Duane L. Donovan

Controlled delivery of vascular endothelial growth factor promotes neovascularization and maintains limb function in a rabbit model of ischemia

PURPOSE Vascular endothelial growth factor (VEGF) modulates new blood vessel development and growth and has been suggested as a potential therapeutic agent that could alleviate debilitating claudication in patients. The objective of this study was to determine whether controlled, local delivery of a low dose of VEGF from an osmotic pump could promote neovascularization, limb perfusion, and functional improvements in the hind limbs of rabbits rendered partially ischemic by surgery. The effects of VEGF were compared with those of the vasodilator nitroglycerin (NTG) and to saline administered similarly. METHODS Thirty rabbits were randomly assigned to either VEGF (n = 10), NTG (n = 10), or saline (n = 10) treatment groups. Partial ischemia was induced in each left hind limb by surgical ligation of the common and superficial femoral arteries, leaving the internal iliac artery intact. The right limb of each animal served as a nonischemic control. Immediately after vessel ligations, a 28-day osmotic pump was implanted to deliver VEGF (0.22 microg/kg/day), NTG (17.8 microg/kg/day), or saline solution into the common iliac artery just proximal to the ligation site. Comparative vascularity between ischemic and nonischemic limbs within treatment groups and between groups was evaluated by (1) capillary counts from representative fields of hematoxylin and eosin stained muscle tissue taken from hind limbs at day 40; (2) digitized arteriograms of ischemic legs at day 40, which were used to quantify the complexity of vascular branching (fractal dimension index) and the total extent of vascularization (vascular density index); (3) measuring capillary refill times in ischemic limbs; and (4) observations of functional and trophic changes in ischemic limbs. Statistical differences between treatment groups were evaluated by one-way ANOVA. RESULTS Complexity of vascular branching and vascular density were significantly greater (p < 0.001) in VEGF-treated ischemic limbs compared with NTG- and saline-treated ischemic limbs. By postoperation day 14, all VEGF-treated ischemic limbs had restored capillary refill (p < 0.001), new hair growth, and greatly improved limb function and appearance. Saline-treated limbs exhibited ischemic changes, with poor capillary refill and negligible limb function. Capillary refill in NTG-treated ischemic limbs did not differ significantly from saline-treated limbs. Ischemic VEGF-treated limbs had significantly more capillaries compared with both ischemic and nonischemic limbs in saline-treated animals (p < 0.05). Ischemic NTG-treated limbs also had significantly more capillaries compared with ischemic limbs in saline-treated animals (p < 0.05). Because of high variability, however, capillary counts in VEGF-treated ischemic limbs did not differ significantly from those of contralateral nonischemic limbs, or from capillary counts in either ischemic or nonischemic limbs of NTG-treated rabbits. CONCLUSIONS Controlled release of microgram quantities of VEGF significantly enhanced neovascularization and vascular perfusion in ischemic limbs compared with controls in this rabbit model of partial ischemia. In addition, VEGF-treated ischemic limbs demonstrated near-normal function and appearance, whereas NTG- and saline-treated ischemic controls remained noticeably impaired. This novel approach of VEGF delivery may prove clinically useful either alone or combined with revascularization procedures.

Small-diameter vascular prostheses: two designs of PTFE and endothelial cell-seeded and nonseeded Dacron

Despite numerous advances in biomaterials design and utilization, the perfect artificial small-vessel substitute has yet to be developed. Dacron and expanded polytetrafluoroethylene (PTFE) are two materials potentially appropriate for use as small-vessel prostheses. We report the patencies of endothelial cell-seeded and nonseeded 4 mm I.D. Dacron grafts and two designs of nonseeded 4 mm I.D. PTFE (Gore-Tex and Impra) in the carotid position in dogs. All graft lengths exceeded the calculated maximum critical length for the material being tested. Dacron grafts, both endothelial cell-seeded and nonseeded, achieved higher patencies than both designs of PTFE. Endothelial cell-seeded Dacron grafts achieved the highest patencies. Endothelium was present to a significant extent only on endothelial cell-seeded Dacron grafts. There was little pannus ingrowth or midgraft pseudointima on nonseeded Dacron or on patent PTFE grafts although thrombus-free surface areas of patent PTFE grafts were high. These comparative data support the utility of endothelial cell seeding in achieving high patencies of small-diameter vascular grafts.

Material and structural characterization of human saphenous vein

Saphenous vein patch rupture after carotid endarterectomy is an infrequent but devastating complication. This study was undertaken to evaluate the material and structural properties of fresh human saphenous veins to understand the causes of this complication. Segments of saphenous veins were obtained from 22 patients from vein harvested during coronary artery bypass surgery. Ninety-three specimens, oriented in both circumferential (n = 45) and longitudinal (n = 48) directions, were prepared from the available vein segments for testing. Specimens were mounted on specially designed grips and then subjected to uniaxial tension testing. For each specimen the following material and structural parameters were determined: vessel diameter, tensile stiffness, failure and ultimate forces, and tensile modulus, failure stress, and strain. The physical properties of specimens evaluated in longitudinal orientations and thus limit the inherent strength of the vein. The physical properties of circumferentially tested vein specimens were negatively correlated to age, female gender, diabetes, and hypertension. The data obtained in this investigation suggest that age, hypertension, as well as diabetes and gender may adversely influence the circumferential tensile strength of human saphenous veins used as patch grafts.

Modern autotransfusion: Experience with a washed red cell processing technique

The technique of intraoperative autotransfusion utilizing the Haemonetics Cell Saver is described. This device separates and washes red blood cells removed from the surgical field. Advantages of this unit over others are (1) no systemic anticoagulation is required, (2) circulatory fibrin debris is removed, (3) plasma hemoglobin is removed, and (4) any circulating anticoagulant is removed. Experience with 136 consecutive cases. 101 elective and 35 emergency, is reported. The only complication was coagulopathy, which occurred in 5.1 percent of the cases. It is easily treated with blood component therapy and occurs in those patients in whom greater then 3,500 cm3 of blood is autotransfused. The Cell Saver has proven an important adjunct in surgical patients in whom greater than 1,000 cm3 of blood will be lost, as well as in emergency patients. It provides an efficient, economical and safe method to autotransfuse blood.

Management of intra-abdominal aneurysms associated with periarteritis nodosa

Periarteritis nodosa is a disease of small and medium-sized arteries, frequently associated with multiple visceral artery aneurysms. Infrequently, these aneurysms rupture, usually with fatal results. A case of spontaneous rupture of a middle colic artery aneurysm in a patient with periarteritis nodosa is reported, and similar cases in the literature are reviewed. Treatment of a ruptured visceral artery aneurysm requires ligation or resection of the aneurysm without delay. Residual aneurysms are treated with cyclophosphamide and/or prednisone in an attempt to induce regression of the aneurysms. An arteriogram performed after 3 to 4 months of medical therapy determines the need for further surgical intervention.

Prostaglandin biochemistry of seeded endothelial cells on Dacron prostheses

The beneficial effect of seeding endothelial cells on synthetic vascular conduits has been well established. The biochemical production and interaction of the prostaglandins, prostacyclin (PGI2) and thromboxane (TxA2), were studied on Dacron vascular grafts that were seeded with autogenous venous endothelial cells. Seventy-three seeded and nonseeded grafts were implanted into the carotid arteries of dogs. Animals were medicated with either cyclooxygenase inhibitors (aspirin and dipyridamole, or ibuprofen, or U-53,059), or dipyridamole alone, or a thromboxane synthase inhibitor, U-63557A. All animals were killed at 5 weeks and analyzed for patency, thrombus-free surface (TFS), and PGI2 and TxA2 production from mid-graft punch biopsies. PGI2 and TxA2 identifications were made by radioimmunoassay determination of 6-keto PGF1 alpha and TxB2, respectively. Results of the study demonstrated in nonmedicated animals a slightly increased patency rate in seeded vs. nonseeded grafts (50% vs. 40%) and a more significant difference in TFS (49% vs. 24%). The addition of cyclooxygenase inhibitors or TxA2 synthase inhibitors significantly improved both patency (90% vs. 47%) and TFS (87% vs. 9%) in seeded vs. nonseeded grafts. PGI2 production was decreased in seeded grafts with the use of cyclooxygenase inhibitors in all cases. It is concluded that seeded endothelial cells on Dacron velour grafts can synthesize PGI2; these PGI2 levels are far less than PGI2 levels produced by endothelial cells from the adjacent carotid artery; and TxA2 synthase inhibitors best improve thromboresistance of seeded grafts without significant reduction in PGI2 production.

Experimental pharmacologic cerebroprotection

In the first part of this experiment, the effects of pharmacotherapy on the neurologic consequences of transient global ischemia were examined in Wistar rats. The control and four experimental groups each contained six rats. In comparison to the control group receiving normal saline (NS) solution, in which no rats survived, all rats given naloxone (Nx) (23 mg/kg), superoxide dismutase (SOD) (10,000 U/kg), or allopurinol (APL) (35 mg/kg), 15 minutes before interruption of cerebral blood flow, survived the 20-minute period of global ischemia (p less than 0.01, p less than 0.01, p less than 0.01, respectively). No rat receiving deferoxamine (DEF) (20 mg/kg) survived the same ischemic period. In the second part of the experiment, the arachidonic acid (AA) content of brain samples was determined by gas chromatography and was used as an indicator of cerebral ischemia. Two control and four experimental groups consisted of six rats each. An ischemia control group received NS, whereas experimental groups were given Nx, SOD, APL, or DEF with the same previous dosage schedule. The animals were decapitated 15 minutes after drug infusion and cerebral ischemia was simulated by incubation of the heads in a 37 degrees C water bath for 60 minutes. AA content of ischemic brain treated with NS was markedly elevated (60.0 +/- 24.1 micrograms/gm of brain tissue), whereas in comparison the AA content of brain treated with Nx (5.1 +/- 3.0 micrograms/gm of brain tissue, p less than 0.05), SOD (3.5 +/- 2.7 micrograms/gm of brain tissue, p less than 0.05), or APL (2.9 +/- 1.5 micrograms/gm of brain tissue, p less than 0.05) all demonstrated much lower levels.(ABSTRACT TRUNCATED AT 250 WORDS)

A Rare Complication in Elective Repair of an Abdominal Aortic Aneurysm: Multiple Transmural Colonic Infarcts Secondary to Atheroemboli

Numerous complications may occur during elective and emergency repair of abdominal aortic aneurysms. The following report will document a rare complication in a patient with chronic renal failure. Multiple atheroemboli were found to produce transmural infarction of the left colon after elective repair of an abdominal aortic aneurysm. The pathologic process and the proposed mechanism of injury are also discussed.

Angioscope-assisted occlusion of venous tributaries with prolamine in in situ femoropopliteal bypass: Preliminary results of canine experiments

Ten mongrel dogs underwent left lower extremity in situ femoropopliteal bypass with femoral vein. A 20 to 25 cm myocutaneous bridge was left between femoral and popliteal anastomoses. A 2.8 mm angioscope was introduced intraluminally to visualize venous tributaries (VT). A balloon occlusion catheter was placed alongside the angioscope and directed in each VT. Prolamine was injected into each VT to effect occlusion. Seven dogs were followed up for 1 week and three dogs for 1 month. A total of 34 VT (range one to five per dog) were available for attempted occlusion. Twenty-nine of 84 (85%) VT were able to be occluded based on comparison of pre-VT and post-VT occlusion angiograms. Poor visualization of VT or VT too small to admit the 5F catheter were reasons for failure. We conclude that (1) in the canine model studied angioscope-assisted occlusion of femoropopliteal during bypass is technically feasible, (2) this technical detail makes unnecessary medial thigh dissection for exposure of the vein graft, and (3) during short-term observation prolamine appeared to be a suitable occluding substance.

A canine model of intimal hyperplasia (IH) in autogenous vein grafting: a preliminary report.

High failure rates (10-40% at 1 year and 2-6% per year thereafter) of autologous vein grafts in peripheral bypass surgery due to progressive intimal hyperplasia (IH) have prompted researchers to search for an animal model that develops IH in a relatively short period of time. This study summarizes our experiences in promoting IH in a canine model. Eight to ten centimeters of both jugular veins were exposed in 40 adult mongrel conditioned dogs. After division into 4-5-cm lengths, the segment of jugular vein most proximal was ballooned at 800-900 mm Hg with a modified 8F Fogarty catheter to induce intimal and medial layer injury. The distal segment was left nonballooned. Segments of these autologous vein grafts, 1.5 cm in length, both ballooned and nonballooned, were then anastomosed, end to end, into the carotid and femoral circulations. Six weeks postoperatively the grafts were perfusion-fixed, harvested, and histologically processed, and the amount of the lumen in midgraft sections occupied by IH was determined by image analysis. In all dogs, the degree of IH was significantly greater in the balloon catheterized vs noncatheterized graft segments. IH was more severe in the femoral than in the carotid arteries. The grafts that developed the most severe intimal hyperplasia were femoral grafts that had been balloon catheterized. We conclude that these protocols are effective in inducing IH in a canine model in short postoperative times.