Microvascular blood flow and stasis in transgenic sickle mice: utility of a dorsal skin fold chamber for intravital microscopy.
Vascular inflammation, secondary to ischemia-reperfusion injury, may play an essential role in vaso-occlusion in sickle cell disease (SCD). To investigate this hypothesis, dorsal skin fold chambers (DSFCs) were implanted on normal and transgenic sickle mice expressing human alpha and beta(s)/beta(s-Antilles) globin chains. Microvessels in the DSFC were visualized by intravital microscopy at baseline in ambient air and after exposure to hypoxia-reoxygenation. The mean venule diameter decreased 9% (P < 0.01) in sickle mice after hypoxia-reoxygenation but remained constant in normal mice. The mean RBC velocity and wall shear rate decreased 55% (P < 0.001) in sickle but not normal mice after hypoxia-reoxygenation. None of the venules in normal mice became static at any time during hypoxia-reoxygenation; however, after 1 hr of hypoxia and 1 hr of reoxygenation, 11.9% of the venules in sickle mice became static (P < 0.001). After 1 hr of hypoxia and 4 hr of reoxygenation, most of the stasis had resolved; only 3.6% of the subcutaneous venules in sickle mice remained static (P = 0.01). All of the venules were flowing again after 24 hr of reoxygenation. Vascular stasis could not be induced in the subcutaneous venules of sickle mice by tumor necrosis factor alpha (TNF-alpha). Leukocyte rolling flux and firm adhesion, manifestations of vascular inflammation, were significantly higher at baseline in sickle mice compared to normal (P < 0.01) and increased 3-fold in sickle (P < 0.01), but not in normal mice, after hypoxia-reoxygenation. Plugs of adherent leukocytes were seen at bifurcations at the beginning of static venules. Misshapen RBCs were also seen in subcutaneous venules.
Kalambur
Venkatasubramaniam S
VS
Department of Mechanical Engineering, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Mahaseth
Hemchandra
H
Bischof
John C
JC
Kielbik
Miroslaw C
MC
Welch
Thomas E
TE
Vilbäck
Asa
A
Swanlund
David J
DJ
Hebbel
Robert P
RP
Belcher
John D
JD
Vercellotti
Gregory M
GM
D000755:Anemia, Sickle Cell / Q000097:blood / Q000150:complications / Q000235:genetics / Q000503:physiopathology*; D000818:Animals; D001772:Blood Cell Count; D002448:Cell Adhesion / Q000502:physiology; D005260:Female; D006400:Hematocrit; D036904:Leukocyte Rolling; D007962:Leukocytes / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008833:Microcirculation / Q000502:physiology; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D015427:Reperfusion Injury / Q000097:blood / Q000150:complications / Q000503:physiopathology*; D012156:Reticulocytes / Q000166:cytology; D012867:Skin / Q000098:blood supply*