Introduction
Toothache is considered to be the highest in the degree of pain and often leads to some emotional symptoms such as anxiety- and depression-related behaviors [
1]. Many studies have confirmed that the neural conduction pathway of the toothache is the trigeminal nerve-thalamo-cortical circuitry, which included the brainstem trigeminal complex, thalamus, and cerebral cortex [
2,
3]. The signal of nociceptive stimulus in the dental pulp is transmitted from the periphery to the central nervous system through the trigeminal ganglion, and then converges to the trigeminal spinal nucleus, which relays nociceptive information to the thalamus, and further transduces nociceptive information to the cerebral cortex [
2‐
4]. However, the relationship between toothache and emotional reaction is not clear. The dynamic spatio-temporal changes of brain function involved in the natural process of toothache also have not been clearly studied. Here, for the first time to our knowledge, we non-invasively investigated the dynamic metabolic changes of brain activities longitudinally using PET molecular imaging approach [
5] in a rat model of dental pulp injury (DPI), and further explored the potential mechanisms underlying the aberrant affective behaviors during the toothache natural course.
Discussion
The behavioral, histological, and metabolic dynamic changes of toothache were longitudinally evaluated for up to 14 days in a rat DPI model. Interestingly, a perfect correlation among the behavioral, histological examinations and PET imaging was identified during the natural process of toothache, and therefore, our study provided a comprehensive understanding for the neurofunctional patterns underlying toothache. More importantly, the nerve conduction pathways related to toothache, especially the emotional reaction to toothache are identified. The related features provide rich resource to inspect toothache-induced brain changes.
In this study, we chose the DPI model as the suitable model to investigate the natural course of toothache. The exposure in the pulp of first molars could lead to local pulp tissue infection by normal flora of the oral cavity, the increase of tissue pressure in the pulp cavity, and the release of inflammatory mediators and neuropeptides, which directly or indirectly acted on the nerve endings of the pulp tissue and resulted in the trigeminal pain symptoms. During this process, the inflammatory response was limited to the pulp cavity and root canal, and did not affect the apical tissue, trigeminal nerve, and related cerebral cortex beyond the apical foramen [
13,
14]. If the DPI model is locally anesthetized, the nerve endings of the rat dental pulp tissue might be blocked and cannot receive nociceptive signals and reduce a series of pain responses. Therefore, locally anesthesia assay could be helpful to further strength our conclusions observed in DPI model.
The upload of toothache stimulation signal caused transient and large-scale reactive activation of rat brain nuclei, and there was a relationship between the functional connectivity of each nucleus and the strength of pain signals. According to previous literatures [
15] and the results of the current study (Fig.
2F and
G), the ascending pathway of toothache is postulated to be conducted through the trigeminal nerve complex to PAG, cc and VPM, and then to the subcortical area (nucleus accumbens, striatum, hippocampus, and amygdala) and cerebral cortex (S1&S2, M1&M2, PFC, Cg, RSD). Among them, pain signals cross cc and PAG, which activate most of the brain regions bilaterally, thus regulating the transmission of pain in the left maxillary teeth. Different from previous studies [
16], the visual signals related to VPM, superior colliculus and LGN, as well as RSD may also be involved in the transmission of toothache. Among them, the superior colliculus and LGN can make a visual reflex response to tactile information [
17], thus reducing perception and emotion caused by pain, suggesting that the signals of toothache might activate the eye branch when ascending through the trigeminal ganglion, which explains why some patients with acute toothache often experience symptoms of eye discomfort and even tears [
18]. The RSD is involved in the control of spatial navigation, episodic memory, and pain-related emotional behaviors [
19]. It is speculated that RSD might be related to the cognitive and spatial memory of toothache in rats. Here, PET showed, for the first time, that the degree of activation and inhibition of the 12 brain regions involved in toothache is closely related to the changes of its pain intensity and may also be related to the cognition and memory of toothache in rats. The changes of brain network caused by toothache are very complex and elusive and need to be further investigated.
The pons was selected as the intensity normalization region to ensure the consistency of the data. It is reported that the pons is the best brain region for the normalization of intensity on the
18F-FDG-PET scans of the brain [
20], which can be adopted for visual and semiquantitative analysis of brain
18FDG-PET imaging associated with other diseases in the absence of any pathological involvement [
21]. However, the current international stable and reliable region for brain PET image analysis related to endodontic pain still needs further research.
Toothache induces anxiety- and depression-related symptoms, thus may cause the patients to miss the best time window to get treatments. Therefore, it is of great significance, to understand how the toothache-related emotion was regulated in order to alleviate the negative emotions. When ectopic pain occurs, synaptic responses in the ACC are enhanced in rodent models of neuropathic pain, which is induced by long-term potentiation (LTP). And the development of LTP is relied on the NMDA receptors and their signaling pathway. APV, as a selective NMDA receptor antagonist, could prevent NMDA receptor activation-induced LTP and inhibit synaptic plasticity [
9,
12]. APV administration reversed the emotional response related to toothache. In our study, the results from experimental group (DPI + APV) and control groups (DPI, DPI + NS) indicated that the changes of behavior and glucose metabolism in the cACC brain area may be due to APV acting on NMDA receptors in the cACC brain area and reducing the expression of NR2B total protein production. The decrease of total NR2B in DPI + APV group might be induced by APV administration, followed by the decrease of p-NR2B in our study. Meanwhile, we also observed that NR2B expression level was increased in DPI group compared with sham group. In a previous study, in a rat model of F-CPA (formalin-induced conditioned place avoidance), unilateral intraplantar (i.pl.) injection of dilute formalin also induced NR2B expression in rACC brain region compared with normal saline group [
9]. Meanwhile, APV administration in sham rats might have no effect on the synaptic response, behavioral, and metabolic changes. Adkins et al. reported that APV administration in vehicle-treated rats had no effect on fear generalization when compared with vehicle-vehicle animals [
22]. Therefore, these results suggest that cACC but not rACC may mediate the emotional reaction to toothache by NR2B receptor, which is the difference between emotional regulation of toothache and somatic pain. This might provide a novel insight for relieving the pain affect induced by toothache.
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