Tomohiro Fukunaga

CTGF and Apoptosis in Mouse Osteocytes Induced by Tooth Movement

It is known that experimental tooth movement stimulates the gene expression of connective tissue growth factor (CTGF) and induces apoptosis in osteocytes in rats. We hypothesized that there is a relationship between CTGF expression and the induction of apoptosis in osteocytes, to play a significant role in triggering bone remodeling during experimental tooth movement. In this study, CTGF mRNA expression was detected at 2 hours in osteocytes on the pressure side, followed by apoptosis at 6 hours after tooth movement in mice. The number of empty lacunae significantly increased on day 1 after mechanical stimulation. Thereafter, the number of osteoclasts significantly increased on the pressure side of the alveolar bone on day 3. Tooth movement increased rapidly on day 10. These findings suggest that CTGF expression, followed by apoptosis in osteocytes in response to mechanical stimulation, might play a significant role in triggering bone remodeling during tooth movement.

Reduction of orthodontic tooth movement by experimentally induced periodontal inflammation in mice.

Orthodontic therapy is known to have an aggravating effect on the progression of destructive periodontitis if oral hygiene is not maintained. However, it is largely unknown how active periodontitis affects the velocity of orthodontic tooth movement. In this study, we examined the effect of periodontal inflammation on orthodontic tooth movement using a mouse model. Orthodontic force was applied on the maxillary first molar of mice, with or without ligature wire to induce experimental periodontitis. The distance moved by the first molar was significantly reduced by the ligature-induced experimental periodontitis. Tartrate-resistant acid phosphatase staining revealed that the number of osteoclasts present during orthodontic treatment was lower in the pressure zone of alveolar bone in the presence of periodontal inflammation. Consistently, the expression level of receptor activator of nuclear factor-kappaB ligand (RANKL) in the pressure zone was decreased in the ligature group. By contrast, experimental periodontitis increased the expression of cyclooxygenase-2 mRNA in the periodontal tissues, while in vitro treatment with prostaglandin E(2) decreased extracellular signal-regulated kinase phosphorylation and RANKL expression induced by mechanical stress in osteoblasts. Taken together, these results suggest that the orthodontic force-induced osteoclastogenesis in alveolar bone was inhibited by the accompanying periodontal inflammation, at least partly through prostaglandin E(2), resulting in reduced orthodontic tooth movement.

Mandibular deviation and canted maxillary occlusal plane treated with miniscrews and intraoral vertical ramus osteotomy: Functional and morphologic changes

Conventionally, the combination of mandibular and maxillary osteotomy is used to correct both mandibular deviation and maxillary canted occlusal plane. This case report describes functional and morphologic changes after an alternative treatment with miniscrew anchorage instead of maxillary osteotomy. A boy, aged 16 years 5 months, had temporomandibular joint disease symptoms, facial asymmetry with mandibular deviation, and a maxillary canted occlusal plane. The maxillary molars on the right side were extruded 3 mm compared with those on the left side. In addition, there were functional problems: lopsided chewing and asymmetric motion range of the bilateral condyles. After correction of the maxillary canted occlusal plane by intrusion of the maxillary right molars with miniscrew anchorage, the mandibular deviation was corrected with mandibular osteotomy. As the therapeutic result, functional symmetry was achieved in addition to symmetric morphologic changes, suggesting that combination treatment of miniscrew anchorage and mandibular osteotomy is effective for treating patients with mandibular deviation and maxillary canted occlusal plane.

Expression of Ten-m/Odz3 in the fibrous layer of mandibular condylar cartilage during postnatal growth in mice

It has been speculated that the mandibular condyle develops via the differentiation of the fibroblast‐like cells covering the condyle into chondrocytes; however, the developmental mechanisms behind this process have not been revealed. We used laser‐capture microdissection and cDNA microarray analysis to elucidate the genes that are highly expressed in these fibroblast‐like cells. Among these genes, the transcription of Ten‐m/Odz3 was significantly increased in the fibroblast‐like cells compared with other cartilage tissues. For the first time, we describe the temporal and spatial expression of Ten‐m/Odz3 mRNA in relation to the expression of type I, II, and X collagen mRNA, as determined by in‐situ hybridization in mouse mandibular condylar cartilage and mouse femoral cartilage during the early stages of development. Ten‐m/Odz3 was expressed in the fibrous layer and the proliferating and mature chondrocyte layers, which expressed type I and II collagen, respectively, but was not detected in the hypertrophic chondrocyte layer. Furthermore, we evaluated the in‐vitro expression of Ten‐m/Odz3 using ATDC5 cells, a mouse chondrogenic cell line. Ten‐m/Odz3 was expressed during the early stage of the differentiation of mesenchymal cells into chondrocytes. These findings suggest that Ten‐m/Odz3 is involved in the differentiation of chondrocytes and that it acts as a regulatory factor in the early stages of the development of mandibular condylar cartilage.

Effects of Mandibular Advancement plus Prohibition of Lower Incisor Movement on Mandibular Growth in Rats

INTRODUCTIONnTo test the hypothesis that mandibular advancement with the use of a fixed functional appliance combined with prohibition of labial movement of the lower incisors will have no effect on mandibular growth in growing rats.nnnMATERIALS AND METHODSnFifteen 4-week-old male rats were divided into fixed, unfixed, and control groups (n = 5, each). Bite-jumping appliances were used in the fixed and unfixed groups. Sites of bone perforation and the lower incisors were connected with ligature wires in the fixed group. The ramus height, mandibular length, and inclination of lower incisors were examined for 4 weeks, and those values were compared among five intervals and three groups by through one-way analysis of variance models and the Bonferroni multiple comparison test for post hoc comparison.nnnRESULTSnIncreases in ramus height and mandibular length during the experimental period were 1.5 mm and 2.5 mm in the fixed group, 1 mm and 1.5 mm in the unfixed group, and 1.2 mm and 1.9 mm in the control group, respectively. Growth of ramus height and growth of mandibular length in the fixed group were greater than in the unfixed and control groups during the experimental period. The inclination of lower incisors in the unfixed group was increased 8.0 degrees throughout the experimental period, which differed from results obtained in the other groups.nnnCONCLUSIONSnMandibular growth was accelerated effectively before and during the pubertal period in rats by mandibular advancement with a fixed functional appliance combined with prohibition of labial movement of the lower incisor.

Sox9 Expression during Fracture Repair

The molecular and cellular mechanisms involved in bone development provide an insight into the nature of bone regeneration. Sox9 is a key transcription factor for chondrogenesis and is also expressed in osteochondroprogenitors during embryonic bone development. However, it has not been determined whether Sox9-expressing cells appear during fracture repair other than in the cartilaginous callus. On the other hand, the difference between bone development and repair is that the motion of the fractured segments is associated with the subsequent fate decision of osteochondrogenic precursors between osteogenesis or chondrogenesis, but the underlying mechanism of this still has to be elucidated. We herein evaluate whether Sox9-expressing cells appear during osseous regeneration in the initial stages of fracture healing in vivo. We also investigated the association between Sox9 induction and mechanical stress and the role of Runx1 expression. As a result, Sox9- and Runx1-expressing cells were detected in the periosteal callus together with Runx2 expression. Their expression levels were significantly downregulated during its ossification, as observed in embryonic bone development. The application of cyclic tension to isolated and cultured stromal cells resulted in the upregulation and maintenance of Sox9 mRNA expression in vitro. These results showed that as in early skeletal development, Sox9- and Runx1-expressing precursor cells first appear in the periosteal callus as an early fracture repair response. Our findings also suggested that the mechanical environment modulates Sox9 expression levels in osteochondrogenic precursors and consequently influences their fate decision between osteogenic and chondrogenic lineage commitment.

A Case Report of Multidisciplinary Treatment of an Adult Patient with Bilateral Cleft Lip and Palate

This is a case report about the successful orthodontic treatment of a bilateral cleft lip and palate patient by using a combination of bone grafting and subsequent prosthodontic rehabilitation. An adult patient with a bilateral cleft lip and palate presented with a concave profile, anterior and lateral crossbite, a markedly deep overbite, and residual bilateral alveolar clefts. His jaw movement patterns were unstable and irregular due to his collapsed bite. Orthodontic treatment with bilateral bone grafting improved his concave profile by downward and backward rotation of the mandible within the freeway space, and optimum occlusion and functionally stable and smooth jaw movements were obtained. After a 6-year retention period, no skeletal relapse could be detected, and his occlusal stability was satisfactory.

Dental and craniofacial characteristics in a patient with leprechaunism treated with insulin-like growth factor-I.

Leprechaunism is an autosomal recessive disease characterized by elfin-like faces, loss of glucose homeostasis, and severe insulin resistance. This disease is caused by inherited defects of the insulin receptor and is lethal early in life. Perhaps for this reason, the teeth and craniofacial features of patients with leprechaunism have never been reported. In the present case, the patient had been diagnosed with leprechaunism with mutation in the insulin receptor gene and had treatment with recombinant human insulin-like growth factor I (IGF-I) starting at the age of 1 year 7 months. It is of interest that all of his teeth were extremely large and subsequently showed severe crowding in the dental arches. He also showed a large tongue with an anterior open bite. He had a convex facial profile with a remarkably steep mandibular plane angle and large gonial angle. This is the first report of the characteristic phenotypes of the teeth and craniofacial morphology of a patient with leprechaunism treated with IGF-I. In addition, the possible association between these features and long-term IGF-I treatment is discussed.

Biosafety, stability, and osteogenic activity of novel implants made of Zr 70 Ni 16 Cu 6 Al 8 bulk metallic glass for biomedical application

Superior mechanical and chemical properties of Zr70Ni16Cu6Al8 bulk metallic glass (BMG) demonstrate its promise as a novel biomaterial for fabrication of implants. The aim of the present study was to validate mechanical, chemical, and biological properties of Zr70Ni16Cu6Al8 BMG through comparison with titanium (Ti). Our data indicated higher tensile strength, lower Youngs modulus, and reduced metal ion release of Zr70Ni16Cu6Al8 BMG compared with Ti. Biosafety of bone marrow mesenchymal cells on Zr70Ni16Cu6Al8 BMG was comparable to that of Ti. Next, screw-type implant prototypes made of Zr70Ni16Cu6Al8 BMG were fabricated and inserted into rat long bones. Zr70Ni16Cu6Al8 BMG implants indicated a higher removal-torque value and lower Periotest value compared with Ti implants. In addition, higher amounts of new bone formation and osseointegration were observed around Zr70Ni16Cu6Al8 BMG implants compared with Ti implants. Moreover, gene expression analysis displayed higher expression of osteoblast- and osteoclast-associated genes in the Zr70Ni16Cu6Al8 BMG group compared with the Ti group. Importantly, loading to implants upregulated bone formation, as well as osteoblast- and osteoclast-associated gene expression in the peri-implant area. No significant difference in concentrations of Ni, Al, Cu, and Zr in various organs was shown between in the Zr70Ni16Cu6Al8 BMG and Ti groups. Collectively, these findings suggest that Zr70Ni16Cu6Al8 BMG is suitable for fabricating novel implants with superior mechanical properties, biocompatibility, stability, and biosafety compared with Ti.nnnSTATEMENT OF SIGNIFICANCEnTitanium is widely used to fabricate orthopedic and dental implants. However, Titanium has disadvantages for biomedical applications in regard to strength, elasticity, and biosafety. Recently, we developed a novel hypoeutectic Zr70Ni16Cu6Al8 BMG, which has superior mechanical and chemical properties. However, the validity of Zr70Ni16Cu6Al8 BMG for biomedical application has not been cleared. The aim of the present study was to validate the mechanical, chemical, and biological properties of Zr70Ni16Cu6Al8 BMG for biomedical applications through comparison with Titanium. The present study clarifies that Zr70Ni16Cu6Al8 BMG has good mechanical properties, corrosion resistance, and osteogenic activity, which are necessary features for biomedical applications. The present study provides for the first time the superiority of Zr70Ni16Cu6Al8 BMG implants to Titanium implants for biomedical applications.

Synergistic acceleration of experimental tooth movement by supplementary high-frequency vibration applied with a static force in rats

Several recent prospective clinical trials have investigated the effect of supplementary vibration applied with fixed appliances in an attempt to accelerate tooth movement and shorten the duration of orthodontic treatment. Among them, some studies reported an increase in the rate of tooth movement, but others did not. This technique is still controversial, and the underlying cellular and molecular mechanisms remain unclear. In the present study, we developed a new vibration device for a tooth movement model in rats, and investigated the efficacy and safety of the device when used with fixed appliances. The most effective level of supplementary vibration to accelerate tooth movement stimulated by a continuous static force was 3 gf at 70u2009Hz for 3u2009minutes once a week. Furthermore, at this optimum-magnitude, high-frequency vibration could synergistically enhance osteoclastogenesis and osteoclast function via NF-κB activation, leading to alveolar bone resorption and finally, accelerated tooth movement, but only when a static force was continuously applied to the teeth. These findings contribute to a better understanding of the mechanism by which optimum-magnitude high-frequency vibration accelerates tooth movement, and may lead to novel approaches for the safe and effective treatment of malocclusion.