This article details a method for delivering a microporous hydrogel to a post-stroke mouse model. The hydrogel serves as a biomaterial scaffold to promote cellular infiltration and new blood vessel formation.
Secure an anesthetized post-stroke mouse on a stereotaxic frame.
Reopen the scalp incision and clean the exposed skull with saline-soaked cotton.
Identify the white-yellow necrotic core in the brain tissue and a burr hole that confirms the stroke site.
This necrotic core contains damaged neurons and non-functional vasculature.
Attach a microporous hydrogel particle-loaded syringe to the pump and set the injection coordinates.
Lower the syringe above the burr hole, then insert it into the stroke site.
Inject the hydrogel at a controlled flow rate to ensure gradual diffusion without brain tissue damage.
Incubate to allow the hydrogel to crosslink, forming a biomaterial scaffold.
Slowly retract the syringe, ensuring hydrogel retention without leakage.
Close the incision with surgical glue and return the mouse to its cage.
The hydrogel scaffold mimics the extracellular matrix, inducing cellular infiltration and new blood vessel formation, indicating the hydrogel's potential in neurotherapeutics.
Reopen the incision over the brain, and use saline soaked cotton to clean any debris from the skull.
A white, yellow circle of tissue, and the burr hole from the drill will be visible. Lock the injection pump at 90 degrees over the mouse.
Load the syringe onto the pump, and move the pump and the X and Y directions to orient the syringe over the burr hole. Move the pump in the Z direction until the needle touches the top of the hole.
And reset the Z on the digital display console. Move the syringe 0.75 millimeters in the Z direction and press start on the pump console to inject four to six microliters of the hydrogel at a one microliter per minute flow rate. When all of the hydro gel has been delivered, set a five minute timer to allow the gel to begin cross-linking.
After five minutes, turn the Z knob to slowly pull up the syringe. When the needle is far enough away from the skull, unlock and remove the pump. Then close the skin over the skull, and place the mouse in a clean cage with monitoring until it has recovered from the anesthesia.
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