Materials Research Express

PAPER

Composite hydrogels for applications in sodium heparin controlled

release systems
To cite this article: Caroline Pigatto et al 2019 Mater. Res. Express 6 125406

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 Mater. Res. Express 6 (2019) 125406 https://doi.org/10.1088/2053-1591/ab562f

PAPER

Composite hydrogels for applications in sodium heparin controlled

RECEIVED
27 August 2019
release systems
REVISED
4 November 2019
ACCEPTED FOR PUBLICATION
Caroline Pigatto1, Júlio Cesar Colpo1, Nayrim Brizuela1, Markus Berger2 and
11 November 2019 Luis Alberto Loureiro dos Santos1
1
PUBLISHED Materials Engineering Department, Federal University of Rio Grande do Sul, Porto Alegre/RS, Brazil
21 November 2019 2
Laboratório de Bioquímica Farmacológica, Centro de Pesquisa Experimental, Hospital de Clínicas de Porto Alegre (HCPA/UFRGS),
Brazil
E-mail: carolpigatto@gmail.com

Keywords: hydrogel, biomaterial, drug delivery system

Abstract
Hydrogels are important in biomaterials due to their similar physical properties to living tissue, such
as high water content and soft and elastic consistency. Poly (N-vinyl-2-pyrrolidone) (PVP) forms
polymer complexes with acrylic acid through hydrogen bonds and electrostatic interactions. The
combination of this hydrogel with hydroxyapatite results in an even more biocompatible biomaterial
due to the contributions of its components, making its use more advantageous with additional sites for
drug attachment. The aim of this work was to obtain and characterize composite PVP and
hydroxyapatite hydrogels with incorporated sodium heparin for application in controlled release
systems. For characterizing the obtained hydrogels, FTIR, swelling and SEM techniques were used.
Hydrogels with hydroxyapatite were used as vehicles in the study of sodium heparin release, as a new
potential material for stent coating, and as an anticoagulant and antithrombotic agent. There was a
controlled release of the drug and this system is suitable for these applications, with more potential for
the new composite hydrogel developed. From the data obtained in the release profiles, mathematical
treatments were used to determine the release kinetics. The Peppas-Shalin and Ritger-Peppas models
were the most adequate in this study.

Introduction

Hydrogels are cross-linked polymeric materials that retain a significant fraction of water in their structure
without dissolving and are composed of a three-dimensional network formed by cross-linked homopolymers or
copolymers [1]. They are biocompatible materials presenting an elastic consistency due to the amount of water
they can retain in their structure with low surface tension, thus reducing the irritation produced by friction on
the tissues they contact [2].
However, hydrogels still exhibit limitations due to uneven distribution of crosslinks in their three-
dimensional structure, resulting in materials with low strength and stability, mainly in the swollen state. To solve
these problems, chemical and physical crosslinking methods are used. However, by increasing the amount of
crosslinking, there is a reduction in swell ability and drug delivery. The presence of particles or nanoparticles of
bioactive inorganic constituents, such as, metal oxides [3], black phosphorus nanosheets [4], silicates [5] or
hydroxyapatite [6] in the hydrogel structure can act as physical crosslinking points by improving the final
properties. Hydroxyapatite can act both for physical crosslinking and as a site for drug attachment, with a
potential increase in drug delivery.
The importance of hydrogels as biomaterials is due to the similarity of their physical properties with living
tissues, such as the high water content and the soft and elastic consistency. In recent years, hydrogels have
become promising candidates for tissue engineering scaffolds because they have hydrophilic porous structures
and have advantages for cell adhesion, nutrient delivery, drug release, and tissue regeneration [7]. In controlled

© 2019 IOP Publishing Ltd

Mater. Res. Express 6 (2019) 125406 C Pigatto et al

Table 1. Hydrogel compositions (%).

Coding of
samples PVP AA HA MBAM TEMED AIBN

VP 30.0 — — 2.0 0.025 3.0

VPAA 30.0 0.55 — 2.0 0.025 3.0
VPHA 30.0 — 1.0 2.0 0.025 3.0
VPAAHA 30.0 0.55 1.0 2.0 0.025 3.0

release systems, hydrogels have been proposed for a number of bioactive agents, such as contraceptives,
ophthalmic, antibiotics, antiarrhythmics, anticancer, antibodies, and anticoagulants [8, 9].
Heparin is the most widely used anticoagulant and antithrombotic agent for treating thrombosis (venous,
pulmonary and cerebral) and myocardial infarction and preventing cerebral vessel obstruction in patients with
atrial fibrillation (a type of cardiac arrhythmia). Suitable anticoagulation control is a very delicate and important
process because it is not only restricted to protection against thromboembolic phenomena, but it is also
responsible for reduced triggering of the systemic inflammatory response by decreasing the hemolysis and
platelet degradation rates. To minimize discomfort and local bruising, an alternative is to use HP with hydrogels
in controlled release systems. These systems improve stability, absorption, and therapeutic concentration over a
pre-established period by reducing dosage frequency and patient discomfort [10, 11].
Poly (N-vinyl-2-pyrrolidone) (PVP) forms polymer complexes with poly (acrylic acid) through hydrogen
bonds and electrostatic interactions and, depending on the acrylic acid composition, these hydrogels can
respond to pH stimuli. The combination of these hydrogels with hydroxyapatite result in an even more
biocompatible biomaterial due to the contribution of its components, making its use more advantageous. PVP,
being a material with high biocompatibility and non-toxicity, has numerous applications in the medical field as
blood-compatible hydrogels and as a vehicle for drug release due to its ability to absorb large amounts of water
without dissolving [12, 13].
In controlled release, the development of a system that maintains the concentration of the drug in the
bloodstream within the therapeutic range for a prolonged time and with well-established kinetics is sought. This
system offers some advantages over conventional methods, such as maintaining constant drug levels, thus
resulting in greater efficiency in drug use as it is applied directly to the site of action. Models that are based on
studies of release kinetics allow conclusions regarding the dissolution process of a given formulation since it is
possible to know the process speed, the maximum amount dissolved, and the points at which significant changes
can occur [14]. The models proposed by Higuchi [15], Korsmeyer-Peppas [16], Peppas-Shalin [17], Ritger-
Peppas [18] and Linder-Lippold [19] were evaluated. Therefore, the objective of this work was to obtain and
characterize a new system of composite hydrogels from PVP, poly (acrylic acid) and hydroxyapatite with
incorporated sodium heparin for controlled release system. To date no studies of sodium heparin release from
these systems have been reported and due to the anticoagulant and antithrombotic characteristics of heparin, the
studied hydrogels are intended to be evaluate in the future as coatings of metallic stents.

Methodology

Synthesis of hydrogels
Hydrogels of poly (N-vinyl-2-pyrrolidone) and poly (N-vinyl-2-pyrrolidone–co–acid acrylic) were prepared by
free-radical polymerization using ultraviolet (UV) light. Azo-bis-isobutyronitrile (AIBN) (Sigma-Aldrich,
Brazil), was used to initiate the reactions, N,N′-methylenebisacrylamide (MBAM) (Sigma-Aldrich, Brazil), was
used as a crosslinking agent and N,N,N′,N′-tetramethylethylenediamine (TEMED) (Sigma-Aldrich, Brazil) was
used as a catalyst for the reaction. The solutions were prepared using 70 wt% distilled water and 30 wt% total
monomers. The hydrogel compositions used throughout this investigation are listed in table 1. The mixture was
stirred for 24 h; afterward, the solution was cured under UV irradiation for 6 min at room temperature. The
hydrogels were washed using distilled water to remove residual monomers. Hydroxyapatite was synthesized
using the wet chemical method from Ca(OH)2 (Dinâmica, Brazil) and H3PO4 (Dinâmica, Brazil), both at 0.5 M
concentration, a Ca/P ratio of 1.67, and particle size of about 35 nm.

The drug doped hydrogels

To obtain the drug doped composite hydrogels, we used VPHA and VPAAHA composite hydrogels and
Hepamax-S 5000 UI/ml (Blau Farmacêutica S.A., Brazil) as the drug. The amount of sodium heparin in each
sample was 26.5 mg (1 ml of Hepamax-S), to which 1.5 ml of the hydrogel solution was added. The mixtures
were stirred for 1 h at room temperature and later placed in a rectangular- silicone mold to obtain the specimens.

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Table 2. Mathematical equations applied to the

kinetic study.

Models Equations

Higuchi Mt/M∞=Kt1/2
Korsmeyer-Peppas Mt/M∞=Ktn
Ritger-Peppas Mt/M∞=K1t1/2+K2t
Peppas-Shalin Mt/M∞=K1tn+K2t2n
Linder-Lippold Mt/M∞=Ktn+b

Table 3. Interpretation of the release mechanisms

through the value of the release exponent.

Release Exponent (n) Release Mechanism

0.5>n Pseudo-Fickian Diffusion

0.5 Fickian Diffusion
0.5<n<1.0 Anomalous Diffusion
1.0 Transport Case II

The irradiation time under UV lamp for curing the hydrogels was 7 min; after that, the hydrogels were washed
using distilled water to remove residual monomers.

Characterization

Fourier transform infrared spectroscopy (FTIR)

FTIR was used to characterize the functional groups present in the hydrogels. KBr pellets containing the sample
were prepared and characteristic absorption peaks were detected in the 4000 to 400 cm−1 range using Spectrum
100-PerkinElmer equipment.

Swelling test
The swelling kinetics were measured using hydrogel water absorption as a function of time. Hydrogel samples
were placed in distilled water at room temperature. The swollen gel mass was measured at different time
intervals until equilibrium was reached. Water absorption was calculated according to the equation: Wat
=(Wst−W0)/W0, where Wat is the weight of the water absorbed by the hydrogel at time t, Wst is the weight of
the swollen hydrogel at time t, and W0 is the initial weight of the hydrogel sample.

Scanning electron microscopy (SEM)

SEM analysis was performed to analyze the hydrogels’ microstructure and to verify the hydroxyapatite
incorporation. The equipment used was a scanning electron microscope, Jeol JSN 6060 model. The analysis
conditions were 15 keV and 400 times magnification.

Drug release
During the experiment to verify drug release, we used test tubes containing the hydrogel specimen and a buffer
solution, which were placed in an Erlenmeyer flask under stirring and heated to about 37 °C to simulate the
blood system. To analyze the solution aliquots, they were collected with a syringe in the first 8 h and then at
determined times for 30 days. The aliquots were analyzed using UV-vis Spectroscopy and the turbidimetric
method, which was performed in triplicate for each hydrogel.

Release kinetics
The data obtained in the release profiles were submitted to mathematical treatments for determining the release
kinetics; five mathematical models (table 2) were applied, Higuchi, Korsmeyer-Peppas, Ritger-Peppas, Peppas-
Shalin, and Linder-Lippold.
Where Mt/M∞ is the fraction of drug released at time t, K is the release rate constant, and n is the exponent
that indicates the mechanism in front of which the release occurs (table 3).
When the value of n is 0.5, the drug release follows the diffusion mechanism is Fickian. An anomalous
diffusion occurs when the values of n are greater than 0.5 and smaller than 1.0. In cases where the release system
is a porous material, the constant n may result in values smaller than 0.5 and a pseudo-Fickian diffusion

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mechanism is observed. When the release process is controlled by polymer chain relaxation, the value of n is 1.0
and the mechanism is transport Case II [20].

Turbidimetric analysis
The drug release were tested using the turbidimetric method. Turbidimetric measurements were carried out by
adding 0.5 ml of the sample containing heparin, 0.5 ml of the buffer solution (Neon, Brazil), 2 ml of a 0.1 wt%
cetylpyridinium chloride (Sanofi-Aventis Farmacêutica, Brazil) and 0.94 wt% NaCl (Sigma-Aldrich, Brazil)
solution. A blank sample was prepared following the same procedure, using 0.5 ml of the buffer, except for the
addition of the drug. The bottles were stirred at 37 °C for 1 h. All analyses were performed on a Varian Cary 100
UV-vis Spectrophotometer at 290 nm [21].

Hemolysis assay
A hemolysis assay was used to verify the release of hemoglobin resulting from premature destruction of red
blood cells by rupturing the plasma membrane and to assess the effect of sodium heparin (HP) on the blood-
reacting HPHA hydrogel. Metallic stents coated with the VPHA hydrogel (with and without heparin) and a non-
covered stent as control (CTRL) were analyzed. To evaluate only the effect of the hydrogel, we used a 24-well
plate coated or not (control). Human blood (10 ml) was centrifuged at 1500 × g for 20 min and the obtained
plasma was discarded. The red cell concentrate was washed 3 times, and after the last wash, a 10% red cell
suspension was prepared in 0.9% NaCl (Sigma-Aldrich, Brazil). 2 ml of this red cell suspension was added
directly to the polypropylene tubes containing the stents and 2 ml/well on the plate containing the hydrogels.
Both plates and tubes were maintained under mild agitation throughout the assay period at room temperature.
After different incubation times (60, 120, 240, 360, and 1440 min), 0.1 ml of the red blood cell suspension was
collected, centrifuged (1500 × g for 20 min), and the free hemoglobin content was measured in the supernatant
by reading absorbance at 540 nm. The concentration (μg/ml) of hemoglobin released during the assay was
calculated with the aid of a standard curve of purified human hemoglobin.

Coagulation assay
The effects of hydrogels or hydrogels plus heparin on blood coagulation was tested in vitro in 24-well plates or
stents covered or not with the hydrogels. For this purpose, human citrated plasma (2 ml) was obtained from the
blood bank of Hospital de Clínicas de Porto Alegre (ethical approval number: XXX) and incubated with the
stents or plates covered with the hydrogel in the presence or absence of heparin. Aliquots (0.03 ml) were taken
after 60, 120, 240, 360, 480, or 1440 min, mixed with 50 mM Tris-HCl (Sigma-Aldrich, Brazil) at pH 7.4, and
coagulation was triggered by the addition of 10 mM CaCl2 (Synth, Brazil). The recalcification time was
determined following the kinetics of clot formation at 650 nm using a microplate reader spectrophotometer
(SpectraMAX, Molecular Devices, USA).

Results

Fourier transform infrared spectroscopy (FTIR)

Figure 1 shows the main bands of hydrogels and sodium heparin. In the PVP spectrum, the characteristic
absorption bands at 3448 cm−1 were associated with the stretching frequency of CN and OH, at 2954 cm−1 with
the CH2, and at 1666 cm−1 with the C=O. The acrylic acid characteristic absorption bands are at 3435 cm−1,
2958 cm−1, and 1729 cm−1 with the OH, CH2 and C=O, respectively. The bands at 1462 cm−1 and 1420 cm−1
correspond to the CH strain. Hydroxyapatite was observed by the presence of the peaks at 1091 cm−1,
1032 cm−1, 601 cm−1, and 564 cm−1, referring to the PO−3 4 [22, 23].
In the heparin spectrum, peaks from the functional groups present in the polymer chain, such as OH, CH,
NH and C=O [24], were observed. The main peaks were found at 3459 cm−1, which is the binding stretching
peak of OH; 2937 cm−1 represents the stretching peak of CH; 1630 cm−1 is the characteristic peak of C=O;
1434 cm−1 is the peak relating to the axial deformation of the C–O group; 1256 cm−1 is from the asymmetrical
deformation of the C–O–C group; 1054 cm−1 is from the symmetrical axial deformation of the C–O–C group;
and the peaks at 827 cm−1, 720 cm−1, and 608 cm−1 correspond to the N–H group stretching out of the plane.

Swelling test
Figure 2 shows the swelling behavior of the hydrogels as a function of the immersion time, and it is possible to
observe different percentages of swelling, this being influenced by the composition of the hydrogels. The
hydrogel swelling increases with increasing immersion time. However, the hydrogels without acrylic acid
present a higher percentage of water absorption, almost double.

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Figure 1. FTIR spectra of hydrogels.

Figure 2. Variation in hydrogel water uptake versus time.

Scanning electron microscopy (SEM)

Figure 3 shows the micrographs of the hydrogels (VP and VPAA), the composite hydrogels (VPHA and
VPAAHA), and the composite hydrogels with incorporated sodium heparin (VPHAHA and VPAAHAHP). The
acrylic acid hydrogel, figure 3(b), has a rough surface. In figures 3(d) and (f) surface porosity can be observed.
The hydroxyapatite is also dispersed more uniformly, showing a more homogeneous surface, in figures 3(c)
and (e).

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Figure 3. SEM images of the hydrogels: (a) VP, (b) VPAA, (c) VPHA, (d) VPAAHA, (e) VPHAHP e (f) VPAAHAHP.

Figure 4. Cumulative percentage sodium heparin release. Mean values of the release percentage up to 24 h of VPHAHP and
VPAAHAHP.

Drug release
The release profiles of the composite hydrogels were obtained through serial collections at predefined time
intervals. Figure 4 shows the graph of the percentage released versus time. The first 8 h the VPHAHP hydrogel
showed the highest drug release, about 80 wt%. The hydrogel containing acrylic acid initially presented a slower
release and, after 48 h, approached the amount released by the other hydrogel. The amount of drug released in
both hydrogel systems decreases with time, reaching a constant value after about 50 h.

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Table 4. Correlation coefficient (r) of the mathematical models evaluated during the first 8 h.

Models Ritger Peppas Peppas Shalin Korsmeyer Peppas Linder Lippold Higuchi

Composite r
VPHAHP 0.926 74 0.995 09* 0.973 04 0.978 78 0.589 33
VPAAHAHP 0.992 61* 0.968 80 0.970 74 0.966 66 0.689 68

Table 5. Peppas-Shalin and Ritger-Peppas equations adjustments at the end of the first 8 h.

VPHAHP VPAAHAHP
Composite
Peppas-Shalin Ritger-Peppas
Models
K1 (min−1) K2 (min−1) n K1 (min−1) K2 (min−1) n

49.9±0.9 −7.6±0.5 0.54±0.03 27.9±0,9 3.5±0.2 0.26±0.03

Figure 5. Concentration of hemoglobin released as a function of time: (A) hydrogels and (B) stents (CTRL: well without hydrogel;
CTRL-STENT: non-hydrogel stent).

Kinetics release analyses

The data obtained from the release profiles were submitted to mathematical treatments to determine their
release kinetics. In this study, five mathematical models were applied, Higuchi, Korsmeyer-Peppas, Ritger-
Peppas, Peppas-Shalin, and Linder-Lippold. The obtained correlation coefficient (r), which is a measure of fit
quality that describes the most suitable model to represent the sodium heparin release, are shown in table 4. The
kinetic models that best suited the hydrogel formulations were Peppas-Shalin to VPHAHP and Ritger-Peppas to
VPAAHAHP.
Table 5 shows the kinetic parameters calculated for the systems involved. The diffusion coefficient value
obtained for the VPHAHP hydrogel, n=0.54, confirms that the diffusion process of sodium heparin in the first
8 h is Fickian. For the VPAAHAHP hydrogel, the Ritger-Peppas model presented a better mathematical
adjustment, and the diffusion coefficient value was 0.26.

Hemolysis assay
Figure 5 shows graphs of the hemoglobin concentration released (μg/ml) as a function of time for hydrogels and
coated stents, with and without heparin. Statistical analyses were performed with two-way ANOVA followed by
the Tukey post-hoc-test. In hydrogel-covered plates, there was a significant increase in hemolytic effect after 360
and 1440 min of incubation (figure 5(A)). The same was not observed with the hydrogel-coated stents. There
was no significant hemolytic effect induced by coated or uncoated stents (figure 5(B)).

Coagulation assay
The effects of hydrogels on blood coagulation were tested in an in vitro coagulation assay using plates or stents
covered with hydrogels and heparin. As depicted in figure 6, both plates or stents covered with the hydrogels
(without heparin) were able to increase the coagulation time of human plasma. VPAAHA hydrogel was able to
completely inhibit plasma coagulation (coagulation time more than 1000 s) after 240 min incubation

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Figure 6. Coagulation time as a function of time: (A) hydrogels and (B) stents (CTRL: well without hydrogel; CTRL-STENT: non-
hydrogel stent).

(figure 6(A)). In the presence of heparin, coagulation was blocked even in the initial incubation times for both
plate or stents (figure 6(B)).

Discussion

The peaks between 1666 cm−1 and 1729 cm−1 refer to the C=O group for both polymers, which according to Jin
et al [25], indicates the presence of intermolecular hydrogen bonds due to the carbonyl groups. As reported by
Sohail et al [12], the carbonyl complex bands are wider than in the pure monomers of PVP and AA, hence
evidencing the intermolecular hydrogen bonds. The N–H stretch is between 3330 and 3060 cm−1 and the C–N
stretch is at 1650 cm−1, indicating the presence of the crosslinking agent (MBA).
According to Jovanovic et al [26], the main characteristic of the PVP spectrum is that the monomer N-vinyl-
2-pyrrolidone contains the carbonyl amide group corresponding to the peak at 1666 cm−1, and the peaks at
3441 cm−1 and 2953 cm−1 refer to OH and CH2, respectively. As reported by Bajpai et al [27] and Jankaew et al
[28], the characteristic peak for acrylic acid is at 1759 cm−1 and relates to the C=O of the carboxylic group
(COO−), and the presence of a wide band at 3220–3600 cm−1 is characteristic of both polymers due to the
hydroxyls linked to hydrogen. Abu-Seman et al [29] identified a new peak at 1637–1640 cm−1, representing the
group C=CN, whereas the peaks between 1670–1678 cm−1 were related to the amide stretching of the PVP
carbonyl group.
The hydrogel swelling behavior has an important role in controlling drug release. In general, the greater the
swelling behavior, the higher the drug release. In addition to the peculiar characteristics of hydrogels, some
factors may change their behavior, such as pH, temperature, irradiation time, the addition of other components,
etc [12]. In this study, the behavior of the obtained curves can be attributed to the interpolymer complex formed
between PVP and AA through the intermolecular coupling of hydrogen bonds, as previously described and
presented in the FTIR spectra. The three-dimensional network formed results in a mobility restriction in the
polymer chains, which directly influences the swelling behavior of the hydrogels. Therefore, water diffusion
becomes harder in these hydrogels, resulting in a lower swelling percentage [13].
Jin et al [30] reported that in these complexes formed by PVP/AA, there is also a significant hydrophobic
interaction due to the inherent hydrophobicity of the C–C structure. Therefore, this may contribute to the lower
percentages of swelling for the hydrogels with acrylic acid. The presence of hydroxyapatite particles has
interfered with the swelling behavior of the hydrogels, i.e., the small particles may have hindered the water
absorption process by the locking the polymeric structure, which is beneficial in a controlled drugs release
system.
The morphological analysis of the hydrogels structure is also an important parameter for controlled drug
release systems. The porosity could be directly related to three factors, molecular weight and chain size,
interpolymer complex formation (crosslinking), and drug interaction with the polymers [12, 24]. PVP forms
larger pores, although in lower amounts due to its high molecular weight and chain length [12]. As already
verified in the FTIR analysis, the hydrogels with acrylic acid form an interpolymer complex through the
intermolecular bonds of hydrogen (crosslinking increasing). This results in the chains forthcoming and,
consequently, in reducing free spaces, increasing the number of smaller pores. There are several works that
justify the formation of smaller pores, due to the greater extent of crosslinking in the biomaterials
manufacturing process [31, 32].

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Regarding the drug interaction with the polymers, depending on the nature of the functional groups of both,
these interactions may indeed exist. As already described in the literature, heparin can be conjugated with
organic compounds to become a more hydrophobic drug [21]. Therefore, for these hydrogel compositions,
sodium heparin has a strong tendency to form interactions with the carboxyl groups of acrylic acid.
The profiles obtained present a controlled release drug profile, and this behavior offers advantages over
conventional methods, such as maintaining constant drug levels in the body for a certain period and,
consequently, increasing patient comfort and treatment effectiveness.
This can be explained by three factors already described above, porosity; forming the interpolymer complex
(crosslinking), drug interaction with the polymers and hydroxyapatite. The VPHA composite hydrogel presents
a lower degree of crosslinking and larger pores, hence facilitating the release of heparin, which occurs faster in
the first few hours because it is closer to the surface. For the hydrogel with acrylic acid (VPAAHA), the presence
of the interpolymer bonds increases chain crosslinking and decreases the pore size, making the drug release
process more difficult. It is also worth mentioning that the interaction of heparin with the carboxyl groups of
acrylic acid delays its release [24, 33], and the addition of HA also contributes to the reduction of reduced water
absorption and swelling, as seen in figure 3, and the interaction of heparin on surface of hydroxyapatite.
This rapid release rate of heparin may be advantageous, as it would ensure a high drug concentration at the
beginning of the implantation. The hydrogels work as a matrix system where the drug may or may not be
homogeneously dispersed in the polymer matrix, and their release involves physical and chemical processes,
including water intake in the matrix that causes the swelling, chain relaxation, drug dissolution, and water
diffusion through the matrix pores [34, 35]. Therefore, the release profiles of these systems are characterized by a
decrease in the release rate over time due to the longer distance the drug must travel from the interior to the
matrix surface.
The kinetic parameters were determined from the obtained dissolution profiles, which relate the dissolved
percentage versus time and produce a comparative analysis concerning the in vitro behavior of these materials.
The kinetics evaluation allows conclusions regarding the formulation dissolution process. The releasing stage of
the active substance from a hydrophilic matrix system results in a complex interaction between swelling,
dissolution, diffusion and erosion mechanisms. This complexity happens because hydrogel molecules, while in
contact with water, acquire micro and macro structures because of the ‘dry’ transition process to the malleable
state, depending on the water exposure time [36]. Consequently, the mathematical treatment covering all these
mechanisms becomes much more complex.
Table 5 shows the kinetic parameters calculated for the systems involved, and it can be observed that the
contribution of Fickian diffusion (K1) predominates over the contribution of polymer chains relaxation (K2),
therefore K1>K2. This is also justified by the obtained values of the diffusion coefficient. Thus, for the
composite hydrogel VPHAHP, the most adequate model to quantify the relative contributions of the two
phenomena responsible for releasing the drug was that of Peppas-Shalin. For the VPAAHAHP, the Ritger-
Peppas model presented a better mathematical fit. In this case, the drug release kinetics were slower, resulting in
an overlap of both mechanisms, Fickian and Case II transport processes, and the value of the diffusion
coefficient indicated the existence of other phenomena, such as the presence of pores resulting in values of ‘n’
less than 0.5 due to a partial diffusion process through the swollen polymer matrix and through the pores formed
in the biomaterial.
According to Lopes et al [37], in systems with controlled release, the rate of drug liberation results from the
combination of diffusion and Case II transport. In these cases, the diffusion follows Fick’s laws and the Case II
transport reflects the influence of the polymer relaxation on the mobility of the matrix molecules.
The contact between blood and the biomaterials used in the devices can trigger an increase in the systemic
inflammatory response, already activated by the hemodynamic condition of the patients. That is, the
biomaterials can cause hemolysis, especially the stents because they are introduced directly into the blood
vessels, where they remain. Therefore, from the hemolysis test, it was observed that the biomaterial did not cause
hemolysis (did not interfere with the physiological hemostasis) and was suitable for its application.

Conclusion

According to this study, it was possible to obtain and characterize composite hydrogels using PVP for
application in controlled heparin release systems. Through FTIR, swelling, and SEM analyses, it is possible to
observe that hydrogels without acrylic acid show less degree of crosslinking and form larger pores; therefore,
they swell more. However, hydrogels with acrylic acid develop interpolymer complexes that increase the degree
of crosslinking, resulting in smaller pores and less degree of swelling.
Hydroxyapatite did not interfere in the wave spectrum of the corresponding hydroxyapatite-free
formulations. However, there was heparin binding to the carboxylic groups of acrylic acid; the formulations

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containing the drug presented changes in the drug release profile and pore formation. The presence of the
hydroxyapatite particles prevented the hydrogel swelling behavior, acting as a physical agent to block this
swelling and as a site for heparin attachment, thus promoting a drug-controlled release.
The composite hydrogel VPHA was a suitable system for releasing sodium heparin. Besides having a
controlled release profile, it showed an accelerated drug release in the first hours, which was an important and
decisive point to avoid complications for the post-implant patient, and a constant release throughout 30 days.
From the hemolysis test, the biomaterial did not cause hemolysis and was suitable for its application.

ORCID iDs

Luis Alberto Loureiro dos Santos https://orcid.org/0000-0002-9099-9748

References
[1] Ahmed E M 2015 J. Adv. Res. 6 121
[2] Güven O, Sen M, Karadag E and Saraydm D 1999 Radiat. Phys. Chem. 56 386
[3] Pasqui D, Atrei A, Giani G, De Cagna M and Barbucci R 2011 Mater. Lett. 65 392
[4] Huang K, Wu J and Gu Z 2019 ACS Applied Materials & Interfaces 11 2908
[5] Gaharwar A K, Rivera C P, Wu C J and Schmidt G 2011 Acta Biomater. 7 4139
[6] Gaharwar A K, Dammu S A, Canter J M, Wu C and Schmidt G 1641 Biomacromolecules 2011 12
[7] Zhang S, Ou Q, Xin P, Yuan Q, Wang Y and Wu J 2019 Biomater. Sci. 7 4230
[8] Hoffman A S 2002 Adv. Drug Deliv. Rev. 43 12
[9] Deligkaris K, Tadele T S, Olthuis W and Berg A V D 2010 Sens. Actuators B 147 774
[10] Demoré B, Benoit E, Maincent P, Hoffman M and Bessièrre J 1998 J. Clin. Pharm. Ther. 23 384
[11] Briger I, Dubernet C and Couvreur P 2002 Adv. Drug Deliv. Rev. 54 651
[12] Sohail K, Khan I U, Shahzad Y, Hussain T and Ranjha N M 2014 Braz. J. Pharm. Sci. 50 184
[13] El-Rehim H A, Hegazy E A, Khalil F H and Hamed N A 2007 Nucl. Instrum. Methods Phys. Res. B 254 112
[14] Siegel R A and Rathbone M J 2012 Overview of Controlled Release Mechanisms (Springer Series in Fundamentals and Applications of
Controlled Release Drug Delivery XIV) (US: Springer)
[15] Fernández J A, Santos R G, Guerra N B and Valdés L O 2009 Rev. Iberoamericana de Polím. 10 130 http://www.ehu.eus/reviberpol/
pdf/MAR09/aragon.pdf
[16] Korsmeyer R W, Gurny R, Doelker E, Buri P and Peppas N A 1983 Int. J. Pharm. 15 35
[17] Peppas N A and Sahlin J J 1989 Int. J. Pharm. 57 172
[18] Ritger P L and Peppas N A 1987 J. Control. Release 5 42
[19] Lindner W D and Lippold B C 1785 Pharm. Res. 1995 12
[20] Cirillo G, Spataro T, Curcio M, Spizzirri U G, Nicoletta F P, Picci N and Iemma F 2015 Mater. Sci. Eng. C 48 510
[21] Oliveira S S M and Marchetti J M 2011 Química Nova 34 1067
[22] Can H K 2005 Radiat. Phys. Chem. 72 710
[23] Chen K S, Ku Y A, Lin H R, Yan T R, Sheu D C, Chen T M and Lin F H 2005 Mater. Chem. Phys. 91 489
[24] Coelho T C 2004 Diploma de Graduação Universidade Federal de Santa Catarina http://repositorio.ufsc.br/xmlui/handle/
123456789/94557
[25] Jin S, Liu M, Zhang F, Chen S and Niu A 2006 Polymer 47 1532
[26] Jovanovic Z, Radosavljevic A, Siljegovic M, Bibic N, Stankovic V M and Popovic Z K 2012 Radiat. Phys. Chem. 81 1728
[27] Bajpai S K and Dubey S 2005 React. Funct. Polym. 62 104
[28] Jankaew R, Rodkate N, Lamlertthon S, Rutnakornpituk B, Wichai U, Ross G and Rutnakornpituk M 2015 Polym. Test. 42 36
[29] Abu-Seman M N, Khayet M and Hilal N 2012 Desalination 287 29
[30] Jin S, Liu M, Chen S and Chen Y 2005 Eur. Polym. J. 41 2415
[31] Huang J, Huang K, You X, Liu G, Hollett G, Kang Y, Gu Z and Wu J 2018 Journal of Materials Chemistry B 6 1328
[32] Schoenmakers D C, Rowan A E and Kouwer P H J 2018 Nat. Commun. 9 3
[33] Lunelli C E 2013 Dissertação de Mestrado Universidade Federal do Paraná http://hdl.handle.net/1884/35343
[34] Lyra M A M, Soares-Sobrinho J L, Brasileiro M T, Roca M F, Barraza J, Viana O S and Rolim-Neto P J 2007 Lat. Am. J. Pharm. 26 793
http://www.latamjpharm.org/trabajos/26/5/LAJOP_26_5_5_1_5NH237W57Y.pdf
[35] Pezzini B R, Silva M A S and Ferraz H G 2007 Rev. Bras. Cienc. Farm. 43 501
[36] Manadas R, Pina M E and Veiga F 2002 Rev. Bras. Cienc. Farm. 38 399
[37] Lopes C M, Lobo J M S and Costa P 2005 Rev. Bras. Cienc. Farm. 41 154

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