Effects of Enoxaparin on Intravascular Sclerostin Release in Healthy Men.

Sclerostin (Scl) is implicated in vascular calcification and angiogenesis and localizes within vasculature. Its molecule incorporates a heparin-binding site that implies also binding to endothelial glycocalyx. We preliminary tested whether intravenous (IV) low-molecular-weight heparin enoxaparin can stimulate intravascular release of this calcification inhibitor in humans. Sixteen male volunteers were injected with a bolus of 1 mg/kg body weight of enoxaparin. After 10 minutes, plasma immunoreactive Scl levels increased uniformly by a mean of 184% versus baseline level of 0.56 ± 0.17 ng/mL ( P = .0004). Plasma Scl levels were found still elevated after 2 and 6 hours (with a median of 20.9% and 8.69%, respectively) and became normal after 24 hours. The percentage of increase (Δ) in plasma Scl after 10 minutes was directly correlated with enoxaparin dose per kg/m2 of body mass index (ρ = 0.587, P = .017) and strongly inversely correlated with the preinjection Scl levels (ρ = -0.747, P = .0008). A robust negative association between the ΔScl increase after 10 minutes and the ΔScl decrease after 2 hours versus 10 minutes was observed (ρ = -0.835, P < .0001). Complementary in vitro spiking experiment showed no effects of enoxaparin addition and whole blood incubation on plasma Scl levels when measured with the immunoassay. This study shows that enoxaparin has a stimulating effect on the intravascular release of calcification inhibitor Scl in healthy men. This novel pharmacological action of the popular anticoagulant drug seems important in cardiovascular medicine.

Unexpected and striking effect of heparin-free dialysis on cytokine release.

Heparin (both unfractionated and low molecular weight) is not only a potent anticoagulant but also has many pleiotropic effects, some of which are mediated by cytokine release. We compared the effect of hemodialysis (HD) with enoxaparin as an anticoagulant and without systemic anticoagulation (heparin-grafted membrane-Evodial) on the release of monocyte chemoattractant protein 1 (MCP-1), endostatin (ES) and activin A (Act-A). Nineteen stable HD patients were dialyzed with or without heparin, and plasma levels of MCP-1, ES and Act-A were measured after such a dialysis. During HD with Evodial, the intradialytic levels of all three cytokines were 2-3 folds lower. The between-anticoagulant differences were significant over time for all three cytokines: MCP-1 (P < 0.001), ES (P < 0.001) and Act-A (P < 0.001). This striking effect of heparin-free dialysis with Evodial membrane may be beneficial not only because it reduces the possibility of bleeding complications but also because it might reduce proinflammatory cytokine concentration and therefore contribute to the improvement in endothelial function. Further studies are needed to determine whether it has a positive effect on morbidity and mortality of maintenance HD patients.

Effects of enoxaparin on myeloperoxidase release during hemodialysis.

Myeloperoxidase (MPO) is a proteolytic and prooxidant enzyme largely assembled with the vascular wall, and a heparin-binding protein. We studied if low-molecular-weight heparin enoxaparin administered for hemodialysis (HD) anticoagulation causes systemic MPO activation. Plasma MPO levels were measured in patients undergoing maintenance HD with an intravenous bolus of enoxaparin. Patients were retested during HD employing dialyzers with heparin-grafted polyacrylonitrile membrane and no systemic enoxaparin administration. During enoxaparin-anticoagulated HD plasma MPO levels strikingly increased in all patients (8.6-fold at 10 minutes and 3.3-fold at 120 minutes, both P < 0.0001). The increments were directly associated with the enoxaparin dosage and strongly inversely with the predialysis levels of the enzyme. The increase in plasma MPO during systemic heparin-free HD was significantly less pronounced. Enoxaparin administered for HD anticoagulation induces a marked and dose-dependent increase in plasma MPO as a plausibly favorable result of the liberation of the enzyme from the vascular wall.

Hepatocyte growth factor in saliva of patients with renal failure and periodontal disease.

BACKGROUND: Hepatocyte growth factor (HGF), endogenous cytokine with pleiotropic repairing and regeneration properties in relation to most tissues and organs, contributes to the progression of periodontal disease (PD). Furthermore, PD is a significant health problem in patients with chronic renal failure (CRF). The role of HGF in the development of PD in this specific population was not a subject of research so far. MATERIAL AND METHODS: The following groups were enrolled in the study: (1) 26 chronic hemodialysis (HD) subjects, (2) 26 patients treated by continuous ambulatory peritoneal dialysis (CAPD), (3) 28 predialysis CRF patients, (4) 26 subjects with advanced PD (without coexisting diseases), and (5) 20 healthy subjects without PDs. HGF level in saliva was measured using the immunoenzymatic method. Gingival index, papillary bleeding index, plaque index, and the loss of clinical attachment level were evaluated. RESULTS: The HGF level in saliva of HD patients was twice higher than in that of subjects with healthy periodontium. Direct relationships between proper HGF level in saliva and the indices GI, PBI, and PI in CAPD-treated patients and with more severe PD were shown. It was found that PD is most advanced in HD patients, moderately in CAPD-treated patients and to the smallest extent in predialysis CRF patients. CONCLUSIONS: The HGF level in mixed saliva is the index of PD progression in subjects without renal failure and in CAPD-treated patients. PD is common in renal failure patients and is a significant problem concerning general health status.

Insulin sensitivity, plasma adiponectin and sICAM-1 concentrations in patients with subclinical hypothyroidism: response to levothyroxine therapy.

Subclinical hypothyroidism is associated with an increased risk of atherosclerosis. The aim of this study was to investigate the concentration of plasma soluble intercellular adhesion molecule-1 and adiponectin in relation to insulin sensitivity in patients with subclinical hypothyroidism and to estimate if L-thyroxine treatment had an influence on these parameters. 13 women with subclinical hypothyroidism and 14 euthyroid controls were included in the study. A physical examination was conducted, hyperinsulinemic euglycemic clamp and plasma soluble intercellular adhesion molecule-1, adiponectin and lipids profiles were measured at baseline in both groups and in the group with subclinical hypothyroidism the above procedures were performed after L-thyroxine therapy (mean time of treatment 5.0 months) in stable euthyroid state. Insulin sensitivity and adiponectin were not different at baseline in the two studied groups. Plasma soluble intercellular adhesion molecule-1 concentration was significantly higher in the patients with subclinical hypothyroidism (P = 0.011). The comparison of lipids profiles revealed that only LDL-cholesterol concentration was higher (P = 0.011) in the group with subclinical hypothyroidism. After therapy, we observed an improvement of insulin sensitivity (P = 0.012) and a decrease of plasma glucose (P = 0.019) and soluble intercellular adhesion molecule-1 (P = 0.01), whereas adiponectin concentration remained unchanged. We concluded that L-thyroxine treatment in patients with subclinical hypothyroidism might exert a beneficial effect by reducing cardiovascular risk factors.

[Salivary phosphate-binding chewing gum--an integrative approach to chronic kidney disease and oral diseases]. / Guma do zucia wiazaca fosforany--jednoczesne leczenie przewleklej choroby nerek i chorób jamy ustnej.

The course and treatment of chronic kidney disease (CKD) is adversely affected by numerous metabolic disarrangements, comorbid states and general diseases, i.e. hyperphosphatemia and metabolic bone disease, chronic inflammation, accelerated atherosclerosis (partly of infectious etiology) and devastating cardiovascular disease. Furthermore, CKD patients are usually afflicted by multiple oral abnormalities, including troublesome oral dryness and the very aggressive form of periodontal disease. The use of chitosan-containing chewing gum in CKD subjects seems to offer a novel therapeutic approach of interdisciplinary importance: the chitosan binds salivary phosphates and, when swallowed, likely phosphates in the alimentary tract, thus beneficially lowering blood phosphate levels, while the gum itself increases impaired salivary flow and has a potent oral antimicrobial activity. Thus, it effectively improves general oral hygiene and prevents progression of periodontal disease being (besides of hyperphosphatemia) one of the established causes of atherosclerosis development/progression. The undemanding maneuver of chewing the chitosan-containing, phosphate-binding gum has a potential to diminish excessive morbidity in CKD patients.

Effect of sulodexide on plasma transforming growth factor-beta1 in healthy volunteers.

It is unknown whether the glycosaminoglycan drug sulodexide interferes with transforming growth factor-beta1--a member of heparin-binding family and a potent regulator of human biology and diseases. Hence, a 2-week pilot study was performed in 11 healthy men. Sulodexide was initially administered intravenously in a single dose, then--orally for 12 days and--again intravenously on study completion. Initial injection had no effect on activated form of the growth factor measured in plasma after 10 and 120 min; no change was also observed after 120 min from drug ingestion on day 7. On final intravenous administration, the growth factor levels increased by almost 60% after 10 min and remained elevated; the 120-min levels directly correlated with sulodexide dosage. Baseline cytokine levels decreased during the 2-week trial by more than 50%. In conclusion, transforming growth factor-beta1 release and likely downregulation of its expression may constitute novel pharmacological effects of sulodexide.

Hepatocyte growth factor/activin A/follistatin system activation during hemodialysis with different low molecular weight heparins.

Hepatocyte growth factor (HGF), activin A (Act A), and follistatin (FS) compose an organotrophic system; interestingly it is modified by heparin. To understand if LMWHs (considered distinct drugs) have different clinical profiles regarding the above growth factors, we studied the effects of enoxaparin, nadroparin, and dalteparin on their plasma levels. Seventeen chronic HD patients completed this prospective, crossover trial. They were randomized into six groups: each patient was administered enoxaparin (effective dose of 0.75 mg/kg), nadroparin (70.4 IU/kg) and dalteparin (78.6 IU/kg) in three time periods of two months each. At the end of this period, the cytokine's plasma levels were measured by immunoassays at the start and at 10 min and 180 min of the HD procedure. At 10 min, we observed a striking increase in plasma HGF (32-fold), Act A (4-fold), and FS (53%), all p = 0.0003. The levels of HGF and Act A remained markedly elevated after 180 min (by 295% and 87%, respectively; both p < 0.002), while those of FS returned to baseline. There were no differences in cytokine profile comparing both their peak concentrations and the areas under the curve. Enoxaparin, dalteparin, and nadroparin are seemingly not different considering the release of HGF/Act A/FS during HD procedures; this may reflect their similar profile in other aspects. Moreover, the concentrations of HGF/Act A/FS are close to therapeutic ones, which may partly explain the mechanisms underlying some of the emerging extra-anticoagulant effects of LMWHs.

Pleiotropic effects of heparin and heparinoids in peritoneal dialysis.

Unfractionated heparin, low-molecular-weight heparins, and sulodexide belong to the family of glycosaminoglycans. Recent studies report on properties other than anticoagulant activities of these medications. They include modulation of cell growth and proliferation via actions on numerous growth factors affecting the immune system and matrix molecules production and degradation. Long-term peritoneal dialysis remarkably influences peritoneal cavity homeostasis by mechanisms mediated by growth factors. They initiate progression of pathological processes and further account for morphological and functional alterations of the peritoneal membrane. The best-recognized pathologies in peritoneal cavity under these conditions encompass inflammation, fibrosis, and vasculopathy, often leading to fatal encapsulating peritoneal sclerosis. Intraperitoneal heparin and its derivatives, by their pleiotropic actions, may influence these crucial processes and improve the peritoneal dialysis technique survival in a complex and so far understudied way. These issues, novel medical approaches, and their likely mechanisms have been reviewed.

Chronic viral hepatitis C, oxidative stress and the coagulation/fibrinolysis system in haemodialysis patients.

INTRODUCTION: The aim of the present study was to establish whether the presence of chronic viral hepatitis (PVH) could be implicated in the elevation of oxidative stress (SOX) and haemostasis system in haemodialysis (HD) patients. MATERIALS AND METHODS: In HD patients with and without PVH and in controls we compared the markers of: coagulation pathway- tissue factor (TF) and its inhibitor (TFPI), prothrombin fragment F (1+2) (F (1+2)); fibrinolysis: tissue-type plasminogen activator (tPA), urokinase-type plasminogen activator (uPA) and its soluble receptor (suPAR), plasminogen activator inhibitor 1 (PAI-1), plasmin/antiplasmin complexes (PAP); and a marker of SOX-Cu/Zn superoxide dismutase (Cu/Zn SOD) levels. RESULTS: Patients, particularly those with PVH, showed a significant increase in the markers of the coagulation, fibrinolysis and oxidative status as compared to controls. All parameters of coagulation/fibrinolysis system were directly associated with the PVH and Cu/Zn SOD levels, and there was a relationship between the PVH and Cu/Zn SOD levels. Multivariable analysis showed that PVH and increased SOX were identified as independent variables significantly associated with the disturbances of coagulation/fibrinolysis system in these patients. CONCLUSIONS: We concluded that PVH is a novel determinant of the increased oxidative stress as well as the disturbances of coagulation/fibrinolysis system in haemodialysis patients.

Sulodexide for hemodialysis anticoagulation in heparin-induced thrombocytopenia type II.

Heparin-induced thrombocytopenia type II (HIT II) is an immune-mediated prothrombotic state. It requires cessation of all forms of heparin exposure. In maintenance hemodialysis (HD) patients, alternative anticoagulants (i.e. bivalirudin, danaparoid, fondaparinux) may be tried for HD procedure anticoagulation. Sulodexide (SLX) - a purified glycosaminoglycan preparation (80% heparan sulfate and 20% dermatan sulfate) - is not neutralized by platelet factor 4 and may be useful in HIT II. A 32-year-old man on continuous ambulatory peritoneal dilaysis (CAPD) and with protracted atrial fibrillation was given enoxaparin prophylaxis. On day 4, his platelets dropped from 119,000/micronL to 27,000/micronL and HIT II was diagnosed by positive heparin-induced platelet aggregation. While enoxaparin was withdrawn, the platelet count increased and remained stable. In the meantime, atrial fibrillation subsided but the patient developed pseudomonal peritonitis; the catheter was removed and the patient was switched to HD with SLX as an anticoagulant (bolus of 30 mg at HD onset). He was uneventfully treated with HD for 6 weeks and then reverted to CAPD. The widely available and inexpensive SLX may be a new, effective and potentially promising alternative anticoagulant in HD patients with HIT II.

Enoxaparin but not unfractionated heparin causes a dose-dependent increase in plasma TGF-beta 1 during haemodialysis: a cross-over study.

BACKGROUND: Transforming growth factor-beta1 (TGF-beta1) is a multi-functional cytokine that presents as a mediator of the heparin's pleiotropic action. In this cross-over study, we compared the effects of enoxaparin and unfractionated heparin (UFH) used as anticoagulants during haemodialysis (HD) on plasma TGF-beta1 levels and some platelet activation markers: platelet-derived growth factor-AB (PDGF-AB), beta-thromboglobulin (beta-TG) and platelet factor-4 (PF-4). METHODS: Plasma immunoreactive markers (in 22 chronically HD patients) were quantified at the start, at 10 and 180 min of HD session. Enoxaparin was administered as a single dose of 0.67 +/- 0.14 mg/kg at the onset of HD, while UFH was given as a bolus of 1500 (500-3500) IU followed by an infusion of 2750 (1500-6500) IU. The time of evaluation for each heparin was 3 months. RESULTS: Pre-dialysis, TGF-beta1 levels tended to be lower in patients anticoagulated with enoxaparin compared with UFH [6.9 (3.3-21.9) ng/ml vs 8.4 (3.8-30.2) ng/ml, respectively; P = 0.05]. Overdialysis, TGF-beta1 levels showed a significant 44.8% increase to 10.0 (2.9-28.0) ng/ml after 10 min (P = 0.002) and to 9.32 (5.3-23.7) ng/ml after 180 min (P = 0.016) of enoxaparin-anticoagulated HD and remained stable during UFH administration [9.4 (3.9-25.3) ng/ml after 10 min, 8.1 (4.1-21.9) ng/ml after 180 min; P = 0.385]. The 35% increase in plasma TGF-beta1 after 180 min of HD positively correlated with the enoxaparin dose/kg (r = 0.553, P = 0.008) and, interestingly, negatively with the baseline level of the cytokine (r = -0.544, P = 0.009). Despite a positive correlation between TGF-beta1 and PDGF-AB during HD, there were no associations between TGF-beta1 and beta-TG or PF-4 regardless of the type of anticoagulation. CONCLUSION: Enoxaparin, compared with UFH, induces a rapid overdialytic but not sustained increase in plasma TGF-beta1 levels. The effect is closely dose-dependent and may reflect systemic activation of this multi-potential cytokine.

Sulodexide induces hepatocyte growth factor release in humans.

Heparin influences numerous pleiotropic growth factors, including hepatocyte growth factor (HGF), partially by their release from endothelial and extracellular matrix stores. The effects of sulodexide, a heparin-like glycosaminoglycan medication of growing importance in medicine, on HGF liberation are not known. We performed a 2-week open-label sulodexide trial in healthy male volunteers. The drug was initially administered intravenously (i.v.) in a single dose of 1200 Lipoprotein Lipase Releasing Units (LRU), then -- orally for 12 days (500 LRU twice a day), and -- again by i.v. route (1200 LRU) on day 14. Intravenous sulodexide injections were repeatedly found to induce marked and reproducible increases in immunoreactive plasma HGF levels (more than 3500% vs baseline after 10 min, and more than 1200% after 120 min), and remained unchanged when measured 120 min following oral sulodexide administration. The percentage increments in plasma HGF evoked by i.v. sulodexide at both time points and on both days inversely correlated with baseline levels of the growth factor. On day 14, the HGF levels after 120 min and their percentage increase vs baseline were strongly and directly dependent on i.v. sulodexide dose per kg of body weight. This study shows that sulodexide has a novel, remarkable and plausibly biologically important stimulating effect on the release of pleiotropic hepatocyte growth factor in humans.