Improvement of hemostatic function by natural substance in cooperation with recombinant batroxobin
Gyeung Mi
Seon1, 2,
Byeong-Ju
Kwon1, 2,
Min Sung
Kim1, 2,
Min-Ah
Koo1, 2,
Mi Hee
Lee1,
Young
Seomun3,
Jong-Tak
Kim3 and
Jong-Chul
Park1, 2
-
1
Yonsei University College of Medicine, Cellbiocontrol Laboratory, Department of Medical Engineering, Korea
-
2
Yonsei University College of Medicine, Brain Korea 21 PLUS Project for Medical Science, Korea
-
3
BioBud Inc., Korea
Introduction: Uncontrolled hemorrhaging is the leading preventable cause of infectious complications in accidents, surgical procedures and battle fields. Therefore, topical hemostats and sealants are required that they can assist the effective treating arrest bleeding and stabilize the casualty before definitive care. In many type of hemostatic agents, collagen and chitosan are one of the natural substance agents which have been widely used in many biomedical applications as biomaterials for surgical suture materials and wound dressing because of their high biocompatibility and antimicrobial properties. In generally, collagen and chitosan play an important role for clot formation and platelet activation and aggregation in blood coagulation. Additionally, chitosan adheres to red blood cell in blood coagulation. However, some studies have been reported natural substances were not adequately quick to hemostasis within relevant time. In this study, we used batroxobin, a venomous component from the snake Bothrops atrox moojeni, which is catalyzes fibrinogen conversion to form fibrin clot in blood coagulation for functional improvement of natural substance as hemostatic pad. So we expect that batroxobin would support natural substances to synergetic effect of hemostatic properties.
Materials and Methods: In order to fabrication of hemostatic pad, collagen and chitosan was dissolved with batroxobin in acetic acid followed by lyophilization, respectively. The hemostatic pad was characterized using scanning electron microscope (SEM) to determine the structure and pore size of sponge. We executed hemostatic mechanism studies by platelet activation assay and fibrinogen assay. For platelet study, sponge incubated with platelet were fixed and viewed under the SEM. In order to fibrinogen activation assay, batroxobin released on sponge by barbital buffer were added to fibrinogen solution and solution turbidity were detected at 405nm. In whole blood assay, blood of SD rats spread to sponge directly and determinated hemoglobin concentration. For animal experiments, blood loss was measured using the femoral artery wound model of SD rats.
Results and Discussion: The prepared hemostatic pad images shown regular and high porous morphology and the average pore size. Also, batroxobin was not affected on porous of hemostatic pad. In platelet activation assay, it can be seen that the activated platelet spread morphology on the sponge surface even ternary composite sponge. As the concentration of batroxobin added to the sponge was increased, the turbidity of the solution was also increased in fibrinogen assay. Both results confirmed that substances fabricated with batroxobin and natural polymers maintained to their hemostatic properties. In whole blood assay, clot formation rate increased by high concentration of batroxobin with mixed natural substances. Animal studies were also demostrated that fabricated hemostatic pad effectively reduced blood bleeding loss and shortened bleeding time by addition of batroxobin.
Conclusion: Based on the result shown above, the novel hemostatic pad is reasonable that a great potential for excessive bleeding injury and improve effects of natural substance hemostatic pad when it is cooperated with batroxobin.
Keywords:
biomaterial,
Scaffold,
biomedical application,
biofunctional molecule
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
Poster
Topic:
Biomaterials in thrombosis and hemostasis
Citation:
Seon
G,
Kwon
B,
Kim
M,
Koo
M,
Lee
M,
Seomun
Y,
Kim
J and
Park
J
(2016). Improvement of hemostatic function by natural substance in cooperation with recombinant batroxobin.
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.02317
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.