Immobilization of protein on Fe3O4 nanoparticles for magnetic hyperthermia application.

We report a facile approach for the preparation of protein conjugated glutaric acid functionalized Fe3O4 magnetic nanoparticles (Pro-Glu-MNPs), having improved colloidal stability and heating efficacy. The Pro-Glu-MNPs were prepared by covalent conjugation of BSA protein onto the surface of glutaric acid functionalized Fe3O4 magnetic nanoparticles (Glu-MNPs) obtained through thermal decomposition. XRD and TEM analyses confirmed the formation of crystalline Fe3O4 nanoparticles of average size ~5 nm, whereas the conjugation of BSA protein to them was evident from XPS, FTIR, TGA, DLS and zeta-potential measurements. These Pro-Glu-MNPs showed good colloidal stability in different media (water, phosphate buffer saline, cell culture medium) and exhibited room temperature superparamagnetism with good magnetic field responsivity towards the external magnet. The induction heating studies revealed that the heating efficacy of these Pro-Glu-MNPs was strongly reliant on the particle concentration and their stabilizing media. In addition, they showed enhanced heating efficacy over Glu-MNPs as surface passivation by protein offers colloidal stability to them as well as prevents their aggregation under AC magnetic field. Further, Pro-Glu-MNPs are biocompatible towards normal cells and showed substantial cellular internalization in cancerous cells, suggesting their potential application in hyperthermia therapy.

A pH-responsive folate conjugated magnetic nanoparticle for targeted chemo-thermal therapy and MRI diagnosis.

Polyacrylic acid functionalized Fe3O4 nanoparticles (PAA-MNPs) of average size of 10 nm are prepared by a simple soft chemical approach. These PAA-MNPs are conjugated with folic acid through peptide bonding between the carboxylic group on the surface of PAA-MNPs and the amine group of folic acid. The good colloidal stability of FA conjugated MNPs makes it a promising candidate for targeted drug delivery, hyperthermia and as a MRI contrast agent with a transverse relaxivity R2 value of 105 mM(-1) s(-1). Folic acid conjugated magnetic nanoparticles (FA-MNPs) achieved ∼ 95% loading efficiency of doxorubicin (DOX) which could be due to strong electrostatic interaction of highly negatively charged FA-MNPs and the positively charged DOX. The drug release study shows a pH-dependent behavior and is higher in acidic pH (4.3 and 5.6) as compared to the physiological pH (7.3). Flow cytometry and confocal microscopic image analysis reveal that around 75-80% of HeLa cells undergo apoptosis due to DNA disintegration upon incubation with DOX-MNPs for 24 h. DOX-MNPs exhibit the synergistic effect due to the combination of DOX induced apoptosis and magnetic hyperthermia treatment (MHT) which enhance the cell death ∼ 95.0%. Thus, this system may serve as a potential pH sensitive nanocarrier for synergistic chemo-thermal therapy as well as a possible MRI contrast agent.

Tocotrienols potentiate lovastatin-mediated growth suppression in vitro and in vivo.

3-Hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase is the rate-limiting enzyme in the mevalonate pathway that provides essential intermediates for the membrane anchorage and biologic functions of growth-related proteins. Contrary to preclinical studies showing the growth-suppressive activity of statins, competitive inhibitors of HMG CoA reductase, clinical application of statins in cancer is precluded by their lack of activity at levels prescribed for the prevention of cardiovascular disease and by their dose-limiting toxicities at high doses. The dysregulated and elevated HMG CoA reductase activity in tumors retains sensitivity to the isoprenoid-mediated posttranscriptional down-regulation, an action that complements the statin-mediated inhibition and may lead to synergistic impact of blends of isoprenoids and lovastatin on tumor HMG CoA reductase activity and consequently tumor growth. d-gamma- and d-delta-tocotrienols, vitamin E isomers containing an isoprenoid moiety, and lovastatin-induced concentration-dependent inhibition of the 48-hr proliferation of murine B16 melanoma cells with IC50 values of 20 +/- 3, 14 +/- 3, and 1.5 +/- 0.4 microM respectively. A blend of lovastatin (1 microM) and d-gamma-tocotrienol (5 microM) totally blocked cell growth, an impact far exceeding the sum of inhibitions induced by lovastatin (12%) and d-gamma-tocotrienol (8%) individually. Synergistic impact of these two agents was also shown in human DU145 prostate carcinoma and human A549 lung carcinoma cells. C57BL6 mice were fed diets supplemented with 12.5 mg lovastatin/kg body weight, 62.5 mg d-delta-tocotrienol/kg body weight, or a blend of both agents for 22 days following B16 cell implantation; only the latter had significantly lower tumor weight than those with no supplementation. Co-administration of isoprenoids that posttranscriptionally down-regulate tumor reductase may lower the effective dose of statins and offer a novel approach to cancer chemo-prevention and/or therapy.