Atipamezole hydrochloride

Surgical procedures for BCAS were described previously [4 (link)]. Briefly, mice were habituated in the surgery room with free access to food and water for 3–5 days before surgery at 26 °C under a 12-h/12-h light/dark cycle. Nine- to eleven-week-old wild-type mice and Btg2^−/− littermates (body weight, 22–28 g) underwent BCAS or sham operation. Before surgery, mice were intraperitoneally injected with a combination of three anesthetics: 0.3 mg/kg medetomidine hydrochloride (Nippon Zenyaku Kogyo, Fukushima, Japan), 4.0 mg/kg midazolam (Astellas, Tokyo, Japan), and 5.0 mg/kg butorphanol tartrate (Meiji Seika Pharma, Gifu, Japan). For sham-operated mice, the common carotid artery (CAA) was exposed and carefully freed from its sheath. For BCAS-treated mice, an 0.18-mm inner diameter micro-coil (Samini, Shizuoka, Japan) was then attached bilaterally to the CAA. After surgery, 3.0 mg/kg atipamezole hydrochloride (Nippon Zenyaku Kogyo), an antagonist of medetomidine hydrochloride, was injected intraperitoneally to promote safe awakening. To maintain body temperature after surgery, the mice were warmed with a heating pad at 37 °C.

We used a triple anaesthetic mixture of 75 µg/mL medetomidine hydrochloride (Nippon Zenyaku Kogyo, Tokyo, Japan), 400 µg/mL midazolam (Sandoz, Tokyo, Japan) and 500 µg/mL butorphanol tartrate (Meiji Seika Pharma, Tokyo, Japan). For awakening from anesthesia, an anesthetic antagonist (75 μg/mL atipamezole hydrochloride) adjusted with antisedan (Nippon Zenyaku Kogyo) and saline solution (Otsuka Pharmaceutical) was used.
Ligation and removal of silk threads were performed under deep anesthesia by intraperitoneal administration of 0.2 mL of the three anesthetic mixtures. After each procedure, the mice were awakened by intraperitoneal administration of 0.2 mL of the anesthetic antagonist. The mouse ligature-induced periodontitis model was performed according to Abe et al.^{52 (link)}. The silk threads used were 5-0 PERMA-HAND SILK BLACK BRAIDED (Ethicon, Raritan, NJ, USA) and ligated to the maxillary bilateral second molars upon visualization under a binocular stereomicroscope.