Carprofen rimadyl

Carprofen (Rimadyl) was purchased from Zoetis (Parsippany, NJ, USA). Tritiated E2, dissolved in ethanol, was purchased from American Radiolabeled Chemicals (St. Louis, MO, USA). Solvable Solution and Hionic-Fluor scintillation cocktail were purchased from PerkinElmer (Waltham, MA, USA). Randomly methylated β-CD, 2HP-β-CD, β-CD, γ-CD, polyethylene glycol 300 (PEG300), heparin sodium salt, and non-radioactive E2 were purchased from Sigma-Aldrich (St. Louis, MO, USA), and ethanol was purchased from VWR (Radnor, PA, USA). Non-toxic black paint was purchased from Blick (Highland Park, IL, USA). Additional chemicals and solutions, including sodium chloride, 30% hydrogen peroxide, monobasic sodium phosphate, and dibasic sodium phosphate, were purchased from Fisher Scientific (Waltham, MA, USA).

All animal experiments were carried out in accordance with the guidelines of the German Animal Welfare Act and approved by the local authorities (Landesamt für Natur, Umwelt und Verbraucherschutz Nordrhein-Westfalen). Animals were socially housed under a fixed 12:12 h light/darkness cycle with ad libitum access to food and water.
A total of 15 adult, male NMRI-Foxnl^nu/nu mice (12–14 weeks old, 32–37 g; Janvier, France) were used in this study. All surgical experiments were performed under anesthesia with 1.5–2% Isoflurane in a mixture of 70/30% N₂O/O₂ atmosphere. Analgetic treatment included subcutaneous injection of 4 mg/kg Carprofen (Rimadyl, Zoetis, Berlin, Germany) twice a day, for 3 days following surgery. Two days after stroke induction by the filament occlusion method (18 (link), 23 (link)), the animals received an intracortical stem cell implantation on the ipsilateral hemisphere. Resting state fMRI was performed 1 week before, as well as 1, 2, 4, 8, and 12 weeks after stroke induction and cell implantation (2 days after stroke induction), while vitality of the transgenic graft was monitored by BLI at weeks 2, 3, 4, 5, 7, 9, 10, and 12 after stroke.

For both the sham operation and CFL transection groups, mice were anesthetized in a chamber using 4% isoflurane and kept under anesthetization using a mask. Prior to the surgery operation, the lateral side of the right ankle was shaved and cleaned with 75% alcohol for both groups. A small incision was made longitudinally on the anterolateral side of the right ankle. Under the anatomical microscope, the CFL was transected using a purpose-made hook tool with sharpened edges (Kim et al., 2008 (link); Wikstrom et al., 2018 (link)). After the operation was finished, the wound was sutured with polyamide monofilament (6/0, B. Braun Surgical, S.A. Rubi, Spain). Mice received subcutaneous injection of 5.0 mg·kg-1 Carprofen (Rimadyl; Zoetis, Parsippany, NJ), one injection per day for 3 days to prevent wound inflammation. For the sham operation group, the mice were anesthetized following the same procedure as the CFL transection group. The same incision as the CFL transection group was made, but no other surgical operations were performed. After the incision, the wound was sutured with the same procedure as the CFL transection group, following the same wound care protocol.