Time-Controlled Drug Delivery in EAE Model

Alzet miniosmotic pumps (Durect, Cupertino, CA) were used for time-controlled drug delivery. Fourteen-day pumps (rate of infusion 0.25 μL/h, 100 μL total volume) were filled with either vehicle or 500 μM tuftsin (Sigma) (23 (link)–26 (link)), 18 μg/μl benztropine (Sigma), or both tuftsin and benztropine at the same doses and incubated overnight at 37°C before use. The pumps were implanted subcutaneously in the back of the mice under anesthesia 14-days post-MOG immunization. For experiments where tuftsin was administered prior to addition of benztropine, 7-days pumps containing vehicle or tuftsin alone were implanted on Day 14 post-MOG immunization and were replaced with 7-days pumps containing either vehicle or both tuftsin and benztropine at Day 21.