Effect of the distraction rate on the activity of the osteoblast lineage in distraction osteogenesis of rat's tibia: Immunostaining study of the proliferating cell nuclear antigen, osteocalcin, and transglutaminase C

The purpose of this study was to investigate kinetics of the osteoblast lineage in the periosteum and endosteum according to different distraction rates in distraction osteogenesis of rat's tibia. An osteotomy was performed on 144 rats at the proximal diaphysis of the left tibia. The lengthening process was started after a latency period of 3 days, with varying distraction rates of 0.25 mm (group I), 0.5 mm (group II), 0.75 mm (group III), 1.0 mm (group IV) and proceeded until a 3.5 mm length gain was achieved. The animals that had an osteotomy alone, without lengthening, served as a control (group V). Immunohistochemical staining of proliferating cell nuclear antigen (PCNA), osteocalcin, and transglutaminase C (TGase C) were done on the four animals in each group sacrificed at post-distraction days 1, 3, 5, 7, 14, and 28 in order to observe the temporal changes among the experimental and control groups. Also, in order to compare the staining rates at a given length gain among the groups, animals in each group were additionally sacrificed 2 days post-distraction in group II: 2 and 4 days in group III; and 1.5, 2 and 2.5 days in group IV. The results of the expression rates of PCNA, osteocalcin, and TGase C in each group were analyzed quantitatively. The immunohistochemical study on callotasis of rat's tibia revealed that the osteoblast lineage in the periosteum is more activated than that in the endosteum for proliferation and differentiation by distraction, suggesting that the periosteum plays a more important role in neo- osteogenesis in the distraction gap. Daily distraction rates ranging from 0.25 mm to 0.75 mm in two increments is appropriate for successful distraction osteogenesis of rat's tibia, but the rate of 0.25 mm a day is significantly better than that of 0.75 mm as was made evident in the immunohistochemical observations.