Composites Science and Technology 123 (2016) 232e240

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Composites Science and Technology

journal homepage: http://www.elsevier.com/locate/compscitech

Synergistic reinforcement of polydopamine-coated hydroxyapatite

and BMP2 biomimetic peptide on the bioactivity of PMMA-based
cement
Ting Kang a, 1, Xiaoli Hua b, 1, Peiqing Liang a, Minyu Rao a, Qin Wang a,
Changyun Quan a, c, *, Chao Zhang a, c, **, Qing Jiang a, c
a
Biomedical Engineering, School of Engineering, Sun Yat-sen University, Guangzhou, 510006, China
b
Department of Pharmacy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan, 430022,
China
c
Guangdong Provincial Key Laboratory of Sensor Technology and Biomedical Instruments, Sun Yat-sen University, Guangzhou, Guangdong, 510006, China

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

Article history: To be better used as implant materials in bone regeneration, bioactivity and osteogenesis of poly(methyl
Received 3 November 2015 methacrylate) (PMMA)-based bone cement need to be further enhanced. In this study, bioactive hy-
Received in revised form droxyapatite (HA) nanoparticle decorated with polydopamine (pDA) and BMP2 mimicking peptide
30 December 2015
(serine-serine-valine-proline-threonine, SSVPT) was incorporated into PMMA-based cement (px-HA/
Accepted 4 January 2016
PMMA). The presence of pDA layer on HA surface could keep the mechanical property of px-HA/PMMA
Available online 5 January 2016
cements comply with the requirement of clinical application. More importantly, the px-HA/PMMA
cement exhibited excellent biomechanical property with enhanced interfacial adhesion between cement
Keywords:
Poly(methyl methacrylate)
and swine femur. Biomineralization and cell experiment in vitro showed that the incorporation of pDA-
Hydroxyapatite coated HA and SSVPT can synergistically enhance the bioactivity of px-HA/PMMA cement for regener-
Polydopamine ation and provide an attractive strategy to reconstruction of load-bearing bone.
BMP-2 biomimetic peptide © 2016 Elsevier Ltd. All rights reserved.
Bioactivity

1. Introduction of PMMA bone cement. However, the incompatibility between

these inorganic particles and MMA/PMMA may deteriorate the
Poly(methyl methacrylate) (PMMA) bone cement has been mechanical property and injectability of the cements. The intro-
widely used in repair of vertebral fracture caused by osteoporosis as duction of siloxane to HA surface could increase the compatibility
well as in stabilizing prosthetic implants, and PMMA cement has between the two phases and help enhance the interfacial adhesion
several advantages such as good mechanical properties, fast self- followed by the improvement of mechanical property [11,12].
curing, and so on [1,2]. However, due to the bio-inertness, i.e. the Many efforts have been devoted to the modification of HA sur-
lack of bioactivity, of PMMA, the poor interfacial integration be- face to enhance the surface biomineralization and bioactivity such
tween the host bone tissue and the cement brings a major chal- as osteoblast adhesion and new bone formation [13]. Dopamine
lenge in the clinical practice and may cause interfacial loosening, (DA), a mussel inspired molecule that can self-polymerize into
fatigue, fracture, and ultimately failure [3,4]. Incorporation of some polydopamine (pDA) and adhere onto HA surface without any
bioactive ingredients such as hydroxyapatite (HA) [5,6], b-TCP [7,8], pretreatments [14,15], can be used to strengthen the interaction
and bioglass [9,10] has proved to effectively enhance the bioactivity between cement and host bone tissue. It has been reported that the
pDA can concentrate Ca2þ and serve as surface-anchor to induce
mineralization and deposition of bioactive HA on its surface [16,17];
* Corresponding author. Biomedical Engineering, School of Engineering, Sun Yat- for similar reason, pDA can be used to immobilize HA nanoparticles
sen University, Guangzhou, 510006, China. [18], which would improve the mechanical strength of nano-HA
** Corresponding author. Biomedical Engineering, School of Engineering, Sun Yat-
[19]. In addition, pDA has also been proved to significantly pro-
sen University, Guangzhou, 510006, China.
E-mail addresses: [email protected] (C. Quan), [email protected]
mote the adhesion and proliferation of osteoblasts [20].
(C. Zhang). The bone morphogenetic proteins (BMPs) have been identified
1
These authors contribute equally to this work.

http://dx.doi.org/10.1016/j.compscitech.2016.01.002
0266-3538/© 2016 Elsevier Ltd. All rights reserved.
 T. Kang et al. / Composites Science and Technology 123 (2016) 232e240 233

as therapeutic proteins that can accelerate ectopic bone formation purchased from Amresco (USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-
[21]. BMP2 is the most powerful osteogenic factor to promote diphenyltetrazolium bromide (MTT) was purchased from GBCBIO
proliferation, migration, and the differentiation of mesenchymal Technology Inc and used as received.
stem cells (MSCs) into osteoblasts [22e24]. Functional oligopeptide
sequence derived from BMP2, such as serine-serine-valine-proline- 2.2. Surface modification of HA
threonine (SSVPT) has been harnessed to fabricate biomaterials
that can facilitate biomineralization and osteogenesis [25,26]. The surface modification of nano-HA by pDA was carried out
In this study, pDA was polymerized in situ to form covalent according to literature [31]. In detail, DA (33 g) and hydroquinol
coating on the surface of the HA nanoparticles; and to the pDA (0.19 g) were dispersed in 50 mL of ethanol/water (9:1, v/v) mixture
coating was conjugated SSVPT peptide via Michael-type addition at 50  C for one h and the pH was adjusted to 3.5e4.0 before adding
[27]. A bioactive PMMA-based bone cement was fabricated from nano-HA (8 g) under stirring. After 48 h reaction, the reaction
this SSVPT/pDA-coated HA nanoparticles at content as high as mixture was centrifued at 10,000 rpm to isolate the pDA-coated HA
40 wt%; the mechanical properties, biomineralization, and cyto- nanoparticles (d-HA). The solid was resuspended-centrifued in THF
toxicity of this cement were evaluated in detail (Fig. 1). The syn- for three times. And then, the product was dried at 60  C for 48 h
ergistic effect of pDA-coated HA and SSVPT on cell culture was under vacuum.
studied in vitro. To conjugate the BMP-2 mimicking peptide onto the surface of
d-HA, d-HA (0.5 g) was dispersed in 50 mL of distilled water under
sonication for 10 min, to which the BMP-2 mimicking peptide
2. Materials and experimental
(20 mg) in 200 mL of PBS was added. After the reaction was
vigorously stirred at 25  C for 24 h, the mixture was centrifuged and
2.1. Materials
washed using distilled water for three times to remove physically
adsorbed peptide. The final product (p-HA) was dried at 60  C for
Methyl methacrylate (MMA) and poly(ethylene glycol) 2000
48 h under vacuum.
(PEG 2000, Mn 2000 g/mol) were purchased from Damao Chemical
Reagent Factory (Tianjin, China). MMA was used after distillation
2.3. Characterization of HA nanoparticle
under reduced pressure. Poly(vinyl alcohol) (PVA, Mw
30,000e50,000 g/mol) and benzoyl peroxide (BPO) were pur-
The successful conjugation of pDA and SSVPT onto the surface of
chased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai,
HA was validated by Fourier-transformed infrared spectroscopy
China). Dopamine (DA) and dimethyltryptamine (DMT) were pur-
(FT-IR, Bruker, Germany). The HA (0.002 g) was pressed into KBr
chased from Aladdin Reagent Company (Shanghai, China) and used
(0.15 g) pellets at 25  C and scanned for 16 times in the range of
as received without further purification. HA nanoparticles (average
400e4000 cm1.
length 30 nm, diameter 8 nm) [28] and BMP2 mimicking peptide
The morphology of d-HA and p-HA were observed under a
(SSVPT-NH2) [29] were prepared according to the reported proce-
transmission electron microscope (TEM, FEI Tecnai G2 Spirit,
dure with minor modifications. Simulated body fluid (SBF) was
Netherlands) at an acceleration voltage of 80 kV. A droplet of HA
prepared following reported protocol [30]. All other reagents were
suspension was placed on a copper grid and allowed to dry before
of analytical grade and used as received.
the observation.
Human fetal osteoblast (hFOB) was purchased from the cell
The modification degree, i.e. the content, of pDA and SSVPT on
bank of Chinese Academy of Sciences at Shanghai, China. Dubeco's
the surface of HA was measured thermogravimetrically on a TGA/
Modified Eagle's Medium (DMEM) and fetal bovine serum (FBS)
DSC1 instrument (Meteler, USA). In detail, 8 mg of HA, d-HA, and p-
were purchased from Hyclone (USA). Geneticin Selective Antibiotic
HA were heated respectively from 30  C to 700  C with a heating
(G-418 Sulfate) was purchased from Notlas (Beijing, China). N-2-
rate of 10  C/min under nitrogen atmosphere. The weight per-
Hydroxyethylpiperazine-N-ethanesulphonic acid (HEPES) was
centage of pDA coating was calculated by the formula as follows:

Weight Percentage (wt%) ¼ (Wo/WHA)  100% (1)

where Wo is the weight loss of the sample, and WHA is the mass of
residual nanoparticles at 700  C.

2.4. Preparation of the bone cement

Bone cements were prepared by mixing solid and liquid com-

ponents at a powder (P) to liquid (L) ratio of 1:3 (w/v). The liquid

Table 1
Feed capacity of the solid component.

Bone cement PMMA(g) Inorganic nanoparticles (g) BPO (g)

BaSO4 HA d-HA p-HA

PMMA 1 0 0 0 0 0.04
BaSO4/PMMA 0.56 0.4 0 0 0 0.04
p/PMMA 0.56 0.3 0 0 0.1 0.04
HA/PMMA 0.56 0 0.4 0 0 0.04
d-HA/PMMA 0.56 0 0 0.4 0 0.04
Fig. 1. Schematic illustration of synergistic mechanism of reinforcing bioactivity of
px-HA/PMMA (x ¼ 1) 0.56 0 0 0.3 0.1 0.04
PMMA-based cement with polydopamine-coated hydroxyapatite and BMP2 bio-
px-HA/PMMA (x ¼ 2) 0.56 0 0 0.2 0.2 0.04
mimetic peptide.
234 T. Kang et al. / Composites Science and Technology 123 (2016) 232e240

presented as mean ± STD.

The interfacial adhesion of the cement to bone tissue was
investigated using fresh swine femur tissue. The sample was pre-
pared as illustrated in Fig 2. Bone cements were poured into the
mould and reacted for four hour at 25  C. The ultimate flexural
strength of each specimen was determined through four-point
bending test according to ISO 5883 standard. Five duplicates were
measured for each samples, the results were presented as
Fig. 2. Schematic illustration of sample size prepared from fresh swine femur segment mean ± STD.
and bone cement for biomechanical test.

component contained two compounds: MMA (monomer, 3 mL)

2.6. Morphology of fracture surface of PMMA-based bone cement
and DMT (accelerator, 0.14 mL). The formulation of the solid
component was listed in Table 1. In order to study the synergistic
The fracture surface of the cement after flexural experiment was
effect of HA an SSVPT on its bioactivity, BaSO4 and BMP2-mimicking
examined on a JSM-6330F scanning electron microscope (SEM,
peptide (SSVPT) were respectively incorporated with PMMA pow-
Japan). The surface morphology of composite bone cement samples
der to prepare the cement (define as BaSO4/PMMA, and P/PMMA)
was observed after being soaked in PBS for 3 days.
and used as controls.

2.5. Mechanical properties

2.7. Biomineralization of PMMA-based bone cement
Mechanical properties of the bone cement samples were ob-
tained on a universal material testing machine (Lloyd, UK) Specimens of the bone cement with 2  6  1 mm size were
following the ISO5833 standard. The samples for the compressive soaked in 1 mL of SBF at 37  C for 0, 3, 7, and 14 days for biomin-
test were cast into a cylindrical mould (length  diameter: eralization. The specimens were washed with distilled water and
12  6 mm). The four-point flexural strength, with a supporting then dried at room temperature. The morphology of mineral layer
span of 34 mm, was measured using rectangular shaped samples on the surface of PMMA-based bone cements was characterized by
(length  width  thickness: 75  10  3.3 mm). All the tests were thermal field emission environmental SEM (Quanta 400F,
carried out at a crosshead speed of 5 mm/min after the samples Netherlands).
were being soaked in PBS for 3 days at room temperature. Five The Ca/P ratio of the mineral layer was determined by energy
duplicates were measured for each sample, the results were dispersive X-ray (EDX) spectrometry (Inca X-act, OXFORD, UK).

Fig. 3. (a) FTIR spectra, (b) TGA, and (cee) TEM images of HA nanoparticles.
 T. Kang et al. / Composites Science and Technology 123 (2016) 232e240 235

2.8. In vitro cytotoxicity containing 1 mL of growth medium in each well. After incubation
for 24 h (34  C, 5% CO2), the medium was removed. 100 mL of the
The cytotocixity of the bone cements towards hFOB was assayed extract of the bone cement was added to each well. After incubation
using the extracts of the cements in DMEM. The extracts of PMMA- for 24 h, 60 mL of MTT solution (5 mg/mL) in DMEM was added to
based bone cements were prepared according to requirement of each well. After incubation for additional four h, the MTT medium
ISO10993 standard. After being pretreated by 75% ethanol and PBS was removed and 200 mL of DMSO was added to dissolve the for-
for 24 h respectively, the bone cements (diameter  thickness: mazan. The optical density of each well was measured at 570 nm on
6  5 mm) were subsequently soaked in DMEM (the ratio of cement a plate reader (BioTek Synergy4, USA). DMEM without extract was
to medium was 1/5 (w/v)) at 34  C for 24 h. The medium was then used as control. The cell viability was calculated by the following
used as the extract (marked as “extract”) in the following cell cul- equation:
ture without further filtration.
hFOBs were seeded into a 48-well plate (5.0  104 cells/well) Cell viability% ¼ (ODcement/ODcontrol)  100 (2)

Fig. 4. (a) Compressive strength, (b) flexural strength, and (c) flexural modulus of PMMA-based bone cement after soaked in PBS for 3 days at 25  C. (deg) SEM image of fracture
surface of PMMA-based bone cement after bending test.
236 T. Kang et al. / Composites Science and Technology 123 (2016) 232e240

where ODcontrol and ODcement were the optical densities of corre- (Fig. 3e) nanoparticles were observed as compared with HA
sponding samples. (Fig. 3c).

2.9. Cell adhesion

3.2. Mechanical property of PMMA-based bone cement
hFOBs (2.0  104 cells/well) were seeded on the surface of bone
cement (diameter  thickness: 6  5 mm) in a 48-well plate. After 7 The mechanical property (Fig. 4aec) of bone cement is one
days' incubation, the surface of the cement was rinsed with PBS to essential criteria for clinical repair of vertebra. It was found that
remove non-adherent cells. The cement specimens were fixed by the compressive strength, flexural strength, and flexural modulus
2.5% of glutaraldehyde in cacodylate buffer and then washed in of HA/PMMA and BaSO4/PMMA were above 67 ± 8.2 MPa,
cacodylate buffer. After dehydration with gradient alcohol, the 50 ± 3.5 MPa, and 2 ± 0.18 GPa, respectively. The BaSO4/PMMA
samples were sputter-coated with gold and subjected to SEM (Fig. 4d) and HA/PMMA (Fig. 4e) cements displayed smooth
(Quanta 400, Philips) observation. fracture surfaces with only minor cracks, agreeing with the fact
The attachment of hFOBs in term of culture time on the sur- that the mechanical properties of HA/PMMA and BaSO4/PMMA
face of the bone cements was also quantified. The cells were cement is higher than that of other cements. The uniform/ho-
incubated on the surface of the bone cement mogenous dispersion of nano-sized filler in the polymer matrix is
(diameter  thickness: 6  5 mm) in 48-well plate for 1, 3, and 5 critical to the reinforcement of the polymer nanocomposite. The
days; after that, the medium was removed, then 40 mL of MTT above observations on BaSO4/PMMA and HA/PMMA cements
solution (5 mg/mL) in DMEM and 360 mL of DMEM were added clearly demonstrated that the dispersion of HA nanoparticle in
to each well and incubated with the cells for additional four h, the PMMA matrix may greatly affect the mechanical property
then the medium was removed and 200 mL of DMSO was added thereof.
to dissolve the formazan. Tissue culture plate was used as the The compressive strength, flexural strength, and flexural
control. The cell viability at each time point was measured as modulus of d-HA/PMMA cement were 50 ± 1.6 MPa,
described above to quantitatively assess the cell adhesion. Two- 26 ± 3.2 MPa, and 1.52 ± 0.02 Gpa, respectively; and those of p2-
way ANOVAs analysis was performed to examine differences in HA/PMMA were 58 ± 2.5 MPa, 43 ± 1.7 MPa, and 1.92 ± 0.03 GPa,
cell adhesion on the cement surface, using material groups and respectively. The much lower mechanical properties than those of
co-cultured time as factors. HA/PMMA and BaSO4/PMMA cements can be attributed to the
aggregation of d-HA or p-HA in the cement matrix, which was
3. Results and discussion confirmed from the observation of the microstructure of the
fracture surface after flexural test (Fig. 4def). In the case of d-HA/
3.1. Characterization of nano-HA particles PMMA cement, catecholamine in pDA on the surface of HA
nanoparticles could conjugate with each other and facilitate the
The surface modification of HA was clearly evidenced by the aggregation of HA. Consequently, the d-HA/PMMA cement
FTIR spectra (Fig. 3a). The strong absorbance at 563 cm1 and exhibited rather rough fracture surface with more cracks,
602 cm1 can be assigned to the PeO bending of the phosphate implying the decreased uniformity of the dispersion of nano-
group, and the peak at 1038 cm1 can be attributed to stretching fillers in the cement which would in turn initiate more cracks
band of OePeO phosphate ions. After the pDA was coated onto the (Fig. 4f); and the formation and propagation of cracks would
surface of HA, the peak at 1038 cm1 in the pristine HA (curve of finally lead to inferior mechanical property. The conjugation of
HA) shifted to 1032 cm1 (curve of d-HA), which was attributed to SSVPT peptide to the surface of d-HA would consume catechol-
the formation of strong adhesion between the HA and the pDA amine in pDA, leading to weakened inter-particle interaction and
coating on nanoparticle surface [14]. The increasing intensity at consequently less aggregation, i.e., better dispersion of the nano-
3433 cm1 can be assigned to the stretching and bending vibrations
of eOH group in pDA. As for p-HA, the intensity of the peak at
1620 cm1 was significantly enhanced as compared with the peak
at 1503 cm1, this could be attributed to the superimposed vibra-
tion of amide II (ıNH) and H2O.
The thermal stability of the modified HA as well as the content
of pDA/peptide on the surface of HA was also investigated. As
shown in Fig. 3b, the slight weight loss of pristine HA at 0e370  C
can be attributed to the desorption of physically adsorbed water.
The rapid weight loss of d-HA and p-HA at 370e700  C can be
assigned to the decomposition of pDA or oligopeptide on the sur-
face of HA, indicating that pDA and/or oligopeptide have been
successfully coated on the sruface of HA. The modification degree of
pDA and SSVPT on the HA surface was found to be 8.95%, 13.09%
respectively.
The morphologies of pristine nano-HA and modified HA were
observed under TEM (Fig. 3cee). All HA nanoparticles exhibited
typical rode-like shape in the distilled water with the length
about 30 nm and the diameter around 8 nm. Very minor change
in size and shape of these nanoparticles was observed after
surface modification, indicating that the surface modification did
not significantly change its morphology and would guarantee Fig. 5. Flexural strength of biomechanical samples that composed of swine femur
good performance/property. Due to the strong interfacial adhe- segment and PMMA-based after soaked in PBS for 3 days at 25  C. (results are the
sion of pDA [12,15], aggregation of d-HA (Fig. 3d) and p-HA mean ± SD, n ¼ 4, t-test; **P < 0.001).
 T. Kang et al. / Composites Science and Technology 123 (2016) 232e240 237

filler in the PMMA matrix. The better dispersion of p-HA in the interact with Ca2þ in the mineral of swine femur and strengthen
PMMA cement matrix in turn resulted in moderately improved the interfacial adhesion [16], this might become an innovative
mechanical properties (Fig. 4g). strategy for clinical bone repair.
Although the d-HA aggregates in PMMA matrix and deteriorates
the bulk mechanical properties of the cement, the presence of d-HA
may significantly enhance the interfacial adhesion between fresh 3.3. Biomineralization
swine femur segment and bone cement. The adhesion forces of d-
HA/PMMA, p1-HA/PMMA, and p2-HA/PMMA cement with swine Another key point of bone cement for clinical application is to
femur were 4.5 ± 0.2 MPa, 4.71 ± 0.32 MPa, and 4.72 ± 0.28 MPa induce the formation of apatite [32]. Similar to that on the surface
(Fig. 5), which were much higher than those of HA/PMMA of PMMA cement in our previous report [33], no discernible
(3.5 ± 0.8 MPa) and BaSO4/PMMA (3.3 ± 0.4 MPa) cement. Taking mineralization can be found on the surface of BaSO4/PMMA cement
advantage of the surface-anchored catecholamine moieties in pDA, (Fig. 6, image a1ea3) after 14 days of immersion in SBF. Upon the
it is reasonable to speculate that the pDA moiety in the cement may addition of SSVPT (image b1eb3) or nano-HA particles (image
c1ec3) alone to the PMMA matrix, needle-like crystallite was found

Fig. 6. SEM images of PMMA-based bone cement soaked in SBF solution at 37  C for 0 (a1ef1), 7 (a2ef2), and 14 (a3ef3) days. EDX spectra of the surface of PMMA-based bone cement
after immersion in SBF at 37  C for 14 days (a4ef4). (Scale bar represents 5 mm).
238 T. Kang et al. / Composites Science and Technology 123 (2016) 232e240

to deposit on the surface of cements and the amount of crystallites attachment is one of the key challenge for vertebral repair. The
increased with time, demonstrating that the bioactive SSVPT or HA effect of pDA-coated HA and SSVPT peptide on the bioactivity of
in the PMMA-based bone cement can induce the formation of cement was evaluated in terms of the cell morphology and
mineral [34,35]. In the case of d-HA/PMMA cement, the mineral adherent number on the surface of cements. Our previous study
(image d3) was much thicker than that on HA/PMMA cement (im- [33] had revealed that the introduction of bioactive HA into PMMA-
age c3) after soaking in SBF for 14 days. The much thicker mineral based cement would promote cementecell interaction and induce
layer on d-HA/PMMA cement can be attributed to the abundant deposition of extracellular matrix (ECM) proteins to offer a good
catecholamine moieties in pDA layer that could induce nucleation substrate for cell adhesion [36]. After 3-day incubation, hFOBs
of hydroxyapatite by concentrating Ca2þ at the interface [16]. showed preferential anchorage to the surface of HA/PMMA cement
The introduction of BMP2 biomimetic peptide can further (Fig. 8b) compared with that of BaSO4/PMMA cement (Fig. 8a).
accelerate the formation of mineral. As shown in image e1ee3 Multi-layer cells were observed on the surface of px-HA/PMMA
(Fig. 6), the crystallite layer became increasingly thicker on p1-HA/ cement (Fig. 8c and d) and hFOBs displayed typical polygonal
PMMA surface. Compared with that of d-HA/PMMA cement (image morphology with more prominent filopodia (insets in Fig. 8c and
d1ed3), higher amount of mineral on the surface of p1-HA/PMMA d). The phenomenon revealed that the presence of pDA, HA, and
cement was observed, suggesting that the introduction of SSVPT SSVPT can improve the cement-cells interfacial bioactivity and
could further accelerate the formation of mineral. Additionally, the promote the adhesion of hFOB cell on cement surface [14,37].
7 days' mineralization on the surface of p2-HA/PMMA cement The number of hFOBs on the cements was also quantified
(images f2) was similar to that on surface of p1-HA/PMMA cement (Fig. 8e). In all cases, the number of cells on the cements increased
in 14 days (image e3); after 14 days' mineralization in SBF, the with time, indicating the proliferation of cells on the cements; and
mineral on p2-HA/PMMA cement surface aggregated to form big
blocks (images f3), indicating that increasing the content of SSVPT
peptide could also promote the formation of mineral.
The Ca/P ratio of the newly formed mineral on the surface of
cement was detected by EDX (image a4ef4). After 14 days' miner-
alization, the Ca/P ratio of the minerals decreased gradually from
1.89 to 1.75, approaching that of HA (1.67).

3.4. Cell viability

In vitro cytotoxicity of PMMA-based bone cement towards

hFOBs was determined via MTT assay (Fig. 7). The cell viabilities in
the presence of extract of PMMA and BaSO4/PMMA cements were
66% and 72%, respectively. While that of HA/PMMA reached 80%,
suggesting that HA could improve the cytocompatibility of the
PMMA cement. The cell viabilities with px-HA/PMMA cements
were even higher, e.g., approx. 98% for p1-HA/PMMA cement and
>100% for p2-HA/PMMA cement, indicating the incorporation of
pDA and SSVPT peptide can synergistically improve the cyto-
compatibility of the cement.

3.5. Cell attachment

The design of a bioactive bone cement for enhancing cell

Fig. 8. (aed) SEM images of hFOB cells adhesion on the surface of PMMA-based bone
cement after 3 days' incubation. (e) Relative number of cell adhesion on the surface of
PMMA-based bone cement. (results are the mean ± SD, n ¼ 5, *represents significant
difference between px-HA/PMMA cement (x ¼ 1,2) and the other control cement,
Fig. 7. Cell viability of hFOB cells incubated with the “extract” of PMMA-based bone p < 0.05).
cement. (results are the mean ± SD, n ¼ 5, t-test; *p < 0.05; **P < 0.001).
 T. Kang et al. / Composites Science and Technology 123 (2016) 232e240 239

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