Reduced mastication during growth inhibits cognitive function by affecting trigeminal ganglia and modulating Wnt signaling pathway and ARHGAP33 molecular transmission.

Binding of brain-derived neurotrophic factor (BDNF) to its receptor tyrosine kinase B (TrkB) is essential for the development of the hippocampus, which regulates memory and learning. Decreased masticatory stimulation during growth reportedly increases BDNF expression while decreasing TrkB expression in the hippocampus. Increased BDNF expression is associated with Wnt family member 3A (Wnt3a) expression and decreased expression of Rho GTPase Activating Protein 33 (ARHGAP33), which regulates intracellular transport of TrkB. TrkB expression may be decreased at the cell surface and affects the hippocampus via BDNF/TrkB signaling. Mastication affects cerebral blood flow and the neural cascade that occurs through the trigeminal nerve and hippocampus. In the current study, we hypothesized that decreased masticatory stimulation reduces memory/learning in mice due to altered Wnt3a and ARHGAP33 expression, which are related to memory/learning functions in the hippocampus. To test this hypothesis, we fed mice a powdered diet until 14 weeks of age and analyzed the BDNF and TrkB mRNA expression in the right hippocampus using real-time polymerase chain reaction and Wnt3a and ARHGAP33 levels in the left hippocampus using western blotting. Furthermore, we used staining to assess BDNF and TrkB expression in the hippocampus and the number of nerve cells, the average size of each single cell and the area of intercellular spaces of the trigeminal ganglion (TG). We found that decreased masticatory stimulation affected the expression of BDNF, Wnt3a, ARHGAP33, and TrkB proteins in the hippocampus, as well as memory/learning. The experimental group showed significantly decreased numbers of neurons and increased the area of intercellular spaces in the TG. Our findings suggest that reduced masticatory stimulation during growth induces a decline in memory/learning by modulating molecular transmission mechanisms in the hippocampus and TG.

Pre-Adolescent Diet Normalization Restores Cognitive Function in Young Mice.

Mastication is a fundamental function critical for human health. Controlled by the central nervous system (CNS), it influences CNS development and function. A poor masticatory performance causes cognitive dysfunction in both older adults and children. Improving mastication may prevent cognitive decline. However, no study has determined the period of masticatory dysfunction that impairs children's later acquisition of cognitive function. Herein, we developed an animal model wherein a soft diet was switched to a normal diet at early and late time points in young mice. We aimed to investigate the impact of restored mastication on learning and memory function. Behavioral studies were conducted to evaluate learning and memory. Micro-CT was used to evaluate orofacial structural differences, while histological and biochemical approaches were employed to assess differences in the hippocampal morphology and function. Correction to a hard-textured diet before adolescence restored mastication and cognitive function through the stimulation of neurogenesis, extracellular signal-regulated kinases, the cyclic adenosine monophosphate-response element-binding protein pathway, and the brain-derived neurotrophic factor, tyrosine receptor B. In contrast, post-adolescent diet normalization failed to rescue full mastication and led to impaired cognitive function, neuronal loss, and decreased hippocampal neurogenesis. These findings revealed a functional linkage between the masticatory and cognitive function in mice during the juvenile to adolescent period, highlighting the need for adequate food texture and early intervention for mastication-related cognitive impairment in children.

Effect of Obesity on Masticatory Muscle Activity and Rhythmic Jaw Movements Evoked by Electrical Stimulation of Different Cortical Masticatory Areas.

This study investigates rhythmic jaw movement (RJM) patterns and masticatory muscle activities during electrical stimulation in two cortical masticatory areas in obese male Zucker rats (OZRs), compared to their counterparts-lean male Zucker rats (LZRs) (seven each). At the age of 10 weeks, electromyographic (EMG) activity of the right anterior digastric muscle (RAD) and masseter muscles, and RJMs were recorded during repetitive intracortical micro-stimulation in the left anterior and posterior parts of the cortical masticatory area (A-area and P-area, respectively). Only P-area-elicited RJMs, which showed a more lateral shift and slower jaw-opening pattern than A-area-elicited RJMs, were affected by obesity. During P-area stimulation, the jaw-opening duration was significantly shorter (p < 0.01) in OZRs (24.3 ms) than LZRs (27.9 ms), the jaw-opening speed was significantly faster (p < 0.05) in OZRs (67.5 mm/s) than LZRs (50.8 mm/s), and the RAD EMG duration was significantly shorter (p < 0.01) in OZRs (5.2 ms) than LZR (6.9 ms). The two groups had no significant difference in the EMG peak-to-peak amplitude and EMG frequency parameters. This study shows that obesity affects the coordinated movement of masticatory components during cortical stimulation. While other factors may be involved, functional change in digastric muscle is partly involved in the mechanism.

Unilateral nasal obstruction affects development of cortical orofacial motor representation in the cortical masticatory area of growing rats.

The cortical masticatory area (CMA) regulates masticatory movements. However, there is limited information regarding the effect of respiratory disturbances on the functional development of the CMA. This study aimed to examine the effect of chronic unilateral nasal obstruction (UNO) on the CMA during the growth period. Forty-eight 8-day-old male Wistar albino rats were randomized into control (CONT, n = 24) and UNO (n = 24) groups. Arterial oxygen saturation (SpO2) and the number of positive intracortical microstimulation (ICMS) sites in the anterior digastric (AD) muscle were recorded at 5, 7, and 9 weeks of age. ICMS was performed under anesthesia. The CONT group exhibited significantly higher SpO2 values than the UNO group at each age. Furthermore, the number of positive ICMS sites in the AD muscle was significantly higher in the UNO group than in the CONT group at each age. The right AD was more strongly affected by UNO than the left AD. In addition, it was found that the P-area was more strongly affected by UNO than the A-area. These findings suggest that UNO during development induces changes in the motor representation in the CMA, which may affect respiratory and orofacial functions in rats. Therefore, we suggest that the development of respiratory dysfunction needs to be monitored and corrected promptly to avoid complications in physiological function.

Nasal obstruction during the growth period modulates the Wnt/ß-catenin pathway and brain-derived neurotrophic factor production in association with tyrosine kinase receptor B mRNA reduction in mouse hippocampus.

There is accumulating evidence that nasal obstruction induces high-level brain dysfunction, including memory and learning deficits. We previously demonstrated that unilateral nasal obstruction (UNO) during the growth period increases the expression of brain-derived neurotrophic factor (BDNF). The expression of BDNF is regulated by the Wnt/ß-Catenin pathway, which is linked to neuronal differentiation, proliferation, and maintenance. However, little is known about whether Wnt3a protein expression could be an index for modulations analyses in the Wnt/ß-Catenin pathway caused by UNO during the growth period. This study aimed to investigate the effects of UNO during the growth period on the Wnt/ß-Catenin pathway in the hippocampus using combined behavioural, biochemical, and histological approaches. Male BALB/C mice were randomly divided into the control (CONT; n = 6) and experimental (UNO; n = 6) groups. Blood oxygen saturation (SpO2 ) levels were measured, and a passive avoidance test was performed in mice aged 15 weeks. Brain tissues were subjected to immunohistochemistry, real-time reverse transcription-polymerase chain reaction, and western blot analysis. Compared with control mice, UNO mice had lower SpO2 levels and exhibited memory/learning impairments during behavioural testing. Moreover, Wnt3a protein, BDNF mRNA, and tyrosine kinase receptor B (TrkB) mRNA expression levels were significantly lower in the hippocampus in the UNO group than in the CONT group. Our findings suggested that UNO during the growth period appeared to modulate the hippocampal Wnt/ß-catenin pathway and BDNF production in association with TrkB mRNA reduction, thereby resulting in memory and learning impairments.

Effects of low occlusal loading on the neuromuscular behavioral development of cortically-elicited jaw movements in growing rats.

The effect of altered occlusal force on masticatory-related neuromuscular control, which projects from the anterior part of the cortical masticatory area (A-CMA), during growth remains unclear. This study sought to evaluate the effect of occlusal hypofunction on neuromuscular development of jaw muscle activities and cortically-induced rhythmic jaw movements (RJMs) in growing rats. Sixty-four 2-week-old male albino Wistar rats were divided into the control (fed normal diet) and experimental (fed soft diet) groups soon after weaning. Electromyographic activity was recorded at 5, 7, 9, and 11 weeks from the right masseter and anterior digastric along with RJMs. We found a significantly longer onset latency and smaller peak-to-peak amplitude in the experimental group than that in the control group. The RJMs showed an increase in gape size and lateral excursion until up to 9 weeks in both groups. However, both the average gape size and lateral excursion were significantly smaller in the experimental group than that in the control group after 9 weeks. The jaw movement pattern also showed a significant decrease at the maximum opening period in the experimental group. Our findings indicate that inadequate occlusal function during growth alters neuromuscular control of masticatory behaviors and impairs the pattern of RJMs.

Correction of bilateral heavily impacted second molar with improved super-elastic nickel-titanium alloy wires.

Impaction and an abnormal eruption of a second molar occur because of a lack of retromolar space, premature eruption of the mandibular third molar, and ankyloses. The prognosis must be carefully considered when extracting a damaged permanent tooth. In addition to extraction for orthodontics, this enables the establishment of an effective force system to align the impacted tooth. A 21-year-old woman exhibited maxillary anterior crowding, deviation of the maxillary midline, and deeply impacted mandibular second molars bilaterally. The molar relationship was Class II. The cephalometric analysis demonstrated a skeletal Class I relationship (ANB angle, 1.4°); maxillary and mandibular incisors were lingually inclined. Cone-beam computed tomography images indicated that root resorption, caries, and periodontitis were absent in all mandibular molar. Mandibular second premolars were extracted to relieve crowding and achieve Class I molar relationships. The second molars moved mesially on both sides, and there were no signs of ankylosis. We used improved super-elastic nickel-titanium alloy wire (ISW) to upright the mesioangular second molars. We heat-treated the anterior portion of the ISW, including the first molar area, to increase wire stiffness; the posterior portion of the ISW, including the impacted second molar area, remained untreated to ensure that its super-elasticity was preserved. We alleviated crowding, corrected the maxillary midline, and created ideal occlusion with Class I relationship. This case shows that the alignment of a deeply impacted tooth with a heat-treated ISW, combined with voluntary adjustment of wire stiffness, can be a simple and useful treatment option for adult patients.

Functional Analysis of Rhythmic Jaw Movements Evoked by Electrical Stimulation of the Cortical Masticatory Area During Low Occlusal Loading in Growing Rats.

The maturation of rhythmic jaw movements (RJMs) and related neuromuscular control has rarely been studied in animals, though this process is essential for regulating the development of stomatognathic functions. Previous studies have shown that occlusal hypofunction during growth alters masticatory performance. However, little is known about patterns of cortically-induced RJMs under conditions of soft-diet feeding during development. The aim of this study is to clarify the effect of low occlusal loading on the pattern of cortically induced RJMs and related neuromuscular responses in growing rats. Sixty-four 2-week-old male albino Wistar rats were randomly divided into two groups and fed on either a normal diet (control) or soft diet (experimental) soon after weaning. At 5, 7, 9, and 11 weeks of age, electromyographic (EMG) activity was recorded from the right masseter and anterior digastric muscles along with corresponding kinematic images in RJMs during repetitive intracortical microstimulation of the left cortical masticatory area (CMA). Rats in both groups showed an increase in gape size and lateral excursion until 9 weeks of age. The vertical jaw movement speed in both groups showed no significant difference between 5 and 7 weeks of age but increased with age from 9 to 11 weeks. Compared to the control group, the average gape size and vertical speed were significantly lower in the experimental group, and the pattern and rhythm of the jaw movement cycle were similar between both groups at each recording age. EMG recordings showed no age-related significant differences in onset latency, duration, and peak-to-peak amplitude. Moreover, we found significantly longer onset latency, smaller peak-to-peak amplitude, and greater drop-off mean and median frequencies in the experimental group than in the control group, while there was no significant difference in the duration between groups. These findings indicate that a lack of enough occlusal function in infancy impedes the development of patterns of RJMs and delays the neuromuscular response from specific stimulation of the CMA.

Nasal obstruction during adolescence induces memory/learning impairments associated with BDNF/TrkB signaling pathway hypofunction and high corticosterone levels.

The hippocampus is an important brain region involved in memory and learning. Brain-derived neurotrophic factor (BDNF), tyrosine kinase receptor B (TrkB), and phospho-p44/p42 mitogen-activated protein kinase (MAPK) are known to contribute to hippocampal memory/learning. The present study aimed to clarify the effects of nasal obstruction during the growth period on memory/learning in an animal model, using combined behavioral, biochemical, and histological approaches. Male BALB/C mice underwent unilateral nasal obstruction (UNO) by cauterization at 8 days of age and were subjected to Y-maze and passive avoidance tests at 15 weeks of age. The serum corticosterone levels were measured using an enzyme-linked immunosorbent assay, and brain tissues were subjected to hematoxylin-eosin staining and histological analysis or homogenization and Western blot analysis. Compared with control mice, UNO mice had lower blood oxygen saturation levels and exhibited apparent memory/learning impairments during behavioral testing. Additionally, the UNO group had higher hippocampal BDNF levels and serum corticosterone levels, lower hippocampal TrkB and phospho-p44/p42 MAPK levels, and reduced neuron numbers relative to controls. Our findings suggest that UNO during adolescence affects the hippocampus and causes memory/learning impairments.

Unilateral nasal obstruction affects motor representation development within the face primary motor cortex in growing rats.

Postnatal growth is influenced by genetic and environmental factors. Nasal obstruction during growth alters the electromyographic activity of orofacial muscles. The facial primary motor area represents muscles of the tongue and jaw, which are essential in regulating orofacial motor functions, including chewing and jaw opening. This study aimed to evaluate the effect of chronic unilateral nasal obstruction during growth on the motor representations within the face primary motor cortex (M1). Seventy-two 6-day-old male Wistar rats were randomly divided into control (n = 36) and experimental (n = 36) groups. Rats in the experimental group underwent unilateral nasal obstruction after cauterization of the external nostril at 8 days of age. Intracortical microstimulation (ICMS) mapping was performed when the rats were 5, 7, 9, and 11 wk old in control and experimental groups (n = 9 per group per time point). Repeated-measures multivariate ANOVA was used for intergroup and intragroup statistical comparisons. In the control and experimental groups, the total number of positive ICMS sites for the genioglossus and anterior digastric muscles was significantly higher at 5, 7, and 9 wk, but there was no significant difference between 9 and 11 wk of age. Moreover, the total number of positive ICMS sites was significantly smaller in the experimental group than in the control at each age. It is possible that nasal obstruction induced the initial changes in orofacial motor behavior in response to the altered respiratory pattern, which eventually contributed to face-M1 neuroplasticity.NEW & NOTEWORTHY Unilateral nasal obstruction in rats during growth periods induced changes in arterial oxygen saturation (SpO2) and altered development of the motor representation within the face primary cortex. Unilateral nasal obstruction occurring during growth periods may greatly affect not only respiratory function but also craniofacial function in rats. Nasal obstruction should be treated as soon as possible to avoid adverse effects on normal growth, development, and physiological functions.

Effects of increased occlusal vertical dimension on the jaw-opening reflex in adult rats.

OBJECTIVE: Malocclusion with deep overbite and facial esthetics improve when facial height is intentionally increased during orthodontic extrusion of the posterior teeth. Thus, a better understanding of post-treatment stability of increased occlusal vertical dimension (iOVD) in adult patients is important. We focused on the jaw-opening reflex (JOR), which plays an important role in the control of jaw movements during mastication, and investigated the effects of iOVD on the JOR in rats with an electrophysiological technique. DESIGN: One hundred and twenty 13-week-old male Wistar rats were randomly divided into control and experimental groups. Rats in the experimental group received a 2-mm buildup of composite resin on the maxillary molars at 13 weeks of age. The JOR was induced by low-intensity electrical stimulation of the left inferior alveolar nerve. The electromyographic responses were recorded from the digastric muscle at 13, 14, 15, 17, 19, and 23 weeks of age. JOR properties including latency, duration, and peak-to-peak amplitude were measured and compared between the groups. RESULTS: The latency of the JOR was significantly longer and the peak-to-peak amplitude was significantly smaller in the experimental group than in the control group from 14 to 19 weeks of age, while the reflex duration was not significantly different. Intra-group comparisons of the latency and peak-to-peak amplitudes among rats 14-19 weeks of age were significantly different between the experimental group and the control group. CONCLUSIONS: iOVD affected the latency and amplitude of the JOR but not the duration. The JOR adapted after 10 weeks of iOVD.

Biochemical properties in membrane of rat astrocytes under oxidative stress.

Oxidative stress induced by the treatment with 100 µM hydrogen peroxide (H2O2) for 10 min enhances release of cytosolic proteins along with fibroblast growth factor 1 (FGF-1) from rat astrocytes without inducing apoptosis. FGF-1 promotes the generation of apolipoprotein E-containing high-density lipoprotein-like particles (apoE/HDL) in astrocytes, which contributes to cholesterol homeostasis in the brain. In this work, we studied various effects of oxidative stress on rat astrocyte׳s membrane to understand the mechanism underlying release of cytosolic proteins and FGF-1. The oxidative stress using 100 µM H2O2 enhanced lipid release from rat astrocytes in addition to suppression of lipid synthesis. The lipid synthesis, however, was hardly suppressed by H2O2 in the cell lines such as bovine endothelial cells and HepG2 cells from which the release of cytosolic proteins is not increased by H2O2 unlike rat astrocytes. The treatment of rat astrocytes with H2O2 changed the distribution of lipids and proteins in the caveolin-1-rich domain of membrane to the non-raft domain, which was canceled by the pretreatment of cells with low-density lipoproteins (LDL). These findings suggest that oxidative stress induced by H2O2 changes lipid level of the plasma membrane to make the membrane structure fragile in rat astrocytes. The direct treatment with H2O2 of membrane fraction prepared from rat astrocytes did not enhance lipid release from the membrane. The lipid release, however, was enhanced from the isolated membrane fraction, after the cells were treated with H2O2 and incubated in H2O2-free DPBS. Hydrogen peroxide enhanced phosphorylation of protein kinases such as Akt, MEK, and ERK in intact astrocytes without injury and stress. A MEK inhibitor, U0126, suppressed not only the H2O2-induced ERK phosphorylation but also cytosolic protein release from rat astrocytes. These findings suggest that the H2O2-induced release of cytosolic proteins depends on imbalance of lipid level in the membrane through suppressing lipid synthesis and increasing lipid release induced by the intracellular biochemical reaction such as signal transduction generated in intact rat astrocytes treated with H2O2.

Liquid diet induces memory impairment accompanied by a decreased number of hippocampal neurons in mice.

It is suggested that masticatory dysfunction affects the central nervous system; however, the underlying mechanism remains unknown. Brain-derived neurotrophic factor (BDNF) and its receptor, TrkB, are known to play important roles in memory and learning. In this study, we examined the effects of mastication on memory, the expression levels of BDNF and TrkB, and the number of neurons in the hippocampus of mice. Male C57 BL/6J mice (3 weeks old) were randomly divided into the control group (N = 7) fed chow pellets and the experimental group (N = 7) fed a liquid diet, which reduces mastication during eating. At 14 weeks of age, we performed a passive avoidance test and found that memory and learning ability were impaired in the experimental group compared with the control group. After the behavioral experiment, brains were harvested and analyzed morphologically and biochemically. In the hippocampus of the experimental group, the expression levels of BDNF were significantly higher, whereas those of TrkB were lower than those of the control group. In the cerebral cortex, these levels remained unchanged between the two groups. The ratio of phospho-p44/42 ERK/pan ERK, a downstream molecule of BDNF/TrkB signaling, in the experimental group was significantly lower than that of the control group in the cortex and hippocampus. The number of pyramidal neurons in the hippocampus was lower in the experimental group than in the control group. These findings suggest that reduced mastication induced by a liquid diet in early childhood may impair memory and learning ability, accompanied by neuronal loss in the hippocampus.