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An ideal biomaterial for bone regeneration is a longstanding quest nowadays. This study aimed to evaluate the osteogenic potentiality of nano-bioactive glass enhanced biocement based silicate with or without hyaluronic acid seeded in rabbits’ tibial bone defects. For this, 24 male rabbits with two 5 mm defects (1 defect per tibia) were divided into three equal groups. Among the predefined three groups, for the rabbits of group 1(control) bone defects were left untreated while for the members of group 2 defects received nano-bioactive glass enhanced biocement based silicate cement, and group 3 defects received nano-bioactive glass cement mixed with hyaluronic acid. Animals of each group were divided equally for euthanization after 3 and 6 weeks. Bone specimens were processed and examined histologically with histomorphometrically analysis of new bone area percentage. The bone defects in group 3 showed significantly improved osseous healing histologically as compared to the group 1&2. The morphometric analysis also revealed a significant increase in the new bone area percentage in group 3 as compared to the group 1 and 2 (P < 0.05). The results of the present study can be concluded that bone defects could be treated with nano-bioactive glass and hyaluronic acid cement. Although, nano-bioactive glass alone was capable of bone regeneration the combination of both had significant regenerative capacity.