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The purpose of the present study was to evaluate the healing events and compare the effects of guided tissue regeneration (GTR) using 3 different membranes: a weakly cross-linked bioabsorbable bovine collagen membrane; a strongly cross-linked bioabsorbable bovine collagen membrane; and a non-resorbable ePTFE membrane. Twenty adult female beagle dogs with naturally occurring periodontitis were subjected to periodontal probing and scaling. In both right and left mandibular quadrants, mucoperiosteal flaps were raised, and after debridement, the roots of experimental premolar teeth received apical reference notches. The following treatments were randomly assigned: 1) gingival flap curettage (GFC) + ePTFE membrane, control membrane; 2) GFC + slightly cross-linked collagen membrane, 1st test membrane; and 3) GFC + strongly cross-linked collagen membrane, 2nd test membrane. The flaps were sutured in such a manner that the membranes were completely covered. All dogs received a soft diet for a 2-week period and an oral hygiene program until time of sacrifice. The animals were randomly scheduled for sacrifice at 2, 4, 12, and 24 weeks. The ePTFE membranes still in place were removed at 6 weeks. The jaws were dissected and specimens prepared for descriptive histology and histomorphometry. The early resorption of the 1st test membrane was achieved at 4 weeks, and the 2nd test membrane at 12 weeks, both with normal inflammatory reaction. Measurements of epithelium, connective tissue attachment, new bone, and neocementum were compared within an animal (paired t test). Analyses were performed on data at 4, 12, and 24 weeks post-healing; little differences were found between these periods. Limited connective tissue repair was favored by the placement of all the membranes (about 20%), with no statistically significant difference. These findings indicate that bioabsorbable collagen membranes with different cross-linking and ePTFE barriers promote similar new attachment in GTR procedures on naturally occurring periodontal defects in dogs. J Periodontol 1998;69:1218–1228.