2 3 5 triphenyl tetrazolium chloride

After 24 h of reperfusion, the rats were subjected to general anesthesia using halothane and then decapitated. The brain was immediately removed and placed in ice-cold saline. Each brain was then placed in a brain matrix, and coronal sections were cut into 2 mm slices. The brain slices were immediately immersed in 2% 2,3,5-triphenyl tetrazolium chloride (TTC) (Wako Pure Chemicals Industries, Osaka, Japan) at 37°C for 15 min and then in 4% formaldehyde [19 (link), 20 (link)]. Following this, infarction areas were identified by an image analysis system (Scion Image 1.62; Scion Corporation, Frederick, MD, USA) and were combined to obtain the infarction volumes per brain according to the following formula: corrected infarction volume (%) = [left hemisphere volume − (right hemisphere volume − the infarction volume)] × 100/left hemisphere volume.

CIL powder (CIL-MPs) was provided by Otsuka Pharmaceutical Co., Ltd. (Tokushima, Japan), and carboxypolymethylene (carbopol, Carbopol^® 934) was purchased from Serva (Heidelberg, Germany). 2-Hydroxyproplyl-β-cyclodextrin (HPβCD) and methylcellulose (MC) were kindly donated by Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan) and Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan), respectively. Nystatin (CavME inhibitor) and docusate sodium salt were obtained from Sigma-Aldrich Japan (Tokyo, Japan). An MFTM membrane filter (450 nm pore size) was purchased from Merck Millipore (Tokyo, Japan). Dynasore (CME inhibitor) and rottlerin (micropinocytosis inhibitor) were provided by Nacalai Tesque (Kyoto, Japan), and l-menthol, indomethacin, dimethyl sulfoxide (DMSO), isoflurane, 2,3,5-triphenyl tetrazolium chloride (TTC), and cytochalasin D (phagocytosis inhibitor) were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). All other chemicals used were of the highest purity commercially available.