Mandibular Endochondral Growth Is Specifically Augmented by Nutritional Supplementation with Myo-Inositol Even in Rabbits.

Mandibular retrognathism occurs by insufficient mandibular growth and causes several issues, such as respiratory difficulty and diminished masticatory function. At present, functional orthodontic appliances are used for stimulating mandibular growth in pediatric cases. However, the effectiveness of functional appliances is not always stable in daily practices. A more effective, reliable, and safer therapeutic method for mandibular growth promotion would be helpful for growing mandibular retrognathism patients. As we previously discovered that nutritional supplementation of myo-inositol in growing mice specifically increases mandibular endochondral growth, we performed preclinical animal experiments in rabbits in this study. Briefly, six-week-old male Japanese white rabbits were fed with or without myo-inositol supplementation in laboratory chow until 25 weeks old, and 3D image analysis using micro CT data and histological examinations was done. Myo-inositol had no systemic effect, such as femur length, though myo-inositol specifically augmented the mandibular growth. Myo-inositol increased the thickness of mandibular condylar cartilage. We discovered that the nutritional supplementation of myo-inositol during the growth period specifically augmented mandibular growth without any systemic influence, even in rabbits. Our results suggest the possibility of clinical use of myo-inositol for augmentation of the mandibular growth in growing mandibular retrognathism patients in the future.

Maternal Hyperthyroidism in Rats Alters the Composition and Gene Expression of the Matrix Produced In Vitro by Chondrocytes from Offspring with Intrauterine Growth Restriction.

Herein, we aimed to evaluate cultures of femoral chondrocytes from offspring of rats with intrauterine growth restriction (IUGR) induced by maternal hyperthyroidism. Fourteen adult female Wistar rats were divided into two groups, a control group and a group treated with daily L-thyroxine administration using an orogastric tube (50 µg/animal/day) during pregnancy. Three days after birth, the offspring were euthanized for chondrocyte extraction. At 7, 14, and 21 days, viability and alkaline-phosphatase (ALP) activity were assessed using the MTT assay and BCIP/NBT method, respectively, in a 2D culture. Pellets (3D cultures) were stained with periodic acid Schiff (PAS) to assess the morphology and percentage of PAS+ areas. The gene transcripts for Col2, Col10, Acan, Sox9, and Runx2 were evaluated by qRT-PCR. The MTT and ALP-assay results showed no significant differences between the groups. Maternal hyperthyroidism did not alter the chondrocyte morphology, but significantly reduced the percentage of PAS+ areas, decreased the expression of the gene transcripts of Col2 and Acan, and increased Sox9 expression. Maternal hyperthyroidism in rats alters the composition and gene expression of the matrix produced by chondrocytes from offspring with IUGR. This may be one of the mechanisms through which excess maternal thyroid hormones reduce offspring bone growth.

Postnatal development of the largest subterranean mammal (Bathyergus suillus): Morphology, osteogenesis, and modularity of the appendicular skeleton.

BACKGROUND: Subterranean mammals show a suite of musculoskeletal adaptations that enables efficient digging. However, little is known about their development. We assessed ontogenetic changes in functionally relevant skeletal traits, and ossification patterns (periosteal and endochondral bone modules) in a truly subterranean scratch-digging rodent, Bathyergus. We studied 52 individuals (202 long bones) from a wild population by using a multiscale approach involving internal and external morphology. RESULTS: Multivariate analysis showed significant morphological changes during ontogeny. A specialized phenotype is expressed perinatally (eg, greater external robustness and developed olecranon, teres major, and deltoid processes), whereas adults presented slender bones with significantly thicker cross-sections. Ossification modules scaled mostly isometrically with body size parameters. Periosteal modules showed high variability and tended to grow faster than endochondral modules. CONCLUSIONS: Scratch-digging adaptations appear at perinatal age and then specialize in subadults. Early development of agonistic and digging behaviors and onset of sexual maturation seems to contribute to its development, although genetic factors also seem to play an important role. Ontogenetic differences are probably a trade-off to counteract weaker cortical bone properties and poor muscle development in juveniles, whereas slender but thicker cortical bones maximize bone resistance during burrow construction without compromising locomotor performance in adults.

Whole-exome sequencing in a Japanese pedigree implicates a rare non-synonymous single-nucleotide variant in BEST3 as a candidate for mandibular prognathism.

Mandibular prognathism is a phenotype of facial deformity seen in populations around the world, but with higher incidence among East Asian populations. Five genome-wide nonparametric linkage analyses and a genome-wide association study to identify susceptibility loci of the phenotype have shown inconsistent results. To explore variants related to mandibular prognathism, we undertook whole-exome sequencing in a Japanese pedigree. The pedigree was ascertained as mandibular prognathism. The pedigree comprised 15 individuals from 4 generations. Four affected individuals across 2 generations and 5 unaffected individuals were chosen for whole-exome sequencing. Five non-synonymous single-nucleotide variants (SNVs) of UBASH3B, OR6M1, OR8D4, OR8B4, and BEST3 genes were detected in all 4 affected individuals, but in none of the 5 unaffected individuals. A non-synonymous SNV of the BEST3 gene, Chr12(GRCh37):g.70048878G>T, NM_032735.2:c.1816C>A, p.(L606I), was identified as rare missense variant. BEST3 is located on chromosome 12q15 and encodes bestrophin 3 from the bestrophin family of anion channels. The 4 other non-synonymous SNVs of UBASH3B, OR6M1, OR8D4, and OR8B4 were not considered plausible candidates for mandibular prognathism. Our whole-exome sequencing implicates a rare non-synonymous SNV of BEST3 as a candidate for mandibular prognathism in the Japanese pedigree.

Asporin stably expressed in the surface layer of mandibular condylar cartilage and augmented in the deeper layer with age.

Mandibular condylar cartilage (MCC) exhibits dual roles both articular cartilage and growth center. Of many growth factors, TGF-ß has been implicated in the growth of articular cartilage including MCC. Recently, Asporin, decoy to TGF-ß, was discovered and it blocks TGF-ß signaling. Asporin is expressed in a variety of tissues including osteoarthritic articular cartilage, though there was no report of Asporin expression in MCC. In the present study, we investigated the temporal and spatial expression of Asporin in MCC. Gene expression profile of MCC and epiphyseal cartilage in tibia of 5 weeks old ICR mice were firstly compared with microarray analysis using the laser capture microdissected samples. Variance of gene expression was further confirmed by real-time RT-PCR and immunohistochemical staining at 1,3,10, and 20 weeks old. TGF-ß and its signaling molecule, phosphorylated Smad-2/3 (p-Smad2/3), were also examined by immunohistochemical staining. Microarray analysis revealed that Asporin was highly expressed in MCC. Real-time RT-PCR analysis confirmed that the fibrous layer of MCC exhibited stable higher Asporin expression at any time points as compared to epiphyseal cartilage. This was also observed in immunohistochemical staining. Deeper layer in MCC augmented Asporin expression with age. Whereas, TGF-ß was stably highly observed in the layer. The fibrous layer of MCC exhibited weak staining of p-Smad2/3, though the proliferating layer of MCC was strongly stained as compared to epiphyseal cartilage of tibia at early time point. Consistent with the increase of Asporin expression in the deeper layer of MCC, the intensity of p-Smad-2/3 staining was decreased with age. In conclusion, we discovered that Asporin was stably expressed at the fibrous layer of MCC, which makes it possible to manage both articular cartilage and growth center at the same time.

Un modelo matemático de la placa de crecimiento / A mathematical model of growth plate

La placa de crecimiento es una estructura que está conformada por células, denominadas condrocitos, que se ordenan en columnas y confieren el alargamiento del hueso debido a su proliferación e hipertrofia. En cada columna se pueden observar condrocitos en estado proliferativo (que se dividen constantemente), e hipertrófico (que crecen para obtener una forma casi esférica). Estas células expresan diferentes proteínas y moléculas a lo largo de su vida media y tienen un comportamiento especial que puede depender de su entorno local mecánico y bioquímico. En este artículo se desarrolla un modelo matemático que describe la relación entre la geometría, el crecimiento por proliferación e hipertrofia y la invasión vascular con los factores bioquímicos y mecánicos presentes durante el desarrollo endocondral(AU)

The growth plate is a structure composed of cells called chondrocytes arranged in columns and causing the bone lengthening due to its proliferation and hypertrophy. In each column it may observed the presence of chondrocytes in proliferation stage (constantly divided) and hypertrophy stage (growing to obtain a almost spherical shape). These cells express different proteins and molecules during half-life and have a special behavior that may to depend on its mechanical or biochemical local environment. In present paper it is developed a mathematical model describing the relationship among the geometry, proliferation and hypertrophy growth and the vascular invasion by biochemical and mechanical factors present during the endochondral development(AU)

Un modelo matemático de la placa de crecimiento / A mathematical model of growth plate

La placa de crecimiento es una estructura que está conformada por células, denominadas condrocitos, que se ordenan en columnas y confieren el alargamiento del hueso debido a su proliferación e hipertrofia. En cada columna se pueden observar condrocitos en estado proliferativo (que se dividen constantemente), e hipertrófico (que crecen para obtener una forma casi esférica). Estas células expresan diferentes proteínas y moléculas a lo largo de su vida media y tienen un comportamiento especial que puede depender de su entorno local mecánico y bioquímico. En este artículo se desarrolla un modelo matemático que describe la relación entre la geometría, el crecimiento por proliferación e hipertrofia y la invasión vascular con los factores bioquímicos y mecánicos presentes durante el desarrollo endocondral

The growth plate is a structure composed of cells called chondrocytes arranged in columns and causing the bone lengthening due to its proliferation and hypertrophy. In each column it may observed the presence of chondrocytes in proliferation stage (constantly divided) and hypertrophy stage (growing to obtain a almost spherical shape). These cells express different proteins and molecules during half-life and have a special behavior that may to depend on its mechanical or biochemical local environment. In present paper it is developed a mathematical model describing the relationship among the geometry, proliferation and hypertrophy growth and the vascular invasion by biochemical and mechanical factors present during the endochondral development

The effect of Tibial Lengthening on Enchondral Growth of the Tibia in Rabbits: A Roentgenographic, Histomorphometric, and Immunohistochemical Study / 대한정형외과학회잡지

We investigated the effect of tibial lengthening by means of callotasis on enchondral growth of the tibia, Ninety-nine 5-week-old immature rabbits were separated into five groups according to the percentage of lengthening : GroupI(10% lengthening), Group II (20% lengthening), Group III (30% lengthening), Group IV (40% lengthening), Group V (sham operation without lengthening). Osteotomy of the tibia and fibula was made at the proximal metaphysiodiaphyseal junction. Distraction was started at the rate of 0.25mm twice a day from the third postoperative day to simulate the currently accepted callotasis. In Group I, II, and V, there two were no statistically significant differences in the growth rates between the left operated and the right normal tibiae. In contrast, the ratios of left over right tibial length decreased significantly in Group III (4.8%), and Group IV (8.6%) at the completion of lengthening(p < 0.0001), and remained decreased until near skeletal maturity. Histomorphometric study revealed that, in Group III and IV, there were statistically significant decreases in the ratios of the thickness of left over night tibiae at both proximal and distal growth plate(p < 0.0001). Immunohistochemical study also revealed that, in Group III and Group IV, the ratios of left over right bromodeoxyuridine(BrdUrd) labeling percentage decreased significantly (p < 0.0001). In summary, when the tiba was lengthened 30% or more, retardation of enchondral growth was evident, which persisted from the time of completion of lengthening to near skeletal maturity. It is postulated that unphysiologically increased pressure on the growth plate due to excessive bone lengthening may be closely related with the inhibition of enchondral growth.