Time-dependent changes in extra-domain A-fibronectin concentration and relative amounts of fibronectin-fibrin complexes in plasma of patients with peripheral arterial disease after endovascular revascularisation

Abstract

Fibronectin (FN) may be involved in time- and stage-dependent and inter-related controlled processes of inflammation, coagulation, and wound healing accompanying peripheral arterial disease (PAD). In the present study, FN and FN-containing extra-domain A (EDA-FN), macromolecular FN-fibrin complexes, and FN monomer were analysed in the plasma of 142 PAD patients, including 37 patients with restenosis, for 37 months after revascularisation. FN concentration increased significantly in the plasma of PAD patients within 7 to 12 months after revascularisation, whereas the high concentration of EDA-FN was maintained up to 24 months, significantly higher in the group 7 to 12 months after revascularisation with recurrence of stenosis and lower in the PAD groups 1 to 3 months and 4 to 6 months after revascularisation with comorbid diabetes and ulceration, respectively. The relative amounts of FN-fibrin complexes up to 1600 kDa and FN monomer were significantly higher, within intervals of 4 to 24 months and 4 to 6 months after revascularisation, respectively. Moreover, the relative amounts of 750 to 1600 kDa FN-fibrin complexes within 13 to 24 months after revascularisation were higher in comparison with those in the group without restenosis. In conclusion, high levels of EDA-FN and FN-fibrin complexes could have potential diagnostic value in the management of PAD patients after revascularisation, predicting restenosis risk.

Keywords: EDA-fibronectin; FN-fibrin complexes; fibronectin; peripheral arterial disease; restenosis.

Figure 1

Representative immunopatterns of FN‐fibrin complexes in plasma of PAD patients after revascularisation of ischaemic lower limbs. Blood plasma samples of 142 PAD patients and 37 normal individuals were subjected to SDS‐agarose gel electrophoresis under non‐reducing conditions (25). The bands were transferred to a nitrocellulose membrane (Serva Electrophoresis GmbH, Heidelberg, Germany) and developed with anti‐FN monoclonal antibody (Clone FN30‐8, TaKaRa Shuzo Co. Ltd., Shiga, Japan). Lanes 1 to 2, normal plasma; lanes 2 to 7, PAD patients’ plasma

Figure 2

Levels of FN molecular forms in plasma samples of PAD patients with respect to comorbid diabetes. Mean values of data: concentrations (mg/l) of plasma FN (A) and EDA‐FN (B); relative amounts (%) of FN monomer (C) and FN‐fibrin complexes I (D), II (E), III (F), and IV (G) of all PAD patients (—○—), PAD patients with diabetes (—□—), and PAD patients without diabetes (—Δ—) shown in relation to period after revascularisation: 1 to 3 months (diabetes: n = 42, without diabetes: n = 30), 4 to 6 months (diabetes: n = 49, without diabetes: n = 36), 7 to 12 months (diabetes: n = 98, without diabetes: n = 66), 13 to 24 months (diabetes: n = 120, without diabetes: n = 66), and 25 to 37 months (diabetes: n = 30, without diabetes: n = 12). *Significantly different between groups of PAD with comorbid diabetes and PAD with diabetes excluded

Figure 3

Levels of FN molecular forms in plasma samples of PAD patients with respect to ulceration. Mean values of data: concentrations (mg/l) of plasma FN (A) and EDA‐FN (B); relative amounts (%) of FN monomer (C) and FN‐fibrin complexes I (D), II (E), III (F), and IV (G) of all PAD patients (—○—), PAD patients with previous ulcer (—□—), and PAD patients without ulcer (—Δ—) shown in relation to period after revascularisation: 1 to 3 months (ulcer: n = 7, without ulcer: n = 66), 4 to 6 months (ulcer: n = 9, without ulcer: n = 76), 7 to 12 months (ulcer: n = 24, without ulcer: n = 154), 13 to 24 months (ulcer: n = 26, without ulcer: n = 176), and 25 to 37 months (ulcer: n = 0, without ulcer: n = 45). *Significantly different between groups of PAD with comorbid ulcer and PAD with ulcer excluded

Figure 4

Levels of FN molecular forms in plasma of PAD patients with respect to restenosis occurrence. Mean values of data: concentrations (mg/l) of plasma FN (A) and EDA‐FN (B); relative amounts (%) of FN monomer (C) and FN‐fibrin complexes I (D), II (E), III (F), and IV (G) shown with respect to occurrence of restenosis (—□—) and without restenosis (—○—) after revascularisation: 1 to 3 months (without restenosis: n = 54, restenosis: n = 23), 4 to 6 months (without restenosis: n = 66, restenosis: n = 19), 7 to 12 months (without restenosis: n = 155, restenosis: n = 23), 13 to 24 months (without‐restenosis: n = 180, restenosis: n = 22), and 25 to 37 months (without restenosis: n = 43, restenosis: n = 3)