Real-time laser speckle contrast imaging for intraoperative neurovascular blood flow assessment: animal experimental study

The use of various blood flow control methods in neurovascular interventions is crucial for reducing postoperative complications. Neurosurgeons worldwide use different methods, such as contact Dopplerography, intraoperative indocyanine videoangiography (ICG) video angiography, fluorescein angiography, flowmetry, intraoperative angiography, and direct angiography. However, there is no noninvasive method that can assess the presence of blood flow in the vessels of the brain without the introduction of fluorescent substances throughout the intervention. The real-time laser-speckle contrast imaging (LSCI) method was studied for its effectiveness in controlling blood flow in standard cerebrovascular surgery cases in rat common carotid arteries, such as proximal occlusion, trapping, reperfusion, anastomosis, and intraoperative vessel thrombosis. The real-time LSCI method is a promising method for use in neurosurgical practice. This approach allows timely diagnosis of intraoperative disturbance of blood flow in vessels in cases of clip occlusion or thrombosis. Additionally, LSCI allows us to reliably confirm the functioning of the anastomosis and reperfusion after removal of the clips and thrombolysis in real time. An unresolved limitation of the method is noise from movements, but this does not reduce the value of the method. Additional research is required to improve the quality of the data obtained

Novel Hydroxypyridine Compound Protects Brain Cells against Ischemic Damage In Vitro and In Vivo

A non-surgical pharmacological approach to control cellular vitality and functionality during ischemic and/or reperfusion-induced phases of strokes remains extremely important. The synthesis of 2-ethyl-6-methyl-3-hydroxypyridinium gammalactone-2,3-dehydro-L-gulonate (3-EA) was performed using a topochemical reaction. The cell-protective effects of 3-EA were studied on a model of glutamate excitotoxicity (GluTox) and glucose-oxygen deprivation (OGD) in a culture of NMRI mice cortical cells. Ca2+ dynamics was studied using fluorescent bioimaging and a Fura-2 probe, cell viability was assessed using cytochemical staining with propidium iodide, and gene expression was assessed by a real-time polymerase chain reaction. The compound anti-ischemic efficacy in vivo was evaluated on a model of irreversible middle cerebral artery (MCA) occlusion in Sprague-Dawley male rats. Brain morphological changes and antioxidant capacity were assessed one week after the pathology onset. The severity of neurological disorder was evaluated dynamically. 3-EA suppressed cortical cell death in a dose-dependent manner under the excitotoxic effect of glutamate and ischemia/reoxygenation. Pre-incubation of cerebral cortex cells with 10–100 µM 3-EA led to significant stagnation in Ca2+ concentration in a cytosol ([Ca2+]i) of neurons and astrocytes suffering GluTox and OGD. Decreasing intracellular Ca2+ and establishing a lower [Ca2+]i baseline inhibited necrotic cell death in an acute experiment. The mechanism of 3-EA cytoprotective action involved changes in the baseline and ischemia/reoxygenation-induced expression of genes encoding anti-apoptotic proteins and proteins of the oxidative status; this led to inhibition of the late irreversible stages of apoptosis. Incubation of brain cortex cells with 3-EA induced an overexpression of the anti-apoptotic genes BCL-2, STAT3, and SOCS3, whereas the expression of genes regulating necrosis and inflammation (TRAIL, MLKL, Cas-1, Cas-3, IL-1β and TNFa) were suppressed. 3-EA 18.0 mg/kg intravenous daily administration for 7 days following MCA occlusion preserved rats’ cortex neuron population, decreased the severity of neurological deficit, and spared antioxidant capacity of damaged tissues. 3-EA demonstrated proven short-term anti-ischemic activity in vivo and in vitro, which can be associated with antioxidant activity and the ability to target necrotic and apoptotic death. The compound may be considered a potential neuroprotective molecule for further pre-clinical investigation