Histological Changes In The Elongated Bone Affected By Osteodistraction Of The Mandible In The Dog

Baek, Sun Ho; Ahn, Byoung Keun; Park, Young Ju; Park, Hee Geon; Park, Jun Woo; Rhee, Gun Joo; Lee, Yong Chan; Cho, Byoung Ouck

Abstract

PURPOSE: Traditionally, the treatement of choice has been a bone grafting procedure to increase the length of bone in case of actual length discrepancy. But, bone grafting procedure has many disadvantages, for example, graft resorption, donor site morbidity, and so on. So, many trials have been performed to avert the use of autogenous bone graft via introducing new materials or methods. And, one of those trials has been realized by the development of a technique inducing bone lengthening by osteotomy (or corticotomy) and slow gradual distraction of the osteotomized segments. This new technique of bone lengthening dates back to the early 20th century. But, the majority of information concerning the biology of new bone formation during bone lengthening and technical details of the procedure were produced by extensive clinical and experimental studies performed by Ilizarov, a Russian surgeon. According to Ilizarov, with adequate blood supply, preservation of periosteum, rigid fixation of the osteotomized segments, and proper rate and rhythm of distraction, intramembranous bone rapidly develops within the distraction gap in the limb lengthening procedure. In the limb lengthening, many orthopedic surgeons try to observe the biologic and clinical principles recommended by Ilizarov. In the oral and maxillofacial region, however, not a few studies must be performed to apply this surgical technique in the clinical cases. Besides, the mechanism of bone formation in the distraction gap is not clear, yet. The purpose of this experiment was to scrutinize seriallly the histological changes in the elongated bone affected by osteodistraction of the mandibular body in an adult canine model. In addition, it was performed to confirm the presence of specific region(s) which was important in the bone formation in the gap through the observation of the expression pattern of osteocalcin and osteonectin with the immunohistochemical examination.
MATERIALS AND METHODS
The experimental and control specimens were obtained from seven adult male mongrel dogs weighing over 20kg. The distractors were custom-made linear extraoral devices and bicortical fixation screws were 2.3mm in diameter, 50mm in total length, 15mm in screw length. The distractors were devised to produce a linear gap of 0.75mm between two bony segments every 360.turn of the rotation rod of the device. The mandibular body of the right side of each animal was corticotomized perpendicular to the occlusal plane and then two bony segments were separated completely by careful manipulation of the segments with bone forceps. The left side of each animal was left intact. This side was served as control. At sixth day after osteotomy and fixation of the segments were performed, distraction of the segments was commenced with a rate of 1.1mm/day and a rhythm of two/day for ensuing 7 days. the animals were euthanized at the 16th. 29th, and 44th day after the osteotomy. The bony specimens were decalcified, embedded in paraffin, sectioned 5micrometerthick and stained with H&E. The prepared specimens were examined under the light microscope. And, immunohistochemical examinations using anti-osteocalcin antibody (OC1, Biodesign, USA) and anti-osteonectin antibody Haematologic Technologies Inc., Essex, VT) to locate the expressions of osteocalcin and osteonectin, respectively, were performed.
RESULTS
1. New bone was observed already at the 16th. day after osteotomy. This suggests that new bone formation in osteodistraction was commenced at an early stage of the regenerative process. But, radiologically and microscopically, bony union was not completed in the distraction gap at the 44th. day after osteotomy. Therefore, rigid fixation must be maintained between the bony fragments till the complete bony union is confirmed clinically rather than one month or so after the completion of distraction. 2. Intramembranous bone formation predominated the regenerative process in the distraction gap. Some of the regenerated bone, however, was formed by endochondral ossification. 3. Focal islands of cartilage were observed at all experimental periods. 4. Considering the expression pattern of osteocalcin and osteonectin, new bone formation was considered to be commenced at the region adjacent to the central fibrous zone and therefore this region, especially the periosteum of this region, was important in regeneration of the distraction gap. Focus needs be set on the region adjacent to the CFZ and the role of periosteum in performing further experiments to elucidate the mechanism of bone formation in the distraction gap.