Vahid Jahed1, Ali Zarrabi2, and Mohsen Hajisadeghian3
1Faculty of Chemical engineering, Tarbiat Modares University, Tehran, Iran
2Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, 81746-73441, Iran
3Department of Plant Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran
Received: 03 December, 2016; Accepted: 25 January, 2017; Published: 28 January, 2017
Ali Zarrabi, Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, 81746-73441, Iran, Tel: +98 31 37934360; Fax: +98 31 37932342; E-mail:
Jahed V, Zarrabi A, Hajisadeghian M (2017) Two Faces of Curcumin; A Molecular Nutrition and an Anti-Cancer Agent. Int J Nanomater Nanotechnol Nanomed 3(1): 019-022. DOI: 10.17352/2455-3492.000016
Â© 2017 Jahed V, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Curcumin, Aqueous solubility, Cyclodextrin, Inclusion complex
Curcumin, a useful herbal medicine with anti-inflammatory and anti-cancer properties is insoluble in water which restricts its therapeutic properties; its prominent application in cancer treatment is limited due to curcumin sub-optimal pharmacokinetics and poor bioavailability at the tumor site. Cyclodextrins have the ability to increase the solubility of many hydrophobic drugs. Several complexes have been proposed to improve the solubility of curcumin. In this study, we have demonstrated the inclusion complexation of equimolar β-cyclodextrin and curcumin (βCD-CUR) which is confirmed through several analytical methods such as FTIR and DSC analysis. Finally, cell viability test as well as cell morphology assessment were conducted using MTT assay on MCF7, human breast cancer cell line and L929, fibroblastic normal cells. The results showed that the inclusion complex formulation had no significant cytotoxic effect on normal cells, while there was a significant morphological changes and shrinkage in cancer cells treated by βCD-CUR.
Currently, the most common way to treat cancer is chemotherapy. While toxic and expensive, the possibility of treatment with chemotherapy agents is very low [1,2]. Among cancers, breast cancer is the second leading cause of death for women in the world Fateme et al. 2011, and . The use of herbal medicine is one of the easiest and the most inexpensive ways to prevent and treat cancer . Curcumin (CUR), the active ingredient of turmeric spice and famous Hindi; solid gold. Also, it has a broad use as a food habit in Japan and South Asia  is one of the herbal medicines that is recently been highlighted in cancer treatment [5,6,7]. It has been proven that curcumin, as the main component of turmeric, has anti-carcinogenic and anti-proliferative features [8,9,10]. Moreover, it has been reported that telomerase activity in MCF-7 cells was increased by curcumin treatment . Despite many medicinal features of curcumin, its low solubility and instability in aqueous media are major drawbacks which prevent its widespread applications . To improve this limitations, variety of formulations have been proposed .
Cyclodextrins are donut-shaped sugar molecules [14,15,16,17] with hydrophobic cavity and hydrophilic shell, which enables them to increase the aqueous solubility of hydrophobic guests . Thus, cyclodextrins have gained more attention as drug carriers in pharmaceutical industries . According to the internal cavity size , β-cyclodextrins (βCD) are the most suitable cyclodextrins to make inclusion complex with hydrophobic molecules such as curcumin. In previous research, we have shown an increased curcumin solubility when making inclusion complex with βCD . Zhang et al have demonstrate that βCD-CUR inclusion complex and sustained release of curcumin could amend therapeutic efficacy in lung cancer comparing with free curcumin . In this research, molecular inclusion complex of βCD-CUR has been verified by FTIR and DSC. Then, Its cellular effect on MCF-7 and L929 cell lines as cancer and normal cells has been investigated.
Materials and Methods
β- cyclodextrin (βCD, 98%, Sigma Aldrich) and curcumin (CUR, >99%, Sigma Aldrich) were used as received. All solvents (acetone, DMSO-d6 and ethanol) were of high purity and were obtained from Merck Company (Germany).
Inclusion complex formation
βCD- CUR inclusion complex was prepared by freeze dry method . Briefly, 40 mg βCD and 12 mg CUR were dissolved in 8 mL deionized water and 0.5 mL acetone in two different glass vials (Fisher Scientific, Pittsburgh, PA, USA), respectively. These solutions were then mixed and put on the stirrer for 24 h without a cap to evaporate the acetone. Then, the solution was centrifuged (1000 rpm, 5 min) and the supernatant containing βCD–CUR inclusion complex was freeze dried (VaCo5, Zirbus)
FTIR: FTIR spectra of βCD, CUR and βCD-CUR inclusion complexes were recorded using a Smiths Detection FT Infrared Spectroscope, JASCO (Japan). FTIR spectra of samples were acquired by placing fluffy powder on the tip of the ATR. Data was acquired between 4000 cm−1 and 400 cm−1 at a scanning speed of 4 cm−1. The average of 10 scans was presented as FTIR spectra. The KBr disc method was used for solid samples.
DSC analysis: DSC analysis of Curcumin, βCD and βCD-CUR inclusion complex were performed by using Q200 Differential Scanning Calorimeter (TOLEDO, Germany) at Isfahan Science and Technology Town. Heating rate employed was 10 °C/min from 25 to 210 oC under a nitrogen gas constant flow.
MTT assay: MCF7 human breast cancer cell line and L929 fibroblastic normal cells in flasks and vials were taken from Pasteur Institute of Iran. These cell lines were resistant to the drug Cisplatin treatment. In this study, the RPMI 1640 culture with 10% FBS and 1% penicillin (50 U/ mL) -streptomycin (50 μg/ mL) was used and incubated in CO2 5% (N-Biotech) at 37 ºC. The anti-cancer and anti-proliferation effect of βCD-CUR and CUR have been studied on normal cells and breast cancer cell lines (5,000 cells per 100 µl medium) in 96-well plates.
After 24 hr, the medium was replaced with fresh medium containing 10 to 50 µM concentrations of βCD-CUR solution. Equal concentrations of βCD were prepared as control.
After two days, the supernatant medium was discarded and 100 µl of MTT solution was added to each well. After 4 hours of incubation at 37 °C, 100 µl acidic isopropanol was added to each well to lyse cells. Using the ELISA reader, the absorption was read at 570 nm (Ref: 630 nm). All concentrations of curcumin have been used in the blank solution construction, so the yellow color of Curcumin was foiled in the reading MTT result.
All tests were repeated three times.
5ml culture medium containing 1×106 cancer cells was transferred to a T25 flask. After 24 hours, the βCD-CUR solution with a concentration of 50 µM was added to the medium. Then, any significant morphological changes has been studied under inverted microscope (Motic, AE2000) at a magnification of 20X.
One way ANOVA test and LSD test in SPSS software version 17.00 was used for statistical analysis of data and the T test has been used for independent samples.
Results and Discussion
Differential scanning calorimeter (DSC) is useful tool for recognition of host-guest inclusion complexes. It is concluded from DSC analysis that the inclusion complex is formed when exothermic or endothermic peak of the guest molecule is disappeared. The results of this analysis is shown is Figure 1. The curve of CUR shows an endothermic peak at 178 0C (melting temperature of CUR), whereas that of βCD has an endothermic peak at 124oC which is attributed to the removal of water molecules from the cavity. However, inclusion complex is formed when water molecules are replaced by less polar molecule like curcumin. Thus, the melting point of curcumin could be influenced after coverage by βCD.