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.com
Volume 9
Journal of Biotechnology & Biomaterials
ISSN: 2155-952X
Biomaterials 2019
February 25-26, 2019
February 25-26, 2019 | London, UK
4
th
Annual Conference and Expo on
Biomaterials
Argatroban immobilization on Cu-modified PVC and PU
Liana Azizova
1
, Volodymyr Chernyshenko
2
and
Lyuba Mikhalovska
1
1
University of Brighton, UK
2
Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Ukraine
T
hrombosis induced by biomaterials after their contact with blood is a main reason of medical device failure. To make
material surface more thromboresistant different approaches have been undertaken. NO generating biomaterial has
proven to play a crucial role in the prevention of thrombosis by inhibiting the platelets activation/adhesion. However,
immobilization of the direct thrombin inhibitors onto material surface makes material more thromboresistant by preventing
thrombin-mediated blood clotting. The aim of this research was to immobilize argatroban a direct thrombin inhibitor with
reliable and predictable anticoagulant effect onto PVC and PU polymers. Both polymers were first imprinted with Cu ions for
the catalytic generation of NO (this research was reported earlier). Argatroban was immobilized on the Cu-modified PVC and
PU using the polydopamine ad-layer via the Michael addition/Schiff base reaction. The amount of argatroban bound to the
polymer surface was measured (spectrophotometric determination at 334 nm) as 11.92 nmol/cm
2
on PVC and 13.10 nmol/
cm
2
on PU surface. Assay using thrombin-specific chromogenic substrate was performed to evaluate the thrombin inhibition
capacity of argatroban-modified polymers. It was found that both Argatroban-modified polymers inhibit thrombin activity
in PBS. In order to confirm the NO generation catalyzed by Cu/Arg-modified PVC and PU samples after incubation with
100 µM GSNO/GSH in the PBS during 1h was examined using ArrowSTRAIGHT™ nitric oxide measurement system (Lazar
Research Laboratories, Los Angeles, CA, USA). The Cu/Arg-modified PVC and PU generate NO with the rate 1.27-1.66×10
10
mole/cm
2
∙min which is within the physiological level. From the data obtained it’s possible to conclude, that immobilization of
Argatroban to the Cu-modified polymers showed combine abilities: i) generate NO caused by Cu ions and ii) have capacity to
inhibit thrombin formed in the blood via surface immobilized argatroban.
Liana Azizova et al., J Biotechnol Biomater 2019, Volume 9
DOI: 10.4172/2155-952X-C1-112
Figure 1: Generation of pNa from chromogenic
substrate S2238 by 0.4 NIH/ml of thrombin that
was pre-incubated with PU or PU modified with
Argatroban during 20 min. The remnant thrombin
activity is being measured.
Figure 2: NO generation after incubation of Cu/
Arg-modified PVC and PU with 100 µM GSNO/
GSH in the PBS during 1h.