Shed-blood-separation and cell-saver: an integral Part of MiECC? Shed-blood-separation and its influence on the perioperative inflammatory response during coronary revascularization with minimal invasive extracorporeal circulation systems - a randomized controlled trial.

OBJECTIVE: The postoperative systemic inflammatory response after cardiopulmonary bypass (CPB) is still an undesirable side-effect after cardiac surgery. It is most likely caused by blood contact with foreign surfaces and by the surgical trauma itself. However, the recirculation of activated shed mediastinal blood is another main cause of blood cell activation and cytokine release. Minimal invasive extracorporeal circulation (MiECC) comprises a completely closed circuit, coated surfaces and the separation of suction blood. We hypothesized that MiECC, with separated cell saved blood, would induce less of a systemic inflammatory response than MiECC with no cell-saver. The aim of this study was, therefore, to investigate the impact of cell washing shed blood from the operating field versus direct return to the ECC on the biomarkers for systemic inflammation. MATERIAL AND METHODS: In the study, patients with MiECC and cell-saver were compared with the control group, patients with MiECC and direct re-transfusion of the drawn blood shed from the surgical field. RESULTS: High amounts of TNF-α (+ 120% compared to serum blood) were found in the shed blood itself, but a significant reduction was demonstrated with the use of a cell-saver (TNF-α ng/l post-ECC 10 min: 9.5±3.5 vs. 19.7±14.5, p<0.0001). The values for procalcitonin were not significantly increased in the control group (6h: 1.07±3.4 vs. 2.15±9.55, p=0.19) and lower for C-reactive protein (CRP) (24h: 147.1±64.0 vs.134.4±52.4 p=0.28). CONCLUSION: The use of a cell-saver and the processing of shed blood as an integral part of MiECC significantly reduces the systemic cytokine load. We, therefore, recommend the integration of cell-saving devices in MiECC to reduce the perioperative inflammatory response.

Ischemic preconditioning results in an ATP-dependent inhibition of cytochrome C oxidase.

PURPOSE: This study addresses the effect of short myocardial ischemia on inhibitory effect of ATP for mitochondrial cytochrome c oxidase (CytOx) activity in myocardium and subsequent hemodynamic alterations. The activity of CytOx is inhibited by ATP (primary substrate control). This additional mechanism was proposed to be switched off at higher mitochondrial membrane potential values in case of stress. The ATP-dependent allosteric enzyme inhibition (second respiratory control) is suggested to reduce the formation of reactive oxygen species and thus is pivotal for cytoprotection. This report addresses the possible involvement of this mechanism in case of myocardial preconditioning. METHODS: Rat hearts were perfused in a Langendorff system (n = 5 each group). The first two groups underwent short recurrent ischemic periods (three times 5 min) and subsequent high or low reperfusion for 40 min. Besides four control groups, hearts were exposed to an ischemia of 15 min and high flow reperfused for 30 min, in addition. Hemodynamic data were evaluated in parallel. Mitochondria were separated for the polarographic respiration measurements in the presence of ADP or ATP, respectively. Phosphorylation patterns of the CytOx subunits were studied by immunoblotting with P-Ser, P-Thr, and P-Tyr antibodies. RESULTS: Short recurrent episodes of ischemia result in an ATP-dependent inhibition of CytOx. Electrophoretic analysis and blotting techniques reveal different phosphorylation patterns of the enzyme. Frequent short-lasting ischemic impacts and subsequent increased coronary flow seem to be essential for this effect. CONCLUSION: The procedure of preconditioning is likely to be dependent on the mechanism of ATP-dependent inhibition of CytOx activity.

Effects of cyclosporine pretreatment on tissue oxygen levels and cytochrome oxidase in skeletal muscle ischemia and reperfusion.

We hypothesized that pretreatment with single-dose cyclosporine (CsA) prevents alterations and improves tissue oxygen and mitochondrial cytochrome oxidase redox (CytOx) state in skeletal muscle ischemia and reperfusion-reoxygenation (I/R). Latissimus dorsi muscle was prepared and mobilized in New Zealand white rabbits. Ischemia was induced for 4 h, followed by 2 h of reperfusion. The animals were randomized to receive a 60-mg/kg intravenous bolus of CsA (CsA group, n = 10) or physiologic saline (control, n = 10) at 10 min before ischemia onset. Muscle tissue oxygen tension (PtO(2)) and mitochondrial CytOx were measured during I/R simultaneously. High-energy phosphate (HEP) levels were determined using high-field (31)P magnetic resonance spectroscopy. Mitochondrial viability index and wet-to-dry ratio were used to assess the tissue viability between groups. Decreases in tissue oxygen levels and CytOx were slower during ischemia in the CsA group in comparison to control group, also the loss of phosphocreatine and adenosine triphosphate depletion. After ischemia, recovery of tissue oxygen, mitochondrial CytOx, and HEP was delayed in controls. Tissue PtO2 in the CsA group (P < 0.05) was significantly higher compared with that in the control group after I/R. Mitochondrial CytOx was also improved in the CsA group (P < 0.01 vs. control). Muscle HEP levels (phosphocreatine, adenosine triphosphate) were significantly preserved in the CsA group versus the control group (P < 0.01, P < 0.05). Mitochondrial viability index and wet-to-dry ratio confirmed significantly preserved tissue and lower edema formation in the CsA group. The pretreatment with single-dose CsA prevents alterations and improves tissue oxygenation and mitochondrial oxidation in skeletal muscle I/R.

Microvascular tissue oxygenation and oxidative metabolism changes in the pedicled latissimus dorsi muscle during graded hypoxia: correlation between near infrared and 31P nuclear magnetic resonance spectroscopy.

BACKGROUND: In this study, the microvascular tissue oxygenation and oxidative muscle metabolism during graded hypoxia and reoxygenation were examined in a rabbit model by near-infrared (NIR) spectroscopy and correlated with high-energy phosphates measured by (31)P nuclear magnetic resonance (NMR) spectroscopy. MATERIALS AND METHODS: Graded hypoxia was performed in a New Zealand rabbit model (n = 20, 2.0 ± 0.4 kg) by a stepwise reduction of the fraction of inspired oxygen (FiO(2)) from 0.3 to 0.05 (intervention group versus control group). Recovery and reoxygenation were achieved using FiO(2) of 0.3. A noninvasive NIR spectroscopy sensor and NMR probe was positioned on the surface of the prepared pedicled latissimus dorsi muscle. Microvascular tissue oxygenation (oxyhemoglobin, HbO(2); deoxyhemoglobin, HHb) and redox state of cytochrome oxidase (CytOx) were measured by NIR spectroscopy and correlated with standard values of oxidative muscle metabolism (phosphocreatine, PCr; adenosine triphosphate, ATP) measured by time-resolved (31)P NMR spectroscopy (4.7T). RESULTS: Significant correlation was found between PCr and HbO(2) (r = 0.85, P < 0.001) and HHb (r = -0.75, P < 0.001). ß-ATP levels correlated significantly with CytOx (r = 0.87, P < 0.001). CONCLUSIONS: The data suggest that changes in high-energy phosphates (PCr- and ATP-levels) correlate closely with microvascular tissue oxygenation (HbO(2), HHb, CytOx) measured by NIR spectroscopy.

Importance of real-time tissue oximetry: relationship to muscle oxygenation and tissue viability.

BACKGROUND: There is currently no efficient and reliable clinical means of assessing the degree of ischemia- and reperfusion-associated damage in microvascular transplants. The objective was to study correlation of tissue oxygen tension measurements with tissue oxygen saturation, cytochrome oxidase redox state, and tissue viability. MATERIALS AND METHODS: Latissimus dorsi muscle was dissected and mobilized in New Zealand white rabbits (n = 30, 2.5 ± 0.5 kg). Muscles were exposed to warm ischemia in two groups with either 4 or 6 h, followed by reperfusion. Tissue PO(2) was measured with a miniature intramuscular oxygen sensor (Licox microprobe; Integra Neurosciences, Ratingen, Germany) all along with tissue hemoglobin saturation (rSO(2)) and cytochrome oxidase aa3 redox state (CytOx) by in vivo near-infrared spectroscopy. Linear correlation was performed between tissue PO(2) and rSO(2), CytOx and tissue viability. RESULTS: After ischemia and reperfusion, tissue PO(2) and CytOx recovery was significantly decreased in both groups compared with control (4 h: P < 0.05; 6 h: P < 0.01). Significant correlations between changes in tissue PO(2) and rSO(2) (r = 0.92; P < 0.01), CytOx (r = 0.90; P < 0.01), wet-to-dry ratio (r = -0.97; P < 0.01), and mitochondrial viability index (r = 0.97; P < 0.01) were found. CONCLUSIONS: Tissue oxygen tension measured with microprobes correlated closely with tissue oxygenation, cellular oxygen utilization, and the extent of ischemia reperfusion injury.

Epicardial measurement of alterations in extracellular pH and electrolytes during ischemia and reperfusion in cardiac surgery.

Simultaneous measurements of extracellular pH, potassium (K(+)), and calcium (Ca(2+)) activity might be indicative of myocardium vitality or ischemia. Ten consecutive patients undergoing elective coronary artery bypass grafting were studied. Epicardial extracellular pH, potassium, and calcium were measured by a miniaturized disposable multi-sensor probe. Blood gases and electrolytes were derived with measurements of arterial and mixed venous blood samples at intervals during surgery. The mean epicardial baseline levels for pH in all patients were 8.04+/-0.22 arbitrary units (AU) for the right ventricle (RV) and 8.03+/-0.21 AU for the left ventricle (LV); for Ca(2+) 0.23+/-0.07 mmol/l (RV) and 0.20+/-0.10 mmol/l (LV); and for K(+) 4.54+/-1.51 mmol/l (RV) and 4.38+/-0.57 mmol/l (LV). Before ischemia, epicardial pH was moderately (p<0.05), and K(+), and Ca(2+) were closely correlated (p<0.001) with blood values. During reperfusion, epicardial measurements were weakly correlated (p<0.001) with blood values for pH, venous K(+) and Ca(2+), but moderately correlated with arterial K(+) and Ca(2+) (p<0.01). The measurements indicated intraoperative episodes of ischemia and reperfusion with reproducible trends of extracellular pH, K(+), and Ca(2+), which results in electrolyte patterns applicable for detecting inadequate myocardial protection during cardiac surgery in patients.

Direct measurement of myocardial oxygen tension and high energy phosphate content under varying ventilatory conditions in rabbits.

Effective myocardial oxygen supply should not be compromised during cardiac surgery as it is essential to avoid circulatory and cardiac dysfunction. Local measurement of myocardial oxygen partial pressure (pO2) was therefore introduced into the operative monitoring of myocardial ischemia. The aim of the present study was to assess whether myocardial oxygen partial pressure correlates with the content of high energy phosphates (HEPs). Seven male rabbits were examined in parallel with measurement of myocardial pO2 by an implanted Clark electrode and 31phosphorus-NMR spectroscopy. The ventilatory management established hyperoxygenation followed by systemic hypoxia with hypercapnia for 20 min. Additionally, analysis of end-expiratory gas composition in combination with blood gas analysis was performed simultaneously, and hemodynamic parameter was recorded. Under hypoxic conditions the cardiovascular system was severely compromised, whereas the myocardial pO2 was only moderately impaired (pO2M 45.0+/-16.0 mm Hg). Immediately before cardiac arrest, low values of arterial and venous pO2 were found (17.6+/-6.0 and 12.9+/-6.1 mm Hg). In contrast to near normal myocardial pO2, HEP content in the myocardium was considerably reduced and inorganic phosphorus was increased. Artificial ventilation leading to systemic hypoxia and eventually circulatory arrest resulted in almost normal myocardial pO2 but severely compromised HEP content. This somewhat unexpected finding requires further clarification, but is in accordance with findings reported previously where regulatory mechanisms have been shown to play a role in the pathophysiology of severe hypoxic conditions such as those for cellular oxygen delivery and demand, P/O coupling and finally control of HEP production facilitating the interaction between respiratory chain and myoglobin oxygen transport.

Heat stress attenuates ATP-depletion and pH-decrease during cardioplegic arrest.

BACKGROUND: The capacity of heat stress induction to improve myocardial tolerance against ischemia is well known. We investigated cardiac energy metabolism after hsp 72(+)/73(+) induction in isolated perfused neonatal rabbit hearts subjected to prolonged cold cardioplegic ischemia. METHODS: Hearts from neonatal rabbits were excised, isolated perfused and arrested by 2-h cold cardioplegic ischemia. Rectal temperature of eight neonatal rabbits was raised to 42.0 to 42.5 degrees C for heat shock protein expression in a whole body water bath for 15 min before the onset of arrest. Another set of eight rabbits without hyperthermia pretreatment served as control. Recovery of left ventricle function was assessed by aortic flow, cardiac output, and max dP/dt. Status of high-energy phosphates was measured by (31)phosphorus nuclear magnetic resonance-spectroscopy. RESULTS: Immunoblot analysis revealed clear hsp 72+/73+ induction after a brief period of systemic hyperthermia. Heat stress pretreatment resulted in a better recovery of left ventricular function (aortic flow and cardiac output improvement P < 0.05, max dP/dt P < 0.01) than in controls at 60 min after reperfusion. During ischemia and reperfusion, myocardial energy metabolism was better preserved in hearts after hsp induction as a consequence of increased gamma-, alpha-, and beta-ATP as well as phosphocreatine-values over controls. The ischemia-induced pH-decrease was attenuated. CONCLUSION: These data contribute to the evidence of heat stress mediated beneficial effects on functional myocardial recovery and improved cardiac energy metabolism after prolonged cold cardioplegic ischemia. More importantly, the attenuation of ischemic pH reduction and better restoration suggest an involvement of mitochondrial membrane potential alterations.

Large-dose pretreatment with methylprednisolone fails to attenuate neuronal injury after deep hypothermic circulatory arrest in a neonatal piglet model.

Conflicting results have been reported with regard to the neuroprotective effects of steroid treatment with cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA). We evaluated the mode and severity of neuronal cell injury in neonatal piglets after prolonged DHCA and the possible neuroprotective effect of systemic pretreatment (>6 h before surgery) with large-dose methylprednisolone (MP). Nineteen neonatal piglets (age, <10 days; weight, 2.1 +/- 0.5 kg) were randomly assigned to 2 groups: 7 animals were pretreated with large-dose systemic MP (30 mg/kg) 24 h before surgery, and 12 animals without pharmacological pretreatment (saline) served as control groups. All animals were connected to full-flow CPB with cooling to 15 degrees C and 120 min of DHCA. After rewarming to 38.5 degrees C with CPB, animals were weaned from CPB and survived 6 h before they were killed, and the brain was prepared for light and electron microscopy, immunohistochemistry, and TUNEL-staining. Quantitative histological studies were performed in hippocampus, cortex, cerebellum, and caudate nucleus. Systemic pretreatment with large-dose MP lead to persistent hyperglycemia but no significant changes of cerebral perfusion. Necrotic and apoptotic neuronal cell death were detected in all analyzed brain regions after 120 min of DHCA. In comparison to the control group, large-dose pretreatment with systemic MP lead to an increase of necrotic neuronal cell death and induced significant neuronal apoptosis in the dentate gyrus of the hippocampus (P = 0.001). In conclusion, systemic pretreatment with large-dose MP fails to attenuate neuronal cell injury after prolonged DHCA and induces regional neuronal apoptosis in the dentate gyrus.

Muscle tissue oxygen tension and oxidative metabolism during ischemia and reperfusion.

BACKGROUND: Recent studies have shown a relationship between alterations in tissue oxygen metabolism and cellular changes following ischemia and reperfusion, such as energy store depletion and intracellular acidosis. The aim of this study was to evaluate the relationship between tissue energy metabolism and intramuscular tissue oxygen tension in the mobilized latissimus dorsi muscle. MATERIAL AND METHODS: The latissimus dorsi muscle was raised in New Zealand white rabbits (n = 10, 2.5 +/- 0.5 kg). During 4 h of ischemia and 2 h of reperfusion, the intramuscular tissue oxygen tension (Licox PO2-microcatheter probe) and the status of phosphorylated muscle energy metabolites were measured using a high-field 31P-NMR spectrometer. Linear correlation was performed between 31P-NMR data and tissue oxygen tension readings. RESULTS: The tissue oxygen tension (PO2) values correlated significantly with phosphocreatine (PCr) (r = 0.96, P < 0.001), beta-adenosin triphosphate (beta-ATP) (r = 0.64, P <0.01), and intracellular pH (r = 0.82, P <0.001). CONCLUSIONS: On the basis of these findings, we conclude that the data provided by tissue oxygen tension measurement offer a real time minimally invasive estimate of muscle oxidative metabolism during ischemia and reperfusion.

Navigation-assisted sclerotherapy of orbital venolymphatic malformation: a new guidance technique for percutaneous treatment of low-flow vascular malformations.

Percutaneous sclerotherapy of orbital low-flow vascular malformations requires precise procedural guidance. For the treatment of a patient with an orbital venolymphatic malformation, we sought to optimize guidance by combining navigation assistance for needle placement with intralesional contrast medium injection for assessment of venous drainage. By using a surgical navigation system (Vector Vision, BrainLAB, Munich, Germany), multiplanar target lesion visualization was performed after fusion of CT and MR imaging data, which allowed precise puncture planning.

Ossifying fibroma of the skull: interactive image-guided minimally invasive localization and resection.

Ossifying fibroma is a benign fibro-osseous tumor commonly affecting the craniofacial bones. It is considered to be a locally aggressive and quickly expansible bone lesion. Because of its aggressive nature and high recurrence rate, early detection and complete surgical removal are essential. Usually, these lesions are excised extensively by craniectomy, and bone loss is reconstructed by cranioplasty using acrylic resin or titanium implants. Alternatively, in the management of skull-ossifying fibroma, an image-guided technique using surgical navigation may provide precise information about localization, enabling complete removal, thereby operating with minimal exposure and within narrow resection borders and avoiding significant bone deformity. A 39-year-old male patient with a history of renal cell carcinoma was admitted to our hospital because a radionuclide scintigraphic bone scan revealed increased uptake in a small area located at the left lateral skull bone. The high-resolution computed tomography scan showed that the lesion was located inside the diploe, destroying the inner table of the calvarium. The patient underwent minimally invasive bone lesion removal using an interactive image-guided approach. Complete resection of the neoplastic lesion was achieved. The histopathological examination revealed an ossifying fibroma. The postoperative course was uneventful, and the patient was discharged 3 days after intervention. To date, there has been no evidence of local recurrence. Interactive multimodal planning and intraoperative image guidance offer an interesting approach for biopsy and minimally invasive removal of small ossifying fibroma lesions of the skull, especially in less accessible locations.

Detection of anti-hsp70 immunoglobulin G antibodies indicates better outcome in coronary artery bypass grafting patients suffering from severe preoperative angina.

BACKGROUND: Recent findings indicate that molecular chaperones actively participate in myocardial cytoprotection. Moreover, ischemic tolerance can be induced in humans by brief ischemic events. Therefore, we investigated patients with severe angina attacks before coronary artery bypass grafting. We focused on appearance of anti-hsp70 antibodies as an immunologic response to heat shock protein induction by ischemia followed up by hemodynamic measurements perioperatively. We correlated these clinical findings with the presence of antibodies against hsp70 and the antioxidative capacity of patients' sera. METHODS: Thirty-five consecutive patients with coronary artery disease scheduled for coronary artery bypass grafting were included. Seventeen patients had severe angina, and 18 patients suffered from chronic stable angina preoperatively. In the patients' sera, antibodies against hsp70 were detected by enzyme-linked immunosorbent assay, and antioxidative capacity was detected using the chromogen assay. Cardiac output and pulmonary capillary wedge pressure were measured using a thermodilution catheter. We also evaluated C-reactive protein and creatine kinase MB isoenzyme, and performed a conventional leukocyte count. RESULTS: The sera of the 17 patients with severe angina attacks before surgery contained antibodies against hsp70 and a low antioxidative capacity. The interval between a severe angina attack and anti-hsp70 antibody titer are inversely correlated. These patients had better cardiac output and lower pulmonary capillary wedge pressure values after surgery. CONCLUSIONS: Severe angina before cardiac surgery coincided with an improved outcome as measured by hemodynamic variables as compared with chronic stable angina. This finding correlated significantly with a low antioxidative capacity and the presence of antibodies against hsp70. These pathophysiologic mechanisms might therefore play a role in myocardial protection.

Temporary intermaxillary fixation using individualized acrylic splints permits image-data-based surgery of the lower jaw and oropharynx.

OBJECTIVES/HYPOTHESIS: Image-data-based surgical navigation is used as a helpful device in the operating room to localize critical structures with a high degree of accuracy. It also enables physicians to plan therapeutic performance. Because it relies on preoperatively acquired computed tomography (CT) or magnetic resonance imaging (MRI) data, there is restricted access for navigation of surgical instruments in areas that show motion uncorrelated with radiologic data. Thus, in the case of moveable structures, for example the lower jaw, navigational procedures could not yet be applied. STUDY DESIGN: We introduce a new technique using individualized intermaxillary splints that fix the mandible in a reproducible aboccluded position at the time of image-data acquisition and surgery. METHODS: Different manufacturing processes were investigated. The feasibility of uni- and bilateral intermaxillary splints was studied under clinical conditions in four patients during different procedures in the mandibular and oropharyngeal regions. RESULTS: The manufacturing of the splints showed was easily performed in a short time. With bilateral fixation, there was a high anatomic target precision of 1.6 to 2.3 mm. CONCLUSIONS: The use of bilateral intermaxillary splints that fix the patient's mandible in a reproducible aboccluded position permits an image-data-based navigated surgical approach to the oropharyngeal and mandibular regions.

Release patterns of astrocytic and neuronal biochemical markers in serum during and after experimental settings of cardiac surgery.

OBJECTIVE: Brain injury and altered psychomotor development in infants, children and adults after cardiac surgery using cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) is still a matter of concern. Early diagnosis and identification of brain injury that has occurred or is ongoing by measurement of biochemical markers in serum may have diagnostic and prognostic value. The aim of the experimental studies in an animal model was therefore to investigate the release patterns of astroglial and neuronal markers in serum and to determine the morphological and immunohistochemical changes in the brain of animals undergoing similar perfusion conditions of CPB and a period of DHCA. METHODS: Fourteen New Zealand rabbits, (weight, 3.1 +/- 0.25 kg) were anesthetized, intubated and mechanically ventilated. Four animals were sham operated and served as controls. After median sternotomy the animals were connected to CPB by cannulation of the aorta and right atrium. Full flow CPB (200-250 ml/kg/min) was initiated to achieve homogeneous systemic cooling. Circulatory arrest of 60 minutes was induced when rectal and nasopharyngeal temperature of 14 degrees C was achieved. After rewarmed reperfusion and establishment of stable cardiac ejection the animals were weaned from CPB and monitored for 6 hours. Then the animals were killed, the brain was immediately removed and cut in standardized sections. These were fixated, embedded in paraffin and stained for further quantitative histological studies. In the brain astrocyte reactivity for S-100B was assessed immunocytochemically (DPC Immustain Los Angeles, USA). Monoclonal mouse anti-human neurospecific enolase (NSE) antibody was used for the localization of NSE in the fixed and paraffin embedded brain (NSE-DAKO, H14). The concentrations of S-100B protein and neurospecific enolase (NSE) in the serum were analyzed using a commercially available immunoluminometric assay (LIA-mat, Sangtec 100, Byk-Sangtec). Immunospecific monoclonal anti-parvalbumin antibody was used for the detection of parvalbumin in the brain. Serum concentrations of parvalbumin were analyzed using a newly developed ELISA method. RESULTS: In all experimental animals a significant increase of the serum concentration of the astroglial protein S-100B was found immediately after reperfusion and the termination of CPB. In contrast the serum levels of the neuronal proteins parvalbumin and NSE were not increased, but rather decreased. Light microscopy and electron microscopy revealed perivascular astrocytic swelling and minor neuronal cell injury. In comparison to the sham operated animals, increased immunohistochemical staining of S-100B was found. This increased reactivity of S100B antibody was found in the astrocytic processes with immediate connection to the perivascular space and around the perivascular oedema. The immunocytochemical stainings for NSE and parvalbumin in the neuronal cells was not different from that of sham-operated animals and indicated well preserved neurons.