With the aim of providing effective periodontal disease therapeutic method, multilayer membranes which were loaded with drug for guided tissue regeneration were prepared using an immerseprecipitation phase inversion technique. Single layer, bi-layer and tri-layer membranes were fabricated with chitosan used as carrier and tinidazole as medicine model which was loaded on the membrane. The influence of layer on structure and properties of membrane were studied by SEM, UV spectrophotometer and mechanical test. Drug release properties of three types of layer membranes were also investigated. The results showed that release rate could be slown down in both bi-layer and tri-layer membranes (to 11 days and 14 days respectively)and tri-layer membrane lasted the longest. After a process of rapid release, the concentration of tinidazole which was released by the membrane was maintained at an efficient dosage level. Compared with single layer and bilayer membranes, we found tri-layer membrane could play a role in controlling low-rate drug release especially at the early stage of release, and keep an efficient dosage at affected part for a long period of time. The loss of drug which loaded on membrane decreased from 84.6% for single layer to 13.04% for tri-layer. The mechanical strength of three types of membrane were detected and showed that it could meet the requiremens of clinical practice. The membranes especially with tri-layer could be more valuable in application.