Elsevier

Biomedicine & Pharmacotherapy

Volume 77, February 2016, Pages 182-190
Biomedicine & Pharmacotherapy

Original article
Preparation and in vitro evaluation of 5-flourouracil loaded magnetite–zeolite nanocomposite (5-FU-MZNC) for cancer drug delivery applications

https://doi.org/10.1016/j.biopha.2015.12.025Get rights and content

Abstract

In this work, super paramagnetic magnetite nanoparticles were synthesized onto/into zeolite, then loaded with anti-cancer drug 5-fluorouracil (5-FU). The physical properties of the prepared nanocomposite and drug loaded nanocomposite were characterized using different techniques. The drug loading and releasing behavior of the magnetic nanocarrier was investigated and the drug-loaded nanoparticles exhibited a sustained release of drug without any burst release phenomenon. Furthermore, 5-FU loaded MZNC were evaluated for its biological characteristics. The functional 5-FU-MZNC has been triggered intra-cellular release of the cancer therapeutic agent 5-fluorouracil (5-FU). Cytotoxic effects of 5-FU loaded MZNC on human gastric carcinoma (AGS) cells were determined by real time cell analysis and colorimetric WST-1 cell viability assay. Apoptosis of cells was further investigated by Annexin-V staining which indicates the loss of cell membrane integrity. According to our results, 5-FU-MZNC showed a concentration-dependent cell proliferation inhibitory function against AGS cells. Morphologic and apoptotic images were consistent with the cytotoxicity results. In conclusion, 5-FU loaded MZNC efficiently inhibit the proliferation of AGS cells in vitro through apoptotic mechanisms, and may be a beneficial agent against cancer, however further animal study is still required.

Introduction

Nanomaterials can be used in many areas due to their electrical, optical, magnetic, and catalytic properties [1]. When nanomaterials turn into the hybrid material can be used in very different areas including protein/DNA bio-separation, disease diagnostics, cancer therapy, and drug and gene delivery [2], [3], [4], [5].

Among all fields, cancer therapy is undoubtedly the most studied area. Cancer is a kind of complex disease which occurs mainly due to genetic or environmental factors. It is difficult to control growth without cell death. Cancer causes to a solid mass of cells known as a tumor or a liquid cancer [6]. Cancer is the second most common cause of death in Europe and North America [7]. Generally cancer patients are treated with traditional therapies such as surgery, chemotherapy, and radiotherapy. Chemotherapy is a type of systemic therapy affecting the whole body, uses medicines to control, slow or cure the cancer [8]. Chemotherapeutic drugs have a low specificity for the targeted tumor because they are dispersed overall the body [9], so toxic agents cause a serious risk to healthy tissues beside cancer cells.

Anticancer drugs are chemicals that are used for the treatment of cancer. Anticancer drugs can be divided into different categories according to their mechanism of action, such as alkylating agents, anti-metabolites, antitumor antibiotics and topoisomerase inhibitors, hormones and hormone antagonists, herbal remedies and other medications [10]. Cyclophosphamide, doxorubicin, docetaxel, methotrexate and fluorouracil are chemotherapeutic agents. They can be used alone or combination with other agents [11]. In a number of studies in recent years, chemotherapeutic nanoparticles include 5-fluorouracil which is a typical pyrimidine analogue [12], [13], [14]. It is used as cytotoxic agent in cancers of the gastrointestinal tract, breast tumors, and skin cancers [15].

Chemotherapy drugs also have some side effects for instance short blood half-life, elimination by the immune system and nonspecific targeting. In order to eliminate these side effects nanocarriers can be used in drug delivery systems as drug carriers [16]. Existing methods of cancer treatment are improved using nanocarriers. They reduce side effects of cancer drugs [9], [18]. Nanocarriers can be composed of both organic and inorganic materials such as polymers, lipids (liposomes, nanoemulsions, and solid-lipid nanoparticles) self-assembling amphiphilic molecules, dendrimers, metals, and inorganic semiconductor nanocrystals (quantum dots) [19]. Living organisms are composed of cells with a diameter of around 10 μm. Uses of nanoparticles allow the observation of the functioning of the cells [1]. Nanoparticles have permeability and retention effect thus lead to leaky tumor vasculatures for better drug accumulation at the tumor sites [17]. With these advantages, nanoparticles gain importance to replace traditional chemotherapy.

Among nanocarriers, conventional magnetic material, Fe3O4 nanomaterials have been used in many fields such as lithium-ion batteries, wastewater treatment, magnetic resonance contrast media, therapeutic agents in cancer treatment and drug delivery. They have unique electric and magnetic properties. Especially superparamagnetic iron-oxide nanocomposites (SPIONs) are seen as a glimmer of hope for the cancer treatment because they have superparamagnetic behavior as well as surface-modification properties [20]. When they are used in biomedical applications they must have certain requirements. For instance in in vivo applications, encapsulation of magnetic nanoparticles in a biocompatible polymer have to be performed. Otherwise the original structure can change, large aggregates can form, and biodegradation can occur when interacted with biological systems. Thanks to the coating, drugs can bind [21].

Zeolites are a type of aluminosilicate crystal that can occur as natural or can be produced synthetically [22]. AlO4 and SiO4 tetrahedrons form nano size zeolite particles. Zeolites have microporous structure with cages and channels running through them. This architecture makes it as desirable candidates for treatment pollutants, sensing materials, and medical diagnostics [23]. They have somesignificant features such as small size, diverse frameworks, abundant accessible ion-exchanged sites, capacity for ionic exchange and high chemical, thermal, mechanical and radiation stability [24], [22]. Especially advantage of these constructions, drug molecules can diffuse out of the channels by slow degrees so the rate of release is under control [25]. In a recent study, Vilaça et al. used two different zeolite types as drug carrier. They measued the drug release kinetics and found that 80-90% of the drug was released in the first 10 min [26].

In this study the anticancer drug 5-fluorouracil (5-FU) was encapsulated into magnetite–zeolite nanocomposite particles. The purpose of this work is to investigate the shapes of the particles, their size, drug loading and releasing capacity and biological activities in gastric cancer cell line AGS. Various structural (FT-IR SEM, XRD, TG) and the magnetization (VSM) characterizations were performed.

Section snippets

Materials

Ferric chloride (FeCl3·6H2O), ferrous chloride (FeCl2·4H2O), ammonia water (NH4OH), sodium hydroxide, 5-flourouracil and zeolite4A (molecular sieve ∼350 mesh) were purchased from Sigma–Aldrich (St. Louis, MO, USA). All chemicals were of analytical grade and were used without further purification. Deionized milli-Q water was bubbled with high-purity nitrogen for 30 min before use.

Structural and physical characterization of samples

Structural and morphological features and thermal properties of the samples were investigated by Fourier transform

FTIR analysis

The FT-IR spectra of magnetite, zeolite 4A, MZNC, 5-FU and 5-FU-MZNC samples are recorded between 4000 and 400 cm−1. The presence of the magnetite nanoparticles is evidenced by the strong absorption bands υ1, observed at 600 cm−1 (red color) corresponds to intrinsic stretching vibrations of the metal at tetrahedral site (Fetetra–O), for naked magnetite but metal-oxygen band, υ2, octahedral-metal stretching (Feocta–O) disappeared or overlapped for this sample although observed at 445 cm−1, for MZNC

Conclusion

5-Fluorouracil was successfully loaded into magnetic zeolite nanocomposite yielding stable fluorescent hybrid nanoparticles with near-infrared optical properties. The biological assays confirmed the association of the 5-fluorouracil to the MZNC and their functionality in a cell-based assay system. The outcome of the current study is application a simple method for the preparation and characterization of 5-FU loaded superparamagnetic zeolite–magnetite nanocomposites which yielded a significant

Acknowledgement

The author is thankful to the Bezmialem Vakif University Research Project Foundation (Project no.: 9.2013/4) for financial support of this study.

References (32)

  • D.H. Han et al.

    Crystallite size effect on saturation magnetization of fine ferromagnetic particles

    J. Magn. Magn. Mater.

    (1994)
  • O. Salata

    Applications of nanoparticles in biology and medicine

    J. Nanobiotechnol.

    (2004)
  • M. De et al.

    Applications of nanoparticles in biology

    Adv. Mater.

    (2008)
  • G. Braun et al.

    Surface enhanced Raman spectroscopy for DNA detection by nanoparticle assembly onto smooth metal films

    J. Am. Chem. Soc.

    (2007)
  • N.L. Rosi et al.

    Oligonucleotide-modified gold nanoparticles for intracellular gene regulation

    Science

    (2006)
  • Ajit S. Narang et al.

    Anticancer drug development

    Pharmaceutical Perspectives of Cancer Therapeutics

    (2009)
  • Cited by (55)

    View all citing articles on Scopus
    View full text