Clinical Hemorheology and Microcirculation - Volume 69, issue 1-2

Authors: Negrão de Figueiredo, G. | Mueller-Peltzer, K. | Zengel, P. | Armbruster, M. | Rübenthaler, J. | Clevert, D.A.

Article Type: Research Article

Abstract: BACKGROUND: Contrast-enhanced ultrasound (CEUS) has been used as an additional imaging technique to conventional ultrasound in order to clarify gallbladder findings, CEUS is easy and fast to perform, overcomes the limitations of gray-scale ultrasonography and is a safe tool. OBJECTIVE: The aim of this retrospective mono-center analysis study is to evaluate the diagnostic performance of CEUS in the assessment of gallbladder disease by comparing its results with additional CT findings. METHODS: A total of 24 patients between 2009 and 2017 with unclear gallbladder appearance were retrospectively analysed. The contrast agent administered was a second-generation blood pool …agent (SonoVue® , Bracco, Milan, Italy). Examinations were performed and interpreted by a single experienced radiologist with more than fifteen years’ experience. RESULTS: 24 patients were successfully examined without any adverse reaction. CEUS presented a sensitivity of 75%, a specificity of 100.0%, a positive predictive value of 100.0% and a negative predictive value of 95%. CONCLUSIONS: In conclusion, the excellent results in this study acknowledged that CEUS is a feasible alternative tool to differentiate gallbladder pathologic alterations. Show more

Keywords: CEUS, gallbladder, pathologic alterations

DOI: 10.3233/CH-189116

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 83-91, 2018

Authors: Apfelbeck, M. | Clevert, D.-A. | Ricke, J. | Stief, C. | Schlenker, B.

Article Type: Research Article

Abstract: INTRODUCTION: Reduced acceptance of radical prostatectomy in patients with low risk or intermediate risk prostate cancer has significantly changed treatment strategies in prostate cancer (PCa) during the last years. Focal therapy of the prostate with high intensity focused ultrasound (HIFU) is an organ-preserving treatment for prostate cancer with less impairment of health-related quality of life. Follow-up after HIFU therapy by imaging modalities remains a major problem as eg. MRI performs poorly. Contrast enhanced ultrasound (CEUS) allows to monitor the vascular architecture of organs non-invasively. However, only limited data are available using CEUS to define successful and complete HIFU treatment of …the prostate. In this study, we aimed to evaluate short-term image findings using CEUS and image fusion before and after HIFU treatment. MATERIALS AND METHODS: Prospective single arm study in patients with uni- or bilateral, low or intermediate risk prostate cancer or recurrent cancer after radiotherapy treated with HIFU at our institution between October 2016 and November 2017. HIFU hemiablation or whole gland treatment was performed using the Focal One® device. PCa was diagnosed either by multiparametric magnetic resonance imaging (mpMRI) followed by MRI fusion based targeted biopsy combined with 12 core transrectal ultrasound (TRUS) guided biopsy or 12 core random biopsy only. Monitoring of the target region before, immediately and 24 hours after the ablation was done by CEUS in combination with image fusion using an axial T2-weighted MRI sequence. RESULTS: 6 consecutive patients with Gleason score (GS) 6, 5 patients with GS 7a prostate cancer and one patient with biochemical recurrence after radiotherapy were included in the study. Three patients underwent whole gland treatment due to histological proven bilateral PCa or recurrent PCa after radiotherapy. Hemiablation was performed in 9 patients with unilateral tumor and no PIRADS 4 or 5 lesion in the contralateral lobe. Median patient age was 69.8 years and median PSA (prostate-specific antigen) level was 8.4 ng/ml. CEUS showed markedly reduced microbubbles in the ablated area, the prostate capsule still showed signs of perfusion. The study is limited by the short follow up and small number of patients. CONCLUSIONS: CEUS examination showed a reduction of microcirculation in the treated area immediately after the treatment and 24 hours later. The combination of CEUS and image fusion seems to be helpful for detecting the PCa target lesion and monitor the success of HIFU ablation treatment. Evidence for image findings after HIFU-therapy are rare. Further studies on this topic are needed. Show more

Keywords: High intensity focused ultrasound therapy, image fusion, prostate, CEUS, contrast enhanced ultrasound

DOI: 10.3233/CH-189123

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 93-100, 2018

Authors: Tsikouras, Panagiotis | Niesigk, Barbara | von Tempelhoff, Georg-Friedrich | Rath, Werner | Schelkunov, Olga | Daragó, Peter | Csorba, Roland

Article Type: Research Article

Abstract: INTRODUCTION: Recent studies have shown increased RBC aggregation and no difference in plasma viscosity in the presence of markedly lower hematocrit in women at term compared to non-pregnant women. Little is known about the outcome of blood rheological parameters and red blood cell (RBC) deformability particularly in the course of normal pregnancy. METHODS: During a 36 months interval 1.913 blood samples were randomly collected from a total of 945 pregnant women in the course of their pregnancy (n  = 1.259) and during puerperium (upto 1 week; n  = 654). Next to the blood count, hemorheological parameters including red blood cell …(RBC) -aggregation (stasis E0; low shear E1), -deformability (low, moderate and high shear conditions) and plasma viscosity (pv) were assessed. Plasma viscosity (pv) was examined using KSPV 1 Fresenius, RBC aggregation (stasis: E0 and low shear: E1) using MA1-Aggregometer; Myrenne and RBC deformability (def) was determined by Rheodyn SSD Diffractometer, Myrenne, Roetgen, Germany were tested. In some of these women laboratory results prior to pregnancy (n  = 145) were available which were compared with those during pregnancy. RESULTS: Mean maternal pv remained unchanged within each trimester and compared to the values before pregnancy and during early puerperium (Range of means: 1.18–1.20 mPa S). In contrast, RBC agg (E0 and E1) was markedly higher in the 2nd (21.8 ± 7.0 and 28.9 ± 9.4; p  < 0.001) and 3rd trimester (18.74 ± 8.4 and 28.2 ± 9.4; p  < 0.01) compared to the values before pregnancy (16.4 ± 6.4 and 20 ± 7.5) and during 1st trimester (17.49 ± 6.5 and 22.4 ± 7.4). There was a stat. significant temporary reduction in RBC def. under all shear rate conditions during 2nd trimester compared to the values before pregnancy which remained significantly lower during 3rd trimester only under high shear rates. An increase RBC agg was stat. significantly inversely correlated with reduced RBC def being most pronounced under low shear rate conditions. While RBC rigidity was stat. significantly correlated with higher hematocrit values there was only a weak correlation between RBC agg and haematocrit (E0: r  = –0.084; p  = 0.03; E1: r  = –0.06; p  = 0.1). Pv was not correlated with haematocrit or RBC def but stat. significantly correlated with RBC agg. CONCLUSIONS: Blood rheological changes manifest during 1st trimester, and fairly remain unchanged during 2nd trimester until term. Physiologic hemodilution and increasing hypercoagulability is accompanied by high RBC -aggregation and – rigidity during 2nd trimester while plasma viscosity remains nearly unaffected throughout normal pregnancy. Show more

Keywords: Normal pregnancy, haemorheology, red blood cell aggregation, red blood cell deformability

DOI: 10.3233/CH-189104

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 101-114, 2018

Authors: Oelert, Anja | Kittel, Manuela | Hahn, Martin | Haase, Hermann | Jünger, Michael

Article Type: Research Article

Abstract: In five patients who suffered from chronic venous insufficiency clinical stage C4 (n  = 3) and C6 (n  = 2) the capillary blood pressure was measured twice by means of invasive direct cannulation of nailfold capillaries of the toe. During one measurement course the patients wore below knee medical compression stockings (40 mmHg) during the other they did not have compression therapy. With the patient in supine position, the CP was investigated by the servo-nulling technique under resting conditions and under dynamic conditions: the calf-muscle/ankle joint venous pump was simulated by means of inflating a blood pressure cuff, which surrounded the mid …lower leg, to 60 mmHg for 60 s. Results: The simulated calf-muscle contraction induced a steep increase of CP with 5.65 mmHg/s (Q1 5.27 mmHg/s, Q3 5.92 mmHg/s), which was significantly (p  = 0.013) reduced by MCS to 2.47 mmHg/s (Q1 1.65 mmHg/s, Q3 3.0 mmHg/s). Time needed to reach the max. CP was 11.35 s, which was lengthened by MCS to 23.4 s (p  = 0.134). Conclusion: Compression therapy prevents capillary hypertension, the major hemodynamic reason for the development of advanced stages of chronic venous insufficiency which are defined by skin disease like hyperpigmentation, lipodermatosclerosis and ulcer. Show more

Keywords: Chronic venous insufficiency, dermal capillary blood pressure, cutaneous microcirculation, dermal capillary hypertension, pathophysiology of chronic venous insufficiency, servo nulling pressure measurement

DOI: 10.3233/CH-189122

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 115-121, 2018

Authors: Wernly, Bernhard | Lichtenauer, Michael | Vellinga, Namkje A.R. | Boerma, E. Christiaan | Ince, Can | Kelm, Malte | Jung, Christian

Article Type: Research Article

Abstract: BACKGROUND AND PURPOSE: The Microcirculatory Shock Occurrence in Acutely Ill Patients (micro-SOAP) study investigated associations of microcirculation and mortality. Risk stratification in critically ill patients is of utmost interest. Established score such as APACHE2 (Acute Physiology And Chronic Health Evaluation 2) are relatively complex and might therefore be of limited use. Blood urea nitrogen (BUN) was described to be associated with mortality in various diseases. We therefore aimed (i) to evaluate BUN for prediction of mortality in a cohort of critically ill patients and (ii) to investigate associations of BUN with microcirculation. METHODS: 412 patients were included in …our post-hoc analysis of the prospective multicenter microSOAP study. Assesment of the sublingual microcirculation (Sidestream Dark Field (SDF) imaging) and collection of laboratory values were performed on the same day in this point prevalence study. Evaluation of associations with mortality was done by logistic regression analysis. An optimal BUN cut-off was calculated by means of the Youden Index. RESULTS: Median BUN was 9.0 mmol/L. BUN was associated with in-hospital-mortality in a logistic regression analysis (HR 1.03; 95% CI 1.01–1.05; p  < 0.001). Per quartile (BUN 0–5.4 mmol/L, 5.4–9.0 mmol/L, 9.0–15.9 mmol/L and above 15.9 mmol/L) in-hospital mortality increased by as much as 51% (HR 1.51; 95% CI 1.23–1.85; p  < 0.001). ROC analysis was done (AUC 0.63 95% CI 0.58–0.67) and the statistically optimal cut-off calculated by means of the Youden Index: 9.7 mmol/L. This cut-off was associated with a significant 3-fold increase in mortality (HR 2.97 95% CI 1.88–4.70; p  < 0.001) and remained robustly associated with adverse outcome after correction for APACHE2 (HR 2.71 95% CI 1.61–4.59; p  < 0.001), renal function as expressed by creatinine (HR 2.63 95% CI 1.59–4.33; p  = 0.001), as well in an integrative model (MAP<60 mmHg, tachycardia (heart rate >90/min), lactate above 1.5 mmol/L, age above 80 years; HR 2.43 95% CI 1.50–3.92; p  < 0.001). Parameters of microvascular perfusion were associated neither with BUN nor mortality. CONCLUSIONS: BUN is associated with hospital mortality and a combination of BUN and clinical signs might constitute a powerful but easy-to-use tool for risk stratification in critically ill patients and help improve their outcome. BUN was not associated with parameters of microcirculation which were not associated with mortality. Show more

Keywords: Critically ill, BUN, ICU, risk stratification, risk score, microcirculation

DOI: 10.3233/CH-189111

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 123-131, 2018

Authors: Nemeth, Norbert | Deak, Adam | Szentkereszty, Zsolt | Peto, Katalin

Article Type: Research Article

Abstract: In surgical pathophysiology ischemia-reperfusion, inflammatory processes, sepsis, vascular interventions, tissue trauma, shock, all mean conditions in which hemorheological parameters show alterations. Despite of numerous clinical and experimental studies, the in vivo hemorheology is not completely understood yet, and several fundamental questions still need to be answered. Investigating these issues, experimental surgical models are important, in point of view of the translational research as well. In this paper we aimed to make an attempt on summarizing the possible factors and conditions that might have an effect on hemorheological results in experimental surgical studies. Hemorheological parameters show alterations in surgical pathophysiological …processes in a complex way. However, the changes are dominantly non-specific. Standardized experimental conditions, related to the experimental animal (species, animal welfare) anesthesia-medications, operation, sampling and, if applicable, conditions of the postoperative period, are inevitable for a safe assessment of valuable (hemorheological) results. Parallel investigations – such as microcirculatory monitoring, imaging techniques, other laboratory methods, histomorphology– have great importance, together with individual analysis of changes, for a better understanding of the changes and for comparability with clinical results. Show more

Keywords: Hemorheology, experimental surgery, experimental design, inter-species differences

DOI: 10.3233/CH-189105

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 133-140, 2018

Authors: Felthaus, Oliver | Schön, Teresa | Schiltz, Daniel | Aung, Thiha | Kühlmann, Britta | Jung, Friedrich | Anker, Alexandra | Klein, Silvan | Prantl, Lukas

Article Type: Research Article

Abstract: BACKGROUND: Multiple symmetric lipomatosis is a rare disease characterized by the excessive growth of uncapsulated masses of adipose tissue. Although the etiology has yet to be elucidated, a connection to brown adipose tissue has been proposed recently. The mTOR pathway which is found to be regulated in lipomatous tissue as well as associated with brown adipose tissue can be inhibited by a compound called rapamycin. METHODS: We isolated adipose tissue derived stem cells from both affected and unaffected tissue and treated these cells with different concentrations of rapamycin. RESULTS: The differences in both proliferation and differentiation …between adipose tissue derived stem cells (ASCs) from lipomatous and normal tissue decreased after mTOR pathway inhibition. In some patients regulation of mTOR genes was opposed in the ASCs from the two different tissues. CONCLUSIONS: Treatment with rapamycin might be a novel therapeutical approach for patients suffering from multiple symmetric lipomatosis. Show more

Keywords: Multiple symmetric lipomatosis, rare disease, mTOR pathway, rapamycin

DOI: 10.3233/CH-189107

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 141-151, 2018

Authors: Arora, N. | Caldwell, A. | Wafa, K. | Szczesniak, A. | Caldwell, M. | Al-Banna, N. | Sharawy, N. | Islam, S. | Zhou, J. | Holbein, B.E. | Kelly, M.E.M. | Lehmann, Ch.

Article Type: Research Article

Abstract: BACKGROUND/OBJECTIVE: Non-infectious uveitis is an inflammatory disease of the eye commonly treated by corticosteroids, though important side effects may result. A main mediator of inflammation are oxygen free radicals generated in iron-dependent pathways. As such, we investigated the efficacy of a novel iron chelator, DIBI, as an anti-inflammatory agent in local and systemic models of endotoxin induced uveitis (EIU). METHODS: Firstly, the effects of DIBI in systemic EIU in Lewis rats were established. 2 hours post intravenous LPS or LPS/DIBI injections, leukocyte activation and functional capillary density (FCD) were examined using intravital microscopy (IVM) of the iridial microcirculation. …Secondly, the toxicity of DIBI was evaluated in BALB/C mice for both acute and chronic dosages through gross ocular examination, intraocular pressure measurements and hematoxylin-eosin staining of ocular tissue. Lastly, three groups of BALB/C mice, control, LPS or DIBI + LPS, were studied to evaluate the effectiveness of DIBI in treating local EIU. Five hours post-local intravitreal (i.v) injection, leukocyte activation and capillary density were examined via IVM. RESULTS: Treatment of systemic EIU with DIBI resulted in a reduction of leukocyte activation and FCD improvement within the iridial microcirculation. Toxicity studies suggested that acute and chronic DIBI administration had no adverse effects in the eye. In the local EIU model, DIBI was shown to reduce leukocyte activation and restored the FCD/DCD ratio, providing evidence for its anti-inflammatory properties. CONCLUSIONS: Our study has provided evidence that DIBI has anti-inflammatory effects in experimental uveitis. Additionally, no local ocular toxicity was observed. Show more

Keywords: Iron chelation, ocular inflammation, endotoxin, intravital microscopy, microcirculation

DOI: 10.3233/CH-189109

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 153-164, 2018

Authors: Saldanha, Carlota | Freitas, Teresa | Silva-Herdade, Ana Santos

Article Type: Research Article

Abstract: Timolol maleate is a compound used in treatment for reducing increased intra-ocular pressure by limiting aqueous humor production. Decreased erythrocyte deformability (ED), increased activity of erythrocyte acetylcholinesterase (AChE), increased values of nitrosoglutathione (GSNO) and nitic oxide (NO) and decreased plasma levels of NO metabolites, were described in primary open angle glaucoma patients. In healthy human red blood cells (RBCs), timolol is an inhibitor of AChE and induces NO efflux and GSNO efflux from that blood component in lower concentration than those obtained in presence of the natural AChE substrate, acetylcholine (ACh). The signal transduction pathway in RBCs described for NO …in dependence of AChE-ACh active complex involves Gi protein, protein tyrosine kinase (PTK like Syk and p53/56Lyn), protein tyrosine phosphatase (PTP) and adenylyl cyclase (AC). The aim of this in vitro study was to verify the effect of timolol maleate in ED, NO efflux and NO derivatives molecules (NOx) like nitrite (NO2 – ), nitrate (NO3 – , peroxynitrite (– ONOO) and GSNO under the presence of PTK, PTP, AC and guanylyl cyclase (GC) enzyme proteins inhibitors. Blood samples from healthy donors were each one divided and were performed aliquots in absence (control aliquots) and presence of timolol or timolol plus each inhibitor and Gi protein uncoupling. No significant differences in erythrocyte NO efflux, GSNO, peroxynitrite, nitrite and nitrate concentrations in response to timolol when compared with the untreated blood samples aliquots were obtained. It was observed an increase in erythrocyte deformability at high shear stresses induced by the simultaneous presence of timolol and band 3 protein dephosphorylation by PTK syk inhibitor. No significant differences where verified in peroxynitrite levels in the blood aliquots in presence of timolol plus each enzyme inhibitor and Gi protein uncoupling in relation to the control aliquots. No variation of GSNO concentration occurs under the presence of timolol and AMGT (PTK lyn inhibitor) besides the significant higher values observed with each one of the other inhibitors. Nitrate concentration increases significantly in all aliquots with timolol plus each one of the inhibitors. The same was observe with nitrite levels with exception of the aliquots with timolol plus AMGT or timolol plus Gi protein uncoupling showing no significant values in relation to the control aliquots. Besides the changes in NO derivative molecules and NO efflux from RBCs obtained in this study with blood samples of healthy donors under the effect of timolol plus each inhibitor of the proteins participants in NO signal transduction mechanism, further analogue studies must be promoted with blood samples of patients with glaucoma or any other inflammatory vascular disease. Show more

Keywords: Erythrocyte deformability, nitric oxide, acetylcholinesterase, timolol

DOI: 10.3233/CH-189110

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 165-173, 2018

Authors: Kumar, Reddi K. | Heuchel, Matthias | Kratz, Karl | Lendlein, Andreas | Jankowski, Joachim | Tetali, Sarada D.

Article Type: Research Article

Abstract: Remaining uremic toxins in the blood of chronic renal failure patients represent one central challenge in hemodialysis therapies. Highly porous poly(ether imide) (PEI) microparticles have been recently introduced as candidate absorber materials, which show a high absorption capacity for uremic toxins and allow hydrophilic surface modification suitable for minimization of serum protein absorption. In this work, the effects of extracts prepared from PEI microparticles modified by nucleophilic reaction with low molecular weight polyethylene imine (Pei) or potassium hydroxide (KOH), on human monocytic (THP-1) cells are studied. The obtained results suggested that the extracts of Pei and KOH modified PEI absorbers …have no negative effect on THP-1 cell viability and do not initiate the critical differentiation towards macrophages. The extracts did not enhance transcript or protein levels of investigated proinflammatory markers in THP-1 cells, namely, TNFμ, MCP1, IL6 and IL8. Based on these findings such modified PEI microparticles should be qualified for further pre-clinical evaluation i.e. in an in vivo animal experiment. Show more

Keywords: Chronic kidney disease, hemodialysis, Inflammation, Porous poly(ether imide) microparticulate absorbers, THP-1 cells, Uremic toxins

DOI: 10.3233/CH-189112

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 175-185, 2018

Authors: Gramatté, Johannes | Pietzsch, Jens | Bergmann, Ralf | Richter, Torsten

Article Type: Research Article

Abstract: Aspiration of low-pH gastric fluid leads to an initial pneumonitis, which may become complicated by subsequent pneumonia or acute respiratory distress syndrome. Current treatment is at best supportive, but there is growing experimental evidence on the significant contribution of both neutrophils and platelets in the development of this inflammatory pulmonary reaction, a condition that can be attenuated by several medicinal products. This review aims to summarize novel findings in experimental models on pathomechanisms after an acid-aspiration event. Given the clinical relevance, specific emphasis is put on deduced potential experimental therapeutic approaches, which make use of the characteristic alteration of microcirculation …in the injured lung. Show more

Keywords: Acute respiratory distress syndrome, critical care medicine, pneumonitis, pulmonary inflammation, pulmonary blood flow, targeted anti-inflammatory therapies

DOI: 10.3233/CH-189113

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 187-195, 2018

Authors: Khan, Imran | Vishwakarma, Sandeep Kumar | Khan, Aleem Ahmed | Ramakrishnan, Ganesan | Dutta, Jayati Ray

Article Type: Research Article

Abstract: BACKGROUND: Gold nanoparticles (GNPs) are key diagnostic and therapeutic agents in biomedical sciences. Several studies have been carried out in different therapeutic areas such as in cancer treatment, antibacterial topical agents, imaging agents etc. There is a necessity to evaluate the gold nanoparticles cytotoxicity at all fronts. Since blood is the first point of contact in any therapy, it is required to have a thorough in vitro investigation of gold nanoparticles to avoid any adverse effects. OBJECTIVE: The objective of the current study is to evaluate the effect of gold nanoparticles capped with lipase on blood clotting …factors, platelets, coagulation time and blood clotting strength. METHODS: Whole blood samples were drawn from healthy volunteers. Plasma and plasma with platelets were isolated from the blood and all the samples were treated with lipase capped gold nanoparticles, except control. Plasma fibrinogen formed in the blood coagulation process after contacting with nanoparticles was quantitatively evaluated. In addition, platelet aggregation, blood clotting kinetics, strength of the blood clot and time were evaluated post nanoparticle treatment. RESULTS: The work primarily explores the effect of GNPs on blood with changing concentrations of lipase capping. Plasma fibrinogen levels of plasma samples were found to be moderately elevated, however, there is no significant effect on blood clotting kinetics, strength, and platelet aggregation. Also, the study showed that lipase capped GNPs did not result in aggregation upon interaction with plasma components and remained stable for 1 hour after incubation. CONCLUSIONS: Our study revealed that lipase capped GNPs synthesized using NaBH4 approach were stable and hemocompatible. There is an increase in fibrinogen levels after the exposure to nanoparticles, an observation which is consistent with other studies. However, the functional consequences of such increase are unknown. The results of no significant platelet aggregation, change in blood clotting time, kinetics, and clot strength revealed the non-toxic effect of lipase capped GNPs towards blood components, which is essential for any in vivo applications. Show more

Keywords: Hemocompatibility of Au nanoparticles, platelet aggregation, plasma fibrinogen test, blood clotting

DOI: 10.3233/CH-189117

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 197-205, 2018

Authors: Renoux, Céline | Joly, Philippe | Gauthier, Alexandra | Pialoux, Vincent | Romanet-Faes, Camille | Bertrand, Yves | Garnier, Nathalie | Cuzzubbo, Daniela | Cannas, Giovanna | Connes, Philippe

Article Type: Research Article

Abstract: The aim of the present study was to compare blood rheological parameters between children with homozygous sickle cell disease (SS), sickle cell SC disease or S/β+ -thalassemia syndrome, and healthy children (AA) and to test the associations between blood rheology and the clinical severity in S/β+ -thalassemia. Sixty-two SS, 14 SC, 11 S/β+ -thalassemia and 12 healthy children participated in this study. Blood viscosity was measured with a cone-plate viscometer at 225 s–1 . Red blood cell (RBC) deformability was measured by ektacytometry and RBC aggregation, by syllectometry. Nitric oxide and nitrotyrosine levels were determined for each child. While most …of the hematological parameters were not different between SC and S/β+ -thalassemia children, we demonstrated that SC patients had lower RBC deformability and aggregation than S/β+ individuals. Nitrotyrosine level, which indicates peroxynitrite production, was similar and lower in both healthy and S/β+ compared to SS children. However, S/β+ -thalassemia children who experienced vaso-occlusive crises (VOC) in the 2 previous years had lower NOx and higher nitrotyrosine levels than those who never had VOC within the same period. These findings suggest that vascular function could be impaired in the most severe S/β+ -thalassemia children compared to the less severe one. Show more

Keywords: Beta-thalassemia, sickle cell disease, red blood cell deformability, nitric oxide

DOI: 10.3233/CH-189119

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 207-214, 2018

Authors: Blocki, Anna | Beyer, Sebastian | Jung, Friedrich | Raghunath, Michael

Article Type: Research Article

Abstract: Pericytes reside within the basement membrane of small vessels and are often in direct cellular contact with endothelial cells, fulfilling important functions during blood vessel formation and homeostasis. Recently, these pericytes have been also identified as mesenchymal stem cells. Mesenchymal stem cells, and especially their specialized subpopulation of pericytes, represent promising candidates for therapeutic angiogenesis applications, and have already been widely applied in pre-clinical and clinical trials. However, cell-based therapies of ischemic diseases (especially of myocardial infarction) have not resulted in significant long-term improvement. Interestingly, pericytes from a hematopoietic origin were observed in embryonic skin and a pericyte sub-population expressing …leukocyte and monocyte markers was described during adult angiogenesis in vivo . Since mesenchymal stem cells do not express hematopoietic markers, the latter cell type might represent an alternative pericyte population relevant to angiogenesis. Therefore, we sourced blood-derived angiogenic cells (BDACs) from monocytes that closely resembled hematopoietic pericytes, which had only been observed in vivo thus far. BDACs displayed many pericytic features and exhibited enhanced revascularization and functional tissue regeneration in a pre-clinical model of critical limb ischemia. Comparison between BDACs and mesenchymal pericytes indicated that BDACs (while resembling hematopoietic pericytes) enhanced early stages of angiogenesis, such as endothelial cell sprouting. In contrast, mesenchymal pericytes were responsible for blood vessel maturation and homeostasis, while reducing endothelial sprouting. Since the formation of new blood vessels is crucial during therapeutic angiogenesis or during integration of implants into the host tissue, hematopoietic pericytes (and therefore BDACs) might offer an advantageous addition or even an alternative for cell-based therapies. Show more

DOI: 10.3233/CH-189132

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 215-232, 2018

Authors: Caimi, Gregorio | Carlisi, Melania | Montana, Maria | Gallà, Eleonora | Hopps, Eugenia | Lo Presti, Rosalia | Siragusa, Sergio

Article Type: Research Article

Abstract: The aim of this study was the evaluation of the erythrocyte deformability in multiple myeloma (MM). We enrolled 29 MM patients and we evaluated, on fasting venous blood, the erythrocyte deformability, expressed as elongation index, and examined using the diffractometric method. By comparing normal controls and MM patients, a significant decrease in erythrocyte deformability, especially at low shear stresses, was found. In this research paper we evaluated all the hypothesis for a possible explanation of the behaviour of red blood cell deformability in MM, even considering how the alteration of erythrocyte deformability worsens the microcirculatory flow in these patients in association …with the marked increase in plasma viscosity. Show more

Keywords: Multiple myeloma, erythrocyte deformability, microcirculation

DOI: 10.3233/CH-189121

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 233-238, 2018

Authors: Brun, Jean-Frédéric | Varlet-Marie, Emmanuelle | Richou, Marlène | Mercier, Jacques | Raynaud de Mauverger, Eric

Article Type: Research Article

Abstract: Rheological properties of plasma and blood cells are markedly influenced by the surrounding milieu: physicochemical factors, metabolism and hormones. Acid/base status, osmolality, lipid status, plasma protein pattern, oxidative stress induced by increased free radicals production, endothelium-derived factors such as nitric oxide (NO), achidonic acid derivatives modulate both red blood cell (RBC) and white cell mechanics. Therefore, regulatory axes involving liver, endothelium, kidney, pancreas, adrenal gland, endocrine heart, adipose tissue, pituitary gland, and surely other tissues play important roles in the regulation of blood fluidity. A comprehensive picture of all this complex network of regulatory loops is still unavailable but current …progress of knowledge suggest that some attempts can currently be made. Show more

Keywords: Erythrocyte, viscosity, deformability, aggregation, hormones, metabolism

DOI: 10.3233/CH-189124

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 239-265, 2018

Authors: Krüger-Genge, A. | Steinbrecht, S. | Küpper, J.-H. | Lendlein, A. | Jung, F.

Article Type: Research Article

Abstract: In cancer therapy, a number of drugs with different mechanisms of action are in clinical use, which act directly after administration without metabolism, while others only become active in the metabolites produced in the liver. Such drugs/metabolites – especially when administered parenterally – interact in high concentrations with the endothelium. Whether this induces adverse responses of the endothelial cells (EC) is barely studied for many medicaments. This pilot in vitro study revealed that the addition of cyclophosphamide (CPA) to the culture medium (5 or 10 mM, respectively) showed a clear influence on EC compared to non-treated EC: The number …of adherent human vein endothelial cells (HUVEC) decreased by the addition of CPA in a concentration-dependent manner compared to the untreated control, whereby the vitality of adherent cells was not affected. In addition, concomitant with activation of the adherent HUVEC, increased migratory activity occurred. These results are in agreement with clinical events like thromboses in patients in compromised condition under therapy with CPA, as the detachment of EC might induce responses of circulating platelets leading to the adherence and aggregation with the risk of the formation of thrombi. Whether CPA acts directly or via toxic metabolites on EC will be examined in more detail in following studies. Show more

Keywords: Endothelial cells, HUVEC, cyclophosphamide, toxicity

DOI: 10.3233/CH-189125

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 267-276, 2018

Authors: Lee, Jiyun | Lee, Chang Youn | Seo, Hyang-Hee | Bazarragchaa, Badamtsetseg | Batdelger, Gantuya | Choi, Sangho | Hwang, Ki-Chul | Lee, Seahyoung | Lim, Soyeon

Article Type: Research Article

Abstract: Excessive vascular smooth muscle cell (VSMC) proliferation and migration accelerate the development of occlusive vascular disease. Therefore, finding a means to control the aberrant proliferation and migration of VSMCs has own clinical significance. In the present study, we examined the feasibility of using extract from medicinal plant Oxytropis pseudoglandulosa (OG) to control pathologic proliferation and migration of VSMCs, which never have been tested. Our data indicate that the extract of OG significantly suppressed proliferation and migration of VSMCs without cytotoxic effect, suggesting the OG extract may be an alternative agent to effectively control the aberrant VSMC proliferation and migration …without any serious adverse effect. These data suggest that the extract of OG may be a potent therapeutic agent for the treatment of occlusive vascular disease and warrant further studies to identify the major acting ingredient and to validate in vivo efficacy. Show more

Keywords: Oxytropis pseudoglandulosa , vascular smooth muscle cell, proliferation, migration

DOI: 10.3233/CH-189126

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 277-287, 2018

Authors: Fornal, Maria | Piergies, Natalia | Pięta, Ewa | Wrobel, Tomasz P. | Kwiatek, Wojciech M. | Paluszkiewicz, Czesława | Królczyk, Jarosław | Grodzicki, Tomasz

Article Type: Research Article

Abstract: OBJECTIVES: To assess the relationships of clinical and laboratory parameters, with erythrocyte oxygen concentration. METHODS: The study group consisted of 47 healthy adults out of which 16 showed slightly higher blood pressure and were classified as individuals with prehypertension. For each individual, data were obtained on: systolic and diastolic blood pressure (SBP, DBP), blood morphology, lipids profile, fibrinogen, hs-CRP. The erythrocyte oxygen concentration was assessed with the Raman spectroscopy technique. Arithmetic means for all laboratory parameters were estimated by oxygen concentration tertiles and tested for statistical significance of linear trends across tertiles. RESULTS: The multivariate regression …analysis showed statistically significant negative relationship of triglycerides level with oxygen concentration. CONCLUSIONS: A hypothesis has been formulated that triglycerides level is an indicator of the erythrocyte ability to transport oxygen to the tissues. Show more

Keywords: Hemoglobin oxygen-binding properties, oxygen transport, Raman spectroscopy, lipid profile, tryglicerydes level

DOI: 10.3233/CH-189127

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 289-294, 2018

Authors: Richardson, Kieran J. | McNamee, Antony P. | Simmonds, Michael J.

Article Type: Research Article

Abstract: Haemochromatosis remains the most prevalent genetic disorder of Caucasian populations in Australia and the United States, occurring in ∼1 of 200 individuals and having a carrier frequency of 10–14%. Hereditary haemochromatosis is an autosomal recessive condition, that is phenotypically characterised by a gradual accumulation of iron, above and beyond that required for biological function. Once the binding capacity of iron carriers reaches saturation, the highly reactive free iron generates radicals that may lead to widespread cellular dysfunction. Thus, the compounding effects of systemic iron overload and the associated oxidative stress in untreated haemochromatosis patients results in tissue damage precipitating severe …complications, including: liver cirrhosis, hepatocellular cancer, cardiomyopathy, and diabetes. The primary treatment indicated for individuals with haemochromatosis is venesection therapy (i.e., regular bloodletting of ∼450 mL). Given the frequency of venesection required to decrease and normalise the elevated iron levels, this population may serve as a valuable source of blood products which are in short supply. While the complications associated with elevated iron deposits are frequently reported, the influence of haemochromatosis on the rheological properties of blood and red blood cells (RBC) – major determinants of microvascular blood flow and tissue perfusion – are poorly understood. Limited studies investigating haemorheology in patients with haemochromatosis have reported altered physical properties of blood, which may partly explain the comorbidities associated with the disorder. The current review will explore the aetiology, pathology, and clinical implications of haemochromatosis disease and the associated oxidative stress, with particular emphasis on RBC. Show more

Keywords: Hemorheology, iron overload, oxidative stress

DOI: 10.3233/CH-189128

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 295-304, 2018

Authors: Krüger-Genge, A. | Braune, S. | Walter, M. | Krengel, M. | Kratz, K. | Küpper, J.H. | Lendlein, A. | Jung, F.

Article Type: Research Article

Abstract: BACKGROUND: Physical and chemical characteristics of implant materials determine the fate of long-term cardiovascular devices. However, there is still a lack of fundamental understanding of the molecular mechanisms occurring in the material-tissue interphase. In a previous study, soft covalently crosslinked poly(n -butyl acrylate) networks (cPn BA) were introduced as sterilizable, non-toxic and immuno-compatible biomaterials with mechanical properties adjustable to blood vessels. Here we study the influence of different surface treatments in particular oxygen plasma modification and fibrinogen deposition as well as a combinatorial approach on the adhesion and viability of fibroblasts. MATERIAL AND METHODS: Two types of cPn …BA networks with Young’s moduli of 0.19±0.01 MPa (cPn BA04) and 1.02±0.01 MPa (cPn BA73) were synthesized and post-modified using oxygen plasma treatment (OPT) or fibrinogen coating (FIB) or a combination of both (OPT+FIB). The water contact angles of the differently post-treated cPn BAs were studied to monitor changes in the wettability of the polymer surfaces. Because of the key role of vascular fibroblasts in regeneration processes around implant materials, here we selected L929 fibroblasts as model cell type to explore morphology, viability, metabolic activity, cell membrane integrity as well as characteristics of the focal adhesions and cell cytoskeleton on the cPn BA surfaces. RESULTS: Compared to non-treated cPn BAs the advancing water-contact angles were found to be reduced after all surface modifications (p  < 0.05, each), while lowest values were observed after the combined surface treatment (OPT+FIB). The latter differed significantly from the single OPT and FIB. The number of adherent fibroblasts and their adherence behavior differed on both pristine cPn BA networks. The fibroblast density on cPn BA04 was 743±434 cells·mm-2 , was about 6.5 times higher than on cPn BA73 with 115±73 cells·mm-2 . On cPn BA04 about 20% of the cells were visible as very small, round and buckled cells while all other cells were in a migrating status. On cPn BA73, nearly 50% of fibroblasts were visible as very small, round and buckled cells. The surface functionalization either using oxygen plasma treatment or fibrinogen coating led to a significant increase of adherent fibroblasts, particularly the combination of both techniques, for both cPn BA networks. It is noteworthy to mention that the fibrinogen coating overruled the characteristics of the pristine surfaces; here, the fibroblast densities after seeding were identical for both cPn BA networks. Thus, the binding rather depended on the fibrinogen coating than on the substrate characteristics anymore. While the integrity of the fibroblasts membrane was comparable for both polymers, the MTS tests showed a decreased metabolic activity of the fibroblasts on cPn BA. CONCLUSION: The applied surface treatments of cPn BA successfully improved the adhesion of viable fibroblasts. Under resting conditions as well as after shearing the highest fibroblast densities were found on surfaces with combined post-treatment. Show more

Keywords: Biomaterial, poly(n-butyl acrylate), fibroblast, oxygen plasma, fibrinogen, cell adhesion, focal adhesion, actin cytoskeleton, viability

DOI: 10.3233/CH-189130

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 305-316, 2018

Authors: Hiebl, B. | Ascher, L. | Luetzow, K. | Kratz, K. | Gruber, C. | Mrowietz, C. | Nehring, M.E. | Lendlein, A. | Franke, R.-P. | Jung, F.

Article Type: Research Article

Abstract: Laser tissue soldering (LTS) based on indocyanine green (ICG)-mediated heat-denaturation of proteins might be a promising alternative technique for micro-suturing, but up to now the problem of too weak shear strength of the solder welds in comparison to sutures is not solved. Earlier reports gave promising results showing that solder supported by carrier materials can enhance the cohesive strength of the liquid solder. In these studies, the solder was applied to the carriers by dip coating. Higher reliability of the connection between the solder and the carrier material is expected when the solder is bound covalently to the carrier material. …In the present study a poly(ether imide) (PEI) membrane served as carrier material and ICG-supplemented albumin as solder substrate. The latter was covalently coupled to the carrier membrane under physiological conditions to prevent structural protein changes. As laser source a diode continuous-wave laser emitting at 808 nm with intensities between 250 mW and 1500 mW was utilized. The albumin functionalized carrier membrane was placed onto the tunica media of explanted pig thoracic aortae forming an overlapping area of approximately 0.5×0.5 cm2 . All tests were performed in a dry state to prevent laser light absorption by water. Infrared spectroscopy, spectro-photometrical determination of the secondary and primary amine groups after acid orange II staining, contact angle measurements, and atomic force microscopy proved the successful functionalization of the PEI membrane with albumin. A laser power of 450 mW LTS could generate a membrane-blood vessel connection which was characterized by a shear strength of 0.08±0.002 MPa, corresponding to 15% of the tensile strength of the native blood vessel. Theoretically, an overlapping zone of 4.1 mm around the entire circumference of the blood vessel could have provided shear strength of the PEI membrane-blood vessel compound identical to the tensile strength of the native blood vessel. These in-vitro results confirmed the beneficial effects of solder reinforcement by carrier membranes, and suggest LTS with covalently bound solders on PEI substrates for further studies in animal models. Show more

Keywords: Laser tissue soldering, blood vessel anastomosis, laser welding, solder carrier, poly(ether imide) membrane

DOI: 10.3233/CH-189108

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 317-326, 2018

Authors: Nowak, Elisabeth | Kammerer, Sarah | Küpper, Jan-Heiner

Article Type: Research Article

Abstract: BACKGROUND: Drug-induced liver injury (DILI) is the most common reason for withdrawal of anticancer drugs from the market. To prevent adverse side effects of drugs, it is important to investigate potential toxicity in vitro. However, outcome of cytotoxicity screenings can differ remarkably depending on the method used. OBJECTIVE: We aimed to compare XTT, ATP-based CellTiter-Glo® 2.0 and trypan blue exclusion (TBE) assays regarding their sensitivity in detecting acute cytotoxicity on HepG2 cells after incubation with the classical anticancer drugs Taxol and Imatinib or with the proteasome inhibitor MG-132. METHODS: HepG2 cells were treated for 48 h …and cell viability was analysed by XTT, CellTiter-Glo® 2.0 or TBE assay. RESULTS: All tested compounds showed a reduction of viability of HepG2 cells. However, assay results differed significantly: Both ATP-based and TBE assay showed concentration-dependent cytotoxic effects, but outcomes were less pronounced with TBE. In contrast, the widely used XTT assay did not detect any acute cytotoxicity of Taxol and Imatinib. CONCLUSIONS: Acute cytotoxic effects of tested compounds could be revealed. However, results were significantly different from each other with ATP assay being the most sensitive one under the conditions tested. Thus, acute cytotoxicity can be dramatically underestimated if only standard XTT test is used. Show more

Keywords: CellTiter-Glo®2.0, XTT, trypan blue exclusion, cytotoxicity, DILI, HepG2, Imatinib, MG-132, Taxol

DOI: 10.3233/CH-189120

Citation: Clinical Hemorheology and Microcirculation, vol. 69, no. 1-2, pp. 327-336, 2018