Synthesis and characterization of spions-bromelain-folic acid on folic acid receptor positive cancer model

Engineering of a physiologically compatible, stable and targetable delivery vehicle superparamagnetic iron oxide nanoparticles-Bromelain-folic acid (SPIONs-Br- FA) was reported. Initially, the synthesized bare SPIONs were coated with citric acid (CA) in order to increase biocompatibility, stability and solubility of the SPIONs. Moreover, through CA coating, carboxyl functional groups for further reactions were produced. Br (as an anti-cancer agent) and FA (as a targeting agent to the folic acid receptor positive (FAR+) cancer cells) were conjugated to the synthesized nanocarrier through 1-ethyl-3-(3-dimethylaminpropyl)carbodiimide hydrochloride/ Nhydroxysuccinimide (EDC/NHS) click chemistry. Subsequently, characterization and physico-chemical analyses were carried out through methods such as Fourier transform infrared spectroscopy, atomic absorption spectroscopy (AAS), dynamic light scattering, vibrating sample magnetometer, x-ray diffraction, transmission electron microscopy (TEM) and field emission scanning electron microscopy. The in vitro tetrazolium dye (MTT) assay and blood compatibility tests were performed to confirm the biocompatibility of the engineered nano delivery system. High level of SPIONs-FA binding to FAR+ cell lines (HeLa, MDA-MB-231 and 4 T1) compared to folic acid receptor negative (FAR-) cell lines (HSF 1184 and MDA-MB-468) was assured via qualitative and quantitative in vitro binding studies (Prussian blue assay and AAS analysis). The reason may be higher transport of SPIONs-FA through the mechanism of receptor endocytosis pathway into FAR+ cells in comparison with the mechanism of passive diffusion of SPIONs into the FAR- cells. Cytotoxicity studies carried out in human cell lines (HSF 1184, MDA-MB-468, MDA-MB-231 and HeLa) and mouse breast cancer cells (4 T1) showed significant dose advantage with SPIONs-Br-FA in reducing the half maximal inhibitory concentration (IC50) values compared with neat Br. Through morphological observation studies by inverted microscope and acridine orange/ethidium bromide fluorescent staining method, it was disclosed that the cells had undergone apoptosis since the shrinkage as well as the apoptotic bodies were obviously seen. The results showed that SPIONs-Br-FA was a rewarding candidate to suppress the migration of the FAR+ cancer cells as well as to inhibit colony formation of the FAR+ cancer cells compared to neat Br. The percentage of apoptotic cells (apoptotic index) with more condensed and fragmented chromatin increased sharply in SPIONs-Br-FA treated cells compared to the neat Br. Overall, the SPIONs-Br-FA induced higher percentage of apoptotic cells than the neat Br. Moreover, after treatment protocol performance on 4 T1 tumor bearing mice, the qualitative and quantitative biodistribution study were carried out in vital organs and tumor using colorimetric method (AAS) and TEM method which indicate significant tumor targetability of SPIONs-FA. Finally, the tumor volume and inhibition growth rate were measured in 4 T1 tumor bearing mice treated with different SPIONs formulations to investigate the effectiveness of SPIONs- Br-FA. Administration of SPIONs-Br-FA through tail vein (three times a week) during the four-week treatment period reduced the tumor burden of tumor bearing mice and also increased their life-span when compared with SPIONs-Br and neat Br at same concentration of bromelain. In conclusion, the current results indicated the dualfunctional synthesized SPIONs-Br-FA is a promising tool in the field of biomedicine, chiefly cancer therapy

Biodegradable Mg/HA/TiO2 Nanocomposites Coated with MgO and Si/MgO for Orthopedic Applications: A Study on the Corrosion, Surface Characterization, and Biocompatability

In the field of orthopedics, magnesium (Mg) and magnesium-based composites as biodegradable materials have attracted fundamental research. However, the medical applications of magnesium implants have been restricted owing to their poor corrosion resistance, especially in the physiological environment. To improve the corrosion resistance of Mg/HA/TiO2 nanocomposites, monolayer MgO and double-layer Si/MgO coatings were fabricated layer-by-layer on the surface of a nanocomposite using a powder metallurgy route. Then, coating thickness, surface morphology, and chemical composition were determined, and the corrosion behavior of the uncoated and coated samples was evaluated. Field-emission scanning electron microscopy (FE-SEM) micrographs show that an inner MgO layer with a porous microstructure and thickness of around 34 m is generated on the Mg/HA/TiO2 nanocomposite substrate, and that the outer Si layer thickness is obtained at around 23 m for the double-layered coated sample. Electrochemical corrosion tests and immersion corrosion tests were carried out on the uncoated and coated samples and the Si/MgO-coated nanocomposite showed significantly improved corrosion resistance compared with uncoated Mg/HA/TiO2 in simulated body fluid (SBF). Corrosion products comprising Mg(OH)(2), HA, Ca-3(PO4)(2), and amorphous CaP components were precipitated on the immersed samples. Improved cytocompatibility was observed with coating as the cell viability ranged from 73% in uncoated to 88% for Si/MgO-coated Mg/HA/TiO2 nanocomposite after nine days of incubation

Synthesis, functionalization, characterization, and: in vitro evaluation of robust pH-sensitive CFNs-PA-CaCO3

The preparation, characterization, and application of Papain (PA) conjugated CaCO3-coated cobalt ferrite nanoparticles (CFNs-PA-CaCO3) is reported. Papain was covalently attached onto silanized cobalt ferrite nanoparticles (CFNs-APS) and then coated with robust pH sensitive CaCO3. It was characterized by FTIR, XRD, TEM, FESEM, and TGA. The release behaviour of PA from functionalized nanoparticles in acidic and physiological pH was investigated. However, the rate of PA release was slow and enhanced with time at physiological pH; the results illustrate that acidic conditions disassembles the coating, which caused a rapid release of PA. The biocompatibility evaluation of CFNs/CFNs-CaCO3 and the cytotoxicity of free PA/CFNs-PA was carried out in vitro on human breast cancer cells (MDA-MB-231) and human skin fibroblast cells (HSF 1184) by performing an MTT assay. An improved cytotoxic effect was obtained for PA conjugated cobalt ferrites in comparison with free PA. The synthesized delivery system was perfectly tolerable to the blood and cells in concentrations below 200 μg mL-1. Investigations were followed out to measure the time and dose dependent cellular uptake of functionalized nanoparticles via the Prussian blue staining method under an inverted microscope, which indicated that functionalized nanoparticles showed internalization properties. The results conveyed that the synthesized system is a promising biomaterial for future biomedical applications, mainly cancer therapy. The present in vitro findings require comparison with those of in vivo studies

Nanoencapsulation of andrographolide rich extract for the inhibition of cervical and neuroblastoma cancer cells

Andrographis paniculata is traditionally used for many diseases and scientifically proven for anti-cancer property. Andrographolide which is the marker compound is believed to be the main contributor to the pharmacological activities. The poor solubility and bioavailability of this diterpenoid lactone could be overcome by nanoencapsulation. Reflux extraction, and followed by successive Soxhlet fractionation were used to obtain andrographolide rich extract from the herb. Spontaneous emulsion solvent diffusion was used to nanoencapsulate andrographolide using poly(lactic-co-glycolic acid) with 1% polyvinyl alcohol as emulsifier. Nanospheres loaded with andrographolide was found to have the particle size, 163 nm; polydispersity index, 0.26 and zeta potential, - 57.85 mV. The encapsulation efficiency and in vitro drug release were 80.0% and 84.2%, respectively. The andrographolide nanoparticles could inhibit the proliferation of cervical and neuroblastoma cells with no adverse effect on normal human skin cells. Andrographolide rich extract loaded nanoparticles could inhibit the proliferation of HeLa and SH-SY5Y cells, mainly through Bax-induced apoptosis. The result was consistent with the low expression of anti-apoptotic genes (Bcl-2 and Bcl-xL) and prognostic factor (Ki-67). The tumour size of HeLa bearing mice was significantly reduced (73%) after treated with andrographolide rich nanoparticles (10 mg/kg body weight) for a month

Communal microaerophilic-aerobic biodegradation of amaranth by novel NAR-2 bacterial consortium

A novel bacterial consortium, NAR-2 which consists of Citrobacter freundii A1, Enterococcus casseliflavus C1 and Enterobacter cloacae L17 was investigated for biodegradation of Amaranth azo dye under sequential microaerophilic–aerobic condition. The NAR-2 bacterial consortium with E. casseliflavus C1 as the dominant strain enhanced the decolorization process resulting in reduction of Amaranth in 30 min. Further aerobic biodegradation, which was dominated by C. freundii A1 and E. cloacae L17, allowed biotransformation of azo reduction intermediates and mineralization via metabolic pathways including benzoyl-CoA, protocatechuate, salicylate, gentisate, catechol and cinnamic acid. The presence of autoxidation products which could be metabolized to 2-oxopentenoate was elucidated. The biodegradation mechanism of Amaranth by NAR-2 bacterial consortium was predicted to follow the steps of azo reduction, deamination, desulfonation and aromatic ring cleavage. This is for the first time the comprehensive microaerophilic–aerobic biotransformation pathways of Amaranth dye intermediates by bacterial consortium are being proposed

Papain grafted into the silica coated iron-based magnetic nanoparticles 'IONPs@SiO 2-PPN' as a new delivery vehicle to the HeLa cells

The present study aims at engineering, fabrication, characterization, and qualifications of papain (PPN) conjugated SiO2-coated iron oxide nanoparticles 'IONPs@SiO2-PPN'. Initially fabricated iron oxide nanoparticles (IONPs) were coated with silica (SiO2) using sol-gel method to hinder the aggregation and to enhance biocompatibility. Next, PPN was loaded as an anticancer agent into the silica coated IONPs (IONPs@SiO2) for the delivery of papain to the HeLa cancer cells. This fabricated silica-coated based magnetic nanoparticle is introduced as a new physiologically-compatible and stable drug delivery vehicle for delivering of PPN to the HeLa cancer cell line. The IONPs@SiO2-PPN were characterized using FT-IR, AAS, FESEM, XRD, DLS, and VSM equipment. Silica was amended on the surface of iron oxide nanoparticles (IONPs, γ-Fe2O3) to modify its biocompatibility and stability. The solvent evaporation method was used to activate PPN vectorization. The following tests were performed to highlight the compatibility of our proposed delivery vehicle: in vitro toxicity assay, in vivo acute systemic toxicity test, and the histology examination. The results demonstrated that IONPs@SiO2-PPN successfully reduced the IC50 values compared with the native PPN. Also, the structural alternations of HeLa cells exposed to IONPs@SiO2-PPN exhibited higher typical hallmarks of apoptosis compared to the cells treated with the native PPN. The in vivo acute toxicity test indicated no clinical signs of distress/discomfort or weight loss in Balb/C mice a week after the intravenous injection of IONPs@SiO2 (10 mg kg-1). Besides, the tissues architectures were not affected and the pathological inflammatory alternations detection failed. In conclusion, IONPs@SiO2-PPN can be chosen as a potent candidate for further medical applications in the future, for instance as a drug delivery vehicle or hyperthermia agent

Cervicare™ induces apoptosis in HeLa and CaSki cells through ROS production and loss of mitochondrial membrane potential

Cervicare™ is a poly-herbal preparation comprised of a combination of 6 plants; most have demonstrated antimicrobial and anticancer properties in preclinical studies. The effect of the ethanol and aqueous extracts of Cervicare™ on cell proliferation and apoptosis using cervical cancer HeLa and CaSki cells was investigated for the first time in the present study. MTT assay results showed that Cervicare™ extracts exerted time- and dose-dependent inhibition of cell viability. The hallmark properties of apoptosis like cell shrinkage and cytoplasmic condensation were observed using an inverted phase contrast microscope, ethidium bromide/acridine orange and Hoechst 33342/propidium iodide fluorescent staining methods. Furthermore, our results demonstrated that Cervicare™ extracts induced apoptosis in HeLa and CaSki cells by ROS generation and mitochondrial depolarization in a concentration dependent manner. The results showed that Cervicare™ extracts were capable of suppressing cell migration and inhibiting colony formation in a dose-dependent manner. Moreover, western blot analysis demonstrated the involvement of a mitochondria-dependent apoptosis pathway in the apoptosis inducing activity of Cervicare™ ethanol extract in HeLa cells. GC-MS analysis of the ethanolic extract afforded the identification of 40 substances, showing that it was primarily composed of anti-cancerous compounds such as xanthorrhizol (60.40%), octacosane (9.93%) and squalene (1.24%). Together, these results point out the Cervicare™ mediated inhibition of HeLa cell growth via induction of apoptosis and that it may be a potential anticancer agent which deserves further investigation

Fabrication and performance evaluation of blood compatible hemodialysis membrane using carboxylic multiwall carbon nanotubes and low molecular weight polyvinylpyrrolidone based nanocomposites

This study focused to optimize the performance of polyethersulfone (PES) hemodialysis (HD) membrane using carboxylic functionalized multiwall carbon nanotubes (c-MWCNT) and lower molecular weight grade of polyvinylpyrrolidone (PVP-k30). Initially, MWCNT were chemically functionalized by acid treatment and nanocomposites (NCs) of PVP-k30 and c-MWCNT were formed and subsequently blended with PES polymer. The spectra of FTIR of the HD membranes revealed that NCs has strong hydrogen bonding and their addition to PES polymer improved the capillary system of membranes as confirmed by Field Emission Scanning Electron Microscope (FESEM) and leaching of the additive decreased to 2% and hydrophilicity improved to 22%. The pore size and porosity of NCs were also enhanced and rejection rate was achieved in the establish dialysis range (<60 kDa). The antifouling studies had shown that NCs membrane exhibited 30% less adhesion of protein with 80% flux recovery ratio. The blood compatibility assessment disclosed that NCs based membranes showed prolonged thrombin and prothrombin clotting times, lessened production of fibrinogen cluster, and greatly suppressed adhesion of blood plasma than a pristine PES membrane. The results also unveiled that PVP-k30/NCs improved the surface properties of the membrane and the urea and creatinine removal increased to 72% and 75% than pure PES membranes. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 513–525, 2019

In vitro antiproliferative activity of fresh pineapple juices on ovarian and colon cancer cell lines

The main component that contributes to the high value of pineapple is bromelain which is a proteolytic enzyme and has been scientifically identified as a therapeutic agent. This study was conducted to obtain high quantity of bromelain from pineapple and to investigate the anticarcinogenic activity of fresh pineapple juices against A2780 ovarian and HT29 colon cancer cell lines. It was found that homogenization, ultrafiltration, precipitation and dialysis contributed to heavy loss of bromelain. Therefore, fresh pineapple juices from the flesh (PJ-F), core (PJ-C) and stem (PJ-S) were selected as a source of crude bromelain. Various bromelain concentrations of PJ-F, PJ-C and PJ-S (1 mu g/ml, 10 mu g/ml, 100 mu g/ml and 1000 mu g/ml) were exposed to the cancer cells and the cell viability was determined using Methylthiazol Tetrazolium Assay (MTT assay) after 24, 48 and 72 h. Besides, IC50 values were measured. Using normal cell (HSF1184) comparison, it was found that a 100 mu g/ml concentration of bromelain would efficiently inhibit the cancer cells without affecting the surrounding normal cells. Microscopic examinations were carried out to elucidate the modes of cell death on the basis of morphological alterations using florescent and inverted phase contrast microscopes. Furthermore, the colony forming abilities of fresh pineapple juices on A2780 and HT29 cells were examined. The results demonstrated that PJ-F, PJ-C and PJ-S effectively suppressed the colony formation in cancer cells. The findings suggest that PJ-F, PJ-C and PJ-S may have the potential to induce anticarcinogenic effects through an apoptosis to A2780 and HT29 cells in vitro