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. 2018 Jan 10;8(1):233.
doi: 10.1038/s41598-017-18665-4.

Thrombin@Fe3O4 nanoparticles for use as a hemostatic agent in internal bleeding

Emiliya M Shabanova  1 , Andrey S Drozdov  2 , Anna F Fakhardo  1 , Ivan P Dudanov  1   3 , Marina S Kovalchuk  3 , Vladimir V Vinogradov  4
Affiliations

Thrombin@Fe3O4 nanoparticles for use as a hemostatic agent in internal bleeding

Emiliya M Shabanova et al. Sci Rep. .

Erratum in

Abstract

Bleeding remains one of the main causes of premature mortality at present, with internal bleeding being the most dangerous case. In this paper, magnetic hemostatic nanoparticles are shown for the first time to assist in minimally invasive treatment of internal bleeding, implying the introduction directly into the circulatory system followed by localization in the bleeding zone due to the application of an external magnetic field. Nanoparticles were produced by entrapping human thrombin (THR) into a sol-gel derived magnetite matrix followed by grinding to sizes below 200 nm and subsequent colloidization. Prepared colloids show protrombotic activity and cause plasma coagulation in in vitro experiments. We also show here using a model blood vessel that the THR@ferria composite does not cause systematic thrombosis due to low activity, but being concentrated by an external magnetic field with simultaneous fibrinogen injection accelerates local hemostasis and stops the bleeding. For instance, a model vessel system with circulating blood at the puncture of the vessel wall and the application of a permanent magnetic field yielded a hemostasis time by a factor of 6.5 shorter than that observed for the control sample. Biocompatibility of composites was tested on HELF and HeLa cells and revealed no toxic effects.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
XRD diffraction pattern of the material corresponds to magnetite oxide phase, peaks referred to JCPDS file No. 19–0629: (red lines) (a); SEM image shows porous microstructure of the material (b); according to TEM, a magnetite matrix consists of truncated tetrahedron nanoparticles (highlighted by white contour) with narrow size distribution (c); low-temperature N2 physisorption curve shows mesoporous structure of the material (d); pore size distribution of the magnetite xerogel matrix by the BJH method (e); zeta potential of the magnetite matrix at different pH values. At pH = 7.4 the matrix is positively charged (f).
Figure 2
Figure 2
SEM (a,b) and TEM (c,d) images of THR@ferria NPs. Interplane spacing is presented in the insert. Hydrodynamic diameter of the composite NPs (e) and profile of thrombin release from them (f) For materials with thrombin mass fractions of less than 10% minor release is observed.
Figure 3
Figure 3
Cytotoxicity on (a) HeLa and (b) HELF cells after 72 h with THR@ferria NPs. Average values of 3 measurements with standard deviations are shown.
Figure 4
Figure 4
Coagulation time reported as percent of control both in glass and plastic cuvettes at fibrinogen concentrations of 2.8 mg/mL (a) and 3.9 mg/mL (b). The increased concentration of fibrinogen (b) is shown to yield comparable coagulation times for free and entrapped thrombin.
Figure 5
Figure 5
Installation scheme for assessing hemostatic effect using the synthesized THR@ferria NPs. A model stand with an artificial blood vessel is shown, inside which a colloid is introduced, followed by localization of nanoparticles at the site of bleeding due to the application of an external magnetic field.
Figure 6
Figure 6
Time (a) and mass of blood (b) flowed out of the model blood vessel during an experiment on analyzing hemostatic activity. The measurements were carried out with different amounts of fibrinogen. The administration of hemostatic nanoparticles is shown to significantly reduce the time and mass of blood prior to hemostasis compared to the control, especially when an external magnetic field is applied and the nanoparticles are concentrated accordingly.
Figure 7
Figure 7
Analysis of hemostatic activity of THR@ferria: (a) General view of the model vessel with circulating blood and nanoparticles (I), bleeding after puncture (II), Magnet apposition to the bleeding zone (III) and appearance of the formed thrombus on the vessel wall after hemostasis. (b) SEM analysis of the thrombus surface with appropriate image mapping of the Fe content (c). The composite nanoparticles are shown to concentrate in the thrombus.
Figure 8
Figure 8
Scheme of synthesis for the hemostatic THR@ferria colloid. After mixing the Fe2 +/Fe3 + salts, an ammonia solution is added (a) followed by washing the precipitate of the formed magnetite (b) and treating it ultrasonically to yield a stable hydrosol (c). Then, the solution of thrombin (d) is added to the sol and the system is condensed by removal of the solvent and physical gelation. The resulting composite with entrapped thrombin molecules is ground (e) and a hemostatic colloid is prepared by filtering the ground nanoparticles through a filter with a nozzle of 200 nm.
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References

    1. De Jong WH, Borm PJ. Drug delivery and nanoparticles: applications and hazards. International journal of nanomedicine. 2008;3:133. doi: 10.2147/IJN.S596. - DOI - PMC - PubMed
    1. Whang HS, Kirsch W, Zhu YH, Yang CZ, Hudson SM. Hemostatic agents derived from chitin and chitosan. Journal of Macromolecular Science, Part C: Polymer Reviews. 2005;45:309–323. doi: 10.1080/15321790500304122. - DOI
    1. Ostomel TA, Shi Q, Tsung C-K, Liang H, Stucky GD. Spherical bioactive glass with enhanced rates of hydroxyapatite deposition and hemostatic activity. Small. 2006;2:1261–1265. doi: 10.1002/smll.200600177. - DOI - PubMed
    1. Anselmo AC, et al. Platelet-like nanoparticles: mimicking shape, flexibility, and surface biology of platelets to target vascular injuries. Acs Nano. 2014;8:11243–11253. doi: 10.1021/nn503732m. - DOI - PMC - PubMed
    1. Yildirim A, Ozgur E, Bayindir M. Impact of mesoporous silica nanoparticle surface functionality on hemolytic activity, thrombogenicity and non-specific protein adsorption. Journal of Materials Chemistry B. 2013;1:1909–1920. doi: 10.1039/c3tb20139b. - DOI - PubMed

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