Objective: To evaluate the effect of two barrier membranes [multilaminated small intestinal submucosa (mSIS) and bioresorable collagen membrane (Bio-Gide)] combined with deproteinized bovine bone mineral Bio-Oss on guided bone regeneration through a canine extraction sockets model. Methods: The distal roots of 18 premolars of the Beagle’s bilateral maxillary and mandibular were removed, and 18 extraction sockets were obtained. They were randomly divided into 3 groups, and the following procedures were performed on the sockets: (1) filled with Bio-Oss and covered by mSIS (mSIS group), (2) filled with Bio-Oss and covered by Bio-Gide (BG group), (3) natural healing (blank control group). Micro-computed tomograph (Micro-CT) was performed 4 and 12 weeks after surgery to eva-luate the new bone regeneration in the sockets of each group. Results: The postoperative healing was uneventful in all the animals, and no complications were observed through the whole study period. Micro-CT analysis showed that the new bone fraction in the mSIS group and the BG group was significantly higher than that in the blank control group at the end of 4 weeks and 12 weeks (P<0.05), and more new bone fraction was observed in the mSIS group than in the BG group, but the difference was not statistically significant (P>0.05). The new bone fraction of coronal third part of the socket in the mSIS group and BG group at the end of 4 weeks were significantly higher than that of the middle and apical third part of each group (P<0.05). The values of bone mineral density were similar at 4 weeks in all the groups (P>0.05), but were significantly higher than that in the control group at the end of 12 weeks (P<0.05). The bone morphometric analysis showed that the trabecular number and trabecular spacing were significantly better in the mSIS group and the BG group than in the control group at the end of 4 weeks and 12 weeks (P<0.05), while the value in the mSIS group was slightly higher than in the BG group, but the difference was not statistically significant (P>0.05). The difference in trabecular thickness between all the groups was not statistically significant (P>0.05). Conclusion: mSIS membrane as a barrier membrane combined with deproteinized bovine bone mineral can enhance new bone formation in canine extraction sockets, similar to Bio-Gide collagen membrane.