The Application And Outcome Of The Rat Brain Operation Hemostatic Materials

Objective: To make model of rat traumatic cerebral hemorrhage,use thehemostatic materials to intracranial hemorrhage (absorbable gelatinsponge,surgicel) to hemostais. Observe the change of hemostatic materialsunder light microscope and electron microscope, pathological changes ofbrain tissue, the apoptosis of the brain cells after use the hemostatic.Methods:1Making the animal model of traumatic cerebral hemorrhage in rats.Wistar rats (72, either sex, weighing300g-350g) were randomly dividedinto:①the control group24, experimental rats only given oppressionhemostasis without application of hemostatic material after operation.②e xperimental group48, experimental rats given hemostatic material aftraoperation.According to the application of absorbable gelatin sponge orsurgicel can be divided into two groups after operation,24rats in eatch group.After intraperitoneal anesthesia by10%chloral hydrate, the skin of therats’longitudinal roof craniotomy were cut. In the right parietal bone (fromthe sagittal suture, coronal suture of5mm) a bone window of5mm in diameterwas removed by the dental burr, maintaining dural integrity.Use surgical3-0belt line suture needle to cut dura mater to expose brain tissue withoutinvolving the cortex, clip of the cerebral cortex:2mm×2mm×1mm by themicroscopic tumor foreps to make the model of traumatic hemorrhage.Effective homostasis with cotton piece oppression, absorbable gelatin spongeand surgicel according to the groups.Then suture the wounds after completehemostasis.Make the rats eat food and drink water freely after injury.4rats ofeach group were sacrificed on the5days10days,15days,20days,25days,30days.2Observe the change of hemostatic materials groups and the controlgroup in rat brain tissue H-E staining of tissue section under light microscope. Rats of each group were sacrificed on the5days,10days,15days,20days,25days,30days. After Left ventricle-ascending aorta cannulation, therats were injected with normal saline150ml irrigation.,4%paraformaldehyde300ml for2hours Slow down after, then fixed in4%paraformaldehyde24hours or more,take out the brain.Cut the tissue which covered with hemostaticmaterials, then conventional dehydration, transparent, paraffin embedding,serial sections of5μm, hematoxylin and eosin(H-E)staining. Imageacquisition.3Observe the change of hemostatic materials groups and the controlgroup in rat brain tissue under transmission electron microscope and scanningelectron microscope.Rats of each group were sacrificed on the5days,10days,15days,20days,25days,30days. After Left ventricle-ascending aorta cannulation, therats were injected with normal saline150ml irrigation.,4%paraformaldehyde300ml for2hours Slow down after, take out the brain.Cut the tissue whichcovered with hemostatic materials,4%glutaraldehyde fixed24h.Scanningelectron microscopy sample preparation for:3mm×3mm×1mm, TEMspecimen preparation for1mm×1mm×1mm, image acquisition.4Observe the apoptosis of the brain cells after use the absorbable gelatinsponge,surgicel and the blank group.Randomly selected group of sub-three specimens After package waxsliced,dehydrated, hydrated, cell permeability,TUNEL reaction solution, therole of converter-POD, substrate DBA reaction color, take photo and countapoptotic cells at high magnification.Results:1Pathological changes and hemostatic material absorption and outcome oftraumatic brain tissue under light microscope.The brain tissue slice H-E staining In5d,10d,15d showed:Traumaticbrain tissue Inflammatory reaction is obvious visible, around the necrotic braintissue, necrosis, edema zone can be found.Infiltration of mononuclear cells,red blood cell diffuse in the broken brain tissue and hemostatic material gap.Over time, inflammation is reduced gradually, the number of mononuclearcells and red blood cell reduce.Red cell rupture, The control group show nervecell edema, a large number of mononuclear cell infiltration, minority redblood cell deformability and broken. the hemoglobin released then oxidizedinto yellow。Pasting property with the brain tissue the absorbable gelatinsponge is lower than Surgicel, the inflammatory reaction is heavy, andoccupying more space.Compatibility with the brain tissue Surgicel is betterthan that of absorbable gelatin sponge.2Pathological changes and hemostatic material absorption and outcome oftraumatic brain tissue under electron microscopy.2.1Pathological changes and hemostatic material absorption and outcome oftraumatic brain tissue under Scanning electron microscope.Brain edema and softening is obviously found in hemorrhage brain.Boundaries in edema and softening brain with normal brain tissue is obvious.The blank group Surgicel group in the25day specimens visible edemacompared with the previous specimens alleviate absorbable gelatin spongespecimens at25days, there are still significant edema. Few hemorrhagic fociremain visible the not deformed red blood cells. Significant intracranial masseffect absorbable gelatin sponge is still visible at the same time point. TheSurgicel paste the cover of the brain tissue is superior than absorbable gelatinsponge. With the progress of time brain edema gradually reduce.2.2Pathological changes and hemostatic material absorption and outcome oftraumatic brain tissue under transmission electron microscope.Phagocytic cell devour massive degeneration of necrosis nerve fibers5days after operation in brain tissue.Neurons in the different groups havedifferent performance15days,25days after operation. Neuronal cellshrinkage, mitochondria swelling, Golgi expand, nerve fiber edema in thecontrol group. Neurons swelling lighter, partial mitochondrial vacuolization,mitochondria swelling, Golgi expand in absorbable gelatin sponge group.Neurons cytoplasm edema, neuronal edema, mitochondria swelling, Golgiexpand in Surgicel group. 3brain cell apoptosis in eath experimental group after operation.After the TUNEL apoptosis Kit samples, collected images. Determina-tion of Neuron apoptosis-positive cell number ratings and positive cell colorintensity ratings, calculated cell apoptosis of immunohistochemical scoring ofstatistical analysis, experimental data show that:①Hemostatic differently,IHS values differ significantly in different groups (P<0.05), statisticallysignificant, and there are differences between the two groups (P<0.05).②When the experimental influence factors of time, IHS values for each point intime there are differences (P<0.05), when comparing two groups, with theexception of20d and25d specimens of the IHS was no difference, the others’IHS values for each point in time there are differences (P<0.05).③Whenconsider the time and the way to hemostatic, Surgicel in10d apoptosis afterbleeding the highest index, apoptosis index blank group25days minimum.④and interaction with the way to hemostasis and time.(P<0.05).Conclusion:1As hemostatic material in neurosurgery，the histocompatibility ofSurgicel is superior than that of absorbable gelatin sponge.2Application of hemostatic material have different levels of apoptosis.3Guarantee the hemostatic effect, we should minimize the application ofhemostatic material.