Slow release of tetracycline hydrochloride from a cellulose membrane used in guided tissue regeneration

The objective of this study was to evaluate if the biologic membrane utilized for GTR can be impregnated by tetracycline hydrochloride and if the chemotherapeutic agent, once impregnated, can be released in minimal inhibitory concentrations for a period compatible with clinical application. Initially, an in vitro study was done with cellulose membranes cut in pieces measuring 9 cm2. A volume of 100 microliters containing a 72,000 micrograms/ml solution of tetracycline was dispensed onto each fragment, and dried for 70 minutes at 37 degrees C. Four pieces measuring 0.5 cm2 were cut from different points of the 9 cm2 membrane (presumably, containing 400 micrograms of tetracycline), placed in test tubes containing 4 ml of sterile deionized water, and agitated for 2 minutes. A standard curve was made from known concentrations of tetracycline and compared to 10 microliters of the test solutions obtained by the elution of the 0.5 cm2 fragments. The concentrations were determined through the bioassay technique in 3 duplicate experiments. The samples recovered from the membrane fragments had a mean of 101 micrograms/ml of tetracycline liberated, demonstrating that the membrane was impregnated homogeneously by the chemotherapeutic agent. In a second phase, an in vivo study was carried out to determine the length of time the drug was liberated from the membranes and at which concentrations, in the presence of an inflammatory process. Fourteen 0.5 cm2 fragments containing 400 micrograms of tetracycline were placed in 14 polypropylene chambers containing 200 microliters of thioglycolate medium. The chambers were implanted in the peritoneal cavities of 14 mice, one chamber per animal, and left in from 1 to 14 days. They were then removed and the concentrations of tetracycline determined from 20 microliters samples using a bioassay. The results showed that the antibiotic was released slowly from the 1st through the 12th day in decreasing concentrations that varied from 218 to 20.8 micrograms/ml. The impregnated cellulose membrane can probably be used in GTR acting as a membrane and as a slow-release device, liberating the chemotherapeutic agent in concentrations high enough to eliminate periodontopathic microorganisms.