Dental drill

Controlled cortical impact was used to replicate TBI through a well-validated rodent model(16 (link)). Briefly, on day 1 after acclimation, mice were anesthetized with intraperitoneal ketamine (Hospira, Lake Forest, IL), xylazine (Akorn, Decatur, IL), and acepromazine (Boehringer Ingelheim, St. Joseph, MO) (KXA: 100, 10, 1 mg/kg, respectively) and placed prone in a stereotactic device. After scalp exposure, a left-sided, 4-mm craniotomy was created between bregma and lambda sutures using a dental drill (Henry Schein, Melville, NY) without dural violation. The left parietotemporal cortex was injured via a controlled cortical impactor (CCI) (AMS201, AmScien Instruments, Richmond, VA) which resulted in a reproducible injury correlated to severe TBI (3-mm-diameter impactor tip, impact velocity of 6 m/s, and cortical deformation depth of 1 mm).

In vivo pial intravital microscopy assessment of the cerebral microcirculation was conducted as described previously.^{18 (link),19 (link)} Forty-eight hours after CCI, animals were anesthetized as earlier, placed in a stereotactic frame, and a second 2.5-mm craniotomy was created adjacent to the first using a dental drill (Henry Schein, Melville, NY). This craniotomy was then covered with a 5-mm coverslip (Fisher Scientific, Waltham, MA), and animals were then transferred to an intravital microscope (ECLIPSE FN1; Nikon Instruments, Melville, NY) after which they received 50 μL IV of 0.3% rhodamine 6G (Sigma-Aldrich, St. Louis, MO) to fluorescently label circulating leukocytes. Non-branching pial venules (25–50 μm diameter) were randomly selected and a 1-minute footage was recorded under a 590-nm epi-illumination emission filter using a digital camera (QuantEM; Photometrics, Tucson, AR). Intravenous bovine fluorescein isothiocyanate (FITC)–labeled albumin (Sigma-Aldrich, St. Louis, MO—100 mg/kg) was then administered intravenously for visualization of albumin leakage as a surrogate of microvascular permeability. The same pial regions were observed under a 488-nm fluorescent filter for 10 seconds, and images were captured for offline determination of microvascular leakage.

Controlled cortical impact was used to replicate TBI through a well-validated rodent model.^{16 (link),17 (link)} Briefly, on day 1, mice were anesthetized with intraperitoneal ketamine (Hospira, Lake Forest, IL), xylazine (Akorn, Decatur, IL), and acepromazine (Boehringer Ingelheim, St. Joseph, MO) (KXA, 100, 10, and 1 mg/kg, respectively) and placed prone in a stereotactic device. After scalp exposure, a left-sided 4-mm craniotomy was created between bregma and lambda sutures using a dental drill (Henry Schein, Melville, NY) without violating the dura. The left parietotemporal cortex was then injured via a controlled cortical impactor (AMS201; AmScien Instruments, Richmond, VA), which resulted in a reproducible injury previously correlated to severe TBI (3-mm-diameter impactor tip, impact velocity of 6 m/s, and cortical deformation depth of 1 mm).