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Thrombus formation induced by laser in a mouse model.

Pérez P, Alarcón M, Fuentes E, Palomo I - Exp Ther Med (2014)

Bottom Line: A simple epifluorescence magnifier was used to detect the presence of thrombi.The results obtained indicate that using rose bengal at concentrations of 25 and 50 mg/kg and a laser power of 5 mW, thrombus formation occurred.For visualization, platelets were labeled with calcein acetyloxymethyl ester for 1 h, which resulted in improved observation of thrombus formation in real time.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Biochemistry and Immunohematology, Faculty of Health Sciences, Interdisciplinary Excellence Research Program on Healthy Aging (PIEI-ES), University of Talca, Talca, Maule 3460000, Chile.

ABSTRACT
Animal models are used for the development of techniques and/or models that aid the study of thrombosis pathophysiology. The aim of the present study was to modify the technique of in vivo thrombosis induction to make it more accessible. BALB/c mice were intraperitoneally anesthetized with 0.4 ml 2,2,2-tribromoethanol (266.6 mg/kg) and xylazine (13.3 mg/kg), whilst maintaining stable blood pressure and temperature. Through abdominal surgery, the mesentery was identified and isolated for the visualization of the arteries. A simple epifluorescence magnifier was used to detect the presence of thrombi. The results obtained indicate that using rose bengal at concentrations of 25 and 50 mg/kg and a laser power of 5 mW, thrombus formation occurred. In addition, formation of the thrombus occurred ~30 min following induction and the thrombus had a total area of 4,878.3 μm(2), which caused total occlusion of the mesenteric artery. For visualization, platelets were labeled with calcein acetyloxymethyl ester for 1 h, which resulted in improved observation of thrombus formation in real time. Therefore, this technique may be used to perform in vivo studies simply and at low cost, and is suitable for use in a variety of studies of thrombosis.

No MeSH data available.


Related in: MedlinePlus

Thrombus formation induced with a laser and 50 mg/kg rose bengal. (A) Typical tracing representing the development of the thrombus with time, following deep laser injury. (B) Schematic representation of the lesion along the artery (left) and in a transversal section at the centre of the lesion (right).
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f2-etm-08-01-0064: Thrombus formation induced with a laser and 50 mg/kg rose bengal. (A) Typical tracing representing the development of the thrombus with time, following deep laser injury. (B) Schematic representation of the lesion along the artery (left) and in a transversal section at the centre of the lesion (right).

Mentions: Fig. 2A shows thrombus formation over time at a concentration of 50 mg/kg. Following laser irradiation, the thrombus formed within 30 min, causing total occlusion of the vessels. During the 60 min of monitoring, the formation of a stable thrombus and blood vessel occlusion was observed. The thrombus was 4,878.3 pixels in size and occluded the blood vessel by 100% at 30 min (n=6). Fig. 2B schematically shows the incidence of the laser beam causing localized damage to the endothelium. Fig. 3 shows in vivo thrombus formation following irradiation with 50 mg/kg rose bengal and fluorescence detection using calcein-labeled platelets.


Thrombus formation induced by laser in a mouse model.

Pérez P, Alarcón M, Fuentes E, Palomo I - Exp Ther Med (2014)

Thrombus formation induced with a laser and 50 mg/kg rose bengal. (A) Typical tracing representing the development of the thrombus with time, following deep laser injury. (B) Schematic representation of the lesion along the artery (left) and in a transversal section at the centre of the lesion (right).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4061192&req=5

f2-etm-08-01-0064: Thrombus formation induced with a laser and 50 mg/kg rose bengal. (A) Typical tracing representing the development of the thrombus with time, following deep laser injury. (B) Schematic representation of the lesion along the artery (left) and in a transversal section at the centre of the lesion (right).
Mentions: Fig. 2A shows thrombus formation over time at a concentration of 50 mg/kg. Following laser irradiation, the thrombus formed within 30 min, causing total occlusion of the vessels. During the 60 min of monitoring, the formation of a stable thrombus and blood vessel occlusion was observed. The thrombus was 4,878.3 pixels in size and occluded the blood vessel by 100% at 30 min (n=6). Fig. 2B schematically shows the incidence of the laser beam causing localized damage to the endothelium. Fig. 3 shows in vivo thrombus formation following irradiation with 50 mg/kg rose bengal and fluorescence detection using calcein-labeled platelets.

Bottom Line: A simple epifluorescence magnifier was used to detect the presence of thrombi.The results obtained indicate that using rose bengal at concentrations of 25 and 50 mg/kg and a laser power of 5 mW, thrombus formation occurred.For visualization, platelets were labeled with calcein acetyloxymethyl ester for 1 h, which resulted in improved observation of thrombus formation in real time.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Biochemistry and Immunohematology, Faculty of Health Sciences, Interdisciplinary Excellence Research Program on Healthy Aging (PIEI-ES), University of Talca, Talca, Maule 3460000, Chile.

ABSTRACT
Animal models are used for the development of techniques and/or models that aid the study of thrombosis pathophysiology. The aim of the present study was to modify the technique of in vivo thrombosis induction to make it more accessible. BALB/c mice were intraperitoneally anesthetized with 0.4 ml 2,2,2-tribromoethanol (266.6 mg/kg) and xylazine (13.3 mg/kg), whilst maintaining stable blood pressure and temperature. Through abdominal surgery, the mesentery was identified and isolated for the visualization of the arteries. A simple epifluorescence magnifier was used to detect the presence of thrombi. The results obtained indicate that using rose bengal at concentrations of 25 and 50 mg/kg and a laser power of 5 mW, thrombus formation occurred. In addition, formation of the thrombus occurred ~30 min following induction and the thrombus had a total area of 4,878.3 μm(2), which caused total occlusion of the mesenteric artery. For visualization, platelets were labeled with calcein acetyloxymethyl ester for 1 h, which resulted in improved observation of thrombus formation in real time. Therefore, this technique may be used to perform in vivo studies simply and at low cost, and is suitable for use in a variety of studies of thrombosis.

No MeSH data available.


Related in: MedlinePlus