Interdisciplinary treatment of an adult with complete bilateral cleft lip and palate.

Interdisciplinary treatment was used for an adult patient born with complete bilateral cleft lip and palate. He had a severe maxillary deficiency with a wide cleft involving the alveolar and maxillary bone and palate. Reconstruction of the arches and occlusion in patients who missed the optimal treatment time is a difficult task for orthodontists. The clinical examination showed severe hypogenesis of the maxillary bone with a total crossbite. The maxillary dental arch was extremely narrow, and the maxillary incisors showed extensive caries caused by improper oral hygiene. Fixed and removable expansion appliances were used to improve the lateral crossbite. Alveolar bone grafting and unilateral LeFort I maxillary osteotomy were performed on the right side for alignment of the maxillary arch. Mandibular setback with bilateral sagittal split ramus osteotomy was also performed to correct the anteroposterior skeletal discrepancy. After postsurgical orthodontic treatment, prosthetic treatment was carried out for final reconstruction of esthetics and orthognathic function. Interdisciplinary treatment was necessary for this patient to achieve a proper occlusion and better esthetics.

Bone morphogenetic protein-2 functions as a negative regulator in the differentiation of myoblasts, but not as an inducer for the formations of cartilage and bone in mouse embryonic tongue.

BACKGROUND: In vitro studies using the myogenic cell line C2C12 demonstrate that bone morphogenetic protein-2 (BMP-2) converts the developmental pathway of C2C12 from a myogenic cell lineage to an osteoblastic cell lineage. Further, in vivo studies using null mutation mice demonstrate that BMPs inhibit the specification of the developmental fate of myogenic progenitor cells. However, the roles of BMPs in the phases of differentiation and maturation in skeletal muscles have yet to be determined. The present study attempts to define the function of BMP-2 in the final stage of differentiation of mouse tongue myoblast. RESULTS: Recombinant BMP-2 inhibited the expressions of markers for the differentiation of skeletal muscle cells, such as myogenin, muscle creatine kinase (MCK), and fast myosin heavy chain (fMyHC), whereas BMP-2 siRNA stimulated such markers. Neither the recombinant BMP-2 nor BMP-2 siRNA altered the expressions of markers for the formation of cartilage and bone, such as osteocalcin, alkaline phosphatase (ALP), collagen II, and collagen X. Further, no formation of cartilage and bone was observed in the recombinant BMP-2-treated tongues based on Alizarin red and Alcian blue stainings. Neither recombinant BMP-2 nor BMP-2 siRNA affected the expression of inhibitor of DNA binding/differentiation 1 (Id1). The ratios of chondrogenic and osteogenic markers relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, a house keeping gene) were approximately 1000-fold lower than those of myogenic markers in the cultured tongue. CONCLUSIONS: BMP-2 functions as a negative regulator for the final differentiation of tongue myoblasts, but not as an inducer for the formation of cartilage and bone in cultured tongue, probably because the genes related to myogenesis are in an activation mode, while the genes related to chondrogenesis and osteogenesis are in a silencing mode.

Treatment of a horizontal open bite with an invisible multiloop appliance in a girl with tooth trauma.

This case report presents the treatment of a girl with a Class II horizontal open bite and severe crowding with an invisible appliance. She had been in a severe traffic accident at 5 years 10 months of age. Her teeth, especially the mandibular right lateral incisor, canine, and first premolar had ankylosed, and their roots were severely resorbed and fused to the alveolar bone. Orthodontic treatment started at the age of 12 years. The mandibular left first premolar and both maxillary first premolars were extracted to reduce overjet and crowding. The left first premolar was extracted and transplanted to the extracted position of the right first premolar. Because the patient wanted an invisible appliance, we selected the Fujita lingual bracket system for her treatment. During the final stage of treatment, a multiloop mushroom archwire was placed to correct the open bite in the maxillary arch with vertical elastics. Six years after retention, her occlusion remained stable, and the transplanted premolar was functioning normally.

Extracellular matrix-mediated tissue remodeling following axial movement of teeth.

Tooth eruption is a multifactorial process involving movement of existing tissues and formation of new tissues coordinated by a complex set of genetic events. We have used the model of the unopposed rodent molar to study morphological and genetic mechanisms involved in axial movement of teeth. Following extraction of opposing upper molars, lower molars supererupted by 0.13 mm. Labeled tissue sections revealed significant amounts of new bone and cementum apposition at the root apex of the unopposed side following supereruption for 12 days. Newly apposited cementum and alveolar bone layers were approximately 3-fold thicker in the experimental vs the control group, whereas periodontal ligament width was maintained. Tartrate-resistant acid phosphatase staining indicated bone resorption at the mesial alveolar walls of unopposed molars and provided in tandem with new bone formation at the distal alveolar walls an explanation for the distal drift of molars in this model. Microarray analysis and semiquantitative RT-PCR demonstrated a significant increase in collagen I, integrin beta5, and SPARC gene expression as revealed by comparison between the unopposed molar group and the control group. Immunohistochemical verification revealed increased levels of integrin beta5 and SPARC labeling in the periodontal ligament of the unopposed molar. Together our findings suggest that posteruptive axial movement of teeth was accomplished by significant formation of new root cementum and alveolar bone at the root apex in tandem with upregulation of collagen I, integrin beta5, and SPARC gene expression.

Bones, teeth, and genes: a genomic homage to Harry Sicher's "Axial Movement of Teeth".

AIM: The model of the unopposed rodent molar was used to study the morphologic and genetic mechanisms of tooth eruption. METHODS: Left maxillary molar teeth of 12-day-old Swiss-Webster mice were extracted under anesthesia, and mandibular molars were allowed to supererupt. To trace areas of tissue remodeling and to determine areas of new tissue formation, mice were injected with fluorescent dyes, tetracycline, alizarin red, and calcein blue. Subsequent to sacrifice, mandibular tissue blocks were prepared for ultrathin ground sections, fluorescent microscopy, and von Kossa's mineral detection procedure. A second set of specimens was prepared for RNA extraction and microarray analysis. RESULTS: The data established significant eruption of first and second mandibular mouse molars 12 days after complete extraction of antagonists, exceeding the control side by 0.13 mm. Labeled tissue sections revealed significant amounts of new bone and cementum apposition on the unopposed side compared to the control side, as revealed by fluorescent markers and ultrathin ground sections. Microarray transcript level comparisons between the experimental and the control groups demonstrated significant (more than twofold) increase in gene expression of elastin and tenascin C extracellular matrix proteins; brevican, lumican, and biglycan proteoglycans; as well as fibroblast growth factor 9. CONCLUSION: In this study, the authors have established the unopposed mouse molar as a model to study tissue dynamics during the axial movement of teeth. The data indicated significant new formation of bone and cementum in tandem with increased expression of extracellular matrix-related genes.

Changes in triacylglycerol-accumulated fiber type, fiber type composition, and biogenesis in the mitochondria of the soleus muscle in obese rats.

Little is known about the effects of obesity on skeletal muscle consisting of approximately 80% type I (slow) fibers, such as that in the soleus muscle, although type I fibers have an enhanced capacity for mitochondrial respiration and fatty acid oxidation. We investigated the effects of obesity on the soleus muscle in the rat. Rats were fed a high-fat diet (protein:fat:carbohydrate = 20:57:23; 508 kcal/100 g) or a control diet (protein:fat:carbohydrate = 20:10:70; 366 kcal/100 g) for 10 weeks. We analyzed the accumulation of intramyocellular triacylglycerol (IMTG), fiber type composition, and the biogenesis and function of the mitochondria in the soleus muscle of the rat during 10 weeks of feeding, using histochemical and real-time polymerase chain reaction analyses. Obesity increased body weight and markedly elevated IMTG levels in type I, but not in type II, fibers of the soleus muscle throughout the feeding period. Obesity also inhibited the biogenesis and function in the mitochondria and altered the fiber type composition in the soleus muscle. The suppression of biogenesis and function in the mitochondria, and the alteration in the fiber type composition may be attributable to the marked IMTG accumulation in the soleus muscle of the rat.

TGFbeta3 is expressed in differentiating muscle of the embryonic mouse tongue.

The purpose of the present study was to elucidate the involvement of transforming growth factor betas (TGFbetas) in the differentiation of tongue striated muscles by analyzing the expression of TGFbetas, their receptors and factors of TGFbeta signal transduction in the mouse tongue between embryonic days 11 (E11) and E15. The expression levels of TGFbeta3 mRNA and protein were much higher than those of TGFbeta1 and TGFbeta2, and the immunolocalization of TGFbeta3 was more consistent with the differentiating muscle cells in comparison with those of TGFbeta1 and 2 between E12 and E15. TGFbetaRI and II were localized to the differentiating muscle cells between E11 and E15. Phosphorylated-smad2/3 was localized to the nucleus of muscle cells which just began to differentiate. These results suggest that the signal of TGFbeta3, but not that of beta1 or beta2, may be involved in the early stages (particularly the beginning) of differentiation of mouse tongue muscle cells through TGFbetaRI, TGFbetaRII, and smad2/3.

BMP-2 regulates the formation of oral sulcus in mouse tongue by altering the balance between TIMP-1 and MMP-13.

The aim of this study is to investigate whether BMP-2 regulates the oral sulcus formation of mouse embryonic tongue by modifying the expression of TIMP and MMP. The BMP-2 siRNA induced a 180% increase in the depth of oral sulcus cavity (P < 0.01) by stimulating the invagination of oral sulcus into the mesenchymal tissues consisting of tongue floor, whereas the recombinant BMP-2 suppressed the process in the organ culture system of mouse embryonic tongue. The BMP-2 siRNA induced a 60% decrease in the expression of TIMP-1 mRNA (P < 0.05) and a drastic decline in TIMP-1 protein was observed around the oral sulcus in the BMP-2 siRNA treated mandibles. The recombinant BMP-2 induced a 220% increases in the expression of TIMP-1 mRNA and the area of the immunostaining for TIMP-1 around the oral sulcus was larger in the mandibles treated with the recombinant BMP-2 than the vehicle. The BMP-2 siRNA induced a 60% increase in the expression of MMP-13 protein and a marked increase in the staining intensity for MMP-13 was observed in the epithelial region of the BMP-2 siRNA treated mandibles. The recombinant BMP-2 induced a 70% decrease in the expression of MMP-13 mRNA and the decrease was mainly observed in the tissues around oral sulcus. The expressions of BMP-2, TIMP-1, and MMP-13 were verified in the tissues around in vivo developing oral sulcus at E11, 12, and 13 by immunohistochemistry. These results suggest that BMP-2 regulates the formation of oral sulcus by altering the balance between TIMP-1 and MMP-13.

BMP2, BMP4, and their receptors are expressed in the differentiating muscle tissues of mouse embryonic tongue.

To investigate the role of bone morphogenetic proteins (BMPs) in the differentiation process of skeletal muscle, we analyzed the in vivo expression of BMP2 and BMP4, of BMP receptors (BMPR) IA, IB, and II, and of activin receptors (ActR) IA, II, and IIB in mouse tongue muscle between embryonic day 11 (E11) and E17. The mRNA expression levels for BMP2 were 5-fold to 11-fold greater than those for BMP4 between E13 and E17 (P < 0.05-0.01). Expression of the BMP2, BMPRIB, ActRIA, ActRII, and ActRIIB proteins was first observed at E13. Expression of BMP2 and BMPRIB was detected in the whole area of the differentiating muscle tissues identified by immunostaining for fast myosin heavy chain (fMHC), but that of ActRIA, ActRII, and ActRIIB was detected only in the peripheral area of the differentiating muscle tissues. In the E15 tongue, all of the BMPs, BMPRs, and ActRs studied herein were expressed in the whole area of the differentiating muscle tissues identified by immunostaining for fMHC. These results suggest that BMPs play a role in the differentiation of tongue muscle tissues at E15 but have little or no effect at E13.

Restoration of mechanical strength and morphological features of the periodontal ligament following orthodontic retention in the rat mandibular first molar.

Biomechanical properties and morphological features of the periodontal ligament (PDL) in the rat mandibular molars were examined during orthodontic retention. Seventy-three male rats of the Wistar strain, 8 weeks of age, were used for biomechanical analysis and six rats for morphological analysis. An elastic band was inserted between the mandibular first and second molars for 4 days; after removal of the elastic band the interdental space was filled with resin for 4 and 8 days. The maximum shear stress, tangent modulus, and failure strain energy density of the PDL of the first molar in the experimental animals decreased markedly following application of an orthodontic force. They increased rapidly and were restored completely to the control levels by the 8th day after retention. Light microscopy showed severe compression and extension of the PDL in the experimental animals on the 8th day after retention. Birefringent collagen fibre bundles running across the compressed and expanded PDL were observed, although they appeared to be thinner with less insertions into the alveolar bone or cementum in the experimental animals than in the controls. This suggests that the periodontal collagen fibres were partially reorganized and rearranged during retention. The reorganization and rearrangement of periodontal collagen fibres seemed to be partly related to the restoration of mechanical strength of the rat molar PDL during the 8 days of retention.

Exogenous hepatocyte growth factor inhibits myoblast differentiation by inducing myf5 expression and suppressing myoD expression in an organ culture system of embryonic mouse tongue.

We examined the effects of exogenous hepatocyte growth factor (HGF) on the differentiation and proliferation of tongue myoblasts by using an organ culture system of tongue obtained from mouse embryos at embryonic day (E) 13. Exogenous HGF induced reductions in the quantities of muscle creatine kinase and myogenin mRNAs and in the number of fast myosin heavy chain-positive myoblasts and myotubes, suggesting that HGF suppressed the differentiation of myoblasts in the cultured E13 tongues. Exogenous HGF induced no significant changes in the percentage of proliferating cell nuclear antigen (PCNA)-positive cell nuclei to total cell nuclei (labeling index) in the muscle portion of the cultured E13 tongue, suggesting that HGF did not affect the proliferation of myoblasts. Exogenous HGF induced the expression of myf5 mRNA but inhibited the expression of myoD mRNA. Since mouse tongue myoblasts are reported to complete proliferation by E13, it appears that exogenous HGF arrests myoblasts in the cell cycle and does not allow them to enter the differentiation process. This is achieved by controlling the expression of myf5 and myoD mRNAs, thus inhibiting the differentiation of tongue myoblasts.

Change from a hard to soft diet alters the expression of insulin-like growth factors, their receptors, and binding proteins in association with atrophy in adult mouse masseter muscle.

To study the role of insulin-like growth factors (IGFs) in the atrophy of mouse masseter muscle in response to a change from a hard to a soft diet, we analyzed the amounts of mRNA and the immunolocalization for IGF-I, IGF-II, their receptors (IGFRs), and binding proteins (IGFBPs). Sixteen male ICR mice were fed a hard diet after weaning; they were divided into two groups at 6 months of age and fed a hard or a soft diet for 1 week. The soft diet treatment decreased masseter weight by 19% ( P<0.01) and the minimal diameter of masseter myofibers by 19% ( P<0.01), verifying that a soft diet led to atrophy of mouse masseter muscle. The soft diet treatment induced a 30% reduction in the amount of IGF-I mRNA ( P<0.05) in preparations of whole masseter tissues. Immunohistochemical findings suggested that a reduction in the expression of IGF-I protein took place in the neural tissues, not in the masseter myofibers. The soft diet treatment induced a 56% decrease in IGF-II mRNA ( P<0.05), a 21% increase in IGFR2 mRNA ( P<0.01), and a 38% decrease in IGFBP5 mRNA ( P<0.01). Immunohistochemical results suggested that these changes at the protein level occurred in the masseter myofibers. No significant or marked difference in the mRNA amount or immunostaining pattern for IGFR1, IGFBP3, IGFBP4, or IGFBP6 was found between the soft and hard diet groups. No IGFBP1 or IGFBP2 mRNA was detected. Thus, IGF-I, IGF-II, IGFR2, and IGFBP5 seem to play a role in the atrophy of mouse masseter muscle in response to the change from a hard to a soft diet in an autocrine and/or paracrine manner.