Minocycline

Eight-week old C57/Black6 (B6) male mice were purchased from CLEA Japan (Tokyo, Japan). Anesthesia was maintained with 2.5% isoflurane in oxygen for all procedures. A small skin incision was made into the abdomen. After cutting the peritoneum and reaching the abdominal cavity, either 500 μL of low- (2.5 mg/g) or high-dose talc (5 mg/g) (Unitalc; Nobelpharma, Tokyo, Japan), 300 μL of minocycline (2 mg/mL; Pfizer, Tokyo, Japan), 375 μL of picibanil (3.0 KE/kg Chugai Pharmaceutical, Tokyo, Japan), 300 μL of heparin-treated blood from donor mice, or saline solution was injected into the pericardial cavity from the diaphragm side using a 23-gauge needle attached to a 1 mL syringe. Injection volumes and doses were determined by referring to published studies [13 (link), 14 (link)]. At 1, 2, and 4 weeks after operation, mice were sedated with ketamine (0.1 mg/g) and xylazine (0.01 mg/g) to score the strength of adherence and measure the volume of adhesion tissue.

The following antimicrobial agents were tested: cefiderocol, cefepime, ceftazidime, ciprofloxacin, colistin, meropenem, minocycline, tigecycline, levofloxacin, and trimethoprim-sulfamethoxazole. In in vivo studies, meropenem was administered with cilastatin at the same dosage to minimize meropenem degradation by murine dihydropeptidase-1 (DHP-1). Cefiderocol was synthetized by Shionogi & Co., Ltd. (Osaka, Japan) for both in vitro and in vivo studies. For the in vitro and/or in vivo studies, cefepime, ceftazidime, ciprofloxacin, colistin, meropenem, cilastatin, minocycline, tigecycline, and levofloxacin were obtained from commercial sources, and trimethoprim-sulfamethoxazole was synthesized by Shionogi & Co., Ltd. For the in vivo studies, ceftazidime was obtained from GlaxoSmithKline K.K. (London, UK), meropenem from Dainippon Sumitomo Pharma Co., Ltd. (Osaka, Japan), minocycline from Pfizer, Inc. (New York, USA), and trimethoprim-sulfamethoxazole from Taiyo Pharma Co., Ltd. (Tokyo, Japan).