Changes in parathyroid hormone-related protein and 3-dimensional trabecular bone structure of the mandibular condyle following mandibular distraction osteogenesis in growing rats.

PURPOSE: Distraction osteogenesis (DO) is commonly performed for mandibular reconstruction during the growth period. We tested the hypothesis that parathyroid hormone-related protein (PTHrP) in mandibular condylar cartilage and underlying trabecular bone in growing individuals undergo changes in response to distraction forces. MATERIALS AND METHODS: Forty-eight 6-week-old male Sprague-Dawley rats were used. Animals underwent unilateral mandibular distraction using a distractor that we devised, and unoperated animals were evaluated as controls. DO procedure was performed: 3 days' latency period, 0.4 mm/day rate, total 4.0 mm. Changes in cartilage morphology, PTHrP activity, and 3-dimensional trabecular bone structure changes measured by micro-computed tomography were examined at 0, 2, 4, and 6 weeks of consolidation. RESULTS: A marked irregularity was noted in the superior portion of the distracted side's condylar cartilage that resolved after distraction ceased. PTHrP was more strongly expressed in the hypertrophic layer of condylar cartilage on the distracted side than in controls, up to 6 weeks after the end of distraction. Subchondral trabecular bone volume, percent bone volume, and trabecular number in the superior and posterior regions of the condyle decreased significantly by 2 weeks after distraction. These parameters returned to normal in the posterior condyle, but not in the superior part of the condyle by 6 weeks following distraction. CONCLUSION: These results suggest that unilateral mandibular distraction in growing rats causes temporary morphologic alterations of trabecular bone structure on the distracted side accompanied by increased production of PTHrP in the mandibular condyle.

Three-dimensional changes in the condyle during development of an asymmetrical mandible in a rat: a microcomputed tomography study.

A rapidly growing postnatal animal model was used to study changes in the calcified tissue of the mandibular condyle during altered muscle function. A maxillary occlusal splint was designed to shift the mandible laterally (left) during closure. Groups of 5 Wistar rats were killed at 5, 9, 15, 21, 30, and 40 weeks (n = 30), with an equal number of controls. The experimental animals developed shorter, asymmetrical mandibles compared with the control animals. The left condyle became larger and thicker than the right condyle. Microcomputed tomography assessment of the left and right condylar trabecular bone indicated that both had less bone volume than the control condyle. The right masseter muscle significantly lost fiber size and type IIA oxidative fibers, suggesting that the right masseter muscle was used with less tension development. In contrast, the left masseter maintained its fiber size and was similar to the control masseter fiber diameters. Comparison in the sequence of changes indicated that the morphologic changes occurred first in the ramus (age, 5 weeks), before the corpus (age, 15 weeks), and before changes in masseter fiber size and composition (age, 9 weeks). This study showed that both the mandible and the condyle modified their shape and size, as well as the trabecular bone of the condyle, during shifting of the mandible to one side as it closed.

Mechanical strain on the human skull in a humanoid robotic model.

Patterns of strain were analyzed in a dry human skull at 15 different regions on the lateral and medial surfaces of the mandible. The strains were induced with a human robotic system that represented each of 8 bilateral muscles by a DC servomotor connected to a wire and pulley. The tractions of the simulated muscles (masseter, medial pterygoid, anterior temporalis, and posterior temporalis) were increased from 1x to 4x with each representing different levels of traction or force (5, 3, 4, and 4 N, respectively). The study was done with the teeth in maximum intercuspal occlusion. Bite forces were also measured with a transducer and reached a maximum of 40 N on the posterior teeth with less force on the anterior dentition. The smallest traction level (1x) developed some small strains. At 2x, compressive strains developed more on the medial (lingual) side beneath the molars through the corpus and radiated into the anterior ramus. Strains at 3x to 4x significantly increased both the tensile and compressive strains throughout the mandible with more strains developing in the ramus. The increased bilateral traction and loading developed significant compressive forces on both sides of the mandible. Evaluation of disparities between compressive and tensile strains at one site, and comparison between the medial and lateral sides of strain, suggested some visible distortion of portions of the mandible under the higher loads.

Disappearance of epidermal growth factor receptor is essential in the fusion of the nasal epithelium.

Abstract Epidermal growth factor (EGF) and receptor (-R) signaling pathway is required for epithelial cell growth and differentiation such as the degeneration of the medial edge epithelial cells during the fusion process of secondary palate formation. As epithelial fusion takes place during primary palate formation, we investigated the involvement of the EGF-R in fusion of the medial (MNP) and lateral (LNP) nasal prominences of the mouse embryo was examined. Immunoreactivity of EGF-R was investigated in embryonic day 10 embryos (32-37 somite stages). The EGF-R immunoreactivity was observed in the nasal epithelia of the presumptive fusion area before fusion. It became undetectable just prior to the fusion and faintly reappeared at the time of the fusion. In contrast, the non-fusing epithelial cells of the nasal groove maintained the immunoreactivity throughout these stages. In order to elucidate whether the EGF/EGF-R signaling pathway was involved in nasal epithelial fusion, EGF solution was injected into the exocoelum of explanted mouse embryos, and the embryos were cultured for 18-24 h by whole embryo culture (WEC). This exogenous EGF inhibited fusion of nasal prominences in 66.7-81.5% of the embryos. Treatment with EGF for 4-14 h showed that exogenous EGF disturbed the EGF-R disappearance and normal alteration of epithelial cell morphology in the fusion area. These results suggest that temporal disappearance of the EGF/EGF-R signaling from presumptive fusion of the nasal prominences is required for morphological change of the epithelial cells leading to the fusion of MNP and LNP.

Regulation of osteoclast differentiation and function by receptor activator of NFkB ligand and osteoprotegerin.

The differentiation and functions of osteoclasts (OCs) are regulated by osteoblast-derived factors. Receptor activator of NFkB ligand (RANKL) is one of the key regulatory molecules in OC formation. Osteoprotegerin (OPG) is a novel secreted member of the TNF receptor superfamily that negatively regulates osteoclastogenesis and binds to RANKL. We examined the biological actions of macrophage-colony-stimulating factor (M-CSF), RANKL, and OPG on the differentiation of OCs isolated from cocultures of mouse osteoblastic cells and bone marrow cells. Preosteoclasts (pOCs) and OCs were characterized by their ultrastructure and the expression of OC markers such as tartrate-resistant acid phosphatase (TRAP) and vacuolar-type H(+)-ATPase. pOCs formed without any additives expressed TRAP, but showed little resorptive activity on cocultured dentine slices. TRAP-positive pOCs treated with M-CSF began to fuse with each other, but lacked a ruffled border (RB) and showed almost no resorptive activity. pOCs treated with RANKL became TRAP-positive multinucleated cells, which expressed intense vacuolar-type H(+)-ATPase along the RB membranes and exhibited prominent resorptive activity. Such effects of RANKL on pOCs were completely inhibited by the addition of OPG. OPG inhibited RB formation in mature OCs and reduced their resorptive activity, and also induced apoptosis of some OCs. These results suggest that 1) RANKL induces differentiation of functional OCs from pOCs, 2) M-CSF induces macrophage-like multinucleated cells, but not OCs, 3) OPG inhibits RB formation and resorptive activity in mature OCs, 4) OPG also induces apoptosis of OCs, and 5) RANKL and OPG are, therefore, important regulators of not only the terminal differentiation of OCs but also their resorptive function.

Osteoclast induction in periodontal tissue during experimental movement of incisors in osteoprotegerin-deficient mice.

Osteoprotegerin (OPG) is a novel secreted member of the tumor necrosis factor (TNF) receptor superfamily that negatively regulates osteoclastogenesis. The receptor activator of the NFKB ligand (RANKL) is one of the key regulatory molecules in osteoclast formation and binds to OPG. In this study, it was suggested that OPG and RANKL are involved in alveolar bone remodeling during orthodontic tooth movement. We examined RANKL localization and osteoclast induction in periodontal tissues during experimental movement of incisors in OPG-deficient mice. To produce orthodontic force, an elastic band was inserted between the upper right and left incisors for 2 or 5 days, and the dissected maxillae were examined for cytochemical and immunocytochemical localization of tartrate-resistant acid phosphatase (TRAP), vacuolar-type H(+)-ATPase, and RANKL. Compared to wild-type OPG (+/+) littermates, TRAP-positive multinucleated cells were markedly induced in the periodontal ligament (PDL) on the compressed side and in the adjacent alveolar bone of OPG-deficient mice. These multinucleated cells exhibited intense vacuolar-type H(+)-ATPase along the ruffled border membranes. Because of accelerated osteoclastic resorption in OPG-deficient mice, alveolar bone was severely destroyed and partially perforated at 2 and 5 days after force application. In both wild-type and OPG-deficient mice, RANKL expression became stronger at 2 and 5 days after force application than before force application. There was no apparent difference in intensity of RANKL expression between OPG (+/+) littermates and OPG-deficient mice. In both wild-type and OPG-deficient mice, expression of RANKL protein was detected in osteoblasts, fibroblasts, and osteoclasts mostly located in resorption lacunae. These results suggest that during orthodontic tooth movement, RANKL and OPG in the periodontal tissues are important determinants regulating balanced alveolar bone resorption.