The spatial dynamics of fibrin clot dissolution catalyzed by erythrocyte-bound vs. free fibrinolytics.
BACKGROUND
Coupling fibrinolytic plasminogen activators to red blood cells (RBCs) has been proposed as an effective, yet safe method of thromboprophylaxis, because of increased circulation lifetime and reduced propensity to induce hemorrhage by selectivity for nascent thrombi rather than pre-formed hemostatic clots.


OBJECTIVE
We used confocal microscopy of fluorescently labeled fibrin and erythrocytes in plasma-derived clots to study the spatial dynamics of lysis catalyzed by RBC-coupled vs. free plasminogen activators (RBC-PA vs. PA).


RESULTS
Clot lysis catalyzed by free PA progressed gradually and uniformly. In contrast, distinct holes formed surrounding RBC-PA while the rest of the clot remained intact until these holes enlarged sufficiently to merge, causing sudden clot dissolution. Compared with naïve RBCs within clots lysed by free PA, RBC-PA moved faster inside the fibrin network prior to clot dissolution, providing a potential mechanism for spatial propagation of RBC-PA induced lysis. We also showed the focal nature of fibrinolysis by RBC-PA as dense loading of PA onto RBCs initiates more efficient lysis than equal amounts of PA spread sparsely over more RBCs. In an in vitro model of clots exposed to buffer flow, incorporated RBC-PA increased permeability and formed channels eventually triggering clot dissolution, whereas clots containing free PA remained intact.


CONCLUSIONS
Clot lysis by RBC-PA begins focally, has a longer lag phase when measured by residual mass than homogeneous lysis by PA, is propagated by RBC-PA motility and provides more effective clot reperfusion than free PA, making RBC-PA attractive for short-term thromboprophylaxis.
Department of Cell & Developmental Biology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-6058, USA.
K C
KC
Gersh
S
S
Zaitsev
V
V
Muzykantov
D B
DB
Cines
J W
JW
Weisel
D000818:Animals; D055162:Biocatalysis; D004912:Erythrocytes / Q000187:drug effects*; D005337:Fibrin; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D012995:Solubility
D005343:Fibrinolytic Agents; D005337:Fibrin