P. Swartbol

Biological Responses to Endovascular Treatment of Abdominal Aortic Aneurysms

Purpose: To review the findings of two studies investigating the apparent differences in inflammatory responses demonstrated in patients undergoing endovascular as opposed to classic surgical treatment of abdominal aortic aneurysms (AAAs). Methods: The clinical course of seven patients treated with an endoluminal procedure (AAA-E) and seven patients undergoing conventional surgery (AAA-C) were compared (all men; ages 52 to 80 years). Blood samples were taken pre-, intra-, and postoperatively for up to 7 days. Inflammatory responses were assessed from measurement of interleukins (IL)-1 β, IL-6, IL-8, and tumor necrosis factor (TNF-α); complement proteins C1q, C4, C5a, and terminal complement complexes, C5b-C9; and C-reactive proteins. Granulocyte and monocyte surface adhesion molecule expression was determined indirectly using a panel of monoclonal antibodies against CD11a, CD11b, CD11c, CD18, and L-selectin in donor white blood cells exposed to patient plasma. Results: In six of the AAA-E patients, blood pressure decreases were recorded during the introduction of the device. Elevated body temperature was sustained for 2 to 5 days postoperatively in the AAA-E group. IL-6 levels were significantly higher in AAA-C patients (p < 0.0005), while TNF-α release was recorded in the AAA-E group only. CD11b, CD11c, and CD18 molecules on both granulocytes and monocytes were significantly upregulated 60 minutes after the endovascular procedure compared to conventional surgery. Conclusions: Endovascular aortic aneurysm repair apparently induces a significant inflammatory response, mainly involving TNF-α release, which differs from open AAA repair. These inflammatory responses, which may be related to the observed intraprocedural blood pressure decreases, could be caused by cell activation arising from intra-aneurysmal device manipulation.

Adverse reactions during endovascular treatment of aortic aneurysms may be triggered by interleukin 6 release from the thrombotic content.

PURPOSE It has been shown that endovascular aortic aneurysm repair might induce a significant inflammatory response, mainly involving tumor necrosis factor (TNF-alpha) release. This study determined in vitro whether these inflammatory responses could depend on white blood cell (WBC) activation caused by the aneurysmal mural thrombus. METHODS Mural thrombus specimens obtained from 10 different aortic aneurysms were weighed, homogenized, and assayed for interleukin 1beta (IL-1beta), interleukin 6 (IL-6), TNF-alpha, and soluble TNF receptor (sTNFRI). RESULTS Only high amounts of IL-6 (mean, 2973 pg/mL) were found. In contrast, after the addition of healthy donor WBCs to the thrombus mass supernatants, elevated levels of TNF-alpha (mean, 523 pg/mL) were seen. Theoretically, WBCs were stimulated by IL-6, resulting in TNF-alpha release. In additional experiments, it was proven that stimulated WBCs, induced by thrombus mass supernatants, synthesize TNF-alpha (mean, 796 pg/mL), and monoclonal antibodies against IL-6, prevented such TNF-alpha production (mean, 62 pg/mL). CONCLUSION The biologic responses during endovascular repair may be explained by a release of IL-6 from the aneurysmal thrombus, causing WBC stimulation and production of TNF-alpha. More complex processes cannot be excluded, but the present findings suggest that restrictions of manipulations within the aneurysm may be advisable.

Deposition of platelets and neutrophils in porcine iliac arteries after angioplasty and Wallstent placement compared with angioplasty alone.

PurposeThis study was designed to compare deposition of111In-labeled platelets and neutrophils after balloon angioplasty (PTA) alone and PTA plus Wallstents.MethodsHistological investigation was performed with scanning electron microscopy (SEM). Fifty percent stenoses of both iliac arteries was created by resorbable ligature in 13 pigs. After 30 days, PTA was performed bilaterally with an additional stenting procedure done on one side. Autologous platelets were labeled and reinfused before the interventional procedure in six pigs, and labeled neutrophils were used in seven pigs. The deposition of the labeled cells was recorded itin vivo over 270 min using a scintillation camera. The results were correlated within vitro measurements.ResultsScanning revealed significant increase in platelet and neutrophil deposition at the site of the stent compared with the site where PTA alone was undertaken.In vitro measurements confirmed these differences. SEM demonstrated a fibrin lining on the stent surface and numerous adherent platelets. The adjacent arterial lumen was almost completely covered by fibrinous material. The PTA-alone site demonstrated denudation of endothelial cells and less fibrinous material, as well as platelets and leukocytes.ConclusionThe complex interaction in the response of the vessel wall and flowing blood involves both platelet and neutrophil adhesion. The self-expandable vascular endoprosthesis contributes to increased deposition of platelets and neutrophils as seen in this experimental model of nonatheromatous stenosis.

Aortobifemoral surgery induces complement activation and release of interleukin-6 but not tumour necrosis factor-alpha

The aim of the present study was to determine the inflammatory response by an extended analysis of complement in 16 patients undergoing aortobifemoral bypass surgery. The patients were randomized to receive either a bifurcated expanded polytetrafluoroethylene graft (n = 8; group I) or a collagen-impregnated knitted Dacron graft (n = 8; group II) to determine whether differences in graft surface properties might influence the inflammatory response during and after the procedure. The following components of complement: C1q, C4, C3, C3d, C5a and terminal complement complexes were all analysed. C-reactive protein and interleukin-6 were also determined to assess the acute phase response. The complement data were corrected for haemodilution, which was assessed from alpha 2-macroglobulin concentrations. A significant decrease of C1q (P < 0.0001) and an increase in C5a (P < 0.0005) was observed in both groups. C4 and C3 levels showed slight fluctuations in group I, whereas in group II these proteins increased significantly (P < 0.05, P < 0.005, respectively) between 2 and 7 days after surgery. Terminal complement complexes remained unchanged in both groups. Interleukin-6 levels peaked at 12-24 h and the C-reactive protein at 24-72 h. Higher interleukin-6 levels (P < 0.05) were found in group II 6 h after surgery compared with group I; no release of tumour necrosis factor-alpha was identified. An early inflammatory response was found in all patients. The patterns of the complement proteins varied with a C1q depletion and a C5a increase, interpreted as complement activation. Whether the variations between the two graft groups represent any differences in graft surface properties has to be further elucidated.

Tumor necrosis factor-α and interleukin-6 release from white blood cells induced by different graft materials in vitro are affected by pentoxifylline and iloprost

Inflammatory mediators such as cytokines produced by white blood cells (WBCs) at the site of implantation are important for the biocompatibility of vascular grafts. The aim of the present study was to demonstrate the tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) release from WBCs incubated with expanded polytetrafluoroethylene (ePTFE) or woven Dacron grafts. In a second series the effects of pentoxifylline (PTX) and iloprost (ILO), both known to inhibit white blood cell function, on this release were determined. Woven Dacron grafts induced significantly higher release of both TNF-alpha and IL-6 compared to ePTFE. TNF-alpha was detectable first after 2 h, whereas IL-6 was seen after 4 h. Maximum values were reached at 6 and 12 h, respectively. The addition of an endotoxin gave more pronounced patterns of cytokine release not influenced by time. Preincubation with both PTX and ILO at final concentrations of 100 and 10 micrograms/mL, respectively, reduced significantly the TNF-alpha release without differences between the two graft materials, whereas the effect on the IL-6 release varied and was graft material-dependent. In conclusion, graft material-dependent induction of TNF-alpha and IL-6 from WBCs was demonstrated. PTX and ILO influenced the cytokine release. It might be suggested that graft material-induced cytokine production could contribute to intimal hyperplasia in vivo. The present findings encourage further studies regarding graft material-induced WBC alterations and the role of pharmacologic agents influencing this function.

Endovascular abdominal aortic aneurysm repair induces significant alterations in surface adhesion molecule expression on donor white blood cells exposed to patient plasma

OBJECTIVE To determine the response of white blood cells in endovascular aortic aneurysm repair. MATERIALS AND METHODS Seven patients treated with an endoluminal procedure (AAA-E) and seven patients undergoing conventional surgery (AAA-C) were included (all males, aged 52-80 years). A panel of monoclonal antibodies against CD11a, CD11b, CD11c, CD18 and L-selectin was used. To determine the surface receptors on both circulating and sequestered white blood cells, plasma from the patients and cells from healthy donors were combined for flow-cytometry. RESULTS The expression of CD11a adhesion molecules only showed slight variations regarding granulocytes, but was more pronounced on monocytes, however, without significant differences between the two patient groups, CD11b, CD11c and CD18 molecules on both granulocytes and monocytes were significantly upregulated 60 min after the endovascular procedure compared to conventional aneurysm repair, and L-selectin molecules were by this time correspondingly cleaved off. CONCLUSION Endovascular aneurysm repair differed significantly from conventional aneurysm surgery with peak adhesion molecule expression 60 min after balloon deflation, probably caused by release of tumour necrosis factor-alpha (TNF-alpha).

Surface adhesion molecule expression on human blood cells induced by vascular graft materials in vitro

The expression of surface adhesion molecules on granulocytes, monocytes (CD11a, CD11b, CD11c, CD18, L-selectin), and platelets (P-selectin, gpIIb-IIIa) was determined after incubation with different graft surfaces [expanded polytetrafluoroethylene (ePTFE) or woven Dacron]. Woven Dacron grafts upregulated the CD11b and CD11c surface antigens on both granulocytes and monocytes. Both graft materials demonstrated increased expression of CD11a and CD18 adhesion molecules on white blood cells at 30 min, followed by a downregulation. Maximum L-selectin expression was seen at 120 min on granulocytes and at 90 min on monocytes without differences between the graft materials. A rapid downregulation of gpIIb-IIIa complexes on platelets was noticed, while no expression of platelet P-selectin molecules was observed. In conclusion, both graft materials induced alteration of the white blood cell adhesion molecule expression, but the intensity and time course were dependent on the cell type and the graft material, suggesting that different mechanisms might be implicated. The expression of platelet surface antigens was less clearly influenced. The clinical significance of an enhanced cell surface antigen receptor expression caused by woven Dacron (CD11b, CD11c) has to be further studied. However, determination of adhesion molecule expression might offer possibilities to predict biocompatibility.

Flavonoid treatment in patients with healed venous ulcer: flow cytometry analysis suggests increased CD11b expression on neutrophil granulocytes in the circulation

The objective was to determine the activation of white blood cells (WBCs) and endothelial cells in patients with healed venous ulcer and the influence of the standing position and of treatment with flavonoids. Ten patients with a healed venous ulcer were treated with flavonoid substance (90% diosmin), 1000 mg three times daily for 30 days. Blood samples were taken from arm and dorsal foot veins before and after standing for 30 minutes. Blood sampling was performed before treatment, after three days, one month and three months. The activation of WBCs was determined by measuring adhesion molecule CD11b and CD18 expression on the surface of granulocytes and monocytes. In addition, interleukin 6 (IL-6), IL-8, soluble E-selectin (sE-selectin), sL-selectin and sICAM-1 levels in serum were quantified. The results showed that standing did not influence any of the measured parameters significantly. Expression of CD11b adhesion molecules on granulocytes was significantly up-regulated (p = 0.044) after treatment with flavonoids for one month, but this increase was not significant (p = 0.056) two months after the treatment period compared with the baseline level. The expression of CD18 remained unchanged. Baseline expression of CD11b or CD18 on monocytes did not change significantly during the study period. Neither was any significant change observed in the levels of IL-6, IL- 8 or the soluble adhesion molecules. It was concluded that flavonoid treatment for 30 days increased the expression of CD11b adhesion molecules on circulating granulocytes. No general effect on the inflammatory process could be observed as assessed by levels of cytokines and soluble adhesion molecules. Possible explanations for these findings could be that a decreased number of primed granulocytes leave the circulation due to a changed WBC/endothelial cell interaction or that flavonoids have a direct effect on granulocytes. Further studies are needed to clarify the mode of action of flavonoids in chronic venous disease.

Metabolic Response of Blood Cells to Synthetic Graft-Materials with Special Reference to a Fluoromer Passivated Dacron® Graft. An in Vitro Study Using Microcalorimetry

Microcalorimetry was used to study in vitro the metabolic response from human platelets and leukocytes when incubated with three different synthetic graft-materials. The graft to be studied primarily was Fluoromer Passivated Dacron (FPD) which was compared with ePTFE and with a knitted Teflon graft. A rapid increase in the metabolic activity of platelets was observed, followed by a steady-state for more than one hour, while the platelet metabolism did not differ among the various graft-materials. Leukocytes incubated with FPD showed a high initial metabolism, with a peak after about 15 minutes. After 60 minutes the metabolic response had reached control values. ePTFE and Teflon grafts differed significantly from FPD, without causing any peak metabolic activity. It may be concluded that FPD and ePTFE grafts, as evaluated in vitro, activate platelets to the same extent, while FPD causes a more extensive leukocyte activation. Whether these findings can be interpreted as differences in thrombogenicity and inflammatory responses has not been proven, but seems probable. This in vitro method should make it possible to further study human responses to synthetic materials a method possibly more reliable than animal experiments.