Effects of Nasal Obstruction on Maturation of the Jaw-Opening Reflex
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a r c h i v e s o f o r a l b i o l o g y 5 9 ( 2 0 1 4 ) 5 3 0 – 5 3 8 Available online at www.sciencedirect.com ScienceDirect journal homepage: http://www.elsevier.com/locate/aob Effects of nasal obstruction on maturation of the jaw-opening reflex in growing rats Yukiha Funaki, Maya Hiranuma, Mai Shibata, Satoshi Kokai *, Takashi Ono Orthodontic Science, Department of Orofacial Development and Function, Division of Oral Health Science, Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549, Japan a r t i c l e i n f o a b s t r a c t Article history: Objective: Nasal obstruction during growth changes craniofacial morphology and function. Accepted 24 February 2014 However, the etiological mechanisms of these changes are unknown. The aim of the present study was to investigate the effects of nasal obstruction during growth on the maturation of Keywords: the jaw-opening reflex (JOR) using an electrophysiological technique. We focused on the oral Growth sensory receptors that regulate the activities and reflexes of the orofacial muscles. Design: Sixty 6-day-old male Wistar rats were randomly divided into control and experi- jaw-opening reflex mental groups (n = 30 each). The experimental group underwent unilateral nasal obstruc- Nasal obstruction tion at 8 days of age. The JOR was evoked by bilateral, low-intensity electrical stimulation of Oral sensory receptor Rat the inferior alveolar nerve. The electromyographic responses were recorded bilaterally from the digastric muscles at 5, 7, and 9 weeks of age. 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 at each age, while the duration was not significantly different. Intragroup comparison of the latency, peak-to- peak amplitude, and duration at 5, 7, and 9 weeks of age revealed no significant differences in either the control or experimental groups. Conclusions: Unilateral nasal obstruction during growth may have significant effects on maturation of craniofacial function. # 2014 Elsevier Ltd. All rights reserved. harmonious craniofacial structures that allow for adequate 1. Introduction 3 interaction between mastication and swallowing. In orthodontic practice, patients often present with Normal craniofacial development and occlusion depend on maxillary protrusion or an anterior open bite, problems that various factors. For example, genetic factors influence the are frequently associated with nasal obstruction and mouth 1 4 constitution of the facial and occlusal pattern. Acquired breathing. Recent studies have indicated that the number of factors such as caries, oral habits, and nasopharyngeal patients with breathing problems caused by obstructive sleep conditions can also play important roles in craniofacial and apnea syndrome and allergic rhinitis is increasing, especially 2 occlusal development. Mammals, including humans, usually during the growth period of preadolescence and adoles- 5,6 breathe through the nose. Normal respiratory activity favors cence. A previous study in humans indicated that mouth * Corresponding author: Tel.: +81 3 5803 5528; fax: +81 3 5803 5528. E-mail address: [email protected] (S. Kokai). http://dx.doi.org/10.1016/j.archoralbio.2014.02.013 0003–9969/# 2014 Elsevier Ltd. All rights reserved. a r c h i v e s o f o r a l b i o l o g y 5 9 ( 2 0 1 4 ) 5 3 0 – 5 3 8 531 breathing may be due to an anatomical predisposition, nostril, consequently occluding the orifice of the nostril without velopharyngeal insufficiency, habit, or interference of lip mechanical or chemical damage to the olfactory mucosa. After 7 closure. Many clinical studies have also elucidated relation- cauterization, the nostril was coated with 3% chlortetracycline 1 ships between nasal obstruction and craniofacial morphology. (Aureomycin Ointment; Pola Pharma, Tokyo, Japan) to prevent For example, a lower position of the mandible caused by infection. The pups were kept warm (37 8C) for 30 min and then 8 mouth breathing increases the lower facial height and returned to their mothers. The control group underwent a sham 9 adenoidal facies. Changes in the craniofacial pattern might operation in which the cauterizing instrument was placed induce simultaneous changes in occlusion, such as an anterior about 1–2 mm above the left nostril. The body weights were 8,10 10 10 open bite, maxillary protrusion, or posterior crossbite. measured throughout the experimental period. The JOR was On the other hand, some studies have elucidated the effects of then recorded from the Dig muscle in control and experimental nasal obstruction on masticatory function, such as inhibited groups when the rats were 5, 7, and 9 weeks old (n = 10 per group 11 masseter muscle activity and increased suprahyoid muscle per time point). 12 activity during mouth breathing. Moreover, hypotonia of the 13 lips, tongue, and buccinator muscle has been reported, while 2.2. Stimulation and recording mouth breathing was also shown to decrease masticatory 14,15 activities. For electrophysiological recordings, all rats were anesthetized 1 Biochemical studies in rats suggest that nasal obstruction with 60 mg/kg of thiamylal sodium (Isozol ; Yoshitomi is associated with reduced growth of the masseter muscle and Pharmaceutical, Osaka, Japan) administered intraperitoneally. 16 anterior digastric (Dig) muscle. In addition, fatigue-resistant Cannulae were then inserted into the tracheal and femoral myosin heavy chain (MHC) fiber types are increased in veins. The depth of anesthesia was carefully monitored 16 muscles in association with nasal obstruction. In terms of throughout the experiment by checking the pupil size, morphological changes, nasal obstruction results in a signifi- withdrawal and corneal reflexes, and heart rate. To maintain cant reduction in the vertical development of the nasomax- the same anesthetic level in all rats while performing the JOR illary complex and in the skull base along the longitudinal recording, a supplemental dose (5 mg/kg, intravenously) was 17 axis. Although alterations of craniofacial morphology and routinely given 15 min before the start of the recording process function during nasal obstruction have been thoroughly if a withdrawal reflex was elicited by pinching the paws. The investigated, little is known about the effects of nasal rectal temperature was maintained at 37 8C with a heating obstruction on the regulation of jaw movement. It is possible pad. Before an incision was made, the skin was injected with that alterations in jaw movements may modify craniofacial 2% lidocaine to prevent noxious stimulation that could inhibit sensory transmission to jaw motoneurons, since the activity the JOR. of jaw motoneurons is dependent on sensory feedback from To bilaterally stimulate the inferior alveolar nerve (IAN), a 18 craniofacial structures. The jaw-opening reflex (JOR), a pair of stainless-steel wire electrodes (0.1 mm in diameter, 0.5 trigeminal brainstem reflex that can be evoked by both mm in tip exposure) was inserted bilaterally into the 19 nociceptive and non-nociceptive stimulation, plays an mandibular canal through the mental foramen at depths of important role in the regulation of jaw movement during 1 and 3 mm. The bipolar electrode was kept in place by fixing it 19 mastication. Thus, the JOR is a useful tool for investigating onto the adjacent mandibular bone with light-cured dental alterations of sensorimotor processing in masticatory perfor- resin. Expression of the JOR was recorded bilaterally from the mance. Dig muscles using paired stainless-steel wire electrodes with In the present study, we focused on the oral sensory 1-mm exposed tips implanted along the direction of the receptors that regulate the activity and reflexes of orofacial muscle fibers. The interpolar distance was set at 3 mm for muscles. The aim of this paper was to investigate the effects of these recording electrodes (Fig. 1A). To ensure consistent unilateral nasal obstruction during growth on JOR maturation reproduction of this distance, a two-keyhole index, with the using an electrophysiological technique. keyholes 3 mm apart, made of dental acrylic (UNIFAST II; GC Corporation, Tokyo, Japan) was used to lead the paired electrode-inserted needles, thus allowing for placement of 2. Materials and Methods the electrodes at the same interpolar distance for every recording. The animals were then transferred to a stereotaxic 2.1. Animal preparation apparatus (models SN-2 and SM-15 M; Narishige Scientific Instruments, Tokyo, Japan) with their bodies in prone position Sixty 6-day-old male Wistar albino rats were used in this (Fig. 1B). Before the stimulation, Dig electromyographic (EMG) experiment. The rats were weighed and then randomly divided baseline activity was captured for 1 min in each rat. into the control and experimental groups (n = 30 each). At 8 days Next, the right and left IAN were alternately stimulated of age, all rat pups were first anesthetized by hypothermia (10 electrically (single pulse, 0.2 ms in duration) once every 5 s to min at À18 8C). Left-sided nasal obstruction was performed in evoke the JOR. The stimulus intensity was applied gradually 17,20 the experimental group. The selected method involved in increasing in steps of 1 mA, and the lowest intensity that cauterization of the left external nostril, which is the most constantly elicited a Dig EMG response was determined as the common and simplest procedure allowing for the establish- JOR threshold (T). To attain comparable responses, the test ment of nasal obstruction in neonatal animals. The tissue stimulation current was adjusted to 1.5 times the threshold 21,22 surrounding the left external nostril was burned by placing a (1.5 T). An intensity of 2 T or less can be considered as 23 surgical cauterizing instrument (1 mm in diameter) on the non-nociceptive stimulation, which we expected in this 532 a r c h i v e s o f o r a l b i o l o g y 5 9 ( 2 0 1 4 ) 5 3 0 – 5 3 8 Figure 1 – Experimental design.