Kinetic study of immunohistochemical colocalization of antigen-presenting cells and nerve fibers in rat periapical lesions.

Immune and nervous systems play key roles in periapical pathosis; however, their spatial associations, which may be a prerequisite for paracrine interactions in the progression of periapical lesions, remain to be examined. In this study we examined the distribution and frequency of spatial associations between major histocompatibility complex class II molecule-expressing antigen-presenting cells (APCs) and protein gene product 9.5-immunoreactive nerve fibers in experimentally induced rat periapical lesions using double-immunofluorescence staining and confocal laser scanning microscopy. When active lesion expansion started, macrophage-like APCs frequently associated with nerve fibers around the apex. When the lesions were starting to stabilize, however, close associations between APCs with dendritic morphology and nerve fibers were found mostly in the periphery of lesions. CD86+ mature dendritic cells were also observed in this area. These findings suggest that functional interactions between APCs and nerve fibers may play some roles in the development of self-defense reactions in periapical lesions.

Coexpression of vasoactive intestinal polypeptide and substance P in reinnervating pulpal nerves and in trigeminal ganglion neurones after axotomy of the inferior alveolar nerve in the rat.

The effect of this axotomy on the expression of vasoactive intestinal polypeptide (VIP) in trigeminal ganglion neurones and nerve fibres in the first molar was examined immunohistochemically 3 weeks postsurgically in rats. A distinct upregulation of VIP-like immunoreactivity was found in 3 to 4% of the neurones (mean dia., 20.9 +/- 0.45 microns; mean cross-sectional area, 367 +/- 13.2 microns 2) in the mandibular region after axotomy. An almost complete coexpression was registered in neurones upregulated for VIP and growth-associated protein 43. Coexpression of VIP and substance P (SP) was found in a small number of the immunoreactive (IR) small-sized neurones, mainly in those in which VIP and SP were both weakly immunoreactive. In the uninjured ganglion, weakly labelled VIP-positive granules were frequently traced at high magnification in the cytoplasm of small neurones. No VIP-IR fibres were found in the control molar pulp, except for a few in the root pulp near the apex. However, 3 weeks after axotomy, a number of VIP-containing nerve fibres were found in the molar pulp and apical periodontium. Coarse VIP-IR fibres directed towards the odontoblast layer were a common finding. In some of these fibres VIP was shown to colocalize with SP. These results indicate that VIP is clearly expressed and transported in regenerating primary afferent neurones after axotomy of the inferior alveolar nerve.

Immunohistochemical examination of the distribution of macrophages and CGRP-immunoreactive nerve fibers in induced rat periapical lesions.

To clarify the role of calcitonin gene-related peptide (CGRP) in the development of periapical lesions, we examined the distribution of CGRP-immunoreactive (IR) nerve fibers and macrophages, and the behavior of bone tissues in experimentally induced rat periapical lesions by immunohistochemical and quantitative methods. Although no extensive changes were observed at 7 days after pulp exposure, CGRP-IR nerve fibers increased in number until 28 days with a decrease thereafter. These neural changes were closely correlated with the alteration in number of macrophages except on day 7 when macrophages were significantly increased in number as compared with control rats. Tissue repair began to take place and a decrease in number of osteoclasts was observed when the density of CGRP-IR nerve fibers reached a peak. These results suggested that there might be a close relationship between macrophages and CGRP-IR nerve fibers and that CGRP-IR nerve fibers might participate in tissue repair in experimentally induced rat periapical lesions.

Steroids reduce the periapical inflammatory and neural changes after pulpectomy.

Root canal treatment, including obturation with gutta-percha and a zinc oxide and eugenol sealer, was conducted, under general anesthesia, on the canine teeth of 12 young ferrets. Six of the ferrets were given 0.5 mg/kg dexamethasone daily. Three months after the root canal treatment, under general anesthesia, the animals were perfused with fixative and the canine periapical tissues prepared for histological examination. The extent of periapical inflammation was measured and the degree of neural sprouting in the periodontal and subapical regions estimated. Periapical lesions in steroid-treated animals were 30% of the size of those in untreated animals. Innervation density in the subapical region of the steroid-treated animals was lower than that in the animals who did not receive steroids and not significantly different from controls. Reduction in periapical inflammation induced by systemic steroids is accompanied by a reduction in neural sprouting.

Periapical neural changes after pulpectomy.

Pulpectomy and pulpal necrosis result in severance of the nerves that supply the pulp as well as loss of their target organ. Inflammatory changes commonly extend into the periapical region to involve those nerves. The neural response to pulpal loss combined with periapical inflammation is a derangement of the periodontal plexus normally located in the center of the periodontal space around the apical third of the root; the result is the formation of a disorganized group of sprouting and branching axons that have some features in common with neuromas. The inflammatory and neural responses continue for at least a year even when pulpectomy is followed by canal debridement and obturation. Then the responses are reduced but not eliminated by steroids. Root canal therapy with techniques that do not leave residual inflammation still results in increased periapical innervation; the increase seems to be an organized addition to the normal periradicular plexus.

A histological comparison of periapical inflammatory and neural responses to two endodontic sealers in the ferret.

Previous studies have shown that there is a proliferation of nerves beneath the apices of pulpectomized teeth. This may be due to the inflammation induced after the procedure, resulting, perhaps, from the irritant nature of the materials used to fill the root canal. The experiment reported here was conducted to determine whether this inflammation was induced by the sealer rather than arising as a result of tissue damage and whether, if inflammation is eliminated or reduced, the neural changes are also reduced. In 12 young adult ferrets under general anaesthesia the pulps of the lower canine teeth were removed and replaced with gutta percha and sealer. A sealer of Grossman's formulation with eugenol as the liquid phase was used on one side and a calcium hydroxide sealer that contained no eugenol on the other. Three months later the animals were, again under general anaesthesia, perfused with a fixative mixture. The mandibles were removed and the presence and size of any periapical inflammatory lesions and the density of periapical innervation determined histologically. All 12 teeth sealed with the Grossman's sealer had inflammatory lesions at their apices. Three of the 12 teeth treated with the calcium hydroxide sealer showed similar lesions. In all the teeth with inflammatory lesions the normal arrangement of nerves in a periodontal 'plexus' was disrupted but there was no statistically significant difference between the overall innervation density in inflamed and non-inflamed periapical areas nor between areas beneath teeth sealed with Grossman's sealer and with calcium hydroxide. The incidence of periapical inflammation is related to the nature of endodontic sealer used.(ABSTRACT TRUNCATED AT 250 WORDS)

Neural changes in periapical lesions after systemic steroids in the ferret.

This study was intended to clarify the relationship between the neural changes which occur around the apex of the ferret canine after pulpectomy and the inflammatory process induced by the procedure. In 12 young adult ferrets, under general anesthesia, the pulps in the mandibular canine teeth were removed and replaced with gutta percha and Grossman's sealer. Six of the animals were treated with dexamethasone to reduce the inflammatory response. Three months later, the animals, again under general anesthesia, were perfused with a fixative mixture. Three unoperated animals that had not been treated with dexamethasone were also perfused. The mandibular canine teeth and their supporting tissues were removed, processed, and serially sectioned. Three-dimensional reconstructions of the periapical lesions in each animal were assembled and their volumes measured. The density of innervation in the periapical region was estimated. The mean lesion volume in the pulpectomized animals not treated with dexamethasone was 3.54 (+/- 2.27) mm3 and in the dexamethasone-treated animals 1.33 (+/- 1.31) mm3. The differences were statistically significant when tested by the Mann-Whitney U test (p < 0.01). Bacteria were not seen within any of the lesions. The innervation density beneath the canines in the pulpectomized animals not treated with dexamethasone was 164 units per mm2 (+/- 80) and in the steroid-treated animals 151 +/- 68 units per mm2. In the control, untreated animals, the innervation density was 22 +/- 10 units per mm2. The difference between the steroid-treated pulpectomized animals and the untreated pulpectomized animals was not statistically significant (p > 0.5).(ABSTRACT TRUNCATED AT 250 WORDS)

Periapical innervation of the ferret canine one year after pulpectomy.

Previous studies have shown that, following removal of the dental pulp and its replacement with a filling material, the nerves in the periapical region proliferate within an area of chronic inflammation. This study examined these responses in the ferret in more detail, both quantitatively and in three dimensions, 12 months after pulpectomy which was followed by obturation with gutta percha and zinc oxide and eugenol sealer. The results were compared with the responses after three months. The basic patterns, in terms of both inflammation and neural proliferation, were similar at the two time periods. Both the sizes of the inflammatory lesions and the degree of neural proliferation were lower at 12 months than at three, although the differences were not significantly different in the small number of specimens examined. Bacterial stains failed to show bacteria either in the apical root canal delta or in the periapical area. It is possible that the persistent inflammation was due to the irritant nature of the obturating materials used but not due to the original tissue damage, since extraction sites, by comparison, healed very quickly. Apparently, following endodontic treatment, chronic periapical inflammation and concomitant neural proliferation can continue for long periods.

[Innervation of inflammatory periapical lesions. Histopathological aspects]. / Innervazione nelle lesioni infiammatorie periapicali: aspetti isto-patologici.

The authors through a literatures, review, shows the istopathological aspects of the innervation of the inflammatory periapical lesions, emphasizing the most important characteristic.

The effect of pulpectomy on the longitudinal distribution of nerve fibres in the periodontal ligament of the ferret.

The innervation density of the periodontal ligament was measured at a series of levels around the mandibular canines of 6 ferrets, in 5 of the which the pulp of one of each of these pairs of teeth had been removed 3 months earlier. In control (unoperated) teeth the density of innervation was greatest in the apical third and lowest in the coronal third. The pattern around the pulpectomized teeth was similar. There was no statistically significant difference between density values for control and operated teeth. Pulpectomy thus does not affect the incidence or distribution pattern of nerves in the periodontal ligament.

Changes in CGRP-immunoreactive nerve fibres during experimental tooth movement in rats.

Immunohistochemical localization of calcitonin gene-related peptide (CGRP) was used to investigate changes in nerves expressing CGRP in periodontium and pulp during experimental mesial movement of the first maxillary molar in rats. The orthodontic appliance consisted of a coil spring connecting the first maxillary molar on one side to the central incisors. After 5 days with a continuous force of 30-50 g the animals were perfused after an overdose of anaesthetic. Serial sections from the experimental and control jaws were exposed to the avidin-biotin-peroxidase technique for demonstration of CGRP-immunoreactive (IR) nerve fibres. The induced tooth movement caused reproducible changes in the relative number of CGRP-IR nerves as well as morphological alterations within parts of the nerve supply of the experimental teeth and related tissue structures. The majority of the experimental teeth showed increased number of CGRP-IR nerves in the coronal pulp and periapical tissues. The results indicate that peptidergic nerve fibres immunoreactive for CGRP takes an active part in tissue responses in pulp and supporting tissues during experimental tooth movement.

Effects of injury and inflammation on pulpal and periapical nerves.

Several studies dealing with the reactions of dental nerve fibers to injury and inflammation are reviewed in this article. The subgroup of dental nerve fibers that contains calcitonin gene-related peptide (CGRP) was examined by immunocytochemistry at various times (1 to 35 days) after one of three degrees of injury: (a) Mild: Four days after making shallow cavities into cervical dentin of first molars of anesthetized adult rats, we found that CGRP fibers had sprouted into the subjacent odontoblast layer and dentin, and then returned to normal by 3 wk. (b) Intermediate: If the cervical cavities were acid etched, we found damage to the odontoblast layer, microabscess formation, and sprouting of CGRP fibers near the abscess, with subsequent formation of reparative dentin and healing. (c) Severe: If the pulp was exposed, a variety of reactions could occur, the most prevalent of which was a severe necrosis leading to development of periapical lesions. Analysis of the progressive stages of pulpal abscess and necrosis showed sprouting CGRP nerve fibers (a) at the retreating interface between abscess and vital pulp; (b) in periapical areas during onset of lesions; and (c) around chronic abscesses in granulomatous periodontal tissues. These studies are discussed in relation to various dental clinical problems such as hypersensitive teeth, episodic toothache, early onset of periapical lesions, dental anesthesia, and possible roles for sensory fibers and neuropeptides in tissue defense and healing.

Tratamento näo cirúrgico de dentes com lesöes periapicais / Non-surgery treatment of the teeth with periapical lesions

Com base em resultados obtidos em trabalhos experimentais anteriores, estudou-se em dentes humanos portadores de lesöes periapicais o efeito do tratamento endodôntico realizado com hidróxido de cálcio. Os resultados clínico-radiográficos obtidos permitiram concluir-se pela validade do tratamento executado, diante da alta cifra de êxito alcançada

A fine-structural investigation on the extent of perineurial investment of the nerve supply to the pulp in rat molar teeth.

The fine structure of the tissues immediately adjacent to nerve fibres supplying the rat molar pulp has been examined and compared with that of perineurium of other peripheral nerves. In the periapical region, almost all nerve fibres were invested with typical perineurium; only a few Schwann cell units (unmyelinated axons) lacked perineurium. In the pulp at the apex of the distal root, most fibres were invested by less typical perineurium but some were not invested at all. In the pulp at the coronal end of the root, nerve fibres were completely invested or incompletely invested with attenuated perineurium or lacked investment altogether. In the coronal pulp, there was virtually no perineurium at all. Thus rat molar pulp nerve fibres lack perineurium for distances of about 0.5-3.0 mm, distances greater than in situations such as skin or muscle. In normal peripheral nerves, the perineurium behaves as a barrier to the passage of substances from the immediate environment of the nerve fibres (endoneurium) to the surrounding environment (epineurium) and vice versa. Its absence around pulp nerve fibres raises questions about the nature of the pulp environment and may help to explain the sensitivity of pulp nerves to chemical, osmotic and other stimuli.

Stimulus-evoked pain in teeth.

When pain occurs in teeth as a result of external stimulation it is more likely to be due to physical factors affecting the enamel and dentine than to pulp disease. The mechanisms are described, clinical conditions are mentioned and a speculative attempt is made to correlate them by introducing the idea of streaming potentials in the dentine and pulp. The pain perception threshold is considered in relation to different teeth, age, sex, lateral dominance, personality, and the area of electrode contact. The latter has proved interesting, for manipulation of it has led to possibly important findings on the nature of convergence in the trigeminal system, two-point discrimination, referred pain and somatopic representation of dental pulps in the medulla oblongata.