Neuroscience
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A Model for Encephalomyosynangiosis Treatment after Middle Cerebral Artery Occlusion-Induced Stroke in Mice
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Summary June 22nd, 2022
The protocol aims to provide methods for encephalomyosynangiosis-grafting of a vascular temporalis muscle flap on the pial surface of ischemic brain tissue-for the treatment of non-moyamoya acute ischemic stroke. The approach's efficacy in increasing angiogenesis is evaluated using a transient middle cerebral artery occlusion model in mice.
Transcript
A stroke injury destroy vascular network in the brain and early revival of vascular supply is necessary for a stroke recovery. EMS surgery swiftly restores vascular networks by increasing angiogenesis in the brain. EMS surgery offers a safe method of achieving cerebral angiogenesis, which ultimately will improve blood flow to the ischemic tissue.
This method may reduce the need for pharmacological interventions. EMS has therapeutic potential as a treatment for ischemic stroke. In particular, it may be applied in the setting of malignant stroke where hemicraniectomy is performed routinely.
EMS may facilitate prompt stroke recovery. EMS surgery can provide insights into the mechanism of neuroangiogenesis, as well as it application can be extended to a study of neurogenesis, which often follow angiogenesis. Although any surgical procedure needs substantial practice, however, I believe any researcher who has some experience of rodent surgery can learn and perform EMS surgery easily.
Sterilize all instruments by autoclaving prior to the surgery. Sanitize the operating surface with 70%ethanol and warm it to 37 degrees Celsius with an electric heating pad. After isofluorine induced anesthesia, place the mouse on its left side on the operating surface.
Verify the depth of anesthesia by toe pinching with blunt forceps. Then lubricate both eyes with ophthalmic ointment. Next, shave the hair over the surgical field with an electric razor and clean the surgical field with 70%ethanol, followed by providone solution.
Administer a single dose of 0.25%bupivacaine by subcutaneous injection as preoperative analgesia at the surgery site. Then set up a surgical microscope at the desired magnification. After 60 minutes of middle cerebral artery occlusion, randomize the mice into middle cerebral artery occlusion only or middle cerebral artery occlusion along with encephalomyosynangiosis groups.
For groups receiving encephalomyosynangiosis make a 10 to 15 millimeter skin incision with scissors, extending from one to two millimeters rostral to the right ear and one to two millimeters caudal to the right eye. Then retract the skin flaps using clamps and visually identify the temporalis muscle and the skull. Bluntly dissect the temporalis muscle away from the skull using scissors with a spreading technique.
Next, perform a two to three millimeter myotomy directed ventrally along the caudal border of the muscle to facilitate ventral reflection. Afterward, perform a craniotomy of approximately five millimeters in diameter at the skull underneath the reflected temporalis muscle using a micro drill. Next, remove the dura mater with tweezers to expose the pile surface of the brain with extreme caution to avoid accidental injury to the brain.
After suturing the incision as described in the text, place the mouse back into its cage and monitor until recovery from anesthesia. Then return the mouse to its housing facility. The temporalis muscle graft showed transient damage of the muscle cells seven days after surgery in the grafted and control muscle, with a muscle cell survival of approximately 71%and 97%However, this difference between the grafted and control muscle vanished and the muscles recovered completely 21 days after surgery.
In the encephalomyosynangiosis only and middle cerebral artery occlusion model, the temporalis muscle grafts adhered to the cortical surface, suggesting successful surgery, graft implantation, and bonding. The encephalomyosynangiosis significantly increases blood vessel surface area and integrated density in the perilesional cortex after stroke. Protein array results showed a significant increase in the FGF acidic protein levels and a decrease in the osteopontin levels in the middle cerebral artery occlusion with encephalomyosynangiosis group 21 days after the stroke, suggesting improved angiogenesis and neuroprotection.
21 days after the middle cerebral artery occlusion surgery, the mice had 10%to 11%mortality. However, encephalomyosynangiosis after the occlusion surgery did not increase mortality, suggesting its tolerance. Take precautions, number one, when you dissect temporalis muscle to avoid damage.
Number two, with the use of micro drill to open the skull. And number three, during the removal of dura mater with tweezers. EMS can be applied for other acute injuries such as hematic brain injuries.
If successful EMS will not only improve the stroke outcome but also may improve benefits in other acute brain injuries. EMS will open doors to the novel treatment approach for malignant stroke. It will provide novel method for improving angiogenesis and may shed light on adult neurogenesis, which often depends well perfused tissue.
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