Cytotoxic profiles of a nanodrug delivery based on 6-mercaptopurine-coated magnetite-peg nanoparticles towards leukemia (WEHI-3B) cell lines

A drug active, 6-mercaptopurine (MP) was coated on the surface of Fe3O4-PEG nanoparticles using co-precipitation method in order to form a new magnetic nanocomposite (FPEGMP). The physico-chemical properties of the nanocomposite were studied via X-ray diffraction, infrared spectroscopy, magnetic measurements, thermal analysis and transmission electron microscopy. The resulting superparamagnetic nanocomposite has spherical shape with average particle size diameter of 11 nm. Thermal analyses and Fourier transform infrared (FTIR) spectroscopy revealed the formation of PEG-MP on the surface of iron oxide nanoparticles and the enhancement of the thermal stability of the nanocomposite compared to its counterpart, free 6-mercaptopurine. Release behavior of MP from FPEGMP nanocomposite was found to be sustained and governed by pseudo-second order kinetic. The maximum percentage release of MP from FPEGMP nanocomposite reached about 60% and 97% within approximately 92 and 72 hours when exposed to aqueous solutions at pH 7.4 and pH 4.8, respectively. Anti-cancer activity of the nanocomposite shows that the choice of coating material as well as the percentage of loading of the active agent could affect the cytotoxic activity of nanocomposite towards the mouse myelomonocytic leukemic cell line (WEHI-3B)

Preparation and characterization of 6-mercaptopurine-coated magnetite nanoparticles as a drug delivery system

Background: Iron oxide nanoparticles are of considerable interest because of their use in magnetic recording tape, ferrofluid, magnetic resonance imaging, drug delivery, and treatment of cancer. The specific morphology of nanoparticles confers an ability to load, carry, and release different types of drugs. Methods and results: We synthesized super paramagnetic nanoparticles containing pure iron oxide with a cubic inverse spinal structure. Fourier transform infrared spectra confirmed that these Fe3O4 nanoparticles could be successfully coated with active drug, and thermogravimet-ric and differential thermogravimetric analyses showed that the thermal stability of iron oxide nanoparticles coated with chitosan and 6-mercaptopurine (FCMP) was markedly enhanced. The synthesized Fe3O4 nanoparticles and the FCMP nanocomposite were generally spherical, with an average diameter of 9 nm and 19 nm, respectively. The release of 6-mercaptopurine from the FCMP nanocomposite was found to be sustained and governed by pseudo-second order kinetics. In order to improve drug loading and release behavior, we prepared a novel nanocomposite (FCMP-D), ie, Fe3O4 nanoparticles containing the same amounts of chitosan and 6-mercaptopurine but using a different solvent for the drug. The results for FCMP-D did not demonstrate "burst release" and the maximum percentage release of 6-mercaptopurine from the FCMP-D nanocomposite reached about 97.7% and 55.4% within approximately 2,500 and 6,300 minutes when exposed to pH 4.8 and pH 7.4 solutions, respectively. By MTT assay, the FCMP nanocomposite was shown not to be toxic to a normal mouse fibroblast cell line. Conclusion: Iron oxide coated with chitosan containing 6-mercaptopurine prepared using a coprecipitation method has the potential to be used as a controlled-release formulation. These nanoparticles may serve as an alternative drug delivery system for the treatment of cancer, with the added advantage of sparing healthy surrounding cells and tissue

Arginine–chitosan- and arginine–polyethylene glycol-conjugated superparamagnetic nanoparticles: preparation, cytotoxicity and controlled-release

Iron oxide magnetic nanoparticles (MNPs) can be used in targeted drug delivery systems for localized cancer treatment. MNPs coated with biocompatible polymers are useful for delivering anticancer drugs. Iron oxide MNPs were synthesized via co-precipitation method then coated with either chitosan (CS) or polyethylene glycol (PEG) to form CS–MNPs and PEG–MNPs, respectively. Arginine (Arg) was loaded onto both coated nanoparticles to form Arg–CS–MNP and Arg–PEG–MNP nanocomposites. The X-ray diffraction results for the MNPs and the Arg–CS–MNP and Arg–PEG–MNPs nanocomposites indicated that the iron oxide contained pure magnetite. The amount of CS and PEG bound to the MNPs were estimated via thermogravimetric analysis and confirmed via Fourier transform infrared spectroscopy analysis. Arg loading was estimated using UV–vis measurements, which yielded values of 5.5% and 11% for the Arg–CS–MNP and Arg–PEG–MNP nanocomposites, respectively. The release profile of Arg from the nanocomposites followed a pseudo-second-order kinetic model. The cytotoxic effects of the MNPs, Arg–CS–MNPs, and Arg–PEG–MNPs were evaluated in human cervical carcinoma cells (HeLa), mouse embryonic fibroblast cells (3T3) and breast adenocarcinoma cells (MCF-7). The results indicate that the MNPs, Arg–CS–MNPs, and Arg–PEG–MNPs do not exhibit cytotoxicity toward 3T3 and HeLa cells. However, treatment of the MCF-7 cells with the Arg–CS–MNP and Arg–PEG–MNP nanocomposites reduced the cancer cell viability with IC50 values of 48.6 and 42.6 µg/mL, respectively, whereas the MNPs and free Arg did not affect the viability of the MCF-7 cells

Controlled-release formulation of perindopril erbumine loaded PEG-coated magnetite nanoparticles for biomedical applications

Iron oxide nanoparticles (FNPs) were synthesized due to low toxicity and their ability to immobilize biological materials on their surfaces by the coprecipitation of iron salts in ammonia hydroxide followed by coating it with polyethylene glycol (PEG) to minimize the aggregation of iron oxide nanoparticles and enhance the effect of nanoparticles for biological applications. Then, the FNPs–PEG was loaded with perindopril erbumine (PE), an antihypertensive compound to form a new nanocomposite (FPEGPE). Transmission electron microscopy results showed that there are no significant differences between the sizes of FNPs and FPEGPE nanocomposite. The existence of PEG–PE was supported by the FTIR and TGA analyses. The PE loading (10.3 %) and the release profiles from FPEGPE nanocomposite were estimated using ultraviolet–visible spectroscopy which showed that up to 60.8 and 83.1 % of the adsorbed drug was released in 4223 and 1231 min at pH 7.4 and 4.8, respectively. However, the release of PE was completed very fast from a physical mixture (FNPs–PEG–PE) after 5 and 7 min at pH 4.8 and 7.4, respectively, which reveals that the release of PE from the physical mixture is not in the sustained-release manner. Cytotoxicity study showed that free PE presented slightly higher toxicity than the FNPs and FPEGPE nanocomposite. Therefore, the decrease toxicity against mouse normal fibroblast (3T3) cell lines prospective of this nanocomposite together with controlled-release behavior provided evidence of the possible beneficial biological activities of this new nanocomposite for nanopharmaceutical applications for both oral and non-oral routes

Graphene oxide as a nanocarrier for controlled release and targeted delivery of an anticancer active agent, chlorogenic acid

We have synthesized graphene oxide using improved Hummer's method in order to explore the potential use of the resulting graphene oxide as a nanocarrier for an active anticancer agent, chlorogenic acid (CA). The synthesized graphene oxide and chlorogenic acid-graphene oxide nanocomposite (CAGO) were characterized using Fourier transform infrared (FTIR) spectroscopy, thermogravimetry and differential thermogravimetry analysis, Raman spectroscopy, powder X-ray diffraction (PXRD), UV–vis spectroscopy and high resolution transmission electron microscopy (HRTEM) techniques. The successful conjugation of chlorogenic acid onto graphene oxide through hydrogen bonding and π–π interaction was confirmed by Raman spectroscopy, FTIR analysis and X-ray diffraction patterns. The loading of CA in the nanohybrid was estimated to be around 13.1% by UV–vis spectroscopy. The release profiles showed favourable, sustained and pH-dependent release of CA from CAGO nanocomposite and conformed well to the pseudo-second order kinetic model. Furthermore, the designed anticancer nanohybrid was thermally more stable than its counterpart. The in vitro cytotoxicity results revealed insignificant toxicity effect towards normal cell line, with a viability of > 80% even at higher concentration of 50 μg/mL. Contrarily, CAGO nanocomposite revealed enhanced toxic effect towards evaluated cancer cell lines (HepG2 human liver hepatocellular carcinoma cell line, A549 human lung adenocarcinoma epithelial cell line, and HeLa human cervical cancer cell line) compared to its free form

Synthesis, characterization, controlled release and cytotoxic effect of anthranilic acid-loaded chitosan and polyethylene glycol-magnetic nanoparticles on murine macrophage raw 264.7 cells

Magnetic nanoparticles (MNPs) were prepared by the coprecipitation method using a molar ratio of Fe3+:Fe2+ of 2:1. The surface of MNP was coated with chitosan (CS) and polyethylene glycol (PEG) to form CS–MNP and PEG–MNP nanoparticles, respectively. Anthranilic acid (AA) was loaded on the surface of the resulting nanoparticles to form AA–CS–MNP and AA–PEG–MNP nanocomposites, respectively. The nanocomposites obtained were characterized using powder X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), thermogravimetry analysis (TGA), vibrating sample magnetometer (VSM) and scanning electron microscopy (SEM). XRD results showed that the as-synthesized nanocomposites are pure magnetite. FTIR results analysis indicated the existence of two polymers on the particle surface of the MNP and the presence of loaded AA on the surface of CS–MNP and PEG–MNP nanoparticles. Anthranilic acid loading and the release profiles of AA–CS–MNP and AA–PEG–MNP nanocomposites showed that up to 8.8% and 5.5% of the adsorbed drug were released in 670 min and 771 min, respectively. Anthranilic acid release profiles followed a pseudo-second-order kinetic controlled process. The cytotoxicity of the as-synthesized anthranilic acid nanocomposities were determined using MTT assay using murine macrophage RAW 264.7 cells. MTT results showed that the cytotoxic effects of AA–CS–MNP were higher than AA–PEG–MNP against the tested cells as compared to free anthranilic acid. In this manner, this study introduces novel anthranilic acid nanocomposites that can be used on-demand for biomedical applications

Graphene Oxide-Gallic Acid Nanodelivery System for Cancer Therapy

Despite the technological advancement in the biomedical science, cancer remains a life-threatening disease. In this study, we designed an anticancer nanodelivery system using graphene oxide (GO) as nanocarrier for an active anticancer agent gallic acid (GA). The successful formation nanocomposite (GOGA) was characterized using XRD, FTIR, HRTEM, Raman, and UV/Vis spectroscopy. The release study shows that the release of GA from the designed anticancer nanocomposite (GOGA) occurs in a sustained manner in phosphate-buffered saline (PBS) solution at pH 7.4. In in vitro biological studies, normal fibroblast (3T3) and liver cancer cells (HepG2) were treated with different concentrations of GO, GOGA, and GA for 72 h. The GOGA nanocomposite showed the inhibitory effect to cancer cell growth without affecting normal cell growth. The results of this research are highly encouraging to go further for in vivo studies

Development of anti-cancer and anti-hypertensive nanodelivery systems using magnetite iron oxide-polymeric nanoparticles

Nanoscience and nanotechnology have received considerable attention due to their benefits to many areas of research and application such as pharmaceutical industry, medicine, electronics and tissue engineering. Much of nanoscience and nanotechnologies are concerned with producing new materials especially for use as diagnosis, and drug delivery systems. According to the World Health Organization and Cancer Research UK, the top two causes of death in the world are due to the hypertension (one of the primary risks factors for cardiovascular diseases) and different known cancers that affect humans. As such, different types of carriers have been used to design different anti-cancer and anti-heypertensive therapeutic and diagnostic agents. The new developing drug delivery system has capability in which the drugs can be released in a sustained manner over long periods of time into the targeted tissue. Therefore, it enable an almost constant level of drug to be kept in the bloodstream (by injection method) or delivering it to a specific region of the gastrointestinal tract, orally for treatment of cancers and cardiovascular diseases. In order to reduce the toxicity of uncoated magnetite nanoparticles and prevent their aggregation which occurs due to dipole-dipole attraction of magnetic particles different biocompatible polymers were used as a coating material. One of the polymer that was used as a coating material for nanoparticles is a natural polymer, chitosan. Due to NH3+ groups of chitosan, it can be attracted by by –OH- groups of iron oxide nanoparticles to inhibit the nuclear growth of iron oxide. The other polymer known as poly ethylene glycol (PEG) which is soluble in both polar and nonpolar solvents due to the presence of polar oxygen atom and nonpolar (CH2)2 group in it. Also, because of coating the nanoparticles with a neutral and hydrophilic compound such as PEG and polyvinyl alcohol (PVA), the circulatory half-life can be increased from minutes to hours or days. This study aimed at the synthesis and development of several anti-cancer and an anti-hyperthensive nanodelivery formulations using iron oxide nanoparticles (FNPs) coated with different biocompatible polymers such as chitosan (C), PEG and PVA, loaded with different active drugs namely gallic acid (GA), 6-mercaptopurine (MP) and perindopril erbumine (PE). A total of 7 nanocomposites based on the aforementioned anti-cancer drugs; GA and MP and anti-hyperthensive drug; PE were prepared by co-precipitation method to increase the residence time in the body via a sustained release formulation to increase the clinical efficacy. All the three (3) active drugs (GA, MP and PE) were integrated separately into iron oxide-chitosan and iron oxide-PEG to form 6 new nanocomposites; FCG, FCMP-D, FCPE, FPEGG, FPEGMP-2 and FPEGPE, respectively. The active drug gallic acid (GA) was also loaded onto iron oxide nanoparticles-polyvinyl alcohol (FNPs-PVA) to form FPVAG nanocomposite. The release behaviour of the drugs from the nanocomposites in human body simulated phosphate buffer solutions (PBS) of intercellular lysosomal pH 4.8 and human blood pH 7.4 was found to be of sustained manner. The release of the drugs from FCG, FCMP-D, FCPE, FPEGG, FPVAG, FPEGMP-2 and FPEGPE nanocomposites in human body simulated phosphate buffer solutions (PBS) of human blood pH 7.4 is 1600, 6300, 5631, 6905, 6594, 5520 and 4223 minutes respectively, compared to 1300, 2500, 2743, 5775, 3045, 4440 and 1231 minutes respectively, at pH 4.8 (human body simulated PBS of intercellular lysosomal). It was found that all the nanocomposites were more biocompatible compared to free drugs although the choice of coating materials as well as loading percentages of active drugs on the nanocarrier was found to be affected by the activity of the resulting materials. Cytotoxicity study of FCG nanocomposite shows greater anticancer activity as was seen in MCF7 cell lines than in HT29 cell lines. Also, after 72 hours of treatment, the FCG nanocomposite was not toxic to a normal human fibroblast (3T3) cell lines in the tested doses. The FCMP-D nanocomposites, shows better anticancer activity against leukemia cell lines (WEHI-3B) than FCMP and pure drug. The IC50 for the FCMP-D is 1.19 ± 0.45 μg/mL compared to 4.94 ± 0.76 μg/mL for FCMP nanocomposite after 72 hours post treatment exposed to 0.47-30 μg/mL concentrations. It was found that the FPEGG nanocomposite demonstrated higher anticancer effect on the breast cancer cell lines (MCF7) in almost all concentrations tested (0.78-25.0 μg/mL) compared to FPVAG nanocomposite. Anticancer activity of FPEGMP-2 nanocomposite was found to be slightly higher than FPEGMP-0.5 in a dose-dependent manner on the leukemic cell lines (WEHI-3B) after 72 hours of treatment exposed to 1.9-60 μg/mL concentrations. This may be attributed to the differences in the percentage of 6-mercaptopurine between the two nanocomposites. Also, MP which is loaded into the surface of FNPs-chitosan compared to FNPs-PEG nanocarrier with the same molar ratio, shows better cytotoxicity effect which is due to the role of chitosan. The whole study shows that, iron oxide nanoparticles had a negligible effect in normal and all cancerous cell lines tested in this study. It was found that between 70-100% of cells remaining viable from 0.47 μg/mL to 60.0 μg/mL concentrations. Thus, the cytotoxicity to cancerous cell lines are likely attributable to release of active drugs (GA and MP) from the nanocarrier rather than the effect of the carrier itself. Therefore, this study demonstrated that all the new nanocomposites show controlled release property of the active drugs, and therefore can be exploited for drug delivery system. Results from in vitro studies were found to be very encouraging to further conduct the in vivo studies of these novel nanocomposites in the future