Enhancement of neointima formation with tissue-type plasminogen activator.
OBJECTIVE
Indirect evidence suggests that tissue plasminogen activator (tPA) either limits or does not alter restenosis. However, tPA enhances tumor invasiveness through matrix remodeling, and several elements of degraded matrix enhance smooth muscle cell mitogenesis. We use either local adenoviral-mediated overexpression of tPA or systemic infusion of recombinant tPA combined with mechanical overdilation of rabbit common femoral arteries to evaluate the impact of tPA on neointima formation.


METHODS
Left common femoral arteries of New Zealand white rabbits were transfected in situ either with an adenoviral-construct-expressing tPA or a viral control (adenoviral-construct-expressing beta-galactosidase) or nonviral (buffer) control after balloon angioplasty injury. At 7 and 28 days, left common femoral artery segments were harvested (n = 4 for each group and time point). Vessel segments were examined for intimato-media ratio, smooth muscle cell proliferation, extracellular matrix, and inflammatory response. Thrombus formation was evaluated after 3 days (n = 3 for each group). In a second experiment, New Zealand white rabbits (n = 3 per group, per time point) underwent mechanical dilation followed by buffer treatment or systemic tPA infusion according to a widely clinically used accelerated infusion protocol. Treated artery segments were harvested after 7 or 28 days and processed for intima-to-media ratio determination and class-wide histochemical determination of collagenous extracellular matrix and collagen content.


RESULTS
Both rate and degree of neointima formation increase dramatically with overexpression (250%-461% relative to controls at 7 and 28 days). Substantial early matrix degradation is observed in vessels treated with local overexpression of tPA, although no increases in local inflammation or in smooth muscle proliferation occur. Late enhancement of smooth muscle proliferation emerges, consistent with secondary impact of perturbed matrix components. Systemic infusion of tPA according to clinical protocols also results in early and late enhancement of neointima formation in this model (34%-52% relative to controls at at 7 and 28 days), with significant early collagenous matrix degradation. Systemic infusion, although significant, did not attain the degree of neointima formation present with overexpression.


CONCLUSIONS
With some evidence of dose-dependence, tissue plasminogen activator enhances neointima formation after angioplasty in a rabbit model. Early matrix degradation precedes change in rates of proliferation and underlies this effect in spite of several antirestenotic actions including decreased thrombus and decreased macrophage recruitment in this model.
Hilfiker
P R
PR
Stanford Institute of Bioengineering and Molecular Medicine, and Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA.
Waugh
J M
JM
Li-Hawkins
J J
JJ
Kuo
M D
MD
Yuksel
E
E
Geske
R S
RS
Cifra
P N
PN
Chawla
M
M
Weinfeld
A B
AB
Thomas
J W
JW
Shenaq
S M
SM
Dake
M D
MD
D000256:Adenoviridae; D000800:Angioplasty, Balloon / Q000009:adverse effects; D000818:Animals; D018791:CD4 Lymphocyte Count; D002455:Cell Division / Q000187:drug effects; D005109:Extracellular Matrix / Q000473:pathology; D005263:Femoral Artery / Q000187:drug effects / Q000293:injuries / Q000473:pathology; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008264:Macrophages / Q000473:pathology; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects; D009504:Neutrophils / Q000473:pathology; D011817:Rabbits; D011994:Recombinant Proteins / Q000494:pharmacology; D013927:Thrombosis / Q000209:etiology / Q000473:pathology; D010959:Tissue Plasminogen Activator / Q000235:genetics / Q000494:pharmacology*; D017539:Tunica Intima / Q000166:cytology / Q000187:drug effects* / Q000473:pathology
D005343:Fibrinolytic Agents; D011994:Recombinant Proteins; D010959:Tissue Plasminogen Activator