Evaluation of the effects of intramyocardial injection of DNA expressing vascular endothelial growth factor (VEGF) in a myocardial infarction model in the rat--angiogenesis and angioma formation.

OBJECTIVES: The effects of direct intramyocardial injection of the plasmid encoding vascular endothelial growth factor (phVEGF165) in the border zone of myocardial infarct tissue in rat hearts were investigated. BACKGROUND: Controversy exists concerning the ability of VEGF to induce angiogenesis and enhance coronary flow in the myocardium. METHODS: Sprague-Dawley rats received a ligation of the left coronary artery to induce myocardial infarction (MI). At 33.1 +/- 6.5 days, the rats were injected with phVEGF165 at one location and control plasmid at a second location (500 microg DNA, n = 24) or saline (n = 16). After 33.1 +/- 5.7 days, the hearts were excised for macroscopic and histologic analysis. Regional blood flow ratios were measured in 18 rats by radioactive microspheres. RESULTS: phVEGF165-treated sites showed macroscopic angioma-like structures at the injection site while control DNA and saline injection sites did not. By histology, 21/24 phVEGF165-treated hearts showed increased focal epicardial blood vessel density and angioma-like formation. Quantitative morphometric evaluation in 20 phVEGF165-treated hearts revealed 44.4 +/- 10.5 vascular structures per field in phVEGF165-treated hearts versus 21.4 +/- 4.7 in control DNA injection sites (p < 0.05). Regional myocardial blood flow ratios between the injection site and noninfarcted area did not demonstrate any difference between phVEGF,165-treated hearts (0.9 +/- 0.2) and saline-treated hearts (0.7 +/- 0.1). CONCLUSIONS: Injection of DNA for VEGF in the border zone of MI in rat hearts induced angiogenesis. Angioma formation at the injection sites did not appear to contribute to regional myocardial blood flow, which may be a limitation of gene therapy for this application.

Nitric oxide synthase modulates angiogenesis in response to tissue ischemia.

We tested the hypothesis that endothelial nitric oxide synthase (eNOS) modulates angiogenesis in two animal models in which therapeutic angiogenesis has been characterized as a compensatory response to tissue ischemia. We first administered L-arginine, previously shown to augment endogenous production of NO, to normal rabbits with operatively induced hindlimb ischemia. Angiogenesis in the ischemic hindlimb was significantly improved by dietary supplementation with L-arginine, compared to placebo-treated controls; angiographically evident vascularity in the ischemic limb, hemodynamic indices of limb perfusion, capillary density, and vasomotor reactivity in the collateral vessel-dependent ischemic limb were all improved by oral L-arginine supplementation. A murine model of operatively induced hindlimb ischemia was used to investigate the impact of targeted disruption of the gene encoding for ENOS on angiogenesis. Angiogenesis in the ischemic hindlimb was significantly impaired in eNOS-/- mice versus wild-type controls evaluated by either laser Doppler flow analysis or capillary density measurement. Impaired angiogenesis in eNOS-/- mice was not improved by administration of vascular endothelial growth factor (VEGF), suggesting that eNOS acts downstream from VEGF. Thus, (a) eNOS is a downstream mediator for in vivo angiogenesis, and (b) promoting eNOS activity by L-arginine supplementation accelerates in vivo angiogenesis. These findings suggest that defective endothelial NO synthesis may limit angiogenesis in patients with endothelial dysfunction related to atherosclerosis, and that oral L-arginine supplementation constitutes a potential therapeutic strategy for accelerating angiogenesis in patients with advanced vascular obstruction.

Laser-assisted debridement of aortic valve calcium.

Experimental debridement of aortic valve calcium by means of laser phototherapy was investigated in vitro. Near-total debridement of calcific deposits observed on pretreatment x-ray films was accomplished using carbon-dioxide laser phototherapy. Analysis of liberated photoproducts suggests that debridement is effected by thermal degradation of the surrounding connective tissue envelope and thermal expansion of the calcific nodules. These results suggest that in selected patients with calcific aortic stenosis, it may be possible to perform intraoperative laser-assisted debridement of aortic valve calcium in order to preserve the native aortic valve and thus avoid prosthetic valve replacement.

Laser photoablation of pathological endocardium: in vitro findings suggesting a new approach to the surgical treatment of refractory arrhythmias and restrictive cardiomyopathy.

In selected patients, malignant ventricular tachyarrhythmias have been successfully abolished by excision of subendocardial arrhythmogenic foci. Likewise, in certain patients in whom restrictive cardiomyopathy is due to endocardial thickening, endocardial resection has resulted in hemodynamic improvement. The present study was designed to explore the utility, in vitro, of laser photoablation of pathologically thickened endocardium. Endocardial photoablation was easily accomplished regardless of etiological or anatomical variations using either the focused beam of a carbon dioxide laser or argon laser light delivered through a 200-microns optical fiber. Photoablation of areas as large as 3.9 X 1.3 cm was performed within 40 seconds. The extent or depth of endocardial photoablation could be limited to 2 mm2 in area or 1 mm in depth using either form of laser therapy. These in vitro results suggest that either carbon dioxide or argon laser phototherapy can be successfully applied to the surgical treatment of refractory arrhythmias and restrictive cardiomyopathy. Advantages of laser photoablation include speed and precision. Furthermore, laser photoablation obviates the difficulty associated with conventional techniques in establishing tissue planes.

Laser myoplasty for hypertrophic cardiomyopathy. In vitro experience in human postmortem hearts and in vivo experience in a canine model (transarterial) and human patient (intraoperative).

The feasibility of performing a myotomy/myectomy for hypertrophic cardiomyopathy (HC) by means of laser phototherapy was evaluated experimentally in vitro and in vivo, and the procedure then applied to a patient intraoperatively. In vitro experience revealed that the beam of an argon laser, delivered directly or via an optical fiber, could both cut and vaporize myocardium, producing a myotomy/myectomy morphologically similar to that produced by the conventional blade technique. In vivo experiments, in which the beam of an argon laser was delivered via an optical fiber to the ventricular septum of a canine heart, confirmed that a laser myoplasty could be achieved in 4 of 5 dogs by a transarterial approach. Finally, laser myoplasty was performed intraoperatively in a patient with HC, using a 200-mu fiber interfaced with an argon laser. Measured laser power was 1.5 W; cumulative exposure was less than 4 minutes; the myoplasty was 4 X 1 X 0.5 cm. These investigations establish the feasibility of using laser therapy to create a myoplasty trough that is similar in appearance to that typically achieved by the conventional blade technique. Illumination of the intraventricular operative field and precise modeling of the myoplasty trough constitute the principal advantages of laser myoplasty for HC.