A 3D analysis of growth trajectory and integration during early human prenatal facial growth.

Significant shape changes in the human facial skeleton occur in the early prenatal period, and understanding this process is critical for studying a myriad of congenital facial anomalies. However, quantifying and visualizing human fetal facial growth has been challenging. Here, we applied quantitative geometric morphometrics (GM) to high-resolution magnetic resonance images of human embryo and fetuses, to comprehensively analyze facial growth. We utilized non-linear growth estimation and GM methods to assess integrated epigenetic growth between masticatory muscles and associated bones. Our results show that the growth trajectory of the human face in the early prenatal period follows a curved line with three flexion points. Significant antero-posterior development occurs early, resulting in a shift from a mandibular prognathic to relatively orthognathic appearance, followed by expansion in the lateral direction. Furthermore, during this time, the development of the zygoma and the mandibular ramus is closely integrated with the masseter muscle.

Co-Administration of Myostatin-Targeting siRNA and ActRIIB-Fc Fusion Protein Increases Masseter Muscle Mass and Fiber Size.

Myostatin, a member of the TGF-ß superfamily, is a negative regulator of skeletal muscle cell growth and differentiation, and binds with high affinity to the activin type IIB receptor (ActRIIB). The soluble ligand-binding domain of ActRIIB fused to the Fc domain of IgG (ActRIIB-Fc) potently binds and inhibits TGF-ß family members in muscle, leading to rapid and marked muscle growth. The present study was designed to assess the effectiveness of the co-delivery of myostatin-targeting siRNA (Mstn-siRNA) and ActRIIB-Fc into skeletal muscle as a potential treatment of atrophic myopathies. Eleven-week-old, male C57BL/6 mice were injected with atelocollagen (ATCOL)-mediated Mstn-siRNA with/without ActRIIB-Fc locally into the masseter muscle twice a week. Inhibition of myostatin function by the combination of Mstn-siRNA and ActRIIB-Fc increased muscle weight and myofibril size in murine masseter muscle. Real-time RT-PCR analysis revealed significant downregulation of myostatin mRNA expression in both the Mstn-siRNA-treated and the combination treatment group. Furthermore, myogenin mRNA expression was upregulated in the combination treatment group, while MuRF-1 and Atrogin-1 mRNA expression was downregulated compared to administration of each compound alone. These findings suggest that double inhibition of myostatin is a potentially useful treatment strategy to increase muscle mass and fiber size and could be a useful treatment of patients with various muscle atrophies, including muscular dystrophy.

Evaluation of masseter muscles in relation to treatment with removable bite-blocks in dolichofacial growing subjects: A prospective controlled study.

INTRODUCTION: The aim of this prospective study was to evaluate the effects of posterior bite-blocks on masseter muscles and on facial growth in prepubertal dolichofacial subjects. METHODS: The treatment group comprised 21 consecutive prepubertal dolichofacial patients treated with rapid maxillary expansion followed by mandibular removable bite-blocks. Lateral cephalograms and ultrasonographic scans of the masseter muscles were made before (T1) and after (T2) treatment with bite-blocks. The treatment group was compared with a control group of 21 subjects matched for sex, age, and skeletal vertical pattern. An independent samples t test was used to compare the T1 to T2 changes in ultrasonographic scan measurements between the treatment group and the control group, and the T1 to T2 cephalometric changes in the treatment group. Regression analysis was performed to investigate associations between masseter muscle thickness and cephalometric treatment outcomes. RESULTS: Masseter muscle thickness showed a statistically significant decrease (-0.7 mm) in the treatment group compared with an increase (+0.6 mm) in the control group. A significant anterior rotation of the mandibular plane was observed in the treatment group as well as significant increases in overbite (1.8 mm) and total posterior facial height (1.5 mm). No significant associations were found between masseter muscle thickness and treatment outcomes apart from a tendency for overbite to increase more in subjects with thicker muscles. CONCLUSIONS: Treatment with removable bite-blocks produced a decrease in masseter muscle thickness and a reduction in vertical facial dimensions due to upward and forward rotation of the mandible. No significant correlation was found between the pretreatment masseter muscle thickness and the T1 to T2 cephalometric changes in the treatment group.

Genetic response in masseter muscle after orthognathic surgery in comparison with healthy controls - A Microarray study.

One third of adult patients with orthognathic surgery of a prognathic or retrognathic mandible show relapse. The sagittal split osteotomy of the mandible leads to a displacement of both parts up to 10 mm without any changes of muscle attachment. Changed mandible length needs adaptation of muscle capacity because of changed force to moment ratio. The aim of this Microarray study was to analyze the general genetic response of masseter muscle in patients with retrognathism or prognathism of the mandible six months after surgery in comparison with healthy untreated controls. We found in tissue samples from masseter muscle a reduction of different entities between patients and controls but less in retrognathic than in prognathic patients (274/429). The different entities to controls in prognathia were reduced from 1862 to 1749 but increased in retrognathia from 1070 to 1563. We have to consider that the total amount of different entities to the controls is higher in patients with prognathic mandible (7364) because of their strong genetic controlled development compared with that in patients with retrognathic mandible (4126), which is more environmentally influenced. It can be concluded that function follows form after surgical change with high inheritance. In retrognathic patients the adaptation could be delayed or the capacity of regeneration potential is not sufficient.

Postnatal changes in glutamatergic inputs of jaw-closing motoneuron dendrites.

Dendrites of masseter (jaw-closing) motoneurons (MMNs) are well developed and ramify extensively throughout the trigeminal motor nucleus and often extend into the adjacent reticular formation. It is possible that the dendrites have active properties, which are altered with the development of the orofacial musculoskeletal system. Thus, we examined the changes in somatic voltage responses evoked by photostimulation of the MMN dendrites by laser photolysis of caged glutamate from postnatal day (P) 2-5 and 9-12 rats. We photostimulated 39 spots that were arranged around each recorded neuron in a concave shape and found that the dendritic stimulation induced somatic depolarization in the presence of tetrodotoxin in all MMNs. With increasing photostimulation intensity, the responses grew in amplitude up to a certain threshold, where a step-like increase in amplitude occurred. In 75% of P2-5 MMNs, the step-like increase in amplitude, which was blocked by 20µM D(-)-2-amino-5-phosphonovaleric acid application, corresponded to the NMDA spikes/plateau potentials. In contrast, at P9-12 the responses became significantly smaller in amplitude and shorter in duration and only one neuron out of 12 MMNs showed NMDA spikes/plateau potentials. These results suggest that the glutamatergic responses evoked by photostimulation of the MMN dendrites change during the first two postnatal weeks, and these changes may be involved in the transition from suckling to chewing during postnatal development.

Expression of CGRP in embryonic mouse masseter muscle.

Neuropeptide calcitonin gene-related peptide (CGRP) is a mediator of inflammation and head pain that influences the functional vascular blood supply. The CGRP also regulate myoblast and acetylcholine receptors on neuromuscular junctions in development. However, little is known about its appearance and location during mouse masseter muscle (MM) development. We detected the mRNA abundance of CGRP, vascular genesis markers (Vascular endothelial growth factor A (VEGF-A), PECAM (CD31), lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1)) and embryonic and adult myosin heavy chain (MyHCs) (embryonic, IIa, IIb, and IIx) using real-time RT-PCR during development from the embryonic stage to after birth (E12.5, E14.5, E17.5, E18.5, P0, P1 and P5). We also endeavored to analyze the expression and localization of CGRP in situ hybridization in the developing mouse MM during development from the embryonic stage to after birth (E12.5, E14.5, E17.5, and P1). The antisense probe for CGRP was detected by in situ hybridization at E12.5, E14.5 E17.5 and then no longer detected after birth. The CGRP, CD31, embryonic MyHC abundance levels are highest at E17.5 (p<0.001) and they show a pattern similar to that of the other markers from E12.5 to P5. PCA analysis indicates a specific relation between CGRP and embryonic MyHC, CD31, and LYVE-1 in MM development. Cluster analyses identified the following distinct clusters for mRNA abundance in the MM: cluster 1, P5; cluster 2, E12.5, E14.5, E17.5, E18.5, P0, and P1. The positive correlation between CGRP and embryonic MyHC (Pearson's r>0.65; p<0.01) was analyzed. These data suggested that CGRP may have an influence on embryonic MyHC during mouse MM development. CGRP also affects the angiogenesis markers at embryonic stages.

Functions of miR-1 and miR-133a during the postnatal development of masseter and gastrocnemius muscles.

The present study investigated the function of miR-1 and miR-133a during the postnatal development of mouse skeletal muscles. The amounts of miR-1 and miR-133a were measured in mouse masseter and gastrocnemius muscles between 1 and 12 weeks after birth with real-time polymerase chain reaction and those of HDACs, MEF2, MyoD family, MCK, SRF, and Cyclin D1 were measured at 2 and 12 weeks with Western blotting. In both the masseter and gastrocnemius muscles, the amount of miR-1 increased between 1 and 12 weeks, whereas the amount of HADC4 decreased between 2 and 12 weeks. In the masseter muscle, those of MEF2, MyoD, Myogenin, and MCK increased between 2 and 12 weeks, whereas, in the gastrocnemius muscle, only those of MRF4 and MCK increased. The extent of these changes in the masseter muscle was greater than that in the gastrocnemius muscle. The amounts of miR-133a, SRF, and Cyclin D1 did not change significantly in the masseter muscle between 1 and 12 weeks after birth. By contrast, in the gastrocnemius muscle, the amounts of miR-133a and Cyclin D1 increased, whereas that of SRF decreased. Our findings suggest that the regulatory pathway of miR-1 via HDAC4 and MEF2 plays a more prominent role during postnatal development in the masseter muscle than in the gastrocnemius muscle, whereas that of miR-133a via SRF plays a more prominent role in the gastrocnemius muscle than in the masseter muscle.

Effect of occlusal hypofunction and its recovery on the three-dimensional architecture of mandibular alveolar bone in growing rats.

BACKGROUD: Normal occlusion is very important for physiological structure of mandible. However, the details of influences of occlusal hypofunction and its recovery on the three-dimensional architecture of mandibular alveolar bone in growing rats are still lacking. MATERIALS AND METHODS: Forty-eight growing male Sprague-Dawley rats were randomly divided into normal (n = 24), hypofunctional (n = 12), and recovery (n = 12) groups. The hypofunction group was developed by inserting a bite-raising appliance between the maxillary and mandibular incisors of the rats. Two weeks after insertion, the appliance was removed to result in the recovery group; the experiment continued for two additional weeks. The experimental animals and control animals were killed weekly. In addition to measuring the body weight and masseter muscle weight of the rats, the histomorphology and microstructure of the mandibular alveolar bone were scanned using microcomputed tomography. RESULTS: A lighter masseter muscle and a higher and narrower alveolar process were observed in the hypofunction group compared with the control animals (P < 0.05). Mandibular remodeling also occurred in the hypofunctional group, as demonstrated by a smaller trabecular cross-sectional area, looser trabecular bone, decreased bone volume fraction, trabecular thickness, trabecular number, and increased bone surface density and trabecular separation, especially at week 2 (P < 0.05). After removing the anterior bite-opening appliance, the altered masseter muscle weight and architecture of the mandibular alveolar bone were gradually reversed and reached normal levels at the end of the experiment (P > 0.05). CONCLUSIONS: A loss of occlusal stimuli can lead into mandibular alveolar bone remodeling, and the recovery of occlusion can restore the altered mandibular architecture in growing rats.

Effectiveness of cationic liposome-mediated local delivery of myostatin-targeting small interfering RNA in vivo.

This study evaluated the effectiveness of local administration of cationic liposome-delivered myostatin-targeting siRNA. Myostatin (Mst)-siRNA and scrambled (scr)-siRNA-lipoplexes were injected into the masseter muscles of wild type and dystrophin-deficient mdx mice, which model Duchenne muscular dystrophy. One week after injection, the masseter muscles were dissected for histometric analyses. To evaluate changes in masseter muscle activity, masseter electromyographic (EMG) measurements were performed. One week after local administration of Mst-siRNA-lipoplexes, masseter muscles and myofibrils were significantly larger compared to control masseter muscles treated with scr-siRNA-lipoplexes. Real-time polymerase chain reaction (PCR) analyses revealed significant upregulation of the myogenic regulatory factors MyoD and myogenin and significant downregulation of the adipogenic transcription factors peroxisome proliferator-activated receptor-γ (PPARγ) and CCAAT/enhancer binding protein-α (CEBPα) in masseter muscles treated with Mst-siRNA-lipoplexes. The duty times of masseter muscle activity exceeding 5% showed a slight tendency to increase in both wild type and mdx mice. Therefore, cationic liposome-mediated local administration of Mst-siRNA could increase muscular size and improve muscle activity. Since cationic liposomes delivered siRNA to muscles effectively and are safe and cost-effective, they may represent a therapeutic tool for use in treating muscular diseases.

The effects of denervation and formoterol administration on facial growth.

OBJECTIVE: To identify and demonstrate possible alterations of skeletal structures which might follow either unilateral surgical denervation of the masseter muscle, unilateral intramuscular injection of formoterol directly into the masseter muscle, or intramuscular formoterol injection after surgical denervation. MATERIALS AND METHODS: Male Sprague Dawley rats (N = 16; four weeks of age) were prepared as four groups: 1. surgical sham + saline injection into the masseter muscle (sham); 2. surgical denervation of the masseter muscle only (den.); 3. surgical denervation of the masseter muscle plus intramuscular formoterol injection into the affected muscle (den.+form.); 4. intramuscular formoterol injection into the masseter muscle only (form.). The specimens were submitted for CT examination, the skulls and hemimandibles were photographed and measurements of craniofacial bones were made. RESULTS: In this relatively small sample, comparisons between non-experimental and experimental sides revealed differences, both within the groups and for the same measurements between groups, with the den. and den.+form. groups showing the most change. Relative increases in the gonial angle shown in these groups occurred bilaterally, with the change on the experimental side always greater in magnitude than the change on the contralateral side. CONCLUSIONS: Surgical denervation of the masseter muscle leads to an alteration in the size and shape of the skeletal structures close to the zygoma and the mandible. The intramuscular injection of formoterol into denervated masseter muscle seems to limit this skeletal alteration after surgical denervation.

Increased vertical dimension effects on masseter muscle fiber phenotype during maturation.

OBJECTIVE: To determine changes in mouse myosin heavy chain (MyHC) protein expression that may occur with a clinically relevant vertical dimension of occlusion (VDO) increase. MATERIALS AND METHODS: Six CD-1 male mice (age: 6 weeks) underwent a 10% bite opening to replicate the clinical condition using composite on the maxillary molars and were compared to six age-matched controls. Mice were sacrificed at day 7 and 14 after bite opening. A representative masseter transverse cryosection from each animal was examined in selected sampling regions (anterior, posterior, posterior-deep, and posterior-intermediate) to assay fiber phenotype proportions and fiber size. RESULTS: In control masseter muscles, the proportion of muscle fibers containing MyHC IIb increased in the posterior-intermediate and posterior-deep regions between 7 and 14 days (ANOVA, P < .05). The increase in the proportion of MyHC IIb fibers in the bite opening group did not occur when compared to the control group (P < .05). In addition, after 14 days of bite opening, the proportion of fibers positive for MyHC IIa was decreased in the anterior region compared to control masseter muscles. Muscle fiber diameter remained unchanged in both groups (experimental and control) and over time (P > .10). CONCLUSION: These data are consistent with a selective plasticity of the expression of MyHC IIb protein in the deep regions of the male masseter muscle in response to a clinically relevant VDO increase.

Craniofacial growth variations in nasal-breathing, oral-breathing, and tracheotomized children.

INTRODUCTION: Childhood oral breathing can alter muscular balance and lead to facial deformities. No articles in the literature have reported on the alteration of facial growth patterns in patients who have received tracheotomies. The purpose of this study was to evaluate craniofacial developmental consequences originating from variations in breathing mechanisms in children who are nasal breathers or oral breathers, and those who have been tracheotomized. METHODS: The sample was divided into 3 groups of 10 each. The nasal group had a mean age of 13.9 years, the oral group had a mean age of 12.7 years, and the tracheotomy group had a mean age of 12.8 years. The masseter and suprahyoid muscles were evaluated with electromyography. The following measurements were made: facial, maxillary, and mandibular widths; nasion-sella-gnathion angle; and facial index. RESULTS: The tracheotomized group was similar to the nasal group for greater activity of the masseter muscles than of the suprahyoid muscles during mastication, as well as in the measurements of facial, maxillary, and mandibular widths. The oral group showed reductions in each category. The tracheotomized group was similar to the oral group during maximum dental occlusion for significantly higher activity of the suprahyoid muscles compared with the masseter muscles, with reductions in vertical values. CONCLUSIONS: A childhood tracheotomy might affect facial development in a way comparable with that of oral breathers, including abnormal facial growth variations.

Differences in fibre type composition between human masseter and biceps muscles in young and adults reveal unique masseter fibre type growth pattern.

The human jaw system is different from those of other primates, carnivores, ruminants, and rodents in temporomandibular joint and muscle anatomy. In adults, jaw muscles also differ markedly from limb and trunk muscles in composition and distribution of fibre types. It can be assumed that age-related changes between young age to adulthood in terms of craniofacial growth, teeth eruption, and improvement of jaw functions are paralleled by alterations also in composition and distribution of jaw muscle fibre types. To address this question, we have examined the fibre type composition of the human masseter, a jaw closing muscle, at young age. For comparison, the young biceps brachii was examined. The results were compared with previous data for adult masseter and biceps muscles. Young masseter and biceps were similar in that type I fibres outnumbered other fibre types and were of the same diameter. However, they differed in composition of other fibre types. Young masseter contained fibre types I, IM, IIC, IIAB, IIB, and scarce IIA, with regional differences, whereas young biceps showed types I, IIA, IIAB, and few IIB. Young masseter differed from young biceps also by smaller type II fibre diameter and by containing fetal MyHC. In addition, the masseter and biceps differed in age-related changes of composition and distribution of fibre types between young age and adulthood. We conclude that the human masseter is specialized in fibre types already at young age and shows a unique fibre type growth pattern, in concordance with being a separate allotype of muscle.

Masticatory hypermuscularity is not related to reduced cranial volume in myostatin-knockout mice.

It has been suggested recently that masticatory muscle size reduction in humans resulted in greater encephalization through decreased compressive forces on the cranial vault. Following this logic, if masticatory muscle size were increased, then a reduction in brain growth should also occur. This study was designed to test this hypothesis using a myostatin (GDF-8) knockout mouse model. Myostatin is a negative regulator of skeletal muscle growth, and individuals lacking this gene show significant hypermuscularity. Sixty-two [32 wild-type (WT) and 30 GDF-8 -/- knockout], 1, 28, 56, and 180-day-old CD-1 mice were used. Body and masseter muscle weights were collected following dissection and standardized lateral and dorsoventral cephalographs were obtained. Cephalometric landmarks were identified on the radiographs and cranial volume was calculated. Mean differences were assessed using a two-way ANOVA. KO mice had significantly greater body and masseter weights beginning at 28 days compared with WT controls. No significant differences in cranial volumes were noted between KO and WT. Muscle weight was not significantly correlated with cranial volume in 1, 28, or 180-day-old mice. Muscle weights exhibited a positive correlation with cranial volume at 56 days. Results demonstrate that masticatory hypermuscularity is not associated with reduced cranial volume. In contrast, there is abundant data demonstrating the opposite, brain growth determines cranial vault growth and masticatory apparatus only affects ectocranial morphology. The results presented here do not support the hypothesis that a reduction in masticatory musculature relaxed compressive forces on the cranial vault allowing for greater encephalization.

Muscle spindle composition and distribution in human young masseter and biceps brachii muscles reveal early growth and maturation.

Significant changes in extrafusal fiber type composition take place in the human masseter muscle from young age, 3-7 years, to adulthood, in parallel with jaw-face skeleton growth, changes of dentitions and improvement of jaw functions. As motor and sensory control systems of muscles are interlinked, also the intrafusal fiber population, that is, muscle spindles, should undergo age-related changes in fiber type appearance. To test this hypothesis, we examined muscle spindles in the young masseter muscle and compared the result with previous data on adult masseter spindles. Also muscle spindles in the young biceps brachii muscle were examined. The result showed that muscle spindle composition and distribution were alike in young and adult masseter. As for the adult masseter, young masseter contained exceptionally large muscle spindles, and with the highest spindle density and most complex spindles found in the deep masseter portion. Hence, contrary to our hypothesis, masseter spindles do not undergo major morphological changes between young age and adulthood. Also in the biceps, young spindles were alike adult spindles. Taken together, the results showed that human masseter and biceps muscle spindles are morphologically mature already at young age. We conclude that muscle spindles in the human young masseter and biceps precede the extrafusal fiber population in growth and maturation. This in turn suggests early reflex control and proprioceptive demands in learning and maturation of jaw motor skills. Similarly, well-developed muscle spindles in young biceps reflect early need of reflex control in learning and performing arm motor behavior.

Electromyographic study of the masseter muscle after lower third molar surgery / Estudio electromiográfico del músculo masetero tras cirugía de tercer molar inferior

Third molar extraction surgery is one of the most frequently performed procedures in the areas of buccal-maxillofacial traumatology and surgery. The post-surgery evolution was evaluated based on the clinical evidence obtained so far. The objective of this study was to analyze the post-surgery clinical evolution of the masseter muscle by means of surface electromyography, to evaluate muscle activity. Four analyses were performed: one pre-surgery, to register the normal activity, and three in post-surgery: on the 7th, 14th and 21st postoperative days, in a sample of 30 patients. On the 21st day, there was near normal recovery of the electrical signal of the masseter in women, but in men this activity did not reach normal levels. Surface electromyography is a safe and reliable tool for post-surgery evolution control of masseter function.

La cirugía de extracción del tercer molar es uno de los procedimientos realizados con mayor frecuencia en las áreas de traumatología y cirugía buco-maxilofacial. La evolución post-quirúrgica fue evaluada sobre la base de la evidencia clínica obtenida hasta el momento. El objetivo de este estudio fue analizar la evolución clínica post-operatoria del músculo masetero por medio de electromiografía de superficie, para evaluar la actividad muscular. Fueron realizados cuatro análisis: uno previo a la cirugía, para registrar la actividad normal, y tres en la post-quirúrgica: en el día del 7, 14 y 21 postoperatorio, en una muestra de 30 pacientes. El día 21, hubo recuperación cercana a la normal de la señal eléctrica del músculo masetero en las mujeres, pero en los hombres esta actividad no alcanzó los niveles normales. La electromiografía de superficie es una herramienta segura y fiable para el control de la evolución post-quirúrgica de la función del músculo masetero.

The effects of tooth extraction on alveolar bone biomechanics in the miniature pig, Sus scrofa.

OBJECTIVE: This study investigated the role of occlusion in the development of biomechanical properties of alveolar bone in the miniature pig, Sus scrofa. The hypothesis tested was that the tissues supporting an occluding tooth would show greater stiffness and less strain than that of a non-occluding tooth. DESIGN: Maxillary teeth opposing the erupting lower first molar (M(1)) were extracted on one side. Occlusion developed on the contralateral side. Serially administered fluorochrome labels tracked bone mineralisation apposition rate (MAR). A terminal experiment measured in vivo buccal alveolar bone strain on occluding and non-occluding sides during mastication. Ex vivo alveolar strains during occlusal loading were subsequently measured using a materials testing machine (MTS/Sintech). Whole specimen stiffness and principal strains were calculated. RESULTS: MAR tended to be higher on the extraction side during occlusion. In vivo buccal shear strains were higher in the alveolar bone of the occluding side vs. the extraction side (mean of 471 microvarepsilon vs. 281 microvarepsilon, respectively; p=0.04); however, ex vivo shear strains showed no significant differences between sides. Stiffness differed between extraction and occlusion side specimens, significantly so in the low load range (344 vs. 668 MPa, respectively; p=0.04). CONCLUSIONS: Greater in vivo shear strains may indicate more forceful chews on the occluding side, whereas the similarity in ex vivo bone strain magnitude suggests a similarity in alveolar bone structure and occlusal load transmission regardless of occlusal status. The big overall change in specimen stiffness that was observed was likely attributable to differences in the periodontal ligament rather than alveolar bone.

Masticatory biomechanics and masseter fiber-type plasticity.

Compared to force-resisting elements of the mammalian feeding apparatus, data on jaw-muscle plasticity are less common. This hinders our understanding of the role of force-producing structures in craniofacial development and integration. Thus, we investigated fiber-type abundance and cross-sectional area in the masseter muscle of growing rabbits subjected to diet-induced variation in masticatory stresses. Three loading cohorts were obtained as weanlings and raised until adult on different diets. Immediately following euthanasia, left-sided masseters were dissected away, weighed, and then divided into anterior, intermediate and posterior sections for fiber-type immunohistochemistry. These data were compared to mandibular proportions and biomineralization from the same subjects. Results indicate that growing mammals fed a tougher, fracture-resistant diet develop: absolutely and relatively lower numbers of Type I jaw-muscle fibers; absolutely larger fiber cross-sectional areas; and relative increases in the amount of Type II fibers. These analyses indicate that an early postweaning dietary shift can induce significant variation in muscle fiber types. Such norms of reaction are comparable to those observed in bony elements. Functionally, the processing of fracture-resistant foods results in jaw adductors potentially characterized by faster contraction times and higher force production capabilities, which may influence the frequency and amplitude of forces experienced by oral tissues.

Properties of synaptic transmission from the reticular formation dorsal to the facial nucleus to trigeminal motoneurons during early postnatal development in rats.

We previously reported that electrical stimulation of the reticular formation dorsal to the facial nucleus (RdVII) elicited excitatory masseter responses at short latencies and that RdVII neurons were antidromically activated by stimulation of the trigeminal motor nucleus (MoV), suggesting that excitatory premotor neurons targeting the MoV are likely located in the RdVII. We thus examined the properties of synaptic transmission from the RdVII to jaw-closing and jaw-opening motoneurons in horizontal brainstem preparations from developing rats using voltage-sensitive dye, patch-clamp recordings and laser photostimulation. Electrical stimulation of the RdVII evoked optical responses in the MoV. Combined bath application of the non-N-methyl-d-aspartate (non-NMDA) receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), and the NMDA receptor antagonist DL-2-amino-5-phosphonopentanoic acid (APV) reduced these optical responses, and addition of the glycine receptor antagonist strychnine and the GABA(A) receptor antagonist bicuculline further reduced the remaining responses. Electrical stimulation of the RdVII evoked postsynaptic currents (PSCs) in all 19 masseter motoneurons tested in postnatal day (P)1-4 rats, and application of CNQX and the NMDA receptor antagonist (+/-)-3(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) reduced the PSC amplitudes by more than 50%. In the presence of CNQX and CPP, the GABA(A) receptor antagonist SR95531 further reduced PSC amplitude, and addition of strychnine abolished the remaining PSCs. Photostimulation of the RdVII with caged glutamate also evoked PSCs in masseter motoneurons of P3-4 rats. In P8-11 rats, electrical stimulation of the RdVII also evoked PSCs in all 14 masseter motoneurons tested, and the effects of the antagonists on the PSCs were similar to those in P1-4 rats. On the other hand, RdVII stimulation evoked PSCs in only three of 16 digastric motoneurons tested. These results suggest that both neonatal and juvenile jaw-closing motoneurons receive strong synaptic inputs from the RdVII through activation of glutamate, glycine and GABA(A) receptors, whereas inputs from the RdVII to jaw-opening motoneurons seem to be weak.

Age-related changes in craniofacial morphology in GDF-8 (myostatin)-deficient mice.

It is well recognized that masticatory muscle function helps determine morphology, although the extent of function on final form is still debated. GDF-8 (myostatin), a transcription factor is a negative regulator of skeletal muscle growth. A recent study has shown that mice homozygous for the myostatin mutation had increased muscle mass and craniofacial dysmorphology in adulthood. However, it is unclear whether such dysmorphology is present at birth. This study examines the onset and relationship between hypermuscularity and craniofacial morphology in neonatal and adult mice with GDF-8 deficiency. Fifteen (8 wild-type and 7 GDF-8 -/-), 1-day-old and 16 (9 wt and 7 GDF-8 -/-), 180-day-old male CD-1 mice were used. Standardized radiographs were taken of each head, scanned, traced, and cephalometric landmarks identified. Significant mean differences were assessed using a group x age, two-way ANOVA. Myostatin-deficient mice had significantly (P < 0.01) smaller body and masseter muscle weights and craniofacial skeletons at 1 day of age and significantly greater body and masseter muscle weights at 180 days of age compared to controls. Myostatin-deficient mice showed significantly (P < 0.001) longer and "rocker-shaped" mandibles and shorter and wider crania compared to controls at 180 days. Significant correlations were noted between masseter muscle weight and all cephalometric measurements in 180-day-old Myostatin-deficient mice. Results suggest that in this mouse model, there may be both early systemic skeletal growth deficiencies and later compensatory changes from hypermuscularity. These findings reiterate the role that masticatory muscle function plays on the ontogeny of the cranial vault, base, and most notably the mandible.