Yoko Onuki

Effect of differential shear stress on platelet aggregation, surface thrombosis, and endothelialization of bilateral carotid-femoral grafts in the dog

PURPOSE The purpose of this study was to determine the effects of increased shear stress on the aggregability of platelets as they traverse a long, small-caliber (6 mm) Dacron graft in the dog and on the surface thrombosis and endothelialization of such a graft. METHODS Each of nine dogs received bilateral carotid-femoral artery grafts, approximately 75 cm long, for 3 months; one graft of each pair had a distal femoral arteriovenous fistula to produce a higher shear rate than the contralateral graft. Platelet aggregation scores were determined on blood withdrawn from the external jugular vein and from the proximal and distal ends of the grafts in each animal. Graft flow rates, which were used in the computation of shear stress, and luminal pressure gradients through grafts were measured during surgery and specimen retrieval. Specimens were studied with light microscopy after hematoxylin and eosin and immunocytochemical staining and by scanning electron and transmission electron microscopy to evaluate the nature, composition, and thickness of the flow surface lining, as well as the transmural healing. RESULTS Two high-shear stress and two low-shear stress grafts occluded unilaterally; five dogs had bilaterally patent grafts, allowing comparative analyses. All subjects had low platelet aggregability with aspirin. Platelet aggregation scores taken from proximal and distal ends of the grafts were not significantly different. The high-shear stress grafts had significantly more endothelial-like cell coverage (p < 0.0371) than the low-shear stress grafts, less flow-surface thrombus (p < 0.0056), and a thinner surface lining (p < 0.0029), on both the neointima and pseudointima. CONCLUSIONS In subjects with low platelet aggregation scores, long Dacron grafts do not elevate platelet aggregability of blood flowing through them. High-shear stress grafts have less flow surface thrombus, more endothelialization, and a thinner surface lining than do low-shear stress grafts.

Endothelium on the flow surface of human aortic Dacron vascular grafts

PURPOSE We have previously observed endothelium on two human vascular prostheses explanted under optimal conditions for flow surface preservation. In this study we sought to further verify the hypothesis that endothelialization can occur on clinical grafts and that it can be detected in specimens that have been promptly removed and properly preserved in a timely manner. METHODS We studied 29 aortic grafts. Of these, 11 Dacron bypass grafts were in a condition suitable for analysis with light microscopy and immunocytochemistry staining, and scanning and transmission electron microscopy. RESULTS Three grafts had endothelium beyond the pannus, identified by factor VIII/ vWF, Ulex europaeus agglutinin, and collagen IV positivity. Specimen A, a knitted 6-year implant, was preserved by embalming 3 hours after the patients death and had a firmly attached outer capsule with fibroblasts, collagen, giant cells, and microvessels in the interstices. Specimen B, a woven 18-year implant, was retrieved at reoperation and immediately fixed in 10% formalin; it had no outer capsule and no tissue ingrowth. Specimen C, a woven 7-year implant, was removed and fixed 5 hours after the patients death; it had a firmly attached outer capsule but no tissue ingrowth beyond the outer portion of the wall. CONCLUSIONS The rapidity with which the specimens were fixed probably enabled identification of endothelium. These findings suggest that endothelialization of synthetic arterial grafts may occur more frequently in human beings than previously recognized.

Comparison of the effect of monopolar and bipolar cauterization on skeletonized, dissected internal thoracic arteries

The internal thoracic artery is preferable to the saphenous vein for use as a conduit for coronary artery bypass. More extensive use is possible if this artery is mobilized in a skeletonized form to provide greater length. Internal thoracic arteries are usually mobilized with cauterization. This study compared the effectiveness and effects on neighboring areas of division of the branches of the canine internal thoracic artery with bipolar cauterization and monopolar cauterization. Branch closure was significantly more secure in the bipolar cauterization group, with bleeding in 25 (9%) of 279 branches of 15 internal thoracic arteries treated with monopolar cauterization, in contrast to bleeding in 4 (1.3%) of 306 branches in the 15 internal thoracic arteries treated with bipolar cauterization, which were paired with the group treated with monopolar cauterization. The group of internal thoracic arteries treated with monopolar cauterization had a significantly higher prevalence of leakage when luminal pressure was increased from 120 to 160 mm Hg. Scanning electron microscopy demonstrated partial loss of endothelial cells on the flow surface of internal thoracic arteries treated with bipolar cauterization, compared with almost complete loss of endothelial cells around the orificial areas after monopolar cauterization. Secondary bipolar cauterization treatment caused only slightly more damage than primary treatment, but secondary monopolar cauterization was much more severe and extensive than primary treatment. These data suggest that bipolar cauterization is preferable to monopolar cauterization for skeletonized dissection of the internal thoracic artery.

Implant site influence on arterial prosthesis healing: A comparative study with a triple implantation model in the same dog

PURPOSE The purpose of this study was to develop a cost-effective canine graft healing model that gives information on various implant sites and controls for variable factors between graft locations and between animals and to compare the influence of implant site (retropleural, retroperitoneal, and subcutaneous areas) on arterial graft healing in the same subject under such controlled study conditions. METHODS Five mongrel dogs were studied for 8 weeks, and one was studied for 3 years. Each received three porous Dacron grafts during the same surgery: a carotid-femoral bypass (C-FB) and interposition grafts in the descending thoracic aorta and abdominal aorta. To produce comparable shear stress calibers of the C-FB and abdominal aorta grafts were 2 mm less than those of the descending thoracic aorta, and a distal arterio-venous fistula was created to further increase the C-FB flow. For comparable blood aggregation status platelet aggregation was preevaluated and adjusted with antiplatelet agents. Graft flow surfaces were assessed for thrombus-free surface and endothelial-like cell coverage scores. Tissue samples were studied with hematoxylin-eosin, factor VIII/ von Willebrand factor, smooth muscle alpha-actin staining, and scanning electron microscopy and transmission electron microscopy. RESULTS All grafts were patent. Shear stress for the three grafts and platelet aggregation among the study subjects were comparable. Healing of descending thoracic aorta and abdominal aorta grafts was similar, but C-FB healing was slow, incomplete, and uneven, with a high incidence of seroma. Eight-week and 3-year results were comparable. CONCLUSIONS This model gives broad healing information about the areas where grafts are often implanted in humans. Eight weeks appears to be a sufficient period to reflect basic and general healing characteristics. Grafts heal better in the retropleural and retroperitoneal areas than in the subcutaneous tissues.

Effect of altered blood flow on the caliber and morphology of the internal thoracic artery in the dog

OBJECTIVE The purpose of this study was to evaluate in dogs the effect of blood flow alteration on caliber and morphology of in situ internal thoracic arteries. METHODS Six dogs underwent creation of a unilateral distal arteriovenous fistula between the internal thoracic artery and vein at the sixth rib to create high flow, and in six others the internal thoracic artery was unilaterally skeletonized and dissected. For both groups the contralateral internal thoracic artery served as the control; sides were alternated among cases. Blood flow was measured for shear stress calculation before and after surgical alteration. After 2 months, internal thoracic arteries were harvested with the entire anterior chest plate, which was dynamically inflated and fixed with 10% formalin at a controlled pressure of 120 mm Hg after angiography had been done at the same pressure. The luminal diameters were then measured at eight levels on the angiograms. Arterial tissue samples were taken at three levels and embedded, sectioned, and treated with hematoxylin-eosin and Verhoeff-van Gieson stains. Digital imaging analysis was used for quantitative morphometric studies. RESULTS All fistulas remained patent. In comparison with control arteries, high-flow internal thoracic arteries dilated and low-flow internal thoracic arteries narrowed, which was associated with significant change in shear stress for both groups. There were no substantial structural changes in the walls of either group. CONCLUSION In the dog, the luminal diameter of the internal thoracic artery responds to altered blood flow without intimal thickening or other undesirable wall changes.

Histologic observation of continuity of transmural microvessels between the perigraft vessels and flow surface microostia in a porous vascular prosthesis.

To ascertain the histologic relationship between flow surface microostia and perigraft vessels in the healing of a porous vascular prosthesis, a series of careful histologic examinations were carried out on a preclotted, knitted Dacron graft, 8 mm in diameter and 7 cm long, after implantation in the canine infrarenal abdominal aorta for 3 months. Four adjacent longitudinal tissue blocks were taken from the middle for evaluation by means of light microscopy, scanning electron microscopy, transmission electron microscopy, and immunocytochemical staining, and the remainder of the specimen was stained with silver nitrate to allow visualization of microostia and cell borders on the flow surface. Following identification of two microostia adjacent to the area where samples had been taken for general healing evaluation, a 3 × 8 mm full-thickness block containing the microostia was embedded in glycol methacrylate and stained with hematoxylin and eosin. Of 240 serial sections cut from this block, 80 were prepared and examined. Silver staining revealed 42 microostia on the flow surface. Light, scanning, and transmission electron microscopy with endothelial factor VIII/von Willebrand factor stain confirmed a single layer of endothelial cells on the flow surface, and beneath was a well-organized neointima containing fusiform cells confirmed as smooth muscle by α-actin stain. Light microscopy of the serial sections revealed transmural microvessels, lined with endothelium, extending continuously between the flow surface ostia and the perigraft vessels in this porous vascular prosthesis.