Sleep alterations in an experimental orofacial pain model in rats

This study sought to assess sleep patterns in rats injected with Freund's adjuvant (FA) in the temporomandibular joint (TMJ) as a potential experimental orofacial pain model. Pain response to indomethacin was also assessed. Rats were implanted with electrodes to record electrocorticogram and eletromyogram signals. After a baseline (B) recording, they were injected with Freund's adjuvant (orofacial pain group, n=8) or saline (sham group, n=8) in the temporomandibular joint, and their sleep was monitored over two 12-h light periods. In the second phase of the study, after injecting Freund's adjuvant, indomethacin was administered (1 mg/kg p.o.) at 12- intervals, and sleep patterns were recorded for two additional light periods. The orofacial pain group showed a reduction in sleep efficiency during the two light periods compared with the baseline recording and with the sham group (p<0.001). Increases in sleep and paradoxical sleep (PS) latencies of approximately 200% and 420%, respectively, were observed, as well as an increase in the number of awakenings during both periods (p<0.001). Treatment with indomethacin increased sleep efficiency (p<0.001) and paradoxical sleep time (p<0.001). The number of awakenings (p<0.001) and sleep (p<0.001) and paradoxical sleep latencies (p<0.001) were reduced reestablishing the normal sleep pattern. The results showed the reliability and usefulness of the temporomandibular joint pain model to characterize sleep disturbances related to pain and its response to indomethacin.     

正畸牙移动过程中牙周组织白细胞介素8的表达*

背景：白细胞介素8作为一种与骨改建相关的炎性细胞因子,参与牙周组织的改建,促进牙槽骨的吸收,是骨吸收的标志物之一。 目的：观察正畸力对牙周组织改建过程中白细胞介素8表达的影响。 方法：将10周龄雄性SD大鼠上颌一侧第一磨牙与上颌切牙之间安置正畸装置,并施加0.49 N近中向正畸力,在加力后1,3,5,7,14 d取材,进行组织形态分析法、免疫组化染色检测白细胞介素8在牙周组织中表达的阳性细胞的面密度值。 结果与结论：正常大鼠牙周组织内白细胞介素8的表达呈弱阳性;随加力时间的延长大鼠牙周组织牙周膜细胞白细胞介素8表达增强,加力后5 d达到高峰,之后开始下降。实验提示正畸力可引起局部牙周组织炎症及白细胞介素8的释放。正畸牙移动早期,白细胞介素8的释放可能是牙周组织早期炎症反应和牙槽骨改建的触发因素之一。     

Complement activation contributes to leukocyte recruitment and neuropathic pain following peripheral nerve injury in rats

Complement activation triggers inflammation and has been implicated in neurological diseases associated with pain. However, the role of complement in neuropathic pain has not been clearly defined. In this study, we tested whether complement is activated by partial ligation of the rat sciatic nerve, a widely used model of neuropathic pain, and whether complement activation or inhibition in peripheral nerve influences leukocyte recruitment and neuropathic pain. We found that C3 deposition significantly increased from 6 h to 7 days in the injured nerve and was associated with the extent of thermal hyperalgesia and mechanical allodynia. However, no deposition of the membrane attack complex was detected. Complement activation by endoneurial injection of aggregated rat immunoglobulin G into normal sciatic nerve produced significant thermal hyperalgesia and mechanical allodynia of the ipsilateral hindpaw at 2-7 days after injection. This was accompanied by increased deposition of C3 and recruitment of macrophages at 7 days following injection. Complement inhibition using systemic injections of soluble complement receptor 1 (AVANT Immunotherapeutics, Inc., Needham, USA) into rats markedly suppressed C3 deposition and T-cell and macrophage recruitment to the injured nerve, and produced significant alleviation of thermal hyperalgesia and mechanical allodynia. These results demonstrate that C3 activation in the nerve contributes to increased infiltration of inflammatory cells and to neuropathic pain behaviors following peripheral nerve injury. Complement inhibition may be a potential therapeutic treatment for neuropathic pain.     

Medullary monoamine levels during experimental tooth movement

We investigated possible influence of nociception induced by experimental tooth movement on the medullary monoaminergic inhibitory systems. Forty-eight hours after the start of the experimental tooth movement, significant increases in dorsal serotonin (5-HT), dopamine (DA), norepinephrine (NE), 5-hydroxyindoleacetic acid (5-HIAA), and dihydroxyphenylacetic acid (DOPAC) levels were detected with ipsilateral dominance. These results suggest that the nociception induced by experimental tooth movement might be under modulation of serotonergic, noradrenergic, and dopaminergic systems.     

Effects of centrally administered vasopressin on orofacial pain perception in rats

Vasopressin (AVP) appears in the cerebrospinal fluid and plays an important role in nociceptive modulation in the central nervous system. The effect of increased concentration of AVP in the cerebrospinal fluid on the excitability of the hypoglossal nerve nucleus was investigated. The experiments were carried out on rats under chloralose anesthesia. Amplitudes of the retractory evoked tongue jerks (ETJ) of the outstretched tongue during the perfusion of cerebral ventricles with solutions containing AVP or its antagonists and also opioid and serotonin antagonists were recorded. Perfusion of the ventricles with AVP in 100 microM concentration suppressed the ETJ amplitude to 66 +/- 3.83%, and in 200 microM concentration, to 53 +/- 3.18% of the control. V1 vasopressin receptor antagonist, d(CH2)5,Tyr(Me)AVP, blocked the suppressive effect caused by cerebral ventricle perfusion with AVP from 64 +/- 4.11% to 83 +/- 1.58%, whereas V2 vasopressin receptor antagonist, d(CH2)5[Ile2, Ile4]AVP, did not block the antinociceptive effect of AVP. Analgesic effect of AVP was also inhibited by opioid and serotonin receptor antagonists, naloxone and methysergide, respectively. Naloxone blocked the suppressive effect of 100 microM AVP from 64 +/- 5.63% to 92 +/- 3.70% and methysergide from 65 +/- 3.62% to 80 +/- 2.72% of the control. The results indicate that exogenous AVP plays an antinociceptive role in the brain of rats penetrating the lining of the cerebral ventricles into the cerebrospinal fluid and exerting a modulating effect on the tongue motor center situated near III and IV cerebral ventricle. V1 vasopressin receptor, but not V2 vasopressin receptor, is involved in this activity in the CNS. The antinociception of AVP seems to be mediated by opioid and serotonergic pathways.     

Assessment of chronic trigeminal neuropathic pain by the orofacial operant test in rats

The present study examined the effects of Tremella fuciformis (TF) on the learning and memory function and the neural activity in rats with trimethyltin (TMT)-induced memory deficits. The rats were administered saline or TF (TF 25, 50, 100 mg/kg, p.o.) daily for 21 days. The cognitive improving efficacy of TF on the amnesic rats, which was induced by TMT, was investigated by assessing the Morris water maze test and by performing Choline acetyltransferase (ChAT) and cAMP responsive element binding protein (CREB) immunohistochemistry. In order to confirm the underlying mechanisms of the memory enhancing effects of TF, we assessed the neurite outgrowth of PC12 cells. We also administered 18F-fluorodeoxyglucose and performed a PET scan of the frontal lobe.The rats with TMT injection showed impaired learning and memory of the tasks and treatment with TF produced a significant improvement of the escape latency to find the platform in the Morris water maze compared to that of the control group. In the retention test, the TF50 group showed increased time spent around the platform compared to that of the control group. Consistent with the behavioral data, TF50 mg/kg significantly alleviated the loss of ChAT-ir neurons in the hippocampus compared to that of the control group. Treatment with TF significantly increased the CREB positive neurons in the hippocampal CA1 area as compared to that of the control group. In addition, TF treatment (50 mg/kg) increased the glucose uptake approximately sevenfold in the frontal lobe and it significantly promoted neurite outgrowth of the PC12 cells, as compared to that of the controls.These results suggest that TF may be useful for improving the cognitive function via regulation of the CREB signaling pathway and cholinergic system in the hippocampus.     

实验性SAH后CVS大鼠脑组织CGRP mRNA表达研究

目的　探讨降钙素基因相关肽（ＣＧＲＰ）在蛛网膜下腔出血（ＳＡＨ）后脑血管痉挛（ＣＶＳ）发生发展中的作用机制。方法　采用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）技术检测大鼠ＳＡＨ 后ＣＶＳ脑组织、血管ＣＧＲＰ ｍ ＲＮＡ 表达。结果　正常对照组ＣＧＲＰｍ ＲＮＡ 表达水平依次为基底节区、海马、大脑皮层和脑血管;注血后３ｄ 各脑区、血管ＣＧＲＰｍ ＲＮＡ 表达均较正常对照组、注血后３０ｍ ｉｎ、注血后７ｄ 显著下调（Ｐ＜ ０．０１）。结论　ＣＧＲＰ ｍ ＲＮＡ 表达下调、舒缩血管物质失衡是ＳＡＨ 后ＣＶＳ的主要原因之一。     

Persistent Fos protein expression after orofacial deep or cutaneous tissue inflammation in rats: implications for persistent orofacial pain.

This study was designed to systematically examine the effects of persistent orofacial tissue injury on prolonged neuronal activation in the trigeminal nociceptive pathways by directly comparing the effects of orofacial deep vs. cutaneous tissue inflammation on brainstem Fos protein expression, a marker of neuronal activation. Complete Freund's adjuvant (CFA) was injected unilaterally into the rat temporomandibular joint (TMJ) or perioral (PO) skin to produce inflammation in deep or cutaneous tissues, respectively. Rats were perfused 2 hours, 24 hours, 3 days, or 10 days following CFA injection. The TMJ and PO inflammation-induced Fos expression paralleled the intensity and course of inflammation over the 10-day observation period, suggesting that the increase in intensities and persistence of Fos protein expression may be associated with a maintained increase in peripheral input. Compared to PO CFA injection, the injection of CFA into the TMJ produced a significantly stronger inflammation associated with a greater Fos expression. In TMJ- but not in PO-inflamed rats, Fos-like immunoreactivity (LI) spread from superficial to deep upper cervical dorsal horn as the inflammation persisted and there was a dominant ipsilateral Fos-labeling in the paratrigeminal nucleus. Common to TMJ and PO inflammation, Fos-LI was induced in the trigeminal subnuclei interpolaris and caudalis, C1-2 dorsal horn, and other medullary nuclei. Substantial bilateral Fos-LI was found in the interpolaris-caudalis trigeminal transition zone. Further analysis revealed that Fos-LI in the ventral transition zone was equivalent bilaterally, whereas Fos-LI in the dorsal transition zone was predominantly ipsilateral to the inflammation. The differential induction of Fos expression suggests that an increase in TMJ C-fiber input after inflammation and robust central neuronal hyperexcitability contribute to persistent pain associated with temporomandibular disorders.     

Decreases in the expression of CGRP and galanin mRNA in central and peripheral neurons related to the control of blood pressure following experimental hypertension in rats

Calcitonin gene-related peptide (alpha CGRP) and galanin (GAL) are peptides known to participate in central mechanisms of blood pressure control. Nonetheless, variations in the synthesis of the peptides in response to a hypertensive challenge are not well described, specially using a model, which allows acute and chronic analyses. In this study, we have employed in situ hybridization to analyse changes in mRNA expression of alpha CGRP and GAL in the nucleus tractus solitarii (NTS), hypothalamic paraventricular nucleus (PVN) as well as petrosal and nodose ganglia after aortic coarctation-induced hypertension in rats. Acute (2h) and chronic (3 and 7 days) analyses were performed in order to evaluate the involvement of both peptides in different periods of hypertension. The analysis of relative mRNA levels showed significant differences between sham-operated and aortic coarcted hypertensive rats. alpha CGRP mRNA expression was decreased 2h (40%) and 3 days (42%) in nodose and petrosal ganglia, respectively, after coarctation. No changes in CGRP mRNA signal were seen in the NTS and PVN in the analysed periods. GAL mRNA expression was decreased in the NTS (19%) and PVN (55%), 3 and 7 days, respectively, after coarctation-induced hypertension. No changes in GAL mRNA expression were observed in petrosal and nodose ganglia following aortic coarctation. Data suggest that alpha CGRP and GAL may participate in the mechanisms involved in the establishment/maintenance of hypertension induced by aortic coarctation. Acute changes might be involved with the adaptation to the hypertensive state, while changes at the chronic phase might be related to counteraction of hypertension.     

实验性牙移动大鼠外周血内皮祖细胞的培养

背景：观察血管内皮祖细胞在牙周血管改建中的作用规律对于研究正畸牙周改建机制具有重要意义。 目的：对牙移动大鼠外周血内皮祖细胞进行体外分离、培养及鉴定,探讨牙齿移动是否可动员骨髓内皮祖细胞进入外周血。 设计、时间及地点：细胞体外分离培养及生物学指标检测,随机对照动物实验。于2007-09/2008-04在吉林省口腔生物医学工程重点实验室完成。 材料：24只Wistar大鼠,随机分为实验组和对照组,每组12只。 方法：实验组建立大鼠牙移动模型,对照组置牵拉装置但不加力。加力第2天取两组动物外周血制作细胞涂片,检测其中CD133阳性细胞数。同时以Percoll密度梯度分离提取其外周血单个核细胞,培养至第12天,取两组细胞爬片进行相关指标检测。 主要观察指标：以免疫细胞化学染色和免疫荧光化学染色检测细胞表面标志物,荧光化学检测细胞吸收乙酰化低密度脂蛋白及荆豆凝集素1情况,MTT法检测细胞增殖情况。 结果：实验组细胞涂片中CD133阳性细胞率明显高于对照组（P < 0.05）。所培养细胞可同时吸收Dil标记的乙酰化低密度脂蛋白、FITC标记的荆豆凝集素1,呈CD133、Ⅷ因子、CD34阳性。实验组细胞增殖活性高于对照组（P < 0.05）。 结论：实验初步证实了实验性牙移动可动员大鼠骨髓内皮祖细胞进入外周血。     

Neuroplastic changes in the sensorimotor cortex associated with orthodontic tooth movement in rats

Orthodontic tooth movement (OTM) causes transient pain and changes in the dental occlusion that may lead to altered somatosensory inputs and patterns of mastication. This study used intracortical microstimulation (ICMS) and electromyographic (EMG) recordings to test whether neuroplastic changes occur in the ICMS‐defined motor representations of left and right anterior digastric (LAD, RAD), masseter, buccinator, and genioglossus (GG) muscles within the rat's face primary motor cortex (face‐M1) and adjacent face primary somatosensory cortex (face‐S1) during OTM. Analyses included any changes in the number of ICMS sites representing these muscles and in the onset latencies of ICMS‐evoked responses in the muscles. Sprague–Dawley rats were divided into experimental (E), sham (S), and naive (N) groups; OTM was induced in the E group. Statistical analyses involved a mixed model repeated‐measures analysis of variance (MMRM ANOVA). OTM resulted in significant neuroplastic changes in the number of positive sites in the E group for LAD, RAD, and GG muscles in face‐M1 and face‐S1 at days 1, 7, and 28 of continuous orthodontic force application, and in the number of sites in face‐M1 from which ICMS could simultaneously evoke EMG responses in different combinations of LAD, RAD, and GG muscles. However, the onset latencies of ICMS‐evoked responses were not significantly different between groups or between face‐M1 and face‐S1. The neuroplastic changes documented in this study may reflect adaptive sensorimotor changes in response to the altered environment in the oral cavity induced by OTM. J. Comp. Neurol. 523:1548–1568, 2015. © 2015 Wiley Periodicals, Inc.     

Basic fibroblast growth factor-enhanced neurogenesis contributes to cognitive recovery in rats following traumatic brain injury

Stem/progenitor cells reside throughout the adult CNS and are actively dividing in the subventricular zone (SVZ) and the dentate gyrus (DG) of the hippocampus. This neurogenic capacity of the SVZ and DG is enhanced following traumatic brain injury (TBI) suggesting that the adult brain has the inherent potential to restore populations lost to injury. This raises the possibility of developing strategies aimed at harnessing the neurogenic capacity of these regions to repair the damaged brain. One strategy is to enhance neurogenesis with mitogenic factors. As basic fibroblast growth factor (bFGF) is a potent stem cell mitogen, we set out to determine if an intraventricular administration of bFGF following TBI could affect the levels of injury-induced neurogenesis in the SVZ and DG, and the degree to which this is associated with cognitive recovery. Specifically, adult rats received a bFGF intraventricular infusion for 7 days immediately following TBI. BrdU was administered to animals daily at 2-7 days post-injury to label cell proliferation. At 1 or 4 weeks post-injury, brain sections were immunostained for BrdU and neuronal or astrocytic markers. We found that injured animals infused with bFGF exhibited significantly enhanced cell proliferation in the SVZ and the DG at 1 week post-TBI as compared to vehicle-infused animals. Moreover, following bFGF infusion, a greater number of the newly generated cells survived to 4 weeks post-injury, with the majority being neurons. Additionally, animals infused with bFGF showed significant cognitive improvement. Collectively, the current findings suggest that bFGF-enhanced neurogenesis contributes to cognitive recovery following TBI.     

Microvascular damage following experimental sinus-vein thrombosis in rats

We evaluated effects of thrombosis of the superior sagittal sinus, its bridging and cortical veins (SVT) on the cerebral microvasculature in rats. Cryosections of brains (n=7) were examined for venous infarction and microvascular basal lamina damage 3 h after SVT by immunohistochemical staining of microtubule-associated protein 2 and collagen type IV. Microvessels in the infarctions showed a decrease in the number (23.5±6.1%, P<0.002) and the total area (24.9±6.5%, P<0.011) of collagen type IV-positive vessels in contrast to control areas (21.7±12.4%, P<0.007; and 26.3±15.1%, P<0.026 in contrast to control areas of unoperated animals). This study showed a significant alteration of the cerebral microvasculature in SVT, which might contribute to edema and hemorrhagic transformation.     

Spinal glial activation contributes to pathological pain states

Chronic pain, a pathological state, affects millions of people worldwide. Despite decades of study on the neuronal processing of pain, mechanisms underlying the creation and maintenance of enhanced pain states after injury or inflammation remain far from clear. In the last decade, however, the discovery that glial activation amplifies pain has challenged classic neuronal views of "pain". This review focuses on recent developments in understanding that spinal cord glia are involved in pathological pain. We overview the action of spinal glia (both microglia and astrocytes) in several persistent pain models, and provide new evidence that spinal glia activation contributes to the development and maintenance of arthritic pain facilitation. We also attempt to discuss some critical questions, such as how signals are conveyed from primary afferents to spinal glia following peripheral nerve injury and inflammation. What causes glia to become activated after peripheral/central injury/inflammation? And how the activated glia alter neuronal sensitivity and pain processing? Answers to these questions might open a new approach for treatment of pathological pain.     

Cerebral responses to electrical tooth pulp stimulation in man. An objective correlate of acute experimental pain.

The pulp of individual teeth of 17 normal adult volunteers was electrically stimulated via pairs of electrodes implanted into dentine. Computer-summated responses recorded from the surface of the head were composed of two concurrent sequences of events, one of which was seen maximally over midline areas and the other over the lower portions of the postcentral regions. Appropriate tests demonstrated that these wave forms represented cerebral tooth pulp-evoked potentials. Because tooth pulp-evoked potentials represent objective, quantifiable, nonverbal concomitants of central events associated with the perception of noxious stimuli, they may prove helpful in investigating acute experimental pain in man.