Expanded terminal fields of gustatory nerves accompany embryonic BDNF overexpression in mouse oral epithelia.

Brain-derived neurotrophic factor (BDNF) is expressed in gustatory epithelia and is required for gustatory neurons to locate and innervate their correct target during development. When BDNF is overexpressed throughout the lingual epithelium, beginning embryonically, chorda tympani fibers are misdirected and innervate inappropriate targets, leading to a loss of taste buds. The remaining taste buds are hyperinnervated, demonstrating a disruption of nerve/target matching in the tongue. We tested the hypothesis here that overexpression of BDNF peripherally leads to a disrupted terminal field organization of nerves that carry taste information to the brainstem. The chorda tympani, greater superficial petrosal, and glossopharyngeal nerves were labeled in adult wild-type (WT) mice and in adult mice in which BDNF was overexpressed (OE) to examine the volume and density of their central projections in the nucleus of the solitary tract. We found that the terminal fields of the chorda tympani and greater superficial petrosal nerves and overlapping fields that included these nerves in OE mice were at least 80% greater than the respective field volumes in WT mice. The shapes of terminal fields were similar between the two groups; however, the density and spread of labels were greater in OE mice. Unexpectedly, there were also group-related differences in chorda tympani nerve function, with OE mice showing a greater relative taste response to a concentration series of sucrose. Overall, our results show that disruption in peripheral innervation patterns of sensory neurons have significant effects on peripheral nerve function and central organization of their terminal fields.

The involvement of TRP channels in sensory irritation: a mechanistic approach toward a better understanding of the biological effects of local irritants.

Peripheral nerves innervating the mucosae of the nose, mouth, and throat protect the organism against chemical hazards. Upon their stimulation, characteristic perceptions (e.g., stinging and burning) and various reflexes are triggered (e.g., sneezing and cough). The potency of a chemical to cause sensory irritation can be estimated by a mouse bioassay assessing the concentration-dependent decrease in the respiratory rate (50 % decrease: RD50). The involvement of the N. trigeminus and its sensory neurons in the irritant-induced decrease in respiratory rates are not well understood to date. In calcium imaging experiments, we tested which of eight different irritants (RD50 5-730 ppm) could induce responses in primary mouse trigeminal ganglion neurons. The tested irritants acetophenone, 2-ethylhexanol, hexyl isocyanate, isophorone, and trimethylcyclohexanol stimulated responses in trigeminal neurons. Most of these responses depended on functional TRPA1 or TRPV1 channels. For crotyl alcohol, 3-methyl-1-butanol, and sodium metabisulfite, no activation could be observed. 2-ethylhexanol can activate both TRPA1 and TRPV1, and at low contractions (100 µM) G protein-coupled receptors (GPCRs) seem to be involved. GPCRs might also be involved in the mediation of the responses to trimethylcyclohexanol. By using neurobiological tools, we showed that sensory irritation in vivo could be based on the direct activation of TRP channels but also on yet unknown interactions with GPCRs present in trigeminal neurons. Our results showed that the potency suggested by the RD50 values was not reflected by direct nerve-compound interaction.

Peripheral nervous system involvement in primary burning mouth syndrome--results of a pilot study.

OBJECTIVE: The pathophysiology of primary burning mouth syndrome (BMS) has remained enigmatic, but recent studies suggest pathology within the nervous system at multiple levels. This study aimed to investigate in detail the contribution of either focal or generalized alterations within the peripheral nervous system (PNS) in the etiopathogenesis of BMS. SUBJECTS AND METHODS: Intraepithelial nerve fiber density (IENFD) of tongue mucosa was assessed in 10 carefully characterized BMS, and the results were compared to 19 age- and gender-matched cadaver controls, 6 with lifetime diabetes. Extensive neurophysiologic and psychophysical examinations of the trigeminal system and distal extremities were performed to profile PNS function in BMS. RESULTS: Patients with BMS had significantly fewer intraepithelial nerve fibers (0,27, s.e. 0,18 mm(-1); P = 0.0253) than non-diabetic controls (0,92, s.e. 0,15 mm(-1)). In the subepithelial space, the amount of nerve fibers did not differ between the groups. The majority (9/10) of patients with BMS showed neurophysiologic or psychophysical signs of a more generalized PNS dysfunction. CONCLUSIONS: Our results in neurophysiologically optimally characterized BMS patients confirm that pure focal small fiber neuropathy of the oral mucosa has a role in the pathophysiology of primary BMS. Furthermore, BMS may be related to a more generalized, yet subclinical peripheral neuropathy.

A systematic review on the innervation of peri-implant tissues with special emphasis on the influence of implant placement and loading protocols.

OBJECTIVES: To systematically review the available literature on the influence of dental implant placement and loading protocols on peri-implant innervation. MATERIAL AND METHODS: The database MEDLINE, Cochrane, EMBASE, Web of Science, LILACS, OpenGrey and hand searching were used to identify the studies published up to July 2013, with a populations, exposures and outcomes (PEO) search strategy using MeSH keywords, focusing on the question: Is there, and if so, what is the effect of time between tooth extraction and implant placement or implant loading on neural fibre content in the peri-implant hard and soft tissues? RESULTS: Of 683 titles retrieved based on the standardized search strategy, only 10 articles fulfilled the inclusion criteria, five evaluating the innervation of peri-implant epithelium, five elucidating the sensory function in peri-implant bone. Three included studies were considered having a methodology of medium quality and the rest were at low quality. All those papers reported a sensory innervation around osseointegrated implants, either in the bone-implant interface or peri-implant epithelium, which expressed a particular innervation pattern. Compared to unloaded implants or extraction sites without implantation, a significant higher density of nerve fibres around loaded dental implants was confirmed. CONCLUSIONS: To date, the published literature describes peri-implant innervation with a distinct pattern in hard and soft tissues. Implant loading seems to increase the density of nerve fibres in peri-implant tissues, with insufficient evidence to distinguish between the innervation patterns following immediate and delayed implant placement and loading protocols. Variability in study design and loading protocols across the literature and a high risk of bias in the studies included may contribute to this inconsistency, revealing the need for more uniformity in reporting, randomized controlled trials, longer observation periods and standardization of protocols.

Roles of innervation in developing and regenerating orofacial tissues.

The head is innervated by 12 cranial nerves (I-XII) that regulate its sensory and motor functions. Cranial nerves are composed of sensory, motor, or mixed neuronal populations. Sensory neurons perceive generally somatic sensations such as pressure, pain, and temperature. These neurons are also involved in smell, vision, taste, and hearing. Motor neurons ensure the motility of all muscles and glands. Innervation plays an essential role in the development of the various orofacial structures during embryogenesis. Hypoplastic cranial nerves often lead to abnormal development of their target organs and tissues. For example, Möbius syndrome is a congenital disease characterized by defective innervation (i.e., abducens (VI) and facial (VII) nerves), deafness, tooth anomalies, and cleft palate. Hence, it is obvious that the peripheral nervous system is needed for both development and function of orofacial structures. Nerves have a limited capacity to regenerate. However, neural stem cells, which could be used as sources for neural tissue maintenance and repair, have been found in adult neuronal tissues. Similarly, various adult stem cell populations have been isolated from almost all organs of the human body. Stem cells are tightly regulated by their microenvironment, the stem cell niche. Deregulation of adult stem cell behavior results in the development of pathologies such as tumor formation or early tissue senescence. It is thus essential to understand the factors that regulate the functions and maintenance of stem cells. Yet, the potential importance of innervation in the regulation of stem cells and/or their niches in most organs and tissues is largely unexplored. This review focuses on the potential role of innervation in the development and homeostasis of orofacial structures and discusses its possible association with stem cell populations during tissue repair.

[Changes of the innervation in the mucous membrane and glands of the tongue in early and late experimental diabetes mellitus]. / A nyelv nyálkahártya és mirigyek innervációjának változása korai és késoi kisérletes diabetes mellitusban.

The number of the different neuropeptides-containing nerve fibres and immunocompetent cells was changed in diabetes mellitus (DM) in different organs. In this work we investigated the effect of DM on quantitation of the nerve fibres using immunhistochemistry. After two weeks of the DM the quantitiy of the different nerve fibres increased significantly both in the mucous membrane and glands of the tongue. The number of the immunocompetent cells (lymphocytes, plasma cells, mast cells) increased as well significantly. Some of these cells showed also immunoreactivity for substance P and neuropeptide Y. A few substance P cells were in very close relation to the SP immunoreactive nerve fibres. After four weeks of DM the number of the nerve fibres was decreased compared to the 2 weeks treatment, however, the number of them was higher compared to the control. The close correlation between the nerve fibres and immune cells might play a crucial role in maintaining the homeostasis in the mucous membrane and glands of the tongue as well as in the increasing inflammation and elimination of it.

Topographical relationship of the greater palatine artery and the palatal spine. Significance for periodontal surgery.

AIM: The aims of this study were to (1) identify the branching pattern and course of the greater palatine artery (GPA), (2) carry out a morphological analysis of the palatal bony prominence that divides the medial and lateral grooves and (3) characterize the topographical relationships between these two structures. METHODS: Thirty-six hemimaxillae were studied with the aid of a surgical microscope to elucidate the GPA. A further 25 dry skulls were examined to establish the morphology of the palatal spine. RESULTS: The most common GPA branching pattern was type I (41.7%, 15 sides), which gave off the medial and canine branches after the bony prominence. The distances from the CEJ to the lateral branch of the GPA were 9.04 ± 2.93 mm (canine), 11.12 ± 1.89 mm (first premolar), 13.51 ± 2.08 mm (second premolar), 13.76 ± 2.86 mm (first molar) and 13.91 ± 2.20 mm (second molar). The palatal spine was frequently observed as the bony prominence (66.3%, 57 sides), and was located at 6.49 ± 1.76 mm from the greater palatine foramen, with a length of 10.42 ± 2.45 mm. There was no a correlation between the bony prominence shape and the GPA branching pattern. CONCLUSIONS: These results could provide the reference data regarding the topography of the GPA for periodontal surgery.

Immediate analgesic effect of 8% lidocaine applied to the oral mucosa in patients with trigeminal neuralgia.

BACKGROUND: Trigeminal nerve block is widely used for trigeminal neuralgia (TN), though with much painful procedure and potential serious complications. The pain of TN occurs most frequently in the second and the third divisions of the trigeminal nerve, which are distributed in intraoral mucous membrane as well as face skin. Here, we examined the response to intraoral application of 8% lidocaine (LDC) in patients with oral TN pain in a double-blind, placebo (PBO)-controlled crossover study. METHODS: Twenty-four outpatients with oral TN pain were randomized to receive intraoral application of either 8% LDC or saline PBO to the painful area. Following 7-days period, patients were crossed over to receive the alternative treatment. The pain was assessed with a numerical rating scale (NRS) before and 15 minutes after treatment. Patients used a descriptive scale to grade pain outcome and were asked to note any recurrence and the latency for recurrence after therapy. RESULTS: Intraoral LDC, but not PBO, significantly decreased the NRS from 5 (4, 8) (median [25, 75 percentiles]) to 1 (0, 4) (P = 0.001). Of the 24 patients, 19 described marked or moderate relief of pain after LDC but only three described the same after PBO application. The effect of LDC and PBO persisted for 2.8 (0.3, 3.0) and 0 (0, 0) hours, respectively. CONCLUSIONS: Intraoral application of 8% LDC produced prompt analgesia without serious side effects in patients with TN who presented with severe intraoral pain.

Neuroanastomosis and the innervation territory of the mental nerve.

The aim of this study was to clarify the distribution pattern and innervation territory of the mental nerve (MN) in the skin and mucosa by topographic examination by Sihler's staining, thereby providing reference anatomical information for surgical procedures and to enable prediction of regions of sensory disturbance following nerve damage. Ten human specimens were subjected to Sihler's staining, which is a highly accurate method for visualizing the distribution of nerve fibers without altering their topography. Each branch of the MN overlapped adjacent branches (five cases), or else they were distributed individually at the lower lip (five cases). The MN anastomosed with some branches of the facial nerve near the mental foramen. Moreover, some branches of the MN anastomosed with the buccal nerve of the trigeminal nerve, which supplies sensation to the skin and mucosa over the lateral region of the lower lip (six cases). The details of the distribution pattern and innervations territory of the MN presented herein may enable the prediction of a region of sensory disturbance following MN damage. Moreover, knowledge of the pattern of synapses with adjacent branches of other nerves, such as the facial (marginal mandibular and cervical branches) and the buccal nerves, might help to improve our understanding around incomplete anesthesia during the surgical procedures in oral & maxillofacial region.

Phenolic composition and mouthfeel characteristics resulting from blending Chilean red wines.

BACKGROUND: The blending of wine is a common practice in winemaking to improve certain characteristics that are appreciated by consumers. The use of some cultivars may contribute phenolic compounds that modify certain characteristics in blended wines, particularly those related to mouthfeel. The aim of this work was to study the effect of Carménère, Merlot and Cabernet Franc on the phenolic composition, proanthocyanidin profile and mouthfeel characteristics of Cabernet Sauvignon blends. RESULTS: Significant differences in chemical composition were observed among the monovarietal wines. Separation using Sep-Pak C18 cartridges revealed differences in the concentration but not in the proportion of various proanthocyanidins. Blending reduced polyphenol concentration differences among the various monovarietal wines. Although no major overall differences were observed after blending the monovarietal wines, this oenological practice produced clear differences in mouthfeel characteristics in such a way that the quality of the perceived astringency was different. CONCLUSION: This study showed that the use of a particular wine variety (Cabernet Sauvignon) in a higher proportion in wine blending produced blends that were less differentiable from the monovarietal wine, owing to a suppression effect, producing an apparent standardization of the wines regarding chemical composition.

Does wearing dentures change sensory nerve responses under the denture base?

BACKGROUND: The standard textbook on complete dentures suggests the necessity of relief for the incisive and posterior palatine foramina of denture wearers to prevent the impingement of the nerves and vessels passing through these foramina. However, concrete evidence of the effect of dentures on the sensory function of the nerves underlying dentures is lacking. OBJECTIVE: The study aim was to investigate the influence of denture-induced compression on sensory nerve responses to stimulations. MATERIALS AND METHODS: Only partially edentulous patients of maxillary Kennedy class II who wear dentures (WD) or who never wear dentures (ND) were recruited as participants. The WD and ND groups had 15 participants with a mean age of 66.9 years and 22 participants with a mean age of 60.2 years, respectively. Current perception thresholds (CPTs) at 2000, 250 and 5 Hz, corresponding to A-beta, A-delta and C fibres, respectively, were measured by the Neurometer(®) NS3000 device and compared between groups. The data were analysed by the t-test, the paired t-test and analysis of covariance adjusted for age, sex and thickness of the mucosa. RESULTS: The CPTs showed large differences at 2000 Hz but no differences at the other frequencies when the WD and ND groups were compared. Only the CPT of the edentulous side at 2000 Hz was significantly different between groups. CONCLUSION: Wearing removable partial dentures contributes to changes of responses to stimulations in the large fibre of the sensory nerve underlying the maxillary alveolar ridge.

A new protocol to evaluate the effect of topical anesthesia.

This double-blind, placebo-controlled, randomized cross-over clinical experimental study tested the reliability, validity, and sensitivity to change of punctuate pain thresholds and self-reported pain on needle penetration. Female subjects without orofacial pain were tested in 2 sessions at 1- to 2-week intervals. The test site was the mucobuccal fold adjacent to the first upper right premolar. Active lidocaine hydrochloride 2% (Dynexan) or placebo gel was applied for 5 minutes, and sensory testing was performed before and after application. The standardized quantitative sensory test protocol included mechanical pain threshold (MPT), pressure pain threshold (PPT), mechanical pain sensitivity (MPS), and needle penetration sensitivity (NPS) assessments. Twenty-nine subjects, mean (SD) age 29.0 (10.2) years, completed the study. Test-retest reliability intraclass correlation coefficient at 10-minute intervals between examinations was MPT 0.69, PPT 0.79, MPS 0.72, and NPS 0.86. A high correlation was found between NPS and MPS (r = 0.84; P < .001), whereas NPS and PPT were not significantly correlated. The study found good to excellent test-retest reliability for all measures. None of the sensory measures detected changes in sensitivity following lidocaine 2% or placebo gel. Electronic von Frey assessments of MPT/MPS on oral mucosa have good validity.

The effect of sonophoresis on topical anesthesia: a pilot project.

The dental anesthesia sonophoresis device (DASD) is a novel device that is intended to reduce the discomfort associated with intraoral mucosa needle puncture. The DASD produces ultrasonic energy that provides a sonophoretic effect on the oral mucosa, generating microchannels through the lipids between the keratinized cells that make up the stratum corneum. Once the topical anesthetic has permeated the stratum corneum, it quickly diffuses through the soft tissue, desensitizing the nerve endings and reducing the perception of pain caused by needle penetration. The aim of this study is to evaluate whether topical anesthesia applied using the DASD will reduce the discomfort of the needle puncture when compared to the control device. A split-mouth model, using 50 healthy subjects with puncture site at the maxillary canine vestibule, was used for this study. Subjects received a needle puncture on both sides of the mouth. Prior to the needle puncture, there was randomized application of 5% lidocaine with the DASD and a control device. Subjects rated their discomfort after needle punctures utilizing the visual analog scale pain scoring system. There was no statistically significant difference in the pain perception using the DASD versus the control device.

Oral neurovascular hamartoma: a lesion searching for a name.

BACKGROUND: Neurovascular hamartoma (NVH), in particular in the oral cavity, is rarely described in the literature. The low number of cases may reflect a genuine rarity of the lesion, or it may be due to its being unrecognized and/or under-reported. OBJECTIVES: To investigate clinical and microscopic features of oral NVH and to define microscopic diagnostic criteria with emphasis on the differential diagnosis. METHODS: Archival cases diagnosed as oral NVH between 1999 and 2011 were retrieved; clinical and demographic data were collected, and a paired morphometric analysis was conducted, with each case of NVH a case of fibrous hyperplasia (FH) from the same oral location. The nerve bundle and blood vessel density were quantified in five microscopic fields at ×100 magnification. RESULTS: The study group included 25 oral NVH, 11 men and 14 women, aged 6-76 years, (mean 44). The majority occurred in the tongue (54%), followed by the buccal mucosa and lower lip (17% each), clinically presenting as asymptomatic 0.25-2.5 cm exophytic masses. Microscopic characteristics included poorly circumscribed masses of closely packed nerve bundles and blood vessels in a loose matrix, containing minimal or no inflammation. The mean nerve bundle density was significantly higher in NVH (4.28 ± 1.26) in comparison with FH (0.27 ± 0.27), (P < 0.00001), and mean vessel density was significantly lower (5.98 ± 1.4 vs. 7.8 ± 1.9, respectively), (P < 0.0003). CONCLUSION: Oral NVH may not be as rare as previously considered. Morphometric analysis demonstrated that NVH presents a separate distinct entity.

Thermal sensory and pain thresholds in the tongue and chin change with age, but are not altered in burning mouth syndrome.

BACKGROUND: Burning mouth syndrome (BMS) is a chronic orofacial pain syndrome that occurs in middle-aged and postmenopausal women and poses a therapeutic challenge to dermatologists and dentists. It has been suggested previously that BMS is a small-fiber neuropathy. AIMS: This study was designed to examine thermal sensory and pain thresholds in the oral mucosa and chin, both innervated by the trigeminal nerve, in patients with BMS, as well as in healthy controls. In addition, the study proposed to examine whether there are any differences in oral thermal and pain sensations between the advanced age group, where BMS is prevalent and a younger group. RESULTS: Thermal and pain thresholds of BMS patients did not differ significantly from those of healthy subjects. An increased threshold to thermal warmth and a decreased threshold for cold sensation for the tongue and chin were noted in the group over 50 years in comparison with younger subjects, indicating a decreased sensitivity to thermal stimuli. The group over 50 years of age displayed an increased sensitivity to cold pain and a decreased sensitivity to hot pain in the tongue (compared with the chin). CONCLUSION: BMS patients do not demonstrate alterations in thermal and pain detection, thus failing to support a true small nerve neuropathy in this condition.

Merkel cells and permanent disesthesia in the oral mucosa after soft tissue grafts.

Connective tissue grafts are routinely procedures in the treatment of gingival defects. The clinical success of the gingival tissue graft procedures anyway should ensure not only the aesthetic integration between the tissues but also the physiological activity of the graft in terms of sensitivity and immunity because the skin and the mucosae constitute the first natural aspecific borders against pathogens. The aim of this paper was to investigate nervous net recovery after connective graft procedure, in relation with sensorial alteration in the injured area. Results showed that there is a close link among the number of Merkel cells and the alteration of sensations. Merkel cells can be found isolated standing in the basal layer, supposed to have neuroendocrine functions in the epithelia or in larger group not associated with nerves; when found in association with nerves they are named Merkel complexes, acting as slow adapter mechanical receptor. Our data can be explained in two ways: Merkel cells increase as a consequence of tissue injury, a sort of "SOS cells" that secrete neuroendocrine signals to guide tissue healing; as an alternative the presence of the Merkel cells could be read as a derailment of tissue regeneration with the stop of cellular differentiation in the direction of an abnormal proliferation, a sort of mad stem cell.

Sensations evoked by microstimulation of single mechanoreceptive afferents innervating the human face and mouth.

Intraneural microneurography and microstimulation were performed on single afferent axons in the inferior alveolar and lingual nerves innervating the face, teeth, labial, or oral mucosa. Using natural mechanical stimuli, 35 single mechanoreceptive afferents were characterized with respect to unit type [fast adapting type I (FA I), FA hair, slowly adapting type I and II (SA I and SA II), periodontal, and deep tongue units] as well as size and shape of the receptive field. All afferents were subsequently microstimulated with pulse trains at 30 Hz lasting 1.0 s. Afferents recordings whose were stable thereafter were also tested with single pulses and pulse trains at 5 and 60 Hz. The results revealed that electrical stimulation of single FA I, FA hair, and SA I afferents from the orofacial region can evoke a percept that is spatially matched to the afferent's receptive field and consistent with the afferent's response properties as observed on natural mechanical stimulation. Stimulation of FA afferents typically evoked sensations that were vibratory in nature; whereas those of SA I afferents were felt as constant pressure. These afferents terminate superficially in the orofacial tissues and seem to have a particularly powerful access to perceptual levels. In contrast, microstimulation of single periodontal, SA II, and deep tongue afferents failed to evoke a sensation that matched the receptive field of the afferent. These afferents terminate more deeply in the tissues, are often active in the absence of external stimulation, and probably access perceptual levels only when multiple afferents are stimulated. It is suggested that the spontaneously active afferents that monitor tension in collagen fibers (SA II and periodontal afferents) may have the role to register the mechanical state of the soft tissues, which has been hypothesized to help maintain the body's representation in the central somatosensory system.

Functional properties of low-threshold mechanoreceptive afferents in the human labial mucosa.

We used microneurography to investigate the functional properties of low-threshold mechanoreceptive afferents innervating the oral mucosa of the inside of the lower lip. Impulse responses were recorded from the inferior alveolar nerve of four human subjects. The threshold force and receptive field boundaries were identified for 19 single mechanoreceptive afferents using thin filaments (von Frey hairs) that applied known forces to the mucosa. Most of the receptive fields were located close to the corners of the mouth. Twelve of the afferents were slowly adapting (SA) and the remaining seven units were fast adapting (FA). Two types of slowly adapting responses were observed, SA I and SA II. Four of the six SA II units were spontaneously active. The geometric mean value of the receptive field sizes was 4.20 mm(2) for the SA I units, 5.65 mm(2) for the SA II units, and 5.60 mm(2) for the FA I units. None of the FA afferents showed response properties characteristic of Pacinian-corpuscle type afferents (FA II units). All afferents showed low force threshold between 0.06 and 1 mN. The properties of the mechanoreceptors supplying the human labial mucosa appear more similar to those of the vermilion and facial skin of the lower lip than those supplying the mucosa of the dorsal tongue.

A comparative study of children's pain reactions and perceptions to AMSA injection using CCLAD versus traditional injections.

UNLABELLED: Pain control is an important part of dentistry, particularly in pediatric dentistry. Recently, a computer-controlled local anesthetic delivery system (CCLAD) has been developed to reduce pain related to the local anesthetic injection. In conjunction with this technology, a new approach to the anterior and middle superior alveolar nerves (AMSA) has been induced. Studies evaluating the CCLAD in pediatric dentistry showed variable results regarding its use in pediatric dentistry. Further evaluation of this technique is needed to provide sound scientific evidence on the use of the CCLAD at this specific injection site in children. AIM: To assess children's pain reactions and pain perceptions of the AMSA injection using the CCLAD compared to the traditional buccal/palatal injections. MATERIALS AND METHODS: Children's pain reactions and perceptions to both techniques were measured in a group of 40 children who received both anesthetic techniques alternatively on two visits. The pain reactions were scored using the SEM scale, whereas the pain perception was evaluated by the Eland color scale. Statistical analysis was carried out using SPSS version 10.0. RESULTS: The AMSA injection delivered with the CCLAD had significantly lower mean pain reaction scores compared to traditional buccal and palatal injections. The prolonged injection time required for delivering the CCLAD injection had no negative impact on the children. The children's pain perception scores when using the CCLAD were also significantly lower compared to the traditional injection. CONCLUSION: The AMSA injection delivered with the CCLAD was found to be a promising device, and had significantly lower pain reaction and perception scores compared to the traditional buccal and palatal injections.

The involvement of CGRP, adrenomedullin, and sensory nerves in remote vasomotor responses within the hamster cheek pouch microcirculation.

Previous work from our laboratory demonstrated a role for sensory nerves in remote dilations to microapplied methacholine by blocking the response with CGRP8-37 and concluded CGRP was the neurotransmitter. Recently, a more specific CGRP receptor antagonist, BIBN4096BS, was developed. The goals of the present study are to characterize the effects of BIBN4096BS on vasomotor responses in the hamster cheek pouch microcirculation, and to verify the role of CGRP in remote dilations to capsaicin and methacholine and to test adrenomedullin as an alternative neurotransmitter. BIBN4096BS pretreatment inhibits dilation to CGRP while having no significant effect on baseline diameter, it shifts the EC(50) to superfused CGRP from 1.5+/-0.3 pM to 2.5+/-0.6 nM and it shifts the apparent EC(50) to capsaicin from 31.5 nM to 171 nM. Local and remote dilations caused by the microapplication of methacholine are not inhibited by 300 nM BIBN4096BS (Local: 9.7+/-1.2 versus 9.7+/-1.5; 500:5.5+/-0.4 versus 5.7+/-0.5; 1000:4.4+/-0.6 versus 4.8+/-0.5). Remote dilations to methacholine were significantly inhibited however when adrenomedullin receptor antagonist adrenomedullin-(26-52) was microapplied to the remote site. Perivascular neurons containing adrenomedullin can be detected with immunohistochemistry. The results, combined with previous work, suggest that adrenomedullin, and not CGRP, is involved in remote dilations to methacholine.