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An In Vitro Thrombolysis Study Using a Mixture of Fast-Acting and Slower Release Microspheres

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Abstract

Purpose

To test the hypothesis that a mixture combining fast and slower release rate microspheres can restore blood flow rapidly and prevent formation of another blockage in thrombolysis.

Methods

We used polyethylene glycol (PEG) microspheres which provide the release of the encapsulated streptokinase (SK) on the scale of minutes, and Eudragit FS30D (Eud), a polymethacrylate polymer, for development of delayed release microspheres which were desirable to prevent a putative second thrombus. Eud microspheres were coated with chitosan (CS) to further extend half-life. Experiments included the development, characterization of Eud/SK and CS-Eud/SK microspheres, and in vitro thrombolytic studies of the mixtures of PEG/SK and Eud /SK microspheres and of PEG/SK and CS-Eud/SK microspheres.

Results

CS-Eud/SK microspheres have slightly lower encapsulation efficiency, reduced activity of SK, and a much slower release of SK when compared with microspheres of Eud/SK microspheres. Counter-intuitively, slower release leads to faster thrombolysis after reocclusion as a result of greater retention of agent and the mechanism of distributed intraclot thrombolysis.

Conclusions

A mixture of PEG/SK and CS-Eud/SK microspheres could break up the blood clot rapidly while providing clot-lytic efficacy in prevention of a second blockage up to 4 h.

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Abbreviations

CS:

Chitosan

EDC:

1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide

EE:

Encapsulation efficiency

Eud:

Eudragit FS30D

FT-IR:

Fourier transform infrared

MES:

2-(N-morpholino)ethanesulfonic acid

NHS:

N-hydroxysuccinimide

PA:

Plasminogen activator

PAI:

Plasminogen activator inhibitor

PBS:

Phosphate-buffered saline

PEG:

Polyethylene glycol

PPP:

Platelet-poor plasma

PRP:

Platelet-rich plasma

PVA:

Poly(vinyl alcohol)

SEM:

Scanning electron microscope

SK:

Streptokinase

TEM:

Transmission electron microscope

tPA:

Tissue-type plasminogen activator

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ACKNOWLEDGMENTS AND DISCLOSURES

The authors gratefully acknowledge Dr. Preston Larson and Greg Strout from the Samuel Roberts Noble Electron Microscopy Laboratory at the University of Oklahoma for technical assistance with the SEM and TEM experiments. We are also grateful to Evonic Röhm for supplying Eudragit FS30D.

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Authors and Affiliations

  1. School of Chemical, Biological and Materials Engineering, University of Oklahoma, 100 E. Boyd, SEC, T301, Norman, Oklahoma, 73019, USA

    Hoai X. Nguyen & Edgar A. O’Rear

  2. University of Oklahoma Biomedical Engineering Center, Norman, Oklahoma, 73019, USA

    Hoai X. Nguyen & Edgar A. O’Rear

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  1. Hoai X. Nguyen
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  2. Edgar A. O’Rear
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Correspondence to Edgar A. O’Rear.

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Nguyen, H.X., O’Rear, E.A. An In Vitro Thrombolysis Study Using a Mixture of Fast-Acting and Slower Release Microspheres. Pharm Res 33, 1552–1563 (2016). https://doi.org/10.1007/s11095-016-1897-1

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  • DOI: https://doi.org/10.1007/s11095-016-1897-1

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