Colloids and Surfaces B: Biointerfaces
Colloids and Surfaces B: Biointerfaces
a r t i c l e i n f o a b s t r a c t
Article history: Magnetic nanoparticles can transport drug and possibly target cancer. DNA-binding of ligands loaded in
Received 6 January 2015 dextran coated magnetic nanoparticles, could aid their better target-specific binding. In this work, we
Received in revised form 21 March 2015 report the loading of chromenones onto aminoethylamino-modified dextran coated iron oxide nanopar-
Accepted 20 July 2015
ticles, their loading efficiency, and openness for binding to DNA. The magnetic behavior, the size, and
Available online 28 July 2015
the morphology of the nanoparticles are analyzed. The crystallite size of the magnetic nanoparticles is
around 40 nm. The chromenones are present on the surface of the dextran shell, as revealed by their
Keywords:
cyclodextrin-binding characteristics, which is a new approach in comprehending the accessibility of
Dextran
DNA
the surface-bound molecules by macromolecules. The mode of binding of the chromenones to DNA is
Chromenone not altered on surface loading on dextran shell, although the binding strength is generally diminished,
Magnetic nanoparticles compared to the strength of binding of the free chromenones to DNA.
Iron oxide © 2015 Elsevier B.V. All rights reserved.
-Cyclodextrin
http://dx.doi.org/10.1016/j.colsurfb.2015.07.049
0927-7765/© 2015 Elsevier B.V. All rights reserved.
S. Yousuf et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 448–457 449
above binding problem has not been reported in detail. In this dextran containing ferrous-ferric solutions and stirred at ≈0 ◦ C. The
article, we present a detailed spectroscopic exploration of the bind- yellow precipitate turned to green, orange and finally dark brown
ing of a series of chromenone-loaded aminoethylamino modified at the gradual addition of ammonia. Then, it was refluxed at 80 ◦ C
dextran–coated iron nanoparticles (CHR-loaded IO-DX nPs) to DNA. for 3 h, and then cooled to room temperature, and the supernatant
Before studying their interaction with DNA, in order to comprehend solution was decanted. To the dark brown precipitate, 150 ml of
the accessibility of the chromenones, we employ -cyclodextrin as ethanol was added to do the aggregation of the colloidal particles.
a fit-and-find detector of their openness to binding. -Cyclodextrin The precipitate was centrifuged, washed, filtered, and dried to get
(-CD) is a doughnut-shaped cyclic oligosachcharide containing the dextran-coated iron oxide nanoparticles.
a hydrophobic cavity capable of accommodating guest molecules
of appropriate size [9]. The biological activities of chromenones 2.5. Preparation of aminoethylamino modified IO-DX
have been well-documented [10] and the bindings of native nanoparticles
chromenones to -CD and duplex DNA have been reported by
our research group [10–16] (Table 1). This article provides the Dextran coated iron oxide (100 mg) was dissolved in 100 ml
possible modes and strengths of binding of the chromenones water and sonicated for 10 min. 0.035 gm of NaBH4 and 35 ml of
viz., 2 -Hydroxyflavanone (2 HF), Hesperidin (HP), Naringin (NR), 2 M NaOH solution were added to the above solution, sonicated for
7-aminoflavone (7AF), Baicalein (BC), Coumarin153 (C153) and 10 min and then heated to 60 ◦ C with constant stirring. 20 ml of
Coumarin314 (C314) loaded, aminoethylamino-modified dextran- epichlorohydrin was added to the above solution in drops under
coated iron oxide nanoparticles to -CD and DNA. vigorous stirring for half an hour and kept overnight to form a col-
loid. The colloidal solution was centrifuged and the supernatant
2. Materials and methods solution was decanted to get the precipitate. Phosphate buffer
solution (40 ml) was used to disperse the precipitate. Aminoethy-
2.1. Chemicals lamino modified dextran–coated iron oxide (IO-DX) was prepared
by adding 35 ml of ethylene diamine to the above precipitate and
Calf thymus DNA (ctDNA), purchased from Genei (Merck), keeping at 50 ◦ C for 12 h. It was then cooled to room temperature,
India was used without further purification. ctDNA is supplied in centrifuged, filtered, and dried.
10 mM Tris-HCl (pH 8.0) and 1 mM ethylene diamine tetra acetic
acid (EDTA). The presence of 1 mM EDTA with ctDNA is used to 2.6. Preparation of CHR-loaded IO-DX nanoparticles
prevent nucleases from degrading the DNA. -Cyclodextrin and
dextran (MW 20,000) were purchased from Hi-Media, India. The The conjugation of IO-DX and the CHR molecules was carried
chromenones were obtained from Sigma, India. Ferrous chloride out by the addition of the IO-DX nanoparticles (5 mg) to 48, 122,
tetrahydrate, ferric chloride hexahydrate, epichlorohydrin, sodium 116, 47, 54, 62 and 63 mg of 2 -Hydroxyflavanone, hesperidin,
hydroxide, and ethylene diamine (AR, Aldrich) were used without naringin, 7-aminoflavone, Baicalein, Coumarin 153 and Coumarin
further purification. 314 respectively, in ethanol. In a typical procedure, the mixture was
stirred vigorously at 50 ◦ C for 30 min and kept at room temperature.
2.2. Preparation of test solutions. The solution was centrifuged to get the chromenone conjugated
IO-DX nanoparticles and the UV absorption of the supernatant solu-
1 mg/ml ctDNA was dissolved in NaCl solution of 50 mmol dm−3 tion was measured in order to find the non-loaded chromenone
prior to use, to obtain the concentration of 2.27 × 10−4 mol dm−3 molecules left behind in the solution. The photographs of the
which was calculated from the molar extinction coefficient of solutions of CHR-loaded IO-DX nPs are shown in SI2 in the Supple-
6600 dm3 mol−1 cm−1 at 260 nm. The purity of ctDNA sample was mentary data. The chromenone–conjugated IO-DX nanoparticles
confirmed with the yield of A260 /A280 of approximately in the range were filtered, crushed to get fine powder, stored at room tempera-
of 1.8–1.9 (where A represents the absorbance). Stock solutions ture.
of all the compounds were prepared in ethanol and diluted with
doubly-distilled water to prepare test solutions. Double-distilled 2.7. Instrumentation
water was used throughout the experiments. All the experiments
were carried out at an ambient temperature of (25 ± 2) ◦ C. Absorption measurements were done using UV–vis spectropho-
tometer (V-630, Jasco, Japan). Fluorescence spectra were recorded
2.3. Preparation of iron oxide nanoparticles using a spectrofluorimeter (FP750, Jasco, Japan), equipped with a
150 W xenon lamp for excitation. Both the excitation and the emis-
Ferrous chloride (2.6 g, 0.5 mol dm−3 ) and 3.3 g of ferric chloride sion band widths were set up at 5 nm. IR spectra were recorded
(1 mol dm−3 ) were dissolved in double distilled water and stirred with KBr pellets on a Perkin–Elmer spectrometer RXI, USA. Ultra-
for 1 h at 50 ◦ C under nitrogen atmosphere. 40 ml of ammonia solu- sonicator PCI 9L 250H, India was used for sonication. The pH was
tion was added in very small portions (in the range of 0.1 ml) to measured using an Elico LI 120 pH meter, India. The particle size
the above solution slowly until the solution turned dark brown. distribution of IO, IO-DX, and CHR-loaded IO-DX nPs was evalu-
The formation of iron oxide nanoparticles was confirmed using ated using dynamic light scattering measurements with a Malvern
UV–vis absorption, IR, SEM, EDX, Raman spectroscopy (SI 1 in the zetasizer nano ZS90, UK. Raman features of nPs were analysed
Supplementary data) and XRD. by Micro Raman spectrometer-800 (LabRAM HR model), France,
with the Argon Laser (514 nm) of 20 mW. The surface topology of
2.4. Preparation of dextran-coated iron oxide nanoparticles the chromenones, iron oxide nanoparticles, IO-DX nanoparticles
were imaged using JEOL JSM 6360 scanning electron microscope
Dextran (5 g, MW: 20,000) was dissolved in 20 ml double dis- (Japan). A minimum accelerating voltage of 15 KeV was used in
tilled water and it was sonicated for 30 min. Ferric chloride (0.65 g, EDX spectrometry. The diffraction pattern of the samples were
0.2 mol dm−3 ), ferrous chloride (0.51 g, 0.1 mol dm−3 ) were dis- recorded using a Shimadzu XRD 6000 X-ray diffractrometer (Japan)
solved in double-distilled water. The sonicated dextran solution using a monochromatic X-ray beam from CuK␣ radiation, operat-
was added to the ferrous-ferric solution slowly with constant ing at the voltage and current of 40 kV and 30 mA respectively. The
stirring. Liquid ammonia (40 ml) was added drop-by-drop to the magnetization measurements were done using a vibrating sam-
450 S. Yousuf et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 448–457
Fig. 2. Vibrating sample magneto metric spectra of free- and CHR-loaded IO-DX nPs (a) free IO-DX; and IO-DX conjugated with (b) 2’-Hydroxyflavanone, (c) Hesperidin, (d)
Naringin, (e) 7-Aminoflavone, (f) Baicalein, (g) Coumarin 153, (h) Coumarin 314.
Fig. 3. X-ray diffraction pattern of (a) free- and IO-DX nPs loaded with (b) 2’-Hydroxyflavanone, (c) Hesperidin, (d) Naringin, (e) 7-Aminoflavone, (f) Baicalein, (g) Coumarin
153, (h) Coumarin 314.
452 S. Yousuf et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 448–457
are in agreement with the standard card of magnetite PDF No. 01- molecule because the bulky dextran blocks one of the binding sites
071-6337. The noise observed at the background of the figures is of Naringin, i.e., the chromen-4-one moiety, as its carbonyl group
due to the amorphous dextran polymer shell. The crystallite sizes is less available on attachment to dextran for binding to -CD. Only
of the modified IO nPs fall in the range of 20 to 43 nm. Intense the phenolic moiety is available for binding to -CD and hence the
reflections are present at 2 values ≈25◦ corresponding to the 1:1 complex is formed.
experimental d–spacing for iron of ≈ 3.7 Å and hence, it is observed The binding of the IO-DX-attached compounds to -CD is
a high percentage of a single phase of Fe3 O4 is present. The crys- studied also using fluorescence titration (shown in SI10), as flu-
tallite size is found very close to the physical size of the IO nPs, orescence spectroscopy is a highly sensitive technique. The scope
which suggests that polycrystalline nanoparticles are not formed. of the fluorescence titration is to derive information on the stoi-
The Scanning Electron Microscopic images of the freshly prepared chiometry and the binding constant of the host–guest complexes.
IO, IO-D, IO-DX and various CHR-loaded IO-DX nPs are compared The binding plots are also shown in insets of figures in SI10.
with those of the free CHRs and shown in Fig. 4. The colloidal IO In all the cases except Coumarin 314, we observe fluorescence
nPs show randomly oriented irregular shaped structures due to the enhancement when -CD is added. For a simple 1:1 host–guest
anisotropic dipole–dipole interactions [23]. Dextran coating and complex, the I/I0 varies with the added -CD according to the fol-
attachment of compounds increase the net size of the IO-DX struc- lowing Eq. (5) [25]:
tures and, among most of them, quasi-linear cuboid or rod–like
I Imax K[CD]0
structures are observed. Lateral attraction between long chains of =1+ (5)
I0 (I0 − 1) 1 + K[CD]0
folded dextran sub-structures induces a self-organization and an
entropy–minimized, favored pattern formation on the surface of IO where Imax /I0 is the maximum enhancement of fluorescence and K
nPs. The striking differences in the morphology between the free- is the binding constant. A double-reciprocal plot of 1/Imax − I0 vs.
and the CHR-loaded IO-DX samples is an indication that the surface 1/[-CD] shows linearity for a 1:1 complex. In the case of 1:2 com-
of the IO-DX is altered at the addition of various compounds. The plex, the concentration of -CD is squared and 1/[-CD]2 is taken
average diameters of the ordered patterns of CHR-loaded IO-DX along the X axis and the plot is linear if the stoichiometry is 1:2. In
particles are in the range of a few hundreds of nanometers. the fluorescence titration of the CHR-loaded IO-DXs against -CD,
there are wavelength shifts of fluorescence maxima and enhance-
ment of fluorescence. The accommodation of guest molecules in
3.3. Binding of CHR-loaded IO-DX nPs to ˇ-CD
the -CD cavity exerts a restriction on the intra-molecular rota-
tional and vibrational freedom of the guest molecule. Also, the local
In general, between the free and the IO-DX-attached com-
polarity experienced by the guest molecule inside the hydrophobic
pounds, there is no appreciable shift of absorption band at the
cavity is much smaller than that in the aqueous solution. This leads
addition of -CD (The significant absorption spectral data of the
to a significant destabilization of the relative polar S1 ground state
CHR-loaded IO-DX nanoparticles, detailing the wavelengths of
and a smaller destabilization of the less polar S0 ground state. This
spectral bands and the shift of wavelengths on the addition of -CD
effect results further in a significantly larger S1 − S0 energy gap of
are given in SI9). This trend is quite commonly observed in bands
the encapsulated guest molecule [26]. Hence, the fluorescence of
corresponding to →* (shorter wavelength band) and n→* tran-
the guest molecule is blue-shifted. The fluorescence enhancement
sitions (longer wavelength bands). The shifts of absorption bands of
is a consequence of the increased energy gap, which reduces the
the IO-DX-attached compounds from those of the free unmodified
probability of the non-radiative decay and a corresponding increase
compounds, on -CD complex formation, are similar. However, in
in the fluorescence quantum yield [27]. For reasons given in the
the case of Coumarin 314, we observed a remarkable difference.
preceding paragraphs on the absorption spectra of Naringin, the
The 448 nm band of the free Coumarin 314 shifts to 454 nm on -
fluorescence titration of this compound against -CD shows a 1:1
CD complexation, whereas the 444 nm band of the IO-DX-attached
stoichiometry. The stoichiometry and binding constants of all the
Coumarin 314 shifts to 409 nm when -CD is added. These oppos-
host–guest complexes of the CHR-loaded IO-DXs are listed out in
ing red vs. blue shifts suggest that different kinds of interactions
Table 1.
exist between -CD and the free- and IO-DX-attached Coumarin
The stoichiometry is unaltered in all the cases except in
314.
Naringin. From the reported binding constants of the free CHR-
Using the absorption spectral data of the guest molecules (form-
-CD complexes, the binding constants of the CHR-loaded IO-DX
ing the host–guest complexes), the plots of 1/AS –A0 vs. 1/[-CD] of
nPs show a marked decrease in their magnitudes. The binding con-
various complexes are made, following the Eq. (4) [24]:
stant is dependent on the fit of the guest molecule inside the -CD
1 1 1 1 cavity as well as its polarity, and any other type of bonded inter-
= + (4)
As − A0 A − A0 A − A0 K ˇ − CD action in the complex. Since the polarity experienced by the native
compounds is different from that of their IO-DX-attached forms,
in order to determine the stoichiometry and the binding constants there occurs a different rate of enhancement of fluorescence. From
of the complexes. the observations in the absorption and fluorescence titrations, the
In the above equation, A0 refers to the absorbance of each one striking point seen is that when the compounds are loaded onto
of the chromophores (IO-DX-attached compounds) in water, AS IO-DX, they still bind to -CD and hence are locally accessible from
is the absorbance at different concentrations of the added -CD, outside the IO-DX surface. However, the binding constant values
and A’ is the absorbance at the highest concentration of -CD. Lin- are lowered, which implies that the strength of binding is altered.
ear plot of 1/ AS –A0 vs. 1/[-CD], in the case of any of the guest This again is a reflection on the accessibility of IO-DX-attached
molecules, leads to the inference that a 1:1 host–guest complex is compounds to -CD. Spectroscopic evidences suggest that the com-
formed. The binding plots made using absorption spectral data are pounds, when attached to IO-DX, are loosely bound on the surface
shown in the insets of Fig. 5. Indeed, we observed a linearity per- of the dextran shell covering the magnetic nanoparticle.
taining to the formation of 1:1 complexes in all the IO-DX-attached
compounds. A notable result is obtained in a structurally similar 3.4. Interaction of CHR-loaded IO-DX nPs with DNA
molecule, Naringin. The native form of Naringin forms a 1:2 -
CD complex, i.e., two -CD molecules bind one Naringin molecule. The CHR-loaded IO-DX nPs display hyperchromic shifts of
However, the IO-DX-attached Naringin binds to only one -CD absorption bands with little blue shifts (Fig. 6). Single isosbestic
S. Yousuf et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 448–457 453
Fig. 4. SEM images of (a) naked Iron oxide, (b) Iron oxide coated with dextran, (c) aminoethylamino modified dextran-coated iron oxide nanoparticles, (d) dextran, and free
and IO-DX nPs attached chromenones viz., (e–f) 2 -Hydroxyflavanone, (g–h) Hesperidin, (i–j) Naringin, (k–l) 7-Aminoflavone, (m–n) Baicalein, (o–p) Coumarin 153, (q–r)
Coumarin 314.
points are absent in some of the compounds, indicative of the fixed. The quenching of fluorescence is an indication of the bind-
absence just one mode of binding. In general, most of the com- ing interaction between each of these compounds and DNA. The
pounds display absorption characteristics similar to unmodified Stern–Volmer quenching constants (KSV ) are calculated from the
native compounds, on DNA binding. The shifts of absorption bands fluorescence titration data using the Eq. (6) [28]:
are listed out in SI11. There is a general decrease in the binding
strength of the CHR-loaded IO-DXs, from the binding strengths F0
of native chromenones. However, larger binding constant values = 1 + KSV [Q ] = kq 0 [Q ] (6)
F
of IO-DX-attached Naringin, and Coumarin 153 are observed on
DNA binding, compared to their corresponding native forms. An where F0 and F are the intensities of fluorescence of the flu-
increase in the binding constant value is an indication that the IO- orophores in the absence and the presence of DNA, kq is the
DX attachment does not hinder their binding to DNA. The notable bimolecular quenching constant, and 0 is the fluorescence lifetime.
general observations are (i) the IO-DX attachment of the com- The KSV values are listed out in Table 1.
pounds does stop their binding to DNA and (ii) dextran does not trap The fluorescence spectra of the IO-DX-attached Naringin and 7-
the IO-DX-attached compounds into their possibly coiled strands; aminoflavone show insignificant changes at the addition of DNA,
instead, these are available on the surface of the IO-DX and are suggesting that their binding to DNA is weak and it occurs in the
accessible to DNA. The binding of CHR-loaded IO-DXs to DNA is ground state, as shown by the absorption spectra. Baicalein, and
also studied using fluorescence spectroscopy (SI12). Coumarin 153 Coumarin 314 show enhancement of fluorescence at the addition
and Coumarin 314 show quenching of fluorescence on the addition of DNA. These show that the binding of these IO-DX-attached com-
of aliquots of DNA, keeping the concentration of the fluorophores pounds to DNA takes place. This is thought to occur due to the
454 S. Yousuf et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 448–457
Fig. 5. Absorption spectral titrations of the IO-DX-compounds–-CD binding (a) 2 -Hydroxyflavanone, (b) Hesperidin, (c) Naringin, (d) 7-Aminoflavone, (e) Baicalein, (f)
Coumarin 153, (g), Coumarin 314.
S. Yousuf et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 448–457 455
Fig. 6. Absorption spectral titrations of the CHR–loaded IO-DX–DNA binding (a) 2 -Hydroxyflavanone, (b) Hesperidin, (c) Naringin, (d) 7-Aminoflavone, (e) Baicalein, (f)
Coumarin 153, (g) Coumarin 314.
456 S. Yousuf et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 448–457
Table 1
Binding parameters of CHR-loaded IO-DX nPs– and free CHRs-binding to -CD and DNA.
UV Fluorescence UV Fluorescence
St. Binding Constant St. BindingConstant St. Binding Constant St. Binding Constant
2’HF 1:1 23 M−1 1:1 47 M−1 1:1 40.54 M−1 1:1 489 M−1
HP 1:2 6.66 × 104 M−2 1:2 1.46 ×104 M−2 1:2 2.84 × 104 M−2 1:2 9.63 × 103 M−2
NR 1:1 94 M−1 1:1 27 M−1 1:2 3.97 × 106 M−2 1:2 8.68 × 104 M−2
7AF 1:1 278 M−1 1:1 290 M−1 1:1 455 M−1 1:1 150.41 M−1
BC 1:1 69 M−1 1:1 36 M−1 1:1 117 M−1 1:1 291.88 M−1
C153 1:2 9.71 × 103 M−2 1:2 1.49 × 104 M−2 1:2 57.74 × 103 M−2 1:2 95.53 × 103 M−2
C314 1:1 84.49 M−1 1:1 202 M−1 1:1 1895.12 M−1 1:1 Ka =257 M−1
K and KSV values of free CHRs and CHR-loaded IO-DXs on DNA binding
UV Fluorescence UV Fluorescence
K (M-1 ) KSV (M-1 ) K (M-1 ) KSV (M-1 )
Table 2
Particle size, magnetic properties, and crystallite size of free- and CHR-loaded IO-DX nPs.
Sample Particle diameter d (nm) Coercivity Hc (G) MS (emu/g) MR (emu/g) Crystallite size in nm with Strain ()
Scheme 1. Summary of the experimental procedures involved and the various events of binding.
change of micro-environmental polarity through the removal of compounds attached to IO-DX, they cannot interact with DNA and
water molecules by intercalation [29,30]. their fluorescence does not change on the addition of DNA (a few
The binding of ligands to DNA can be tuned by - representative cases are shown in SI13). This is an evidence for the
CD–encapsulation of the compounds [31–33]. When -CD partially availability of the compounds on the surface of IO-DX, which ren-
covers up a guest molecule, the part of the molecule outside the - ders them accessible by DNA for binding, and inaccessible when
CD cavity binds to DNA. We applied this method to the CHR-loaded encapsulated by -CD. The experiments and inferences discussed
IO-DXs in their -CD–bound form. When in -CD–covers up the above are represented in the Scheme 1.
S. Yousuf et al. / Colloids and Surfaces B: Biointerfaces 135 (2015) 448–457 457
4. Conclusions [5] M. Calero, L. Gutiérrez, G. Salas, Y. Luengo, A. Lázaro, P. Acedo, M.P. Morales, R.
Miranda, A. Villanueva, Nanomed. Nanotech. Biol. Med. 10 (733) (2014),
http://dx.doi.org/10.1016/j.nano.2013.11.010
We report the loading of chromenones onto aminoethy- [6] J.M. Perez, A. Asati, S. Nath, C. Kaittanis, Small 4 (552) (2008), http://dx.doi.
lamino modified dextran-coated superparamagnetic iron oxide org/10.1002/smll.200700824
nanoparticles and their binding to -cyclodextrin and ctDNA. [7] K. Li, Y. Chen, J. Phys. Chem. C 118 (2014) 13876, http://dx.doi.org/10.1021/
jp500737c
The chromenones get loaded onto the shell of modified dextran, [8] H. Cho, D. Alcantara, H. Yuan, A.R. Sheth, H.H. Chen, P. Huang, S.B. Andersson,
with the involvement of hydrogen bonding between the modi- D.E. Sosnovik, U. Mahmood, L. Josephson, ACS Nano. 7(2032) (2013),
fied dextran and the chromenones. This conjugation, weaker than doi:10.1021/nn305962n.
[9] L. Feng, W. Li, J. Ren, X. Qu, Nano Res. (2014), http://dx.doi.org/10.1007/
covalent bonding, is best for delivery of drugs. They are acces-
s12274-014-0570-4
sible by -cyclodextrin in the formation of inclusion complexes. [10] I.B. Jaganath, A. Crozier, Dietary Flavonoids and Phenolic Compounds, Plant
The strengths of -CD complexes of CHRs surface-bound to IO- Phenolics and Human Health: Biochemistry, Nutrition, and Pharmacology,
Wiley, New York, 2010, pp. 1–4.
DX nPs are lesser when compared to the free chromenones’ -CD
[11] S. Chandrasekaran, Y. Sameena, I.V.M.V. Enoch, J. Mol. Recognit. 27(640),
complexes. The chromenone-loaded IO-DX nanoparticles are kept (2014), doi:10.1002/jmr.2387.
intact in solution and do not undergo precipitation within the [12] S. Chandrasekaran, Y. Sameena, I.V.M.V. Enoch, V. Santhanam, J. Solution
studied concentration range. The surface–loaded chromenones are Chem. 43 (2014) 1132, doi:10.1007/s10953-014-0187-y.
[13] S. Chandrasekaran, Y. Sameena, I.V.M.V. Enoch, Supramol. Chem. 26 (2014)
able to bind to DNA, although with a lesser binding strength than 740, doi:10.1080/10610278.2013.863313.
the free chromenone–ctDNA binding. Anticancer drug delivery [14] C. Sowrirajan, S. Yousuf, I.V.M.V. Enoch, Aust. J. Chem. 67 (2014) 256, http://
using iron oxide hinges on the possible directed transport of the dx.doi.org/10.1071/CH13364
[15] S. Yousuf, I.V.M.V. Enoch, AAPS Pharma. Sci. Tech. 14 (2013) 770,
magnetic nanoparticles into tumor cells. The drug delivery using doi:10.1208/s12249-013-9963-z.
drug–nanoparticle binding plays a vital mode of drug targeting, [16] S. Yousuf, D. Radhika, I.V.M.V. Enoch, M. Easwaran, Spectrochim. Acta A. 98
it can be utilized applying the idea of loading of DNA-interacting (2012) 405, doi:10.1016/j.saa.2012.08.068.
[17] G. Herzer, J. Magn. Magn. Mater. 112 (1992) 258.
molecules onto magnetic nanoparticles and delivering them into [18] M. Ma, Y. Wu, J. Zhou, Y. Sun, Y. Zhang, N. Gu, J. Magn. Magn. Mater. 268
cancer cells, by directing using an external magnetic field. Our work (2004) 33, doi:10.1016/S0304-8853(03) 426-8.
has explored such an important area in order to comprehend the [19] J.K. Vassiliou, V. Mehrotra, M.W. Russell, E.P. Giannelis, R. McMichael, R.D.
Shull, R.D.R.F. Ziolo, J. Appl. Phys. 73 (1993) 5109.
behavior of drug-loaded IO-DX nanoparticles as binding agents.
[20] A. Kostopoulou, K. Brintakis, M. Vasilakaki, K.N. Trohidou, A.P. Douvalis, A.
Lascialfari, L. Manna, A. Lappas, Nanoscale 6 (2014) 3764,
Acknowledgments doi:10.1039/c3nr06103e.
[21] S.P. Gubin, Magnetic Nanoparticles. Wiley-VCH, Verlag GmBH & Co.,
Weinheim, 2009.
We thank our Vice-Chancellor Dr. S. Sundar Manoharan, for [22] J. Liu, E.M. Lawrence, A. Wu, M.L. Ivey, G.A. Flores, K. Javier, J. Bibette, J.
his efforts in establishing our new research lab. The funding by Richard, Phys. Rev. Lett. 74 (1995) 2828.
[23] J.Y. Lee, Z. Shou, A.C. Balazs, Macromolecules 36 (2003) 7730,
the DST, Government of India, is acknowledged (Project: SR/FT/CS-
doi:10.1021/ma034765d.
062/2009). [24] A.A. Ensafi, H. Reza, E. Sara, J. Braz. Chem. Soc. 20 (2009) 266, http://dx.doi.
org/10.1590/S0103-50532009000200011
[25] A.M.P. Pena, F. Salines, M.J. Gomez, M.A. Acedo, M.S. Pena, J. Incl. Phenom.
Appendix A. Supplementary data
Mol. Rec. Chem. 5 (1993) 131.
[26] B.D. Wagner, In: Cyclodextrin Materials: Photochemistry, Photophysics, and
Supplementary data associated with this article can be found, in Photobiology. Ed.: Douhal A., Elsevier, 2006, p. 41.
the online version, at http://dx.doi.org/10.1016/j.colsurfb.2015.07. [27] R. Englman, J. Jortner, Mol. Phys. 18 (1970) 145.
[28] J. R. Lakowicz, Principles of Fluorescence Spectroscopy, 3rd Ed., Springer,
049 2006, 277–84 pp.
[29] H. Zang, K.S. Gates, Biochemistry 39 (2000) 14968, doi:10.1021/bi001998d.
References [30] N. Chitrapriya, Y.J. Yang, S.K. Kim, H. Lee, J. Inorg. Biochem. 105 (2011) 1569,
doi:10.1016/j.jinorgbio.2011.08.016.
[31] S. Yousuf, N. Sudha, G. Murugesan, I.V.M.V. Enoch, Carbohyd. Res. 365 (2013)
[1] X. Chen, O. Ramström, M. Yan, Nano Res. 7 (2014) 1381 http://dx.doi.org/10. 46, doi:10.1016/j.carres.2012.10.003.
1007/s12274-014-0507-y [32] S. Chandrasekaran, Y. Sameena, I.V.M.V. Enoch, Turk. J. Chem. 38 (2014) 725,
[2] A. Jordan, R. Scholz, P. Wust, H. Schirra, T. Schiestel, H. Schmidt, R. Felix, J. doi:10.3906/kim-1308-11.
Magnet. Magn. Mater. 194 (185) (1999) 7–558, http://dx.doi.org/10.1016/ [33] Y. Sameena, N. Sudha, S. Chandrasekaran, I.V.M.V. Enoch, J. Biol. Phys. 40
S0304-8853(98) (2014) 347, doi:10.1007/s10867-014-9355-y.
[3] V.P. Torchilin, Biopolym. (Pept. Sci.) 90 (2008) 604, http://dx.doi.org/10.1002/
bip.20989
[4] A. Schroeder, D.A. Heller, M.M. Winslow, J.E. Dahlman, G.W. Pratt, R. Langer, T.
Jacks, D.G. Anderson, Nat. Rev. Cancer 12 (39) (2011), http://dx.doi.org/10.
1038/nrc3180