A quadruped study on chitosan microspheres containing atorvastatin calcium: preparation, characterization, quantification and in-vivo application.

Atorvastatin is commonly used as a cholesterol lowering agent in patients. Recently, the neuroprotective effects of atorvastatin became the focus of many research studies. In this study, we have formulated chitosan microspheres containing atorvastatin calcium. In-vitro characterization of chitosan microspheres and quantification of atorvastatin calcium from formulations were also evaluated. The neuroprotective efficiency of atorvastatin calcium was investigated by an experimental spinal cord injury model. Atorvastatin calcium microspheres were implanted at the laminectomy area (1 mg/kg) immediately after trauma. Twenty-four hours after injury, motor functions of animals were scored according to modified Tarlov Scale. In spinal cord tissues tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6 and lipid peroxidation levels were quantified and ultrastructural changes have been investigated. The results of all parameters indicate that microspheres containing atorvastatin calcium were capable of improving functional outcome, attenuating the expression of TNF-alpha, IL-1beta and IL-6; lowering lipid peroxidation levels and maintaining the preservation of the cellular uniformity.

Local administration of chitosan microspheres after traumatic brain injury in rats: a new challenge for cyclosporine--a delivery.

The major aim of this study was to evaluate the efficiency of chitosan microspheres containing cyclosporine-A (Cs-A) on mitochondrial damage in traumatic brain injury (TBI) animal model. Trauma was introduced to male Sprague-Dawley (SD) rats similar to that of modified Feeney Method. Briefly, after craniectomy in the left parietal region (5 mm). Trauma was performed by dropping 24 g metal sterile rods through a teflon guide tube (9.3 cm) on a foot plate placed over the duramater. Just after the trauma, 20 mg/kg Cs-A (Sandimmune) has been administered to the traumatised SD rats intraperitoneally (i.p.). On the other hand, only chitosan microspheres containing 10 mg/kg was implanted at the craniectomy area locally after trauma in Group E. A small piece of surgicell was placed over the craniectomy hole and the scalp incision was sutured. 24 h after injury and the brain tissues were removed intact. The results were evaluated through lipid peroxidation ratio and ultrastructural grading system. The statistical comparisons were evaluated by using Mann Whitney- U test at the significance level p = 0.05. The lipid peroxidation ratios of sham (78.4 +/- 6.0 nmol/g tissue) and vehicle (80.2 +/- 10.6 nmol/g tissue) were significantly increased 24 h after TBI. However, for treatment groups (i.p. Cs-A; 20 mg/kg) and (10 mg/kg Cs-A in microspheres), statistically significant lower lipid peroxidation ratios were determined as 53.5 +/- 9.7 and 47.9 +/- 8.1 nmol/g tissue, respectively (p < 0.05). The mitochondrial damage scores of the treatment groups were recorded as 21.7 +/-2.6 and 19.4 +/- 3.9 for Group D and Group E, respectively. Both of these scores of the treatment groups were found as significantly different from the sham and vehicle groups' scores individually. The implantation of microsphere formulation has provided a better efficiency in keeping the uniformity of mitochondrial structure in this complex cascade of events after TBI.

Immunomodulation of acute experimental spinal cord injury with human immunoglobulin G.

Immunomodulation of acute spinal cord injury may inhibit the activity of specific inflammatory cascades and result in recovery of motor function. In this study, evaluation of the protective effect of a well-known anti-inflammatory immunomodulator, immunoglobulin G (IgG), was conducted in rats after a 50 g/cm contusion spinal cord injury. Following injury, 400 mg/kg of IgG was administered to the treatment group. Twenty-four hours later, animals were assessed functionally via an inclined plane and the Basso-Beattie-Bresnahan motor scale and compared to controls. Tissue was reviewed for myeloperoxidase activiy (MPO) and lipid peroxidation (LPO), and electron microscopy was conducted to assess tissue ultrastructure. Significant functional preservation was observed in the IgG treatment group. In addition, biochemical assays revealed decreased MPO activity, and electron microscopic views of tissue showed preserved ultrastructure. IgG treatment following acute contusion injury to the rat spinal cord confers functional and structural neuroprotection.

Neuroprotective effect of erythropoietin after experimental cold injury-induced vasogenic brain edema in rats.

BACKGROUND: The aims of this study were to evaluate the efficiency of EPO in the treatment of cold injury-induced brain edema, apoptosis, and inflammation and to compare its effectiveness with DSP. METHODS: One hundred fifteen adult male Sprague-Dawley rats weighing between 280 and 300 g were used for the study. Rats were divided into 5 groups. Controls received craniotomy only. The injury group underwent cold injury and had no medication. In the EPO group, a single dose of 1000 IU/kg body weight of EPO was administered. The DSP group received 0.2 mg/kg body weight of DSP. The vehicle group received a vehicle solution containing human serum albumin, which is the solvent for EPO. Brain edema was formed by cold injury using metal sterile rods with a diameter of 4 mm that were previously cooled at -80 degrees C. Twenty-four hours after the injury, animals were decapitated and brain tissues were investigated for brain edema, tissue MPO and caspase-3 levels, and ultrastructure. RESULTS: A significant increase in brain water content was revealed in injury group of rats at 24 hours after cold injury. Injury significantly increased tissue MPO and caspase-3 levels and resulted in ultrastructural damage. Both EPO and DSP markedly decreased tissue MPO and caspase-3 levels and preserved ultrastructure of the injured brain cortex. CONCLUSIONS: Erythropoietin and DSP were found to be neuroprotective in cold injury-induced brain edema model in rats via anti-apoptotic and anti-inflammatory actions.

A comparative study of treatment for brain edema: magnesium sulphate versus dexamethasone sodium phosphate.

Treatments for brain edema are important and one of the major options is corticosteroids. Cell membrane stabilization and prevention of formation of free radicals are the main mechanisms of action of steroids in edema treatment. As an alternative therapeutic agent, magnesium sulphate has been used for its neuroprotective effect in various injury models. In our animal model of brain injury, cold has been used in Sprague-Dawley rats. After brain injury, magnesium sulphate (600 mg/kg) or dexamethasone sodium phosphate (0.2 mg/kg) were administered to experimental groups. The degree of brain edema and lipid peroxidation was evaluated using the wet-dry weight method, the determination of malondialdehyde (MDA) levels and an ultrastructural grading system. Magnesium sulphate treatment was found to be the most effective choice due to the absence of side effects and comparable efficacy to corticosteroids.

Venous outflow of the brain after bilateral complete jugular ligation.

A case of a patient with bilateral internal, external, posterior external and anterior jugular vein ligations and excisions performed in the neck due to a larynx tumor is presented. Radical neck dissection is a standard otorhinolaryngological procedure in the management of head and neck cancer patients with bilateral lymph node metastasis to the neck. Sacrifice of both internal and external jugular veins bilaterally has been recognized as a dangerous approach leading to intracranial hypertension with subsequent neurological sequela and death. In this report, we aimed to demonstrate how venous outflow from the brain diverts after jugular venous system obliteration. After bilateral jugular vein ligations, digital subtraction angiography (DSA) showed that the venous drainage route of the brain had been diverted from the jugular veins to the vertebral venous plexus.

Recombinant human erythropoietin decreases myeloperoxidase and caspase-3 activity and improves early functional results after spinal cord injury in rats.

Inflammatory response and apoptosis have been proposed as mechanisms of secondary injury of the spinal cord after primary insult. Recent studies have shown that erythropoietin (EPO) has neuroprotective properties. In this study, we assessed the efficacy of recombinant human erythropoietin (r-Hu-EPO) in the treatment of acute spinal cord injury (SCI) in rats. Rats were divided into five groups of eight rats each. Controls (Group 1) received laminectomy only. The trauma-only group (Group 2) underwent 40 g/cm contusion injury and had no medication. In group 3, 30 mg/kg of methylprednisolone (MPSS) was administered. Group 4 received 1000 IU/kg body weight of r-Hu-EPO. The vehicle group (Group 5) received a vehicle solution containing human serum albumin, which is the solvent for r-Hu-EPO. Twenty-four hours after trauma, animals were functionally evaluated and a spinal cord samples were obtained for the assessment of caspase-3 and myeloperoxidase (MPO) activities. The results showed that MPO and caspase-3 activities increased to statistically significant higher levels in the spinal cord after contusion injury comparing to the control group. MPO and caspase-3 enzyme activity levels were significantly reduced in animals treated either with r-Hu-EPO or MPSS. In addition, we observed significant early functional recovery in EPO-treated rats. EPO has anti-apoptotic and anti-inflammatory effects, and improves early clinical results after SCI.

Effects of magnesium sulphate following spinal cord injury in rats.

Neutrophil infiltration has been implicated in the secondary destructive pathomechanisms after initial mechanical injury to the spinal cord. Tissue myeloperoxidase (MPO) activity has been shown to be an exclusive indicator of the extent of post-traumatic neutrophil infiltration. We have studied the effect of magnesium sulphate on MPO activity after spinal cord injury in rats. Rats were randomly allocated into 5 groups. Group 1 was control and normal spinal cord samples were obtained after clinical examination. Forty g-cm contusion injury was introduced to Group 2. Group 3 was vehicle, 1 ml of physiological saline was injected post-trauma. Group 4 was given 30 mg/kg methylprednisolone sodium succinate (MPSS) immediately after trauma. Group 5 was given 600 mg/kg magnesium sulphate immediately after trauma. Animals were examined by inclined plane technique of Rivlin and Tator 24 h after trauma. Spinal cord samples obtained following clinical evaluations. Magnesium sulphate treatment improved early functional scores and decreased MPO activity. These findings revealed that magnesium sulphate treatment possesses neuroprotection on early clinical results and on neutrophil infiltration after acute contusion injury to the rat spinal cord.

Multiple isolated spinous process fracture (Clay-shoveler's fracture) of cervical spine: a case report.

Fractures of isolated spinous processes of cervical and thoracic vertebrae are called as Clay shoveler's fracture. In this report, a case of 32-year-old male with multiple isolated spinous process fracture of cervical spine is reported. The patient treated conservatively with a cervical collar. These fractures may be a warning sign of more severe spinal injuries.

Intracranial metastasis of lung adenocarcinoma mimicking colloid cyst of the third ventricle.

A patient with intracranial lung adenocarcinoma metastasis mimicking a colloid cyst of the third ventricle is reported. These tumours may be associated with excessive bleeding and may infiltrate into surrounding structures. Open microsurgery rather than endoscopic surgery should be considered for these cases, particularly a transcortical-transventricular or transcallosal approach, in order to avoid serious complications.

Unilateral proptosis arising from an ectopic lacrimal gland of the orbit.

This 4-year-old boy presented with an ectopic lacrimal gland in the right orbit. Computerized tomography scans revealed a well-encapsulated mass lesion in the right orbit with lateral wall destruction and extension to the subcutaneous tissue. A lateral orbital approach was performed. Although ectopic lacrimal gland tissue is rare in the orbit, this lesion can cause proptosis and orbital bone destruction in childhood.

Magnesium sulfate treatment decreases caspase-3 activity after experimental spinal cord injury in rats.

BACKGROUND: Apoptosis has increasingly been considered as an important factor in secondary injury after spinal cord injury (SCI). Manifestation of apoptotic cell death process involves activation of the caspase-3 apoptotic cascade. The aim of the study was to demonstrate the effect of magnesium sulfate on caspase-3 activity and to compare its effectiveness with methylprednisolone after acute SCI. METHODS: The rats were randomly and blindly allocated into 5 groups of 8 rats each. Spinal cord contusion injury was produced by the weight drop method. The control group consisted of non-injured rats. In the trauma group, no treatment was given, whereas 1 mL saline, 600 mg/kg magnesium sulfate, and 30 mg/kg methylprednisolone sodium succinate (MPSS) were administered in the vehicle and both treatment groups immediately after injury. Twenty-four hours after trauma, spinal cord samples were obtained, and tissue caspase-3 activity levels were examined. A 1-way analysis of variance and the post hoc test were used for statistical analysis. RESULTS: The results showed that caspase-3 activity increased to statistically significantly higher levels in spinal cord after contusion injury than in the control group. Caspase-3 enzyme activity levels were significantly reduced in animals treated either with magnesium sulfate or MPSS. CONCLUSIONS: We have shown that magnesium sulfate decreases caspase-3 activity in rat spinal cord subjected to contusion injury. Magnesium sulfate may have potential therapeutic benefits by reducing apoptotic tissue damage after SCI.

The efficiency of dexamethasone sodium phosphate-encapsulated chitosan microspheres after cold injury.

OBJECTIVE: The aims of this study were to evaluate the efficiency of dexamethasone sodium phosphate (DSP) in the treatment of cold injury-induced brain edema and to compare systemic and topical application of DSP. METHODS: Sprague-Dawley rats weighing nearly 300 g were used in the experiments. Brain edema was formed by cold injury using metal sterile rods with a diameter of 4 mm that were previously cooled at -80 degrees C. Twenty-four hours after the injury, animals were decapitated and brain tissues were investigated by wet-dry weight method, lipid peroxidation ratio, and histological examination. RESULTS: The degree of edema was significantly lowered in groups in which DSP was administered using chitosan microspheres and by intraperitoneal route (P < .05). The statistical evaluation of the experimental results was performed using Mann-Whitney U test. Histological findings transmission electron microscopy (TEM) correlated with the quantitative results. CONCLUSION: Both intraperitoneal- and microsphere-administered DSP were found to be very effective in a cold injury brain edema model. The authors believe that future studies should lead to new applications of the microsphere formulations prepared by chitosan as the matrix material in many other therapies.

Foraminal migration of a lumboperitoneal shunt catheter tip.

Although lumboperitoneal shunts have some advantages over other shunt types, they are also associated with unique complications, including scoliosis, back pain, and sciatica. We report a case of foraminal migration of a lumboperitoneal shunt catheter tip, which resulted in radicular pain and neurological deficit.

Increased xanthine oxidase activity after traumatic brain injury in rats.

Oxidative stress may contribute to many of the pathophysiologic changes that occur after traumatic brain injury (TBI). There are a number of potential sources and mechanisms for oxygen free radical (OFR) production and lipid peroxidation after TBI. In this study, we investigate the time-dependent changes in xanthine oxidase (XO) activity and lipid peroxidation using a focal TBI animal model. We demonstrate that there is an immediate increase in lipid peroxidation by-products and in XO enzyme activity after TBI.

Closed posterior superior iliac spine impeding pediculocorporeal S-1 screw insertion.

OBJECT: Placement of pedicle screws into S-1 is difficult. In cases in which there is a closed posterior superior iliac spine (PSIS), its medial situation prevents lateral oblique placement of the screw inserter sleeve and directing the screw to the anteromedial aspect of S-1. In the present study, the authors discuss anatomical variations of the PSIS and sacrum, and they describe a safe and effective S-1 screw insertion technique. METHODS: The relation of 50 PSISs obtained from 25 dry pelvises (15 male and 10 female cadavers) was examined. The distance from the inferolateral aspect of the S-1 superior articular facet to its promontory was estimated. The relation between the point of anterior penetration of the "screw line" and "safe zone" was analyzed. Penetration of screw lines into the S-1 body was also measured. (An illustrative case of closed PSIS is presented with pre- and postoperative computerized tomography [CT] scan findings.) The authors found that that PSIS was situated in 28% of the specimens. When screws were directed anteromedially, the screw lines failed to penetrate the S-1 body in 24% of the male and in 15% of female specimens. The screw lines deviated from the safe zone anteriorly in 34% of the male and in 20% of the female specimens. When the PSIS was medial to the line that connects the inferolateral aspect of the S-1 superior articular facet to the promontory, a classification of closed PSIS was assigned. CONCLUSIONS: The accuracy of the placement of the screws and their pullout strength are increased when using the present technique. Preoperative CT scanning should be performed to determine the presence of a closed PSIS; in cases in which a closed PSIS is found, the ilium should be resected to enable a greater anteromedial trajectory for placement of S-1 pedicle screws.

Ultrastructural scoring of graded acute spinal cord injury in the rat.

OBJECT: There is a need for an accurate quantitative histological technique that also provides information on neurons, axons, vascular endothelium, and subcellular organelles after spinal cord injury (SCI). In this paper the authors describe an objective, quantifiable technique for determining the severity of SCI. The usefulness of ultrastructural scoring of acute SCI was assessed in a rat model of contusion injury. METHODS: Spinal cords underwent acute contusion injury by using varying weights to produce graded SCI. Adult Wistar rats were divided into five groups. In the first group control animals underwent laminectomy only, after which nontraumatized spinal cord samples were obtained 8 hours postsurgery. The weight-drop technique was used to produce 10-, 25-, 50-, and 100-g/cm injuries. Spinal cord samples were also obtained in the different trauma groups 8 hours after injury. Behavioral assessment and ultrastructural evaluation were performed in all groups. When the intensity of the traumatic injury was increased, behavioral responses showed a decreasing trend. A similar significant negative correlation was observed between trauma-related intensity and ultrastructural scores. CONCLUSIONS: In the present study the authors characterize quantitative ultrastructural scoring of SCI in the acute, early postinjury period. Analysis of these results suggests that this method is useful in evaluating the degree of trauma and the effectiveness of pharmacotherapy in neuroprotection studies.

Antioxidant actions and early ultrastructural findings of thiopental and propofol in experimental spinal cord injury.

Thiopental and propofol are effective antioxidant agents. The current study was undertaken to examine the neuroprotective effects of a single intraperitoneal dose of thiopental and propofol. Effects of the drugs were evaluated by lipid peroxidation and ultrastructural findings. Fifty male Wistar rats were divided into five groups. Group 1 was the control group. Rats underwent laminectomy only, and nontraumatized spinal cord samples were obtained 1 hour after surgical intervention. All other rats sustained a 50-g/cm contusion injury by the weight drop technique. Group 2 rats underwent spinal cord injury alone, group 3 rats received 1 mL intralipid solution intraperitoneally immediately after trauma as the vehicle group, group 4 rats received a 15-mg/kg single dose of thiopental, and group 5 rats received a 40-mg/kg single dose of propofol intraperitoneally following the trauma. Samples from groups 2, 3, 4, and 5 were obtained 1 hour after injury. Lipid peroxidation was determined by measuring the concentration of malondialdehyde in the spinal cord tissue. The ultrastructure of the spinal cord was determined by electron microscopy. The contusion injury was associated with a rise in lipid peroxidation. Compared with the trauma group there was significant attenuation in lipid peroxidation of groups 4 and 5. Ultrastructural findings showed that the rats of group 4 sustained minor damage after spinal cord injury, but there was more evident damage in group 5 rats. These results indicate that thiopental decreases lipid peroxidation and improves ultrastructure, whereas propofol decreases lipid peroxidation without improving ultrastructure 1 hour after spinal cord injury in rats.

Correlation of injury severity and tissue Evans blue content, lipid peroxidation and clinical evaluation in acute spinal cord injury in rats.

OBJECTIVE: To demonstrate the changes in microvascular permeability occurring in association with graded acute spinal cord injury and to determine whether tissue Evans blue content is a useful indicator of the severity of spinal cord injury. The study also aimed to test the ability of the Evans blue method to demonstrate secondary injury after spinal cord contusion. METHODS: In step one of the study, spinal cord lipid peroxidation levels and spinal cord Evans blue content were evaluated at 2 h post-injury in five groups of rats: a control group, a laminectomy-only group and three trauma groups (10, 50, and 100 gcm). In step two, these rats were used for Evans blue assessment following clinical examination at 24 h post-injury. RESULTS: The laminectomy-only group showed no difference from the control group with regard to spinal cord lipid peroxidation levels, tissue Evans blue content, and clinical findings. Increase in spinal cord tissue Evans blue content and lipid peroxidation was correlated with increasing intensity of trauma. There was a negative correlation between trauma intensity and clinical findings, and there was an increase in spinal cord tissue Evans blue content at 24 h compared with that at 2 h. CONCLUSIONS: Determination of spinal cord tissue Evans blue content is a reliable, rapid, simple and inexpensive method that can be used in experimental spinal cord injury to assess the severity of injury and to evaluate neuroprotection studies. The present study is the first to show that the Evans blue technique is a useful method to demonstrate secondary injury of spinal cord tissue and vasculature.

Effect of immunomodulation with human interferon-beta on early functional recovery from experimental spinal cord injury.

STUDY DESIGN: Electron and light microscopic changes, neutrophil infiltration, and lipid peroxidation in the spinal cord and early neurologic examination were studied in rats. OBJECTIVE: To examine the effects of immunomodulator treatment with recombinant human interferon-beta after spinal cord contusion injury. SUMMARY OF BACKGROUND DATA: Immunomodulator treatment with interferon-beta has been the subject of extensive studies, but mainly in relation to multiple sclerosis. Recently, it was reported that interferon-beta possessed significant neuroprotection after experimental transient ischemic stroke. However, to our knowledge, there have been no previous reports about the neuroprotective effect of interferon-beta after spinal cord injury. METHODS: Rats were randomly allocated into 5 groups. Group 1 was control and after clinical examination, normal spinal cord samples were obtained. Group 2 was introduced 50 g/cm contusion injury. Group 3 was vehicle, immediately after trauma 1 mL of physiologic saline was injected. Group 4 was given 30 mg/kg methylprednisolone sodium succinate intraperitoneally immediately after trauma. Group 5 was given 1 x 10(7) IU interferon-beta immediately and 0.5 x 10(7) IU interferon-beta 4 hours after trauma. Animals were examined by inclined plane and Basso-Beattie-Bresnahan scale 24 hours after trauma. Spinal cord samples obtained following clinical evaluations. Neutrophil infiltration was evaluated by myeloperoxidase activity and lipid peroxidation was estimated by thiobarbituric acid test. Electron and light microscopic results were also performed to determine the effects of interferon-beta on tissue structure. RESULTS: Interferon-beta treatment improved neurologic outcome, which was supported by decreased myeloperoxidase activity and lipid peroxidation. Electron and light microscopic results also showed preservation of tissue structure in the treatment group. CONCLUSIONS: Immunomodulator treatment with interferon-beta possesses obvious neuroprotection after acute contusion injury to the rat spinal cord.