See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/270913553

Physical and biological properties of the ion beam irradiated PMMA-based

composite films

Article  in  Applied Surface Science · February 2015

DOI: 10.1016/j.apsusc.2014.12.129

CITATIONS READS

7 216

9 authors, including:

Shanthini Muthukoori Catherine Martin

Anna University, Chennai Anna University, Chennai
6 PUBLICATIONS   32 CITATIONS    4 PUBLICATIONS   9 CITATIONS   

SEE PROFILE SEE PROFILE

Sakthivel Nagarajan Sarath Chandra Veerla

Anna University, Chennai Chosun University
21 PUBLICATIONS   76 CITATIONS    22 PUBLICATIONS   291 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Electronic Devices View project

This work is to understand the influence of divalent substitutions on the electromagnetic properties of Ni-Zn ferrites. View project

All content following this page was uploaded by Shanthini Muthukoori on 19 June 2019.

The user has requested enhancement of the downloaded file.

 Applied Surface Science 329 (2015) 116–126

Contents lists available at ScienceDirect

Applied Surface Science

journal homepage: www.elsevier.com/locate/apsusc

Physical and biological properties of the ion beam irradiated

PMMA-based composite films
G.M. Shanthini a , Catherine Ann Martin a , N. Sakthivel a , Sarath Chandra Veerla a ,
K. Elayaraja a , B.S. Lakshmi b , K. Asokan c , D. Kanjilal c , S. Narayana Kalkura a,∗
a
Crystal Growth Centre, Anna University, Chennai 600025, India
b
Department of Biotechnology, Anna University, Chennai 600025, India
c
Materials Science Group, Inter University Accelerator Centre, Aruna Asaf Ali Marg, New Delhi 110067, India

a r t i c l e i n f o a b s t r a c t

Article history: Polymethyl methacrylate (PMMA) and PMMA-hydroxyapatite (PMMA-HAp) composite films, prepared
Received 1 September 2014 by the solvent evaporation method were irradiated with 100 MeV Si7+ ions. Crystallographic, morpho-
Received in revised form logical and the functional groups of the pristine and irradiated samples were studied using glancing
16 December 2014
incident X-ray diffraction (GIXRD), scanning electron microscopy (SEM) and Fourier transform infrared
Accepted 18 December 2014
spectroscopy-attenuated total reflectance (FTIR-ATR) respectively. SEM reveals the creation of pores,
Available online 26 December 2014
along with an increase in porosity and cluster size on irradiation. Decrease in crystalline nature and crys-
tallite size with an increase in ion fluence was observed from GIXRD patterns. The surface roughness and
Keywords:
SHI irradiation
the wettability of the material were also enhanced, which could favour the cell–material interaction. The
PMMA irradiated samples adsorbed significantly greater amount of proteins than pristine. Also, irradiation does
Hydroxyapatite not produce any toxic byproducts or leachants, and maintains the viability of 3T3 cells. The response of the
Protein adsorption irradiated samples towards biomedical applications was demonstrated by the improved antimicrobial
Cytotoxicity activity, haemocompatibility and cytocompatibility. Swift heavy ion irradiation (SHI) could be an effec-
tive tool to modify and engineer the surface properties of the polymers to enhance the biocompatibility.

© 2014 Elsevier B.V. All rights reserved.

1. Introduction cements and orthopaedic fixations such as hip and knee joints as
it is a thermoplastic polymer possessing good biocompatibility [3].
Hydroxyapatite (HAp – Ca10 (PO4 )6 OH2 ) is used widely for bone The gamma irradiation induces physical and structural changes in
implant and bone cement applications due to its compositional PMMA [4]. Antibiotic loaded PMMA has been successfully demon-
and biological similarities to the native tissues. Both dense and strated as a local drug delivery system [5,6]. The addition of,
porous HAp have been vigorously investigated as implant materials small volume of particulates such as hydroxyapatite, strengthens
for orthopaedic and dental applications [1,2]. Due to the bioactive the material [7,8]. Polymeric biomaterials have good bulk proper-
nature and stability of HAp on contact with the aqueous medium ties, but lack sufficient surface properties in terms of efficient cell
at body temperature, it has been widely accepted as biocompat- response [9–11]. The surface properties such as surface morphol-
ible material. In spite of these advantages, HAp cannot be used ogy, topography, chemical structure, wettability, and interfacial
as such for load bearing applications due to its brittleness. There- free energy play a major role in biocompatibility of the biomaterial
fore, the composites of HAp with polymers can compensate the [12–15]. All these essential surface properties can be attained using
poor mechanical properties of HAp. For long-term clinical applica- a single tool swift heavy ion irradiation.
tions, the HAp-based polymer composite can be prepared using Ion irradiation provides extraordinary effects on a wide range of
a wide range of degradable and non-degradable polymers such materials which is quite unfeasible by any other methods and is of
as polyethylene, polypropylene, PMMA, polyesters, polysaccha- two types, namely low energy (<few tens of MeV) and high energy
rides, etc. Among these polymers, PMMA is used to prepare bone (>few tens of MeV) ion beams, which are classified based upon the
energy at which it travels. Low energy ions lose their energy by elas-
tic collisions with the nucleus of the target atom, while high energy
∗ Corresponding author: Tel.: +91 044 2235 8335. ions lose their energy through inelastic collisions with target elec-
E-mail address: [email protected] (S.N. Kalkura). trons [16–18]. The energy loss with the nucleus and electrons of the

http://dx.doi.org/10.1016/j.apsusc.2014.12.129
0169-4332/© 2014 Elsevier B.V. All rights reserved.
 G.M. Shanthini et al. / Applied Surface Science 329 (2015) 116–126 117

target atom is called as nuclear energy loss (dominant in the case represented as Ppris, P1e11, P5e11 and P1e12. Similarly, PMMA-
of low energy ion implantation and negligible as a result of swift HAp composites will be represented as PHpris, PH1e11, PH5e11
heavy ion (SHI) irradiation) and electronic energy loss (dominant and PH1e12.
in the case of SHI irradiation and negligible as a result of low energy
ion implantation) respectively [19–21]. In general, biomaterials are
fabricated using polymers, ceramics, metals and composites. These 2.3. Characterization
biomaterials possess good bulk properties such as haemocompat-
ibility, cytocompatibility, swelling, dissolution and water uptake Atomic Force Microscopic (AFM) images were recorded using
but lack in their surface properties which could be overcome by Park XE-100 in tapping mode in the scan area of 10 ␮m × 10 ␮m.
means of SHI irradiation. PMMA and HAp based samples were syn- Glancing incident X-ray diffraction (GIXRD) was carried out on
thesized and irradiated with different ranges of ions to study the pristine and irradiated samples using Bruker AXS Diffractometer
effect of irradiation on bioactivity [22] and the dielectric proper- applying the maximum power of 40 kV/40 mA and the X-ray source
ties [23]. Shahabi et al. have reported that the gamma irradiation used was Cu K␣ radiation, with a scan rate of 2◦ min−1 at an incre-
of PLGA-PEG-HAp composite has resulted in enhanced biocompat- ment of 0.02◦ . The glancing angle was fixed at 2◦ . The experiment
ibility [24]. was carried out in detector-scan mode, in the range of 10–60◦ .
In the present study SHI irradiation is used to engineer The surface morphology of the samples before and after irradiation
the surface properties of polymethyl methacrylate (PMMA) and was analyzed using Hitachi S3400-N Scanning Electron Microscopy
PMMA-hydroxyapatite (PMMA-HAp) films. The phenomena such (SEM) with an accelerating voltage of 5 kV and the measurements
as Coloumb expansion [25,26] and thermal spike [27–29] are com- were done using “Image J” software. The functional groups of the
monly employed mechanisms to understand the effect of swift pristine and irradiated samples were analyzed using FTIR Jasco-
heavy ion irradiation on the surface properties [30,31]. 6300 in the Attenuated Total Reflectance mode in the wavenumber
In spite of various researches based on synthetic biomaterials, range of 400–4000 cm−1 . The glass transition temperature of the
the major drawbacks lie in the initial cell adhesion and prolifera- irradiated samples was determined using DSC Exstar-SII DSC 6220
tion due poor surface properties. There are many reports dealing by purging nitrogen gas. DSC measurements were made in 3 mg
with change in surface properties by chemical and biological mod- samples in the temperature range 50–200 ◦ C at 10 ◦ C/min.
ification, changing the surface roughness and wettability, thermal
treatment, etching, etc., and examining its effect on cell adhesion
2.4. Wettability
and proliferation. But very few researchers report the effect of SHI
irradiation on surface properties and its biological applications. In
The static water contact angle measurement of pristine and
this work we report the defect formation caused by the thermal
irradiated PMMA and PMMA-HAp was performed by the sessile
spike due to SHI and its effect on physical properties and initial
drop method using Euromex optical microscope with inbuilt colour
biological evaluation of irradiated films.
Charge Coupled Device (CCD) camera at room temperature. Deion-
ized water with drop size of 10 ␮L was used to measure the
2. Materials and methods wettability. The contact angles of the sessile drop in contact with
air, liquid and material were measured in three samples of each
2.1. Preparation of PMMA and PMMA-HAp composites condition, with the help of “ImageJ” software.

The polymer films were prepared using the solvent cast-

2.5. Antimicrobial activity
ing/evaporation method. HAp was synthesized by the wet chemical
method. 1 M calcium nitrate tetrahydrate (Ca(NO3 )2 ·4H2 O, Merck)
The disk diffusion method was employed to obtain the suscep-
was prepared and added drop wise to 0.6 M diammonium hydrogen
tibility pattern of Escherichia coli and Staphylococcus aureus against
phosphate ((NH4 )2 ·HPO4 , Merck) with continuous stirring. The pH
gentamicin loaded pristine and the irradiated samples. The sam-
of the mixture was maintained at 10, throughout the reaction (using
ples were cut into discs of 5 mm diameter and dispersed in the
ammonia solution). The obtained white precipitate was subjected
gentamicin solution of concentration 2 mg/mL for 24 h. The nutri-
to microwave irradiation (Household microwave oven, 900 W and
ent agar was poured into the petri plate and allowed to solidify at
2.45 GHz) for 15 min. The precipitate was washed repeatedly with
room temperature. About 10 ␮L of E. coli and S. aureus cells, which
deionized water to remove excess ammonia and dried in oven for
were cultured in the nutrient broth were added to the solidified
24 h at 80 ◦ C. For film preparation, PMMA (Alfa Aesar) (10 wt/v%)
agar medium and swabbed uniformly throughout the plate. The
was dissolved in acetone. To the prepared polymer solution, HAp
drug-loaded samples were then blotted on tissue paper to remove
in powder form, was added in 10:1 (PMMA:HAp) ratio and cast in
the excess drug and placed over the bacteria inoculated plate. The
a glass petri plate. HAp powder was thus dispersed in a polymer
plates were incubated at 37 ◦ C for 24 h and the inhibition zones
matrix. PMMA films were also prepared using the same procedure
were measured in all directions and the average was calculated
without the addition of HAp.
and recorded.

2.2. Irradiation experiments

2.6. Haemocompatibility
PMMA and PMMA-HAp composite films were irradiated with
Si7+ ions of energy 100 MeV and current 1pnA in three different flu- 2.6.1. Blood collection and preparation
ences 1 × 1011 ions/cm2 , 5 × 1011 ions/cm2 and 1 × 1012 ions/cm2 . The human blood was collected and protected from clotting by
SHI irradiation was carried out using 15 UD pelletron accelerator mixing the blood with ACD (Acid Citrate Dextrose) in the ratio
at Inter-University Accelerator Centre (IUAC), New Delhi. The sam- of 9:1. The ACD was prepared by mixing anhydrous citric acid,
ples used for beam scan where of 1.6 cm × 1.6 cm dimension and trisodium citrate dihydrate and dextrose monohydrate. Platelet-
300 ␮m thick. The scanning was performed using, magnetic scan- poor plasma (PPP) was separated by subjecting the whole blood for
ner. Hereafter, PMMA pristine and PMMA irradiated with fluences centrifugation at 3000 × g for 15 min and used immediately for the
1 × 1011 ions/cm2 , 5 × 1011 ions/cm2 and 1 × 1012 ions/cm2 will be quantification of protein adsorption.
118 G.M. Shanthini et al. / Applied Surface Science 329 (2015) 116–126

2.6.2. Haemolysis to each sample and control well and incubated for 4 h. The medium
Haemolysis experiments were carried out with human blood by was then removed and 100 ␮L of DMSO was added to dissolve the
a haemolytic assay method. 100 mg of accurately weighed pristine formazan product. MTT is reduced to purple coloured formazan
and irradiated samples were utilized for the assay. The samples and crystals inside the metabolically active live cells. Hence, the colour
PBS (pH 7.4) were sterilized under UV for 30 min and subjected intensity of formazan is directly proportional to cell viability. The
for haemolysis. The sterilized samples were equilibrated in 3 mL absorbance of the formazan was then measured at 595 nm using
of PBS for 24 h at 37 ◦ C, after which the PBS was removed com- Multiskan plate reader, Thermo scientific, USA.
pletely and 250 ␮L of ACD blood was added and incubated at 37 ◦ C
for 20 min. To this, 2 mL of PBS was added and incubated for 1 h to
3. Results and discussion
stop haemolysis. After 1 hour of incubation the samples were cen-
trifuged for 5 min at 1500 rpm and the supernatant was collected
3.1. Energy loss and penetration depth
to determine the percentage haemolysis. Two samples each con-
taining human blood with deionized water and human blood with
On irradiating the PMMA and PMMA-HAp films with 100 MeV,
PBS were served as a positive and negative control respectively. The
Si7+ ions in fluence-range of 1 × 1011 ions/cm2 to 1 × 1012 ions/cm2 ,
optical density (OD) of the supernatants was measured at 545 nm
the samples experienced two types of energy losses, namely,
and the percentage haemolysis was calculated using the formula:
nuclear energy loss (Sn ) and electronic energy loss (Se ). The energy
 OD of test sample − OD of negative control 
loss of Si7+ ions on irradiation with the sample was calculated using
%haemolysis =
OD of positive control − OD of negative control SRIM (Stopping and Ranging of Ions in Matter) 2008 as shown in
Fig. 1a and b. With this simulation program, it was found that,
× 100% (1)
until 70 keV nuclear energy loss was much pronounced. Above
70 keV, electronic energy loss is dominant. The penetration depth
The percentage haemolysis was then compared with ASTM stan- of 100 MeV Si7+ ions, in PMMA and PMMA-HAp was found to be
dards: 59 ␮m and 60 ␮m respectively.

Highly haemocompatible – if less than 5% haemolysis; 3.2. Ion-material interaction

Haemocompatible – if within 10% haemolysis;
Non haemocompatible – if greater than 20% haemolysis [32]. The passage of swift heavy ions vertically through the solid
material (PMMA and PMMA-HAp films), bring about disturbances
2.6.3. Protein adsorption to about few nanometers of diameter in the cylindrical path (Fig. 2)
The samples were cut into 5 mm × 5 mm dimension and were and to the depth of a few microns depending upon the mate-
immersed in 2 mL of Platelet Poor Plasma (PPP) and incubated at rial. Results that the disturbances can make are the formation of
37 ◦ C for 90 min. The excess protein solution was then removed defects and craters, amorphization, chain scission and crosslink-
and the samples were then rinsed with PBS. The concentration ing [34–36]. When an ion with higher energy is traversed through
of the plasma proteins adsorbed to the sample surface was then the material, it undergoes an electronic collision with the atoms
quantified using microBCA assay kit (Thermo scientific) based on in its straight path, leaving behind the charged/ionized state and
the manufacturer’s protocol. The standard used to quantify the comes to rest when it loses all its kinetic energy to the target atoms.
unknown protein concentration is plotted with BSA [33]. The electrons which are directly knocked-off by the incoming ion
undergo sputtering or collision cascade/recoiling. Collision cascade
2.7. Cytotoxicity assay is the successive collisions of the primary electron with other atoms
in the ion track. The atoms involved in the successive collisions are
Viability of 3T3 cell lines (NCCS, Pune) was determined using called as recoil atoms and the electrons ejected from these atoms
MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bro- are secondary electrons. The collision of the impinging ion/primary
mide) assay. The samples of 5 mm diameter were placed in the 48 electrons with the atoms results in the electronic excitation. As
well plates and the cells were seeded to it followed by incubation the cascade proceeds there will be a constant decrease in energy
at 37 ◦ C, 5% CO2 for 24 h in DMEM medium. After incubation, 10 ␮L transfer. When there is no ample energy to excite an electron, the
of 5 mg/mL MTT (Invitrogen) solution prepared in PBS was added remaining energy will be converted to heat instigating the thermal

Fig. 1. (a) Electronic and nuclear energy loss in PMMA and PMMA-HAp films; and (b) penetration depth of silicon ion as a function of energy in PMMA and PMMA-HAp films.
 G.M. Shanthini et al. / Applied Surface Science 329 (2015) 116–126 119

Fig. 2. Mechanism of ion-material interaction (a) unirradiated material; (b) SHI irradiation and thermal spike mechanism: (i) incident ion hits the target atom, (ii) collision
cascade, (iii) thermal spike; (c) nanostructures formed as the effect of SHI interaction with material; (d) SEM images of pores and nanostructures formed in PMMA and
PMMA-HAp.

spike in the local region to about few microns for a few picosec- were ascertained using XRDA software and with the help of these
onds. The heating up of atoms leads to atomic motion/vibration parameters the crystallite size and crystallinity were calculated
until quenching (dissipation of heat to the lattice to attain room using the Scherrer’s formula:
temperature). Heat dissipation to the lattice causes amorphization
K
and the sudden quenching leads to displacement of atoms forming L= (2)
ˇ002 cos 
pores and nanostructures on the surface [37–39].
where L is the crystallite size, K is constant 0.9,  is the wavelength
3.3. Glancing incident X-ray diffraction (GIXRD) of X-rays (1.5405), ˇ002 is Full Width Half Maximum (FWHM) of
(0 0 2) peak. The percentage crystallinity was calculated using:
The GIXRD pattern as shown in Fig. 3, revealed the crystalline
Ic
nature of PMMA-HAp composite. The broad peak in between 10◦ Xc = (3)
Ic + Ia
and 25◦ represents the amorphous PMMA. The addition of PMMA
with the HAp has lead to the peak shift, which might be due to where Xc is the degree of crystallinity. Ic and Ia are the integrated
the interaction of PMMA molecules with HAp particulates. A sim- area of crystalline and amorphous peaks respectively [22].
ilar peak shift in the XRD pattern of the composites of sodium There was a decrease in crystallinity and crystallite size with an
alginate-hydroxyapatite [40], Chitosan phosphate-hydroxyapatite increase in the fluence (Table 1). The variation in crystallinity and
[41] and Chitosan-hydroxyapatite [42] was reported on the addi- crystallite size might be due to the irradiation induced lattice strain.
tion of polymer as composites with the hydroxyapatite due to During the irradiation, the sample surface gets heated and under-
polymer-hydroxyapatite interaction. All the peaks of irradiated goes local melting followed by rapid heat dissipation. The structure
samples are identical to the pristine sample and matches with the disorder due to the increase in temperature, leads to the decreased
pure phase of HAp according to the JCPDS: 09-0432. Also, the pat- crystallinity. When there is sudden heat dissipation, the molten,
tern proved that there was no phase change due to the irradiation. disordered structure tries to recrystallize leading to the consecu-
The major plane was observed in (2 1 1). The lattice parameters and tive increase of crystallinity and crystallite size. The samples were
the FWHM of the peaks for the pristine and the irradiated samples irradiated in the fluences of 1 × 1011 ions/cm2 , 5 × 1011 ions/cm2
and 1 × 1012 ions/cm2 . At lower fluences (≤5 × 1011 ions/cm2 ),
the sample experiences breaking of bonds, which lead to amor-
phization and reduction of crystallite size. In higher fluence
(1 × 1012 ions/cm2 ), the tendency for recrystallization is high lead-
ing to the increase in crystallinity and crystallite size [43,44].

3.4. Fourier transform infrared spectrometer-attenuated total

reflectance (FTIR-ATR)

The FTIR-ATR spectrum of the samples was as shown in Fig. 4a

and b and the assignments of the functional groups are tabulated
(Table 2). The peaks found at 1150–1250 cm−1 is due to the C O C
stretching of PMMA. The intensity of C O C stretching peaks has
decreased significantly, indicating the breakage of these bonds. The
presence of two bands at 1370 and 754 cm−1 could be attributed
to the ␣ methyl group vibration. The intensity of the peaks at
2993, 2950 and 1370 cm−1 has decreased greatly signifying lose of
water molecules and scission of ␣ methyl group. The major peak at
1732 cm−1 is assigned to the acrylate carboxylic groups. The bend-
ing mode of C H of methyl groups is observed at 1444 cm−1 . The
two bands at 2948 and 2997 cm−1 are ascribed to the CH stretching
vibration of methyl groups. Additional peaks of phosphate bend-
ing modes were observed at 564 and 603 cm−1 (Fig. 4b). The broad
peak from 1025 to 1030 cm−1 is originated from the asymmetric
stretching vibration of phosphate, which confirms the presence of
Fig. 3. GIXRD patterns of PHpris, PH1e11, PH5e11, and PH1e12. HAp in pristine and irradiated samples. The shift in C O stretching
120 G.M. Shanthini et al. / Applied Surface Science 329 (2015) 116–126

Table 1
Average crystallite size and crystallinity of pristine and irradiated samples of PMMA-HAp composite.

Samples Lattice parameters (Å) Crystallite size (±1 nm) Percentage crystallinity (±1%)

PHpris a = b = 9.42 ± 0.17; c = 6.87 ± 0.18 35 45

PH1e11 a = b = 9.67 ± 0.26; c = 6.86 ± 0.34 31 42

PH5e11 a = b = 9.54 ± 0.56; c = 6.90 ± 0.99 18 41

PH1e12 a = b = 9.42 ± 0.43; c = 6.87 ± 0.15 25 45

Fig. 4. FTIR-ATR spectra of (a) PMMA and (b) PMMA-HAp composite. (c) Magnified portion of Ppris and PHpris.

and O C stretching towards higher wavenumber on the addition Table 2

FTIR-ATR assignments of functional groups of PMMA and HAp.
of HAp substantiates that, PMMA is strongly bonded with HAp,
which consecutively inhibits the free movement of HAp crystals Wavenumber (cm−1 ) Assignments
and results in PMMA-HAp aggregate formation. The significant red 2948 and 2997 C H stretching
shift from 1232 to 1244 of COC stretching (Fig. 4c) clearly evidences 1732 C O stretching
that HAp is having strong electrostatic interaction with the car- 1444 O CH3 bending
boxylate groups of PMMA. The peak observed at 2300–2400 cm−1 1370 CH2 asymmetric bending
1150–1250 C O C stretching
in irradiated PMMA-HAp samples, is attributed to atmospheric CO2
754 C H out of plane bending
adsorption on the surface of HAp. 564 PO4 3− stretching and bending
603 PO4 3− stretching and bending
1030 Asymmetric PO4 3− stretching
3.5. Scanning electron microscopy (SEM)

Fig. 5 shows the Scanning Electron Micrograph of pristine and

swift heavy ion irradiated polymer (PMMA; Fig. 5a–d) and polymer- respective porosity of PMMA irradiated with 5 × 1011 ions/cm2 and
ceramic composite (PMMA-HAp; Fig. 5e–h). PMMA revealed the 1 × 1012 ions/cm2 is increased to about 95% and 390% compared to
formation of pores and also an increase in number of pores 1 × 1011 ions/cm2 . While, in the composites of PMMA-HAp there
and pore size with an increase in ion fluence (Table 3a). The was formation of clusters with irradiation and there is no significant
 G.M. Shanthini et al. / Applied Surface Science 329 (2015) 116–126 121

Fig. 5. SEM micrographs of PMMA: (a) Ppris, (b) P1e11, (c) P5e11, (d) P1e12 and PMMA-HAp: (e) PHpris, (f) PH1e11, (g) PH5e11, (h) PH1e12.

Table 3a approximately 100% raise, compared to the pristine (Table 4). In the
Average pore size of pristine and Si7+ ion irradiated PMMA films.
case of PMMA, there was no significant change in roughness due
PMMA Average pore size (mean ± SD (nm)) to irradiation. Each ion impinging on the PMMA surface produces
Ppris – a clearly visible hillock (Fig. 6b) that spreads on the surface due to
P1e11 19 ± 4 low fluence of ion irradiation (1011 ions/cm2 ) (Fig. 6b). With the
P5e11 37 ± 4 increase in irradiation fluence, large number of ions hit the sample
P1e12 93 ± 9 surface breaking the hillocks into nanosized particles leading to a
reduction in surface roughness (Fig. 6c and d). The localized heating
Table 3b and quenching, produced by the SHI irradiation, is likely to be the
Average cluster size of pristine and Si7+ ion irradiated PMMA-HAp films.
reason for the formation of the cluster in the irradiated PMMA-HAp
PMMA-HAp Average cluster size (mean ± SD (nm)) composites, leading to the enhanced surface roughness. Lim et al.
PHpris –
reported that the roughness in the range 5–15 nm has enhanced
PH1e11 25 ± 6 the adhesion and proliferation of human fetal osteoblast cells [46].
PH5e11 28 ± 7 Therefore, the roughness in the range of 8–17 nm as observed in the
PH1e12 27 ± 3 irradiated PMMA-HAp could enhance the initial cell adhesion. The
cell–material interaction is mediated by the ECM molecules such as
change of size of agglomerates with increase in fluences (Table 3b). proteins. The nanoscale roughness on the sample surface aids the
The irradiation led to the local heating due to thermal spike and adsorption of these molecules to a greater extent. The active sites
this resulted in pore formation in the case of polymers without on the surface of these molecules facilitate the interaction of cells
any ceramic. In consequence, when HAp is incorporated no pores to the biomaterial. Thus, the improved roughness due to irradia-
were observed, but thermal spike has resulted in the aggregation tion could assist the cell attachment [47–49] and enhances the cell
of HAp, hence forming clusters. Thermal spike for a few picosec- attraction and spreading [50–52].
onds causes short-term motion of PMMA molecules and the surface
of the material remains in the molten state for the moment until 3.7. Differential scanning calorimetry (DSC)
the temperature quenches. The short-term local melting and rapid
quenching has thus, led to the aggregation of HAp [45]. Fig. 7 shows the DSC thermograph of PMMA and PMMA-
HAp composites of pristine and highest fluence samples
3.6. Atomic force microscopy (AFM) (1 × 1012 ions/cm2 ). Ppris and PHpris exhibit an endotherm at
116 ◦ C and 125 ◦ C respectively, which corresponds to the glass
AFM topography images of pristine and irradiated samples transition temperature (Tg ). PHpris shows higher Tg compared
of PMMA and PMMA-HAp were as shown in Fig. 6a–d and e–h to Ppris, which clearly explains that HAp crystals has bound
respectively. The irradiated samples of PMMA-HAp showed an to PMMA molecules, thus preventing the segmental motion of
increase in roughness with an increase in ion fluence, which is PMMA chains on heating cycle. The decrease in Tg from 116 ◦ C

Table 4
Contact angle and surface roughness of pristine and irradiated samples.

Contact angle % enhancement of wettability Surface roughness, Rq in nm

◦
Ppris 68.8 ± 0.8 – 1.3
P1e11 52.5◦ ± 0.6 23.6 4.2
P5e11 51.3◦ ± 1.9 25.4 4.8
P1e12 50◦ ± 2 27.1 2.7
PHpris 80◦ ± 0.3 – 8.2
PH1e11 75.3◦ ± 0.2 6.2 9.5
PH5e11 67.2◦ ± 1.2 16.2 12.3
PH1e12 62.8◦ ± 0.3 21.7 17.7
122 G.M. Shanthini et al. / Applied Surface Science 329 (2015) 116–126

Fig. 6. AFM micrographs of PMMA: (a) pristine, (b) 1 × 1011 , (c) 5 × 1011 , (d) 1 × 1012 ions/cm2 and PMMA-HAp: (e) pristine, (f) 1 × 1011 , (g) 5 × 1011 , (h) 1 × 1012 ions/cm2 .

to 110 ◦ C was detected with irradiation of Ppris in the fluence Irradiation transforms the hydrophobic surfaces into hydrophilic
of 1 × 1012 ions/cm2 , which indicates that the ion irradiation dis- surface. The water contact angle decreased with increase in ion flu-
turbs the intermolecular interaction of PMMA leading to, shift in ence. The hydrophobic and smooth surface has become hydrophilic
Tg towards lower temperature compared to the pristine. Whereas, and rough on irradiation, by 27% and 21% for PMMA and PMMA-
there is no significant change in endotherms of PHpris and PH1e12. HAp respectively (Table 4). This might be due to the increase in
The decrease in Tg as the consequence of irradiation substantiates surface roughness of the samples with respect to, increase in ion
that the PMMA molecules evaporate faster and lead to the forma- fluences. The increase in hydrophilicity on irradiation is due to,
tion of pores. Increase in Tg on incorporation of HAp, validates that, rise in surface charge which is caused by intermolecular damage
HAp crystals might have coordinated with the PMMA network. This of PMMA through swift heavy ion irradiation [53,54]. According
coordination of HAp with PMMA decreased the possibility of evap- to Waugh et al. osteoblast cell response was modulated based on
oration of PMMA from the surface, which on the contrary, induces the wettability of the polymeric material. The threshold window
the formation of PMMA-HAp aggregates. for the enhanced osteoblast response is between 47◦ and 53◦ and
the material should not be super hydrophilic or super hydrophobic
3.8. Wettability [13]. Hence, as the effect of irradiation, the wettability range of the
samples was tuned to match the threshold window of osteoblast
The contact angle of pristine and irradiated samples was as cells. The highly hydrophilic surface will not promote an enhanced
shown in Fig. 8. All the samples were intermediately hydrophobic. osteoblast cell response as hydrophilic surfaces are highly
 G.M. Shanthini et al. / Applied Surface Science 329 (2015) 116–126 123

Fig. 7. DSC thermographs of (a) Ppris, (b) P1e12, (c) PHpris, and (d) PH1e12.
Fig. 9. Haemolytic assay for the pristine and irradiated samples of PMMA and
PMMA-HAp composite.

Table 5
Inhibition zone of gentamicin loaded pristine and irradiated samples.

Samples Diameter of inhibition zone (mm) ± 0.1 mm

E. coli S. aureus

Ppris 10.0 21.0

P1e11 21.5 22.0
P5e11 13.2 24.0
P1e12 18.2 30.0
PHpris 25.0 20.0
PH1e11 37.0 26.0
PH5e11 32.0 28.0
PH1e12 35.0 30.0

the surface charge, accordingly the interaction of the biomolecules

varies. In the case of PMMA samples, there was a slight increase in
haemolysis percentage which is due to the fluctuation in surface
Fig. 8. Wettability of pristine and irradiated PMMA and PMMA-HAp composite.
charge, but the percentage haemolysis of P1E11, P5E11 and P1E12
are not statistically significant compared to the Ppris. The statistical
“cell-repellent” as they hinder the initial protein adsorption needed analysis was performed using Student’s paired t-test by consider-
for the positive cell response. Better the surface wettability of the ing p < 0.05 as the minimum significance level. Thus, the surface
surface, higher will be the adsorption of blood proteins which in modified PMMA and PMMA-HAp samples may be best suitable as
turn will promote cell attachment [55–58]. implants without pronounced haemolysis effect.

3.9. Haemolysis 3.10. Antimicrobial activity

The haemocompatibility of the pristine and the irradiated sam- The antimicrobial efficacy of gentamicin loaded pristine and
ples were estimated by haemolysis. The haemolysis percentage irradiated samples were tested against E. coli and S. aureus. Genta-
shows the extent of lysis of red blood cells when they come in micin binds specifically to the A site of 16S rRNA, which will inhibit
contact with the pristine and irradiated samples. The haemolytic the synthesis of the bacterial proteins [60]. The zone of inhibition
activity of the samples as shown in Fig. 9, suggests that, with formed by gentamicin was found to be in the range of 10–22 mm
reference to ASTM standards, all the samples are highly haemo- for PMMA pristine and irradiated samples (Table 5). Whereas, the
compatible. The haemolysis percentage of all the samples was well inhibition zone of pristine and irradiated PMMA-HAp is observed
within the permissible limit according to ASTM standards. No sig- in between 25 and 37 mm. In the case of S. aureus the inhibition
nificant difference of haemolysis percentage was observed due to zone formed by PMMA and PMMA-HAp does not show any sig-
irradiation. The decrease in contact angle with an increase in ion nificant difference. The gentamicin activity on E. coli and S. aureus
fluence indicates the polarity change of the sample surface. The sur- are significantly lower in the case of pristine films, which, indi-
face charge implies the major role in blood–material interaction. cates that the binding efficiency of gentamicin to irradiated films
The blood cells and other components possess negative surface. have improved widely caused by the change in surface morphology.
The difference in percentage lysis of blood cells might be due to this This study confirmed that the antibiotic loaded samples exhibited
change in surface charge. The surface polarity of the material which increased zone of inhibition in PMMA-HAp composites. The inhi-
comes in contact with the blood plays the major role in blood com- bition zone of PMMA-HAp is comparatively higher than the zone
patibility [59]. The haemolysis percentage of PMMA-HAp is reduced formed by PMMA when tested against both negative strain (E. coli)
after the SHI irradiation. This decrease might be due to changes in and the positive strain (S. aureus). The zone was clear and stable
the surface charge after irradiation. The SHI irradiation modifies for more than 24 h. The result confirms that the antibiotic loaded,
124 G.M. Shanthini et al. / Applied Surface Science 329 (2015) 116–126

Fig. 10. Amount of plasma proteins adsorbed.

Fig. 11. Cytotoxicity of pristine and irradiated PMMA and PMMA-HAp.

irradiated polymer-ceramic composite will be helpful in preventing
the infection, during the implantation in humans.
3.12. Cytotoxicity assay
3.11. Protein adsorption
Quantitative analysis for the cytotoxicity of pristine and irra-
The adsorption of plasma proteins to the material surface is diated samples followed by 24 h incubation with 3T3 cells was
one of the vital process in determining the haemocompatibility of represented in Fig. 11. The cytotoxicity of any biomaterial will be
the biomaterial. The cells adsorb to the biomaterial through the due to leachants and byproducts. Cytotoxicity is one of the initial
plasma proteins. When the biomaterial is placed in the living sys- key evaluations required for any medical device. The potential of
tem, blood is the first fluid which comes in contact with them. The pristine and irradiated samples in inducing toxicity to 3T3 cells was
blood possesses platelets and plasma proteins, apart from blood analyzed with MTT assay and the relative growth rate of cells were
cells and other biomolecules. Next to water molecules present in calculated using:
the blood, plasma proteins compete among themselves to adsorb  OD of test sample 
onto the biomaterial surface. These adsorbed plasma proteins act Relative growth rate(RGR) = × 100% (4)
OD of control
as intermediate molecules, in order to attract ECM proteins such
as fibrinogen, fibronectin, vitronectin, etc., which in turn initiates In accordance to international standard ISO 10993-5:2009 (E),
cell adhesion [61]. The possible interactions of biomaterial with the the cytotoxicity of biological materials are graded from 0 to 1 with
protein molecules are ionic/electrostatic, hydrophobic, hydrogen respect to RGR ≥100% (grade 0), 75–99% (grade 1), 50–74% (grade
bonding and Van der Waals forces [62]. The capability of adsorbing 2), 25–49% (grade 3), 1–24% (grade 4) and 0 (grade 5). Of these gra-
plasma proteins increased with irradiation (Fig. 10). As the effect dations, 0 and 1 are good and biocompatible, 2 is weakly compatible
of irradiation, the enhancement of about 433% protein adsorption and the cytotoxicity in between 3 and 5 are not acceptable [64,65].
was achieved. Surface roughness, wettability and surface chem- Comparing the percentage of viable cells in all the test samples, it
istry of the samples are the key factors in determining the protein was found that the irradiated samples are non-toxic to the seeded
adsorption ability of any surface. More plasma protein is absorbed cells. The samples exhibit the cytotoxicity grade of 1 which rep-
in SHI modified PMMA samples than in Ppris. The carboxyl groups resents these materials can be used as biomaterials. Both pristine
of methacrylic acid monomers undergo an electrostatic interaction and irradiated samples show that all the samples are biocompati-
with proteins. The irradiation has led to increased pore formation. ble, which corroborates that irradiation does not release any toxic
Hence, the increase in surface area, along with the electrostatic byproducts. Besides cytocompatibility, the sample PH5E11 shows
interaction of the irradiated sample, enhances the protein adsorp- increased cell viability to about 4%. Also, comparing the results
tion. Zhu et al. studied the effect of different structures of biphasic obtained for PMMA and PMMA-HAp composites in general, it was
calcium phosphate and proved that the presence of porous struc- confirmed that the presence of HAp does not show any significant
tures significantly assisted the protein adsorption [63]. In the case change in cell viability compared to PMMA samples. Accordingly,
of PMMA-HAp, the thermal spike, which is induced by the irradia- with the results of MTT assay it is concluded that, owing to irra-
tion, rapidly quenches the PMMA molecules and hence, exposes the diation there was no release of toxic leachants from the samples.
buried HAp aggregates (Fig. 5e–h). The exposed HAp clusters expe- More to the point, irradiation helps in maintaining a higher number
rience strong electrostatic interaction with the protein molecules of viable cells to a certain extent which is very much essential for
and results in the increased protein adsorption (≤PH5e11). At their potential application in tissue engineering.
higher fluence (PH1e12), the exposed HAp molecules, undergoes
severe damage due to SHI, resulting in the net change in sur- 4. Conclusions
face charge, and causes electrostatic repulsion with the protein
molecule which further leads to the decrease in protein adsorption. The swift heavy ion irradiation modifies the surface of the sam-
Also, the wettability results support that on increasing the irradi- ples. There was a decrease in the crystallite size and crystallinity
ation fluence, wettability decreases gradually up to 50◦ which is with the increase in irradiation fluences. The surface roughness
highly suitable for protein adsorption [56]. As a consequence, the and wettability of the material surface has improved significantly,
irradiated samples exhibit enhanced protein adsorption compared which in turn will facilitate the protein adsorption. Irradiation
to pristine. resulted in the creation of pores and clusters, which will assist
 G.M. Shanthini et al. / Applied Surface Science 329 (2015) 116–126 125

the proliferation of the attached cells. Haemolysis and cytotoxic- to extreme ultraviolet (EUV) radiation, J Biomed. Mater. Res. A 102 (2014)
ity studies show that there is no formation of toxic byproducts due 3298–3310.
[16] D.K. Avasthi, Some interesting aspects of swift heavy ions in materials science,
to the passage of swift heavy ions through the sample in spite of Curr. sci. 78 (2000) 1297–1303.
changes in functional groups as observed in FTIR. The irradiated [17] A. Kumar, S. Banerjee, Swift heavy ion irradiation induced structural, optical
samples show excellent haemocompatibity and cytocompatibil- and conformational modifications in conducting polymer nanostructures, Adv.
Mater. Lett. 4 (2013) 433–437.
ity. PMMA and PMMA-HAp modified by SHI irradiation augments [18] R. Singhal, A. Tripathi, D.K. Avasthi, Synthesis of carbon nanowires by SHI irra-
the protein adsorption which could favour cell–material interac- diation of fullerene C70 thin film, Adv. Mater. Lett. 4 (2013) 413–417.
tion. The irradiated PMMA-HAp exhibited improved morphology, [19] D. Dunlop, P. Lesueur, H. Legrand, Dammak, Effects induced by high electronic
excitations in pure metals: a detailed study in iron, Nucl. Instrum. Method B 90
enhanced roughness and higher zone of inhibition against E. coli and
(1994) 330–338.
S. aureus compared to the irradiated PMMA. Moreover, the incor- [20] T. Mohanty, N.C. Mishra, F. Singh, U. Tiwari, D. Kanjilal, Swift heavy ion irradi-
poration of HAp followed by SHI irradiation has played a major ation induced modifications in sapphire, Nucl. Instrum. Method B 212 (2003)
179–183.
role in enhancing the physical properties. The observed irradiation
[21] T. Som, B. Satpati, O.P. Sinha, D.K. Avasthi, D. Kanjilal, Role of electronic and
induced morphological changes such as the formation of pores and nuclear energy losses in swift heavy ion beam induced epitaxial crystallization
aggregates and the reduction in contact angle greatly influenced the of a buried Si3 N4 layer, Nucl. Instrum. Method B 245 (2006) 255–259.
protein adsorption which will facilitate; cell attachment, spreading [22] R.V. Suganthi, S. Prakash Parthiban, K. Elayaraja, E.K. Girija, P. Kulariya, Y.S.
Katharria, F. Singh, K. Asokan, D. Kanjilal, S. Narayana Kalkura, Investigations
and proliferation, consequently, resulting in the ideal biomaterial on the in vitro bioactivity of swift heavy oxygen ion irradiated hydroxyapatite,
for tissue engineering. The irradiation of a composite of polymers J. Mater. Sci. Mater. Med. 20 (2008) 271–275.
with lower glass transition temperature and melting temperature [23] N.L. Singh, S. Shah, A. Qureshi, A. Tripathi, F. Singh, D.K. Avasthi, P.M. Raole,
Effect of ion beam irradiation on metal particle doped polymer composites,
will result in surface patterning. Thus a biocompatible material Bull. Mater. Sci. 34 (2011) 81–88.
with patterned surface, improved morphology, topography, wett- [24] S. Shahabi, F. Najafi, A. Majdabadi, T. Hooshmand, M.H. Nazarpak, B. Karimi,
ability can be attained by using a single tool, SHI. S.M. Fatemi, Effect of gamma irradiation on structural and biological prop-
erties of a PLGA-PEG-hydroxyapatite composite, Sci. World J. 2014 (2014)
420616.
[25] I. Last, I. Schek, J. Jortner, Energetics and dynamics of Coulomb explosion of
Acknowledgements highly charged clusters, J. Chem. Phys. 107 (1997) 6685–6692.
[26] P. Karmakar, S. Bhattacharjee, V. Naik, A.K. Sinha, A. Chakrabarti, Coulomb
explosion sputtering of selectively oxidized Si, J. Phys.: Condens. Matter 22
This work was financially supported by the Inter University
(2010) 175005.
Accelerator Centre, New Delhi through project No. UFR-50308. One [27] M. Toulemonde, C. Dufour, Z. Wang, E. Paumier, Atomic and cluster ion bom-
of the authors GMS also acknowledges CSIR, India for the award of bardment in the electronic stopping power regime: a thermal spike description,
Nucl. Instrum. Method B 112 (1996) 26–29.
SRF fellowship (File no.: 09/468 (0474)/2013-EMR-I).
[28] Z.G. Wang, C. Dufour, E. Paumier, M. Toulemonde, The Se sensitivity of metals
under swift-heavy-ion irradiation: a transient thermal process, J. Phys.: Con-
dens. Matter 6 (1994) 6733–6750.
References [29] M. Toulemonde, C. Dufour, E. Paumier, Transient thermal process after high-
energy heavy-ion irradiation of amorphous metals and semiconductors, Phys.
[1] N. Degirmenbasi, D.M. Kalyon, E. Birinci, Biocomposites of nanohydroxyapatite Rev. B 46 (1992) 14362–14369.
with collagen and poly (vinyl alcohol), Colloids Surf. B 48 (2006) 42–49. [30] A. Kumar, S. Banerjee, J.P. Saikia, B.K. Konwar, Swift heavy ion irradiation
[2] M. Boutinguiza, J. Pou, R. Comesana, F. Lusquinos, A.D. Carlos, B. Leon, Biological induced enhancement in the antioxidant activity and biocompatibility of
hydroxyapatite obtained from fish bones, Mater. Sci. Eng. C 32 (2012) 478–486. polyaniline nanofibers, Nanotechnology 21 (2010) 175102.
[3] V.V. Thai, Y.K. Min, B.T. Lee, Fabrication of hybrid composites consists of poly [31] L. Thome, S. Moll, A. Debelle, F. Garrido, G. Sattonnay, J. Jagielski, Radiation
methyl methacrylate and polyvinyl alcohol and hydroxyapatite, Bioceram. Dev. effects in nuclear ceramics, Adv. Mater. Sci. Eng. 2012 (2011) 1–13.
Appl. 1 (2011) 1–4. [32] J.P. Singhal, A.R. Ray, Synthesis of blood compatible polyamide block copoly-
[4] P. Silva, C. Albano, R. Perera, N. Dominguez, Study of the gamma irradia- mers, Biomaterials 23 (2002) 1139–1145.
tion effects on the PMMA/HA and PMMA/SW, Radiat. Phys. Chem. 79 (2010) [33] P.S. Liu, Q. Chen, S.S. Wu, J. Shen, Surface modification of cellulose membranes
358–361. with zwitterionic polymers for resistance to protein adsorption and platelet
[5] P.A. Stone, A.Y. Mousa, S.M. Hass, D.D. Dearing, J.R. Campbell II, A. Parker, S. adhesion, J. Membr. Sci. 350 (2010) 387–394.
Thompson, A.F. Aburahma, Antibiotic-loaded polymethylmethacrylate beads [34] R.C. Ramola, A. Alqudami, S. Chandra, S. Annapoorni, J.M.S. Rana, R.G.
for the treatment of extracavitary vascular surgical site infections, J. Vasc. Surg. Sonkawade, F. Singh, D.K. Avasthi, Effects of swift heavy ions irradiation on
55 (2012) 1706–1711. polypyrrole thin films, Radiat. Eff. Defects solids 163 (2008) 151–159.
[6] D. Neut, H.V.D. Belt, J.R.V. Horn, H.C.V.D. Mei, H.J. Busscher, Residual [35] B.S. Kaith, K. Sharma, V. Kumar, V. Kumar, H.C. Swart, S. Kalia, Effects of swift
gentamicin-release from antibiotic-loaded polymethylmethacrylate beads heavy ion beam irradiation on the structural and morphological properties
after 5 years of implantation, Biomaterials 24 (2003) 1829–1831. of poly(methacrylic acid) cross-linked gum Gatti films, Vacuum 101 (2014)
[7] M.J. Dalby, L.D. Silvio, E.J. Harper, W. Bonfield, Increasing hydroxyapatite incor- 166–170.
poration into poly(methylmethacrylate) cement increases osteoblast adhesion [36] A. Biswas, S. Lotha, D. Fink, J.P. Singh, D.K. Avasthi, B.K. Yadav, S.K. Bose, D.T.
and response, Biomaterials 23 (2002) 569–576. Khating, A.M. Avasthi, The effects of swift heavy ion irradiation on the radio-
[8] R.K. Roeder, M.M. Sproul, C.H. Turner, Hydroxyapatite whiskers provide chemistry and melting characteristics of PET, Nucl. Instrum. Method B 159
improved mechanical properties in reinforced polymer composites, J. Biomed. (1999) 40–51.
Mater. Res. A 67 (2003) 801–812. [37] E. Akcoltekin, T. Peters, R. Meyer, A. Duvenbeck, M. Klusmann, I. Monnet, H.
[9] J.I. Rosales-Leal, M.A. Rodriguez-Valverde, G. Mazzaglia, P.J. Ramon-Torregrosa, Lebius, M. Schleberger, Creation of multiple nanodots by single ions, Nat. Nan-
L. Diaz-Rodriguez, O. Garcia-Martinez, M. Vallecillo-Capilla, C. Ruiz, M.A. otechnol. 2 (2007) 290–294.
Cabrerizo-Vilchez, Effect of roughness, wettability and morphology of engi- [38] D. Fink, L.T. Chadderton, Ion-solid interaction: status and perspectives, Braz. J.
neered titanium surfaces on osteoblast-like cell adhesion, Colloids Surf. A 365 Phys. 35 (2005) 735–740.
(2010) 222–229. [39] Y.Y. Wang, C. Grygiel, C. Dufour, J.R. Sun, Z.G. Wang, Y.T. Zhao, G.Q. Xiao, R.
[10] K. Merrett, R.M. Cornelius, W.G. Mcclung, L.D. Unsworth, H. Sheardown, Surface Cheng, X.M. Zhou, J.R. Ren, S.D. Liu, Y. Lei, Y.B. Sun, R. Ritter, E. Gruber, A. Cas-
analysis methods for characterizing polymeric biomaterials, J. Biomater. Sci. simi, I. Monnet, S. Bouffard, F. Aumayr, M. Toulemonde, Energy deposition by
Polym. Ed. 13 (2002) 593–621. heavy ions: additivity of kinetic and potential energy contributions in hillock
[11] L. Ponsonnet, K. Reybier, N. Jaffrezic, V. Comte, C. Lagneau, M. Lissac, C. Martelet, formation on CaF2 , Sci. Rep. 4 (2014) 5742.
Relationship between surface properties (roughness, wettability) of titanium [40] M. Rajkumar, N. Meenakshisundaram, V. Rajendran, Development of
and titanium alloys and cell behavior, Mater. Sci. Eng. C 23 (2003) 551–560. nanocomposites based on hydroxyapatite/sodium alginate: synthesis and
[12] D.G. Waugh, J. Lawrence, On the use of CO2 laser induced surface patterns to characterization, Mater. Charact. 62 (2011) 469–479.
modify the wettability of poly(methyl methacrylate) (PMMA), Opt. Laser Eng. [41] N. Pramanik, D. Mishra, I. Banerjee, T.K. Maiti, P. Bhargava, P. Pramanik,
48 (2010) 707–715. Chemical synthesis, characterization, and biocompatibility study of hydrox-
[13] D.G. Waugh, J. Lawrence, Wettability and osteoblast cell response modulation yapatite/chitosan phosphate nanocomposite for bone tissue engineering
through UV laser processing of nylon 6,6, Appl. Surf. Sci. 257 (2011) 8798–8812. applications, Int. J. Biomater. (2009) 512417.
[14] D. Aronov, R. Rosen, E.Z. Ron, G. Rosenman, Tunable hydroxyapatite wettabil- [42] M.R. Nikpour, S.M. Rabiee, M. Jahanshahi, Synthesis and characterization of
ity: Effect on adhesion of biological molecules, Process Biochem. 41 (2006) hydroxyapatite/chitosan nanocomposite materials for medical engineering
2367–2372. applications, Composites B 43 (2012) 1881–1886.
[15] I.U. Ahad, A. Bartnik, H. Fiedorowicz, J. Kostecki, B. Korczyc, T. Ciach, D. [43] S. Chandramohan, R. Sathyamoorthy, P. Sudhagar, D. Kanjilal, D. Kabiraj, K.
Brabazon, Surface modification of polymers for biocompatibility via exposure Asokan, V. Ganesan, T. Shripathi, U.P. Deshpande, High-energy heavy-ion
126 G.M. Shanthini et al. / Applied Surface Science 329 (2015) 116–126

induced physical and surface-chemical modifications in polycrystalline cad- [55] B.N. Lourenco, G. Marchioli, W. Song, R.L. Reis, C.A. Van Blitterswijk, M.
mium sulfide thin films, Appl. Phys. A 94 (2009) 703–714. Karperien, A.V. Apeldoorn, J.F. Mano, Wettability influences cell behavior
[44] Vijay Kumar, R.G. Sonkawade, Y. Ali, A.S. Dhaliwal, Study of chemical, optical on superhydrophobic surfaces with different topographies, Biointerphases 7
and structural properties of 120 MeV Ni11+ ions beam irradiated poly (ethylene (2012) 1–11.
terephthalate) film, Int. J. Appl. Eng. Res. 2 (2011) 419–430. [56] Y. Arima, H. Iwata, Effect of wettability and surface functional groups on pro-
[45] I.P. Jain, G. Agarwal, Ion beam induced surface and interface engineering, Surf. tein adsorption and cell adhesion using well-defined mixed self-assembled
Sci. Rep. 66 (2011) 77–172. monolayers, Biomaterials 28 (2007) 3074–3082.
[46] J.Y. Lim, A.D. Dreiss, Z. Zhou, J.C. Hansen, C.A. Siedlecki, R.W. Hengstebeck, [57] L.C. Xu, C.A. Siedlecki, Effects of surface wettability and contact time on protein
J. Cheng, N. Winograd, H. Donahue, The regulation of integrin-mediated adhesion to biomaterial surfaces, Biomaterials 28 (2007) 3273–3283.
osteoblast focal adhesion and focal adhesion kinase expression by nanoscale [58] Y. Zhou, V.E. Gregor, Z. Sun, B.K. Ayida, G.C. Winters, D. Murphy, K.B.
topography, Biomaterials 28 (2007) 1787–1797. Simonsen, D. Vourloumis, S. Fish, J.M. Froelich, D. Wall, T. Hermann, Structure-
[47] Y. Wu, J.P. Zitelli, K.S. TenHuisen, X. Yu, M.R. Libera, Differential response of guided discovery of novel aminoglycoside mimetics as antibacterial translation
Staphylococci and osteoblasts to varying titanium surface roughness, Biomate- inhibitors, Antimicrob. Agents Chemother. 49 (2005) 4942–4949.
rials 32 (2011) 951–960. [59] S.A. Shabalovskaya, D. Siegismund, E. Heurich, M. Rettenmayr, Evaluation of
[48] M.P. Molina, P.G. Moreno, J.E.F. Barbero, F.O. Valle, A.B.J. Reyes, J.L.O. Vin- wettability and surface energy of native Nitinol surfaces in relation to haemo-
uesa, P.J.R. Torregrosa, Role of wettability and nanoroughness on interactions compatibility, Mater. Sci. Eng. C 33 (2013) 127–132.
between osteoblast and modified silicon surfaces, Acta Biomater. 7 (2011) [60] S. Yoshizawa, D. Fourmy, J.D. Puglisi, Structural origins of gentamicin antibiotic
771–778. action, EMBO J. 17 (1998) 6437–6448.
[49] F. Gentile, L. Tirinato, E. Battista, F. Causa, C. Liberale, E.M.D. Fabrizio, P. [61] B.D. Ratner, A.S. Hoffman, F.J. Schoen, J.E. Lemons, Biomaterials Science Intro-
Decuzzi, Cells preferentially grow on rough substrates, Biomaterials 31 (2011) duction to Materials in Medicine, 2nd ed., Elsevier Academic Press, New York,
7205–7212. 2004.
[50] M. Vandrovcova, L. Bacakova, Adhesion, growth and differentiation of [62] J.H. Kim, J.Y. Yoon, Protein Adsorption on Polymer Particles, Encyclopedia of
osteoblasts on surface-modified materials developed for bone implants, Surface and Colloid Science, 2002, pp. 4373–4381.
Physiol. Res. 60 (2011) 403–417. [63] X.D. Zhu, H.S. Fan, Y.M. Xiao, D.X. Li, H.J. Zhang, T. Luxbacher, X.D. Zhang,
[51] T.W. Chung, D.Z. Liu, S.Y. Wang, S.S. Wang, Enhancement of the growth of Effect of surface structure on protein adsorption to biphasic calcium-
human endothelial cells by surface roughness at nanometer scale, Biomaterials phosphate ceramics in vitro and in vivo, Acta Biomater. 5 (2009) 1311–
24 (2003) 4655–4661. 1318.
[52] D.D. Deligianni, N.D. Katsala, P.G. Koutsoukos, Y.F. Missirlis, Effect of surface [64] J. Zhang, D. Zhang, Preparation of a nanosized As2 O3 /Mn0.5 Zn0.5 Fe2 O4 complex
roughness of hydroxyapatite on human bone marrow cell adhesion, prolifera- and its anti-tumor effect on hepatocellular carcinoma cells, Sensors 9 (2009)
tion, differentiation and detachment strength, Biomaterials 22 (2001) 87–96. 7058–7068.
[53] G. Hanly, D. Fornasiero, J. Ralston, R. Sedev, Electrostatistics and metal oxide [65] D. Chen, Q. Tang, X. Li, X. Zhou, J. Zang, W.Q. Xue, J.Y. Xiang, C.Q. Guo, Biocom-
wettability, J. Phys. Chem. C 115 (2011) 14914–14921. patibility of magnetic Fe3 O4 nanoparticles and their cytotoxic effect on MCF-7
[54] L.S. Puah, R. Sedev, D. Fornasiero, J. Ralston, Influence of surface charge on cells, Int. J. Nanomed. 7 (2012) 4973–4982.
wetting kinetics, Langmuir 26 (2010) 17218–17224.

View publication stats