400 μm width, were fabricated using a 3D printer system. The final structure of the scaffolds was obtained by gluing the nanofiber mats between 3D printed micro-strand grids. FESEM images showed the nanofiber mats fully covered the pores of 3D printed grids while the morphology of the fibers remained the same. BET analysis was conducted to calculate the mean pore diameter, total pore volume, and surface area of the scaffolds. Monitoring the in vitro release of PAM showed mainly two stages; water penetration-controlled for early hours and a degradation-controlled release stage up to 28 days. The biocompatibility of the scaffolds was assessed by culturing the MG-63 cell lines. Scaffolds with PAM-LDH containing nanofiber mats highly supported cell adhesion and improved alkaline phosphatase activity, an indicator of osteoconductivity. Given the results, these novel 3D structured scaffolds have the potential application to be used as bone tissue engineering scaffolds.

Knockdown of CD44 inhibits proliferation, migration and invasion of osteosarcoma cells accompanied by downregulation of cathepsin S

BackgroundOsteosarcoma (OS) is a malignant bone tumour of mesenchymal origin. These tumours are characterised by rich vascularisation, therefore promoting rapid proliferation and facilitating metastasis. CD44 has been reported to be involved in OS, but its role and molecular mechanisms in the pathogenesis of the disease are not fully determined.MethodsIn this study, we investigated the antitumor effect of CD44 on the development of OS and further explored the molecular mechanisms. The expression of CD44, cathepsin S and MMP-9 was detected by Western blot (WB) and reverse transcription-polymerase chain reaction (RT-qPCR) in different cell lines (MG63, U2OS OS and hFOB 1.19). To elucidate the role of CD44 in OS, MG63 and U2OS cells were treated with small interference RNA (siRNA) to knock down CD44, and the knockdown efficiency was validated with GFP and RT-qPCR. Furthermore, cell proliferation was assayed using Cell Counting Kit‑8 (CCK-8) and colony formation assays, and cell migration and invasion were assayed by transwell and wound-healing assays.ResultsWe found that CD44 expression in the MG63 and U2OS OS cell lines was markedly increased compared to that of the human osteoblast hFOB 1.19 cell line. Knockdown of CD44 inhibited proliferation, migration and invasion of MG63 and U2OS cells. Cathepsin S expression in the MG63 and U2OS OS cell lines was increased compared to that of the human osteoblast hFOB 1.19 cell line. When CD44 was knocked down, its expression level went down.ConclusionTaken together, our data reinforced the evidence that CD44 knockdown inhibited cell proliferation, migration and invasion of OS cells accompanied by altered expression of cathepsin S. These findings offer new clues for OS development and progression, suggesting CD44 as a potential therapeutic target for OS.

In-vitro cytotoxicity of zinc oxide, graphene oxide, and calcium carbonate nano particulates reinforced high-density polyethylene composite

This research aims to develop and increase the appropriate polymer composite for human implants and to evaluate its mechanical properties. The composites have proved their application in several fields, but still, many types of research are going on to improve their properties and to increase the area of application. The polymer composites have also entered their utility into bone implants as a substitute for metallic implants due to problems like high cost and corrosion resistance. The polymer composites are less in weight, economical, high corrosion resistance and they can be made into any required shape. In this research the Zinc oxide (ZnO), Graphene oxide (GO), and Calcium carbonate (CaCO3) nanoparticles with wt. % reinforced polymer composites provide good mechanical properties and show good tribological properties. Among Zinc oxide, graphene oxide, calcium carbonate nanoparticles the graphene oxide reinforced High-density polyethylene (HDPE) composites have better properties and they can be most suitable for bio implant materials. With the extract-cotton seed oil and extract-Phosphate Buffer Saline (PBS) from graphene oxide (HDPE) composites, the culture medium (cell line MG-63) with direct cytotoxicity reactivity first test on extract-cotton seed oil of 10 μl and 20 μl extract exhibited a maximum of 71% and 57% cell viability and a minimum of 29% and 43% mild cytotoxicity. The culture medium with direct cytotoxicity reactivity second test on extract-Phosphate Buffer Saline (PBS) of 10 μl and 20 μl extract exhibited maximum 61% and 60% cell viability and minimum 39% and 40% cytotoxicity.

Development of Silver Doped Hydroxyapatite Thin Films for Biomedical Applications

Silver doped hydroxyapatite [AgHAp, Ca10−xAg(PO4)6(OH)2], due to its antimicrobial properties, is an advantageous material to be used for various coatings. The AgHAp thin films with xAg = 0.05 and xAg = 0.1 were achieved using the spin-coating method. The resulting samples were examined by X-ray diffraction (XRD), scanning electron microscopy (SEM), atomic force microscopy (AFM), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, and X-ray photoelectron spectroscopy (XPS). XRD analysis revealed that the particles of both samples are ellipsoidal. Also, in agreement with the results obtained by XRD measurements, the results of the SEM studies have shown that the particles shape is ellipsoidal. Optical properties of silver doped hydroxyapatite thin films deposited on Si substrate were investigated through Fourier transform infrared spectroscopy (FTIR) and Raman spectroscopy. The results obtained by the two complementary techniques highlighted that the molecular structure of the studied samples is not influenced by the increase of the silver concentration in the samples. Our studies revealed that the surface morphology of the obtained samples consist of uniform and continuous layers. The biocompatibility of the obtained thin films was also evaluated with the aid of human osteosarcoma MG63 (ATCC CRL 1427) cell line. Moreover, the in vitro antifungal activity against Candida albicans fungal strain of the AgHAp thin films was studied and the obtained results revealed their antifungal effect. The results of the biological assays showed that the AgHAp thin films are a very promising material for biomedical applications.

Fabrication, Characterization and In Vitro Assessment of Laevistrombus canarium-Derived Hydroxyapatite Particulate-Filled Polymer Composite for Implant Applications.

This paper presents the formulation, characterization, and in vitro studies of polymer composite material impregnated with naturally derived hydroxyapatite (HA) particulates for biomedical implant applications. Laevistrombus canarium (LC) seashells (SS) were collected, washed and cleaned, sun-dried for 24 h, and ground into powder particulates. The SS particulates of different weight percentages (0, 10, 20, 30, 40, 50 wt%)-loaded high-density polyethylene (HDPE) composites were fabricated by compression molding for comparative in vitro assessment. A temperature-controlled compression molding technique was used with the operating pressure of 2 to 3 bars for particulate retention in the HDPE matrix during molding. The HDPE/LC composite was fabricated and characterized using X-ray diffraction (XRD), field-emission scanning electron microscopy (FESEM), energy-dispersive X-ray (EDX), differential scanning calorimetry (DSC), and TGA. Mechanical properties such as tensile, compression, flexural, hardness, and also surface roughness were tested as per ASTM standards. Mass degradation and thermal stability of the HDPE/LC composite were evaluated at different temperatures ranging from 10 to 700 °C using thermogravimetric analysis (TGA). The maximum tensile strength was found to be 27 ± 0.5 MPa for 30 wt% HDPE/LC composite. The thermal energy absorbed during endothermic processes was recorded as 71.24 J/g and the peak melting temperature (Tm) was found to be 128.4 °C for the same 30 wt% of HDPE/LC composite specimen. Excellent cell viability was observed during the in vitro biocompatibility study for EtO-sterilized 30 wt% of HDPE/LC composite specimen, except for a report of mild cytotoxicity in the case of higher concentration (50 µL) of the MG-63 cell line. The results demonstrate the potential of the fabricated composite as a suitable biomaterial for medical implant applications.

Development of sodium alginate-pectin/TiO2 nanocomposites: Antibacterial and bioactivity investigations

In this research, sodium alginate-pectin composite (ALG-PEC CS) and nanocomposites (NCs) films with 0.5, 1, and 2 wt% TiO2 nanoparticles (NPs) were prepared using CaCl2 and glutaraldehyde (Glu) as cross-linkers. The cross-linking produces rigid scaffolds for sedimentation of hydroxyapatite (HA), it can also decrease solubility in water and simulated body fluid (SBF) solution to 10% or less. The increase of the adsorbed water and SBF extends the pores and consequently the surface area for HA growth. Bioactive ability was confirmed via HA's presence on the all films. It was revealed that the film containing 2 wt% TiO2 NPs had the best bioactivity without any in vitro cytotoxicity on MG-63 cell line and the best antibacterial performance against Staphylococcus aureus, and after 1 h all the bacteria were killed.

GPR64, Screened from Ewing Sarcoma Cells, Is a Potential Target for Antibody-Based Therapy for Various Sarcomas.

Simple SummaryNew strategies for immunotherapy have led to an increased interest in tumor-specific antigens on the cell surface in the field of oncology. Identifying markers in sarcomas is difficult because their tumor mutation burden is less than that of carcinomas. We assumed that a target protein may be acceptable as a therapeutic target, even if it is only expressed in the epididymis along with the tumor, because the epididymis has special barriers, known as the blood–epididymis barrier (BEB). We identified GPR64 as a therapeutic target for Ewing sarcoma via next-generation RNA-sequencing. GPR64 is located on the apical membranes of efferent ductules and separated from antibodies by the BEB. This study revealed, for the first time, that anti-GPR64 antibodies accumulate in various sarcomas and avoid targeting GPR64 in the epididymis in vivo. Furthermore, GPR64 is widely expressed in various sarcomas and is, therefore, a potential antibody-based therapeutic target for sarcomas.Ewing sarcoma is an aggressive and the second most common bone tumor in adolescent and young adult patients. The 5-year survival rate is 60–70% for localized disease but 30% for patients with metastases. Here, we aimed to identify a therapeutic target for Ewing sarcoma and evaluate antibody-based therapeutic agents using in vitro and in vivo models. We identified G protein-coupled receptor 64 (GPR64) as a therapeutic target for Ewing sarcoma via next-generation RNA-sequencing. GPR64v205 mRNA was expressed in HTB166, A673, MG63, 143B, HS-Sy II, and HT1080 cell lines as well as in Ewing sarcoma, undifferentiated pleomorphic sarcoma, leiomyosarcoma, dedifferentiated liposarcoma, and synovial sarcoma tissues. GPR64 expression was observed in 62.5% of sarcoma cases and was overexpressed in 33.9% cases. GPR64-specific monoclonal antibodies were tested as near-infrared probes for in vivo imaging using subcutaneous tumor mouse xenografts. Fluorescence intensity was stronger for the AF700-labeled anti-GPR64 antibody than that for the AF700-labeled isotype control antibody. GPR64 was detected in engrafted tumors of A673, 143B, HT1080, and the epididymis but not in other resected tissues. The anti-GPR64 antibody showed excellent binding to GPR64-positive tumors but not to healthy tissues. This antibody has potential for drug delivery in the antibody-based treatment of sarcomas.

Microstructural Analysis of Terbium Doped Zirconia and Its Biological Studies

Zirconia has its place in the biomedical industry because of its mechanical strength, bio-inertness, and physiochemical properties. Zirconia was synthesized and doped with Terbium (Tb), a lanthanide that was reported to show a photoluminescence property, which was a major characteristic for carcinogenic studies. Zirconia and Tb doped Zirconia were synthesized using the co-precipitation technique and were sintered at a temperature ranging from 900 to 1200 °C. The Zirconia sample and Tb doped Zirconia were thus studied for structural diversities using the X-ray powder diffraction technique (XRD), FTIR, FE-SEM, and TEM. From XRD, Zirconia phase transformation from monoclinic to tetragonal phase was observed, which signified limited fracture, elasticity, and crack formation. It was evident that Terbium stabilized the tetragonal phase of Zirconia, which reportedly shows mechanical properties, which include fracture toughness and flexural strength. The particle size of the Zirconia was comparatively more than the Tb doped Zirconia. The particle size of Zirconia ranged between 176 nm and 393 nm and the particle size of Tb doped Zirconia ranged between 110 nm and 343 nm. The biocompatibility of both the samples was tested using an Mg-63 cell line, and the cell viability was observed to be higher in Tb doped Zirconia when compared to the undoped Zirconia sample.