Do Microemulsion-based Nanoparticles Affect Blood Coagulation?

Yan Li, M.D., David G. Bjoraker, M.D., Timothy E. Morey, M.D., Donn M. Dennis, M.D.
Department of Anesthesiology, University of Florida, Gainesville, FL

Original Science

Introduction: Each year there are >300,000 emergency room visits due to poisoning. The cost of treatment exceeded $10 billion/year in 2000, not including $80 billion/year attributed to alcohol related treatment. There are few specific antidotes (e.g., naloxone) available to treat patients. Therefore, the mainstay of therapy remains supportive and symptomatic treatment. Microemulsion-based nanoparticles are nanometer-sized droplets formed of biologically compatible oils and surfactants that offer the ability to absorb large quantities of lipophilic drugs and poisons. One synthetic microemulsion, ME-6, absorbs 97% of bupivacaine in an in vitro assay of whole human blood. As part of toxicity evaluation for this microemulsion, global coagulation function was assessed using the thromboelastograph (TEG).

Methods: After IRB approval, informed consent was obtained from 6 healthy volunteers for venipuncture. Two ml of blood was drawn with a butterfly catheter and discarded. Subsequently, another 2 ml of blood was collected for TEG analysis. No anticoagulant was used. Within 6 min from venipuncture, 330 ?l of blood was transferred into TEG cuvettes prepared with either 30 ?l of normal saline (NS) or ME-6 (0.1 ?g/ml final concentration). No activator other than the plastic cuvette and pin surfaces were used. Assays were performed using a thromboelastograph coagulation analyzer (2000 D, Haemoscope, Skokie, IL). The r time, k time, a angle, and maximum amplitude (MA) of both test and control groups were compared and analyzed. In addition, a CBC was performed. A paired Student t test was used for statistical analysis. P<0.05 was considered to be a statistically significant.

Results: In vitro thromboelastograph and blood count values with and without ME-6 treatment (mean±SD).
 

 

R (min)

K (min)

a (º)

MA (mm)

Platelet (106/ml)

Hg  (g/dl)

WBC (103/ml)

Control

22.4± 3.5

9.9± 1.4

29.1± 3.9

51.0± 3.2

175± 28

13.3± 1.2

5.6± 1.7

Microemulsion

19.1± 2.4

19.4± 4.3

13.8± 3.8

16.3± 5.3

182± 23

13.7± 1.8

5.2± 1.3

P value

0.145

0.0013

0.00007

0.00003

0.1225

0.1772

0.206


Discussion: From other studies (data unpublished), ME-6 absorbed 97% of bupivacaine in vitro and, therefore, may be a potential antidote for bupivacaine toxicity. In this study, ME-6 in vitro affected the coagulation profile significantly. The lack of effect on the onset time of clot formation suggests that impairment of coagulation factor biochemistry did not occur. However, the marked loss of maximum amplitude with ME-6 suggests either severe platelet dysfunction or fibrin polymerization interference. Further studies are needed to define its mechanism.


References:
1. Statistical Abstract of the United States, 120th Edition, 2000.
2. Dennis D. et al. Unpublished data from this group.

 

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