Gingival crevicular fluid and plasma levels of neuropeptide Substance-P in periodontal health, disease and after nonsurgical therapy.

BACKGROUND AND OBJECTIVE: The level of Substance-P in gingival crevicular fluid has been found to correlate with clinical measures of periodontal disease. The present study was designed to assess the relationship between clinical parameters and levels of Substance-P in the gingival crevicular fluid from inflamed gingiva, periodontitis sites and after treatment of periodontitis sites, and to correlate them to the Substance-P levels of plasma. MATERIAL AND METHODS: Thirty, age- and gender-matched subjects were divided into three groups (healthy, gingivitis and chronic periodontitis) based on modified gingival index scores and clinical attachment loss. A fourth group consisted of 10 subjects from the periodontitis group, 6-8 wk after initial therapy. Plasma and gingival crevicular fluid samples were collected and quantified for Substance-P using an enzyme immunoassay. RESULTS: The mean concentration of Substance-P, both in gingival crevicular fluid and plasma, was observed to be highest in the periodontitis group (45.13 pg/mL in gingival crevicular fluid and 67.8 pg/mL in plasma) and lowest in the healthy group (6.07 pg/mL in gingival crevicular fluid and below the detection level in plasma). The mean Substance-P concentration in the gingivitis group (11.42 pg/mL in gingival crevicular fluid and 38.8 pg/mL in plasma) and in the after-treatment group (7.58 pg/mL in gingival crevicular fluid and 39.7 pg/mL in plasma) lay between the highest and lowest values. In all groups the gingival crevicular fluid levels showed a statistically significant positive correlation with that of plasma and clinical attachment loss. CONCLUSION: Substance-P levels were highest in the gingival crevicular fluid from sites with periodontal destruction; however, periodontal treatment resulted in the reduction of Substance-P levels. Gingival crevicular fluid and plasma Substance-P levels showed a positive correlation in all of the groups.

Experimental tooth pain elevates substance P and matrix metalloproteinase-8 levels in human gingival crevice fluid.

OBJECTIVE: Tooth pain can induce a neurogenic inflammatory reaction in gingiva in association with local elevations of matrix metalloproteinase (MMP)-8, which is considered the major tissue destructive protease in gingival crevice fluid (GCF). The pro-inflammatory neuropeptides released by sensory nerves coordinate the activities of the immuno-effector cells and may influence the secretion of MMP-8. With this background, we studied whether experimental tooth pain can trigger changes in GCF levels of the neuropeptide substance P (SP) and MMP-8. MATERIAL AND METHODS: The GCF SP levels of stimulated and non-stimulated teeth were analyzed for SP using a competitive enzyme immunoassay (EIA). The GCF MMP-8 levels were determined by quantitative immunofluorometric assay (IFMA). RESULTS: Painful stimulation of the upper central incisor caused significant elevations in GCF SP and MMP-8 levels of the stimulated tooth. At the same time, the GCF SP and MMP-8 levels of non-stimulated control teeth were unchanged. CONCLUSIONS: These data indicate that experimental tooth pain can induce local elevations of SP and MMP-8 levels in GCF simultaneously. This supports the possibility of a local neurogenic spread of inflammatory reactions from intrapulpal to surrounding periodontal tissues.

Capsaicin-sensitive Innervation Modulates the Development of Apical Periodontitis.

INTRODUCTION: Nociceptive neurons play a critical role in the detection of stimuli evoking actual or potential tissue injury. In addition, they are involved in neurogenic inflammation by the peripheral release of neuropeptides such as calcitonin gene-related peptide (CGRP). The dental pulp and periradicular tissues are innervated by capsaicin-sensitive neurons known to release CGRP. However, the role of these capsaicin-sensitive neurons in the development of apical periodontitis is largely unknown. The aim of this study was to evaluate the contribution of peptidergic neurons to the development of apical periodontitis. METHODS: Neonatal Sprague-Dawley rats were injected with vehicle (control group) or a single subcutaneous capsaicin dose to cause the selective ablation of peptidergic neurons (neonatal capsaicin group). Ablation of capsaicin-sensitive neurons was verified with confocal microscopy, capsaicin-induced eye-wipe nocifensive behavior test, and by measurement of immunoreactive CGRP levels in the dental pulp. Five weeks after ablation, standardized pulp exposures were made in the mandibular left first molars. Mandibles were harvested at 7, 14, 21, and 28 days after pulp exposure and imaged with micro-computed tomography (µCT) to quantify apical lesion volume. Data were analyzed by using 2-way ANOVA analysis with Bonferroni post hoc test. RESULTS: Rats in the control group displayed a robust capsaicin-induced nocifensive behavior, which was nearly abolished in the neonatal capsaicin group. In addition, the neonatal capsaicin group showed a significant depletion of susceptible neurons and CGRP in the dental pulp compared with control. Importantly, micro-computed tomography analysis showed larger periradicular lesions at 7 and 14 days after pulp exposure in the neonatal capsaicin group when compared with control. CONCLUSIONS: Results identify a protective role for capsaicin-sensitive neurons in the initial phase of apical periodontitis. Thus, interventions or disorders that alter activity of capsaicin-sensitive fibers are likely to alter the development of apical periodontitis.

Painful tooth stimulation elevates matrix metalloproteinase-8 levels locally in human gingival crevicular fluid.

Recent studies have demonstrated that pulpal pain can induce neurogenic inflammatory reactions in gingiva and the expression of pro-inflammatory neuropeptides in gingival crevicular fluid (GCF). Neuropeptides co-ordinate the activity of immuno-effector cells and may influence the secretion of matrix metalloproteinase (MMP)-8, the major tissue-destructive protease in GCF. With this background, we studied whether experimental pulpal pain can trigger changes in GCF MMP-8 levels. The molecular forms of MMP-8 in the GCF of stimulated and non-stimulated teeth were analyzed by Western immunoblot, and MMP-8 levels by quantitative immunofluorometric assay. Painful stimulation of the upper incisor provoked significant elevations in GCF MMP-8 levels of the stimulated tooth. Western immunoblot revealed elevations in both neutrophil- and mesenchymal-type MMP-8 isoforms. At the same time, the GCF MMP-8 levels of the non-stimulated teeth were not changed. Analysis of these data indicated that pulpal pain can induce local elevations in MMP-8 levels in GCF.

Gingival crevicular fluid levels of neuropeptides following dental restorations.

PURPOSE: Local neuropeptide release has a critical role in the initiation and progression of an inflammatory response. This study investigated the effects of different restorative materials on periodontium in this regard, by evaluating their neuropeptide-producing effects on gingival crevicular fluid (GCF). METHODS: The study included 14 patients suitable for metal-ceramic, composite and amalgam restorations. Four weeks after periodontal therapy, the restorations were performed. Study groups were constituted regarding the tooth/restoration surfaces contacting gingiva in each patient: 1 ceramic surface of a metal-ceramic crown (ceramic group), its opposite metal surface (metal group), 1 composite surface (composite group), its opposite enamel surface (opposite-composite group), 1 amalgam surface (amalgam group), its opposite enamel surface (opposite-amalgam group) and 1 nonrestored enamel surface (enamel group). Four weeks after dental restorations, clinical data and GCF were obtained from the group sites. Clinical data, GCF volume and its proinflammatory cytokine profile were utilized to evaluate the periodontal health. GCF levels of substance P (SP), neurokinin A (NKA) and calcitonin-gene related peptide (CGRP) were determined by ELISA for revealing the neuropeptide levels. RESULTS: GCF volume was found to increase in all groups compared with the enamel group (p<0.05). SP and NKA levels were higher in the ceramic, composite and amalgam groups than those in the enamel group (p<0.05). SP and NKA levels were also higher in the composite and amalgam groups than those in the opposite-composite/amalgam groups (p<0.05). CONCLUSIONS: These results suggest that ceramic, composite and amalgam materials may uniquely trigger local neuropeptide release in periodontium.

Prostaglandin E2 enhances bradykinin-evoked iCGRP release in bovine dental pulp.

Mediators produced during inflammation are responsible for hyperalgesia and expression of neurotransmitters and receptors in the nervous system. The production of bradykinin (BK) and the prostaglandins (PGs) may regulate initiation of pain. This study tested the hypothesis that BK and prostaglandin E2 (PGE2) have a positive interaction in evoking neurosecretion of immunoreactive calcitonin gene-related peptide (iCGRP). Bovine dental pulp was prepared and stimulated by the superfusion method with BK alone and in combination with PGE2. Kinin receptor antagonists to bradykinin-evoked release of iCGRP were also tested. Also tested was the hypothesis that dental pulp contains either the B1 or B2 or both BK receptors. Results showed that PGE2 enhanced BK-evoked iCGRP release by more than 50%. Western immunoblots revealed detectable B2 receptor protein with no detectable B1 receptor protein. We conclude that BK evokes iCGRP release from bovine dental pulp which is enhanced by a positive interaction with PGE2. Neurosecretion is evoked from isolated terminals of dental pulp fibers via the bradykinin B2 receptor-dependent mechanism.

Tissue levels of immunoreactive substance P are increased in patients with irreversible pulpitis.

Odontogenic pain often involves inflammation of dental pulp tissue. Dental pulp is highly innervated with a subpopulation of sensory neurons containing neuropeptides. Substance P, released from afferent fibers (e.g. nociceptors) is associated with the development of neurogenic inflammation. In this study, we tested the hypothesis that irreversible pulpitis is associated with increased activity of peptidergic neurons, as measured by increased pulpal levels of immunoreactive substance P (iSP). We determined in vivo pulpal levels of immunoreactive substance P in human teeth with a diagnosis of normal pulp or irreversible pulpitis using CMA/20 microdialysis probes inserted into vital pulps of 24 teeth from 21 patients. Probes were perfused with a modified Locke-Ringer's buffer and immunoreactive substance P levels in the dialysate were measured using a radioimmunoassay. Mean extracellular levels of immunoreactive substance P were significantly higher (>8-fold) in teeth diagnosed with irreversible pulpitis than immunoreactive substance P levels in dental pulp diagnosed as normal (147.7 +/- 34.0 pM versus 18.2 +/- 6.2 pM). These observations suggest that biochemical measures of inflammatory mediators exhibit significant change during irreversible pulpitis and may contribute to clinical signs and symptoms.

Expression of calcitonin gene-related peptide (CGRP) in irreversible acute pulpitis.

The main goal of this study was to evaluate tissue levels of calcitonin gene-related peptide (CGRP) in human pulpal samples collected from teeth with a clinical diagnosis of acute irreversible pulpitis, normal pulps, and teeth with induced pulpal inflammation. All the pulp tissue was mechanically separated, collagenase digested to release individual cells, and labeled with FITC detection of an anti-CGRP polyclonal antibody. Detection of CGRP was possible in these cells due to a binding of the antibody to CGRP that was itself bound to its cell surface receptor. Flow cytometry analysis indicated that the labeled pulp cells were located in a region of low size and complexity according to their forward (FSC) and side scatter (SSC) properties. Significant statistical differences were found between the percentages of CGRP expression in healthy pulps and pulps with induced inflammation and between healthy pulps and pulps with acute irreversible pulpitis. No significant statistical differences were found between pulps with induced inflammation and pulps with acute irreversible pulpitis. These findings support the hypothesis that the CGRP system is active in human pulpal inflammation and may modulate the inflammatory response.

Substance P expression is elevated in inflamed human periradicular tissue.

Substance P is a neuropeptide believed to be a major mediator of neurogenic inflammation. The aim of our study was to evaluate whether substance P levels are elevated in the clinical biopsies collected from inflamed periradicular or control tissue. In this study, the presence of substance P was examined in infected human periradicular granulation tissue and control tissue. Sections from 19 periradicular granulomas and pulp tissues from two healthy control teeth were examined using the immunohistochemical method. Substance P-expressing neutrophils, macrophages, and plasma cells were found in both acute and chronic periradicular granulomas. In addition, we observed the presence of neutrophils expressing substance P without concurrent clinical symptoms of acute inflammation. Our results are consistent with the hypothesis that substance P may be released from neutrophils in the inflamed region, and thus, substance P may modulate clinical inflammatory response by release from either neuronal or immunocompetent cell populations.

Effect of inflammation on the delivery of drugs to dental pulp.

Although numerous studies have evaluated the effects of drugs on inflammation, comparatively few studies have evaluated the effects of inflammation on drugs. In this study, we have evaluated whether pulpal inflammation alters the delivery of flurbiprofen or Evan's blue, two agents that bind with high affinity to plasma proteins. The results indicate that pulpal inflammation alters the delivery of these agents to inflamed molars, that activation of capsaicin-sensitive nerves increases pulpal content of protein-bound agents, and that reduced pH increases free drug concentrations of flurbiprofen. Thus, alterations in both plasma extravasation and tissue pH seem to be relevant factors regulating the delivery and bioavailability of this nonsteroidal anti-inflammatory drug to dental pulp. Because many drugs used in endodontics (e.g. nonsteroidal anti-inflammatory drugs, clindamycin, bupivacaine, etc.) are heavily bound to plasma proteins, it is likely that the status of pulpal inflammation is a contributing factor in modifying the pharmacological efficacy of these agents.

Relationship between levels of neuropeptide Substance P in periodontal disease and chronic pain: a literature review.

The aim of the current review was to investigate the relationship between levels of neuropeptide Substance P in periodontal disease and chronic pain. Substance P is a neuropeptide that is directly related with pain. In periodontal disease, it is expressed during the inflammatory process, and is one of the factors responsible for bone resorption. Studies have shown that Substance P levels are highest in the gingival crevicular fluid from sites with active periodontal disease and bone loss. The persistence of these substances could be sufficient to stimulate neurogenic inflammation in susceptible tissues, and cause pain. The scientific literature shows that Substance P expressed during periodontal disease can be a risk factor for patients with systemic inflammatory pathologies, such as chronic arthritis or rheumatoid arthritis. Additional research is needed to confirm the participation of this substance in the origin of some types of chronic pain.

Expression of substance P, calcitonin gene-related peptide, ß-endorphin and methionine-enkephalin in human dental pulp tissue after orthodontic intrusion: a pilot study.

OBJECTIVE: To determine the levels of two sensory neuropeptides (substance P [SP] and calcitonin gene-related peptide [CGRP]) and two endogenous opioids (methionine-enkephalin [Met-Enk] and ß-endorphin [ß-End]) in dental pulp tissue samples subjected to controlled orthodontic intrusive forces. MATERIALS AND METHODS: Sixteen healthy premolars were selected from eight patients who were undergoing extraction for orthodontic purposes. Eight were randomly used as controls, and the other eight were assigned to an experimental group (controlled orthodontic intrusive forces applied for 24 hours). After this period, teeth were extracted, and pulp samples were obtained. All samples were processed to quantify the expression levels of SP, CGRP, Met-Enk, and ß-End using commercial radioimmunoassay kits. RESULTS: All samples exhibited basal levels of both neuropeptides and endogenous opioids. After 24 hours of the intrusive stimulus, all patients reported a tolerable discomfort localized at the involved premolar. Only SP was significantly increased (P<.05). For the other molecules, no statistically significant differences were observed (P>.05); however, they expressed important increasing trends. CONCLUSIONS: The expression levels of SP and CGRP in dental pulp samples from the experimental group support the positive correlation between the symptomatic clinical scenario and increased expression levels of neuropeptides, clarifying the role of neurogenic inflammation in early injury response.

Sex differences in neuropeptide content and release from rat dental pulp.

INTRODUCTION: Studies to examine sex differences in response to pain have suggested that females exhibit lower threshold responses to painful stimuli and that threshold response varies greatly at different stages of the menstrual cycle. Additional studies suggest that sex differences may be caused by societal sex roles or differences in anxiety responses by men and women. OBJECTIVE: The purpose of this study was to evaluate biologically evident sex differences in male and female rats chronically treated with a systemic algogen, the nerve growth factor (NGF), by measuring neuropeptides (calcitonin gene-related peptide) content and release from isolated dental pulp. METHODS: Rats were injected subcutaneously every other day with either murine NGF (1 mg/kg) or vehicle for 7 or 13 days. Isolated incisor pulp tissue was evaluated from these male and female rats (n = 96). Capsaicin-evoked neurosecretion of CGRP and tissue content were measured using a previously validated radioimmunoassay. RESULTS: Dental pulp from female rats at 7 days showed significantly increased capsaicin-evoked immunoreactive CGRP release (>50% increase) compared with tissue from male rats. After 13 days, this release was significantly increased only in NGF-treated female rats (3-fold increase) when compared with control females or both male groups. The CGRP content in tissue from both female groups was also significantly increased after 7 days of treatment (>3 fold), but after 13 days this content was only significantly increased in tissue from NGF-treated female rats (P = .0001). CONCLUSIONS: These data suggest that sex differences affect the role of NGF in the modulation of inflammation through the regulation of peripheral neuropeptide release and content.

Dissecting the role of the S1P/S1PR axis in health and disease.

Sphingosine-1-phosphate (S1P) is a pleiotropic sphingophospholipid generated from the phosphorylation of sphingosine by sphingosine kinases (SPHKs). S1P has been experimentally demonstrated to modulate an array of cellular processes such as cell proliferation, cell survival, cell invasion, vascular maturation, and angiogenesis by binding with any of the five known G-protein-coupled sphingosine 1 phosphate receptors (S1P1-5) on the cell surface in an autocrine as well as a paracrine manner. Recent studies have shown that the S1P receptors (S1PRs) and SPHKs are the key targets for modulating the pathophysiological consequences of various debilitating diseases, such as cancer, sepsis, rheumatoid arthritis, ulcerative colitis, and other related illnesses. In this article, we recapitulate these novel discoveries relative to the S1P/S1PR axis, necessary for the proper maintenance of health, as well as the induction of tumorigenic, angiogenic, and inflammatory stimuli that are vital for the development of various diseases, and the novel therapeutic tools to modulate these responses in oral biology and medicine.

Substance P expression by human dental pulp fibroblasts: a potential role in neurogenic inflammation.

Neurogenic inflammation describes the local release of neuropeptides, notably substance P (SP), from afferent neurons and might play a role in the pathogenesis of pulpal disease. The fibroblast is the most numerous cell type in the dental pulp, and recent work has suggested that it is involved in the inflammatory response. Primary pulp fibroblast cell populations were isolated by enzymatic digestion. Whole pulp tissue was obtained from freshly extracted sound (n = 35) and carious (n = 39) teeth. Expression of SP and neurokinin-1 receptor (NK-1) mRNA by pulp fibroblasts was determined by reverse transcriptase polymerase chain reaction (RT-PCR). SP was expressed by pulpal fibroblasts at both mRNA and protein levels. In addition, NK-1 mRNA and protein expression was detected in fibroblast cultures by RT-PCR and Western blotting, respectively. SP levels, determined by radioimmunoassay, were significantly greater (P < .05) in carious compared with sound teeth. These findings suggest that pulp fibroblasts play a role in neurogenic inflammation in pulpal disease.

Calcitonin gene-related peptide receptor expression in healthy and inflamed human pulp tissue.

AIM: To use radioreceptor analysis for comparing calcitonin gene-related peptide (CGRP) receptor expression in human pulp tissue samples collected from teeth having a clinical diagnosis of acute irreversible pulpitis, healthy pulps and teeth with induced inflammation. METHODOLOGY: Six pulp samples were obtained from teeth having a clinical diagnosis of acute irreversible pulpitis. Another eight pulp samples were obtained from healthy premolars where extraction was indicated for orthodontic purposes. In four of these premolars, inflammation was induced prior to pulp collection. All the samples were processed and labelled with 125I-CGRP. Binding sites were identified by 125I-CGRP and standard CGRP competition assays. RESULTS: CGRP receptor expression was found in all human pulp tissue samples. Most receptors were found in the group of pulps from teeth having a clinical diagnosis of acute irreversible pulpitis, followed by the group of pulps having induced inflammation. The least number of receptors was expressed in the group of healthy pulps. The Kruskal-Wallis and Mann-Whitney (post-hoc) tests showed statistically significant differences between the groups (P < 0.05). CONCLUSION: CGRP receptor expression in human pulp tissue is significantly increased during inflammatory phenomena such as acute irreversible pulpitis.

Carboxypeptidase-mediated metabolism of calcitonin gene-related peptide in human gingival crevicular fluid--a rôle in periodontal inflammation?

BACKGROUND: Metabolism by peptidases plays an important rôle in modulating the levels of biologically-active neuropeptides. The metabolism of the anti-inflammatory neuropeptide calcitonin gene-related peptide (GCRP), but not the pro-inflammatory neuropeptides substance P (SP) and neurokinin A (NKA) by components of the gingival crevicular fluid (GCF), could potentiate the inflammatory process in periodontitis. AIMS: To characterise the extracellular hydrolysis of CGRP as a mechanism for the selective inactivation of this neuropeptide in GCF from periodontitis sites. METHODS: Samples of GCF from periodontitis patients and periodontally-healthy subjects were incubated with synthetic human SP, NKA or CGRP. Reaction between the GCF constituents and synthetic peptides was allowed to progress from 0-180 min. Results of neuropeptide metabolism at each time were analysed by matrix-assisted laser desorption/ionisation mass spectrometry. RESULTS: There was no evidence of metabolism of SP, NKA or CGRP by constituents of healthy GCF. Metabolism of synthetic SP and NKA was minimal even after extensive incubation with periodontitis GCF. However, loss of carboxy-terminal amino acids was evident after only 1 min incubation with periodontitis GCF. The pattern of CGRP metabolism, which proceeded from the C-terminus, indicated that the neuropeptide was degraded by a carboxypeptidase. After 180 min, there was extensive carboxypeptidase degradation of CGRP to an 11 amino acid peptide. CONCLUSIONS: It is concluded that carboxypeptidase activity in GCF from periodontitis patients is responsible for rapid breakdown of CGRP but not SP or NKA. The rapid action of this carboxypeptidase on the anti-inflammatory neuropeptide CGRP is suggestive of a pathophysiological rôle for the enzyme in selectively degrading CGRP, thereby potentiating periodontal inflammation.

Changes in substance P and neurokinin A in gingival crevicular fluid in response to periodontal treatment.

BACKGROUND: The study of periodontitis provides a unique model for assessing the involvement of neuropeptides in inflammatory disease. AIM: To investigate the effects of periodontal treatment, resulting in a return to periodontal health, on the levels of substance P (SP) and neurokinin A (NKA) in gingival crevicular fluid (GCF). METHOD: We completed a cause of non-surgical treatment for 8 subjects with periodontitis (6 females 2 males, mean age 45.1, range 38-67 years) started a course of non-surgical periodontal treatment. Clinical indices were measured at 2 periodontitis sites at the initial visit and at 8 weeks after the completion of treatment in each subject. A 30-s sample of GCF was collected from each test site using perio paper strips. Each strip was placed into 500 microl of ice cold 0.1 M PBS, pH 7.4, vortex mixed for 30 s, and then stored at -70 degrees C until analysed by radioimmunoassay. RESULTS: The clinical condition of all test sites improved as a result of the periodontal treatment. The levels (pg/30 s sample) of SP fell from 56.3 (SD 66.0) at the initial visit to 4.2 (3.1) after treatment, p=0.017. The concentration (pg/microl) of SP in GCF fell from 140.6 (175.6) to 24.2 (11.1), p=0.036. The levels of NKA fell from 30.5 (17.1) to 10.6 (4.9), p=0.012 whereas the concentration changed little from 85.4 (43.5) to 61.6 (15.1), p=0.41. CONCLUSION: The reduction in inflammation resulting from effective periodontal treatment is associated with a reduction in the levels of tachykinins in gingival crevicular fluid.

Role of substance P and calcitonin gene-related peptide in the regulation of interleukin-8 and monocyte chemotactic protein-1 expression in human dental pulp.

AIM: To determine whether leucocyte infiltration during neurogenic inflammation in the pulp is regulated by neuropeptides via inducing the release of proinflammatory chemokines interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) from human dental pulp. METHODOLOGY: Cultured primary pulp cells and pulp tissue explants were stimulated with substance P (SP) and/or calcitonin gene-related peptide (CGRP). IL-8 or MCP-1, secreted from cultured cells or produced in pulp explants, was analysed by enzyme-linked immunosorbent assay. RESULTS: Substance P induced IL-8 secretion from cultured pulp cells (approximately threefold increase over control, P < 0.05) and from pulp tissue explants (two- to three fold). SP only minimally to moderately induced MCP-1 (approximately two fold) in cultured pulp cells. While MCP-1 induction in cultured pulp cells was detected after 24 h of SP stimulation, no induction was observed in pulp tissue. CGRP did not induce IL-8, but moderately increased MCP-1 production (approximately three fold) in cultured pulp cells. There was no synergistic induction of MCP-1 by SP plus CGRP stimulation of pulp cells. CONCLUSIONS: Substance P is a stronger inducer of IL-8 production in dental pulp than CGRP. IL-8 is more strongly induced than MCP-1 by SP, suggesting a more important role for IL-8 than MCP-1 in leucocyte infiltration during neurogenic inflammation in dental pulp.

Involvement of substance P, mast cells, TNF-alpha and ICAM-1 in the infiltration of inflammatory cells in human periapical granulomas.

BACKGROUND: Substance P (SP), a potent proinflammatory peptide present in sensory neurons, is believed to be a major mediator of neurogenic inflammation. The aim of this study was to examine the localization and involvement of SP, mast cells and tumor necrosis factor-alpha (TNF-alpha)-positive cells in human periapical granulomas. METHODS: Sections from seven periapical granulomas were stained using a variety of immunohistochemical methods. RESULTS: SP-immunoreactive nerve fibers, mast cells and TNF-alpha-positive cells were found localized in the vicinity of blood vessels in all the samples of periapical granulomas. The vascular endothelial cells stained positively for E-selectin and intercellular adhesion molecule-1. SP, TNF-alpha-positive cells and E-selectin could not be detected in clinically healthy periodontal ligament, and served as a negative control. CONCLUSION: Our findings suggest that SP, mast cells, TNF-alpha-positive cells and E-selectin may modulate the pathogenesis of apical periodontitis and may be responsible for stimulating the formation of granuloma with the resorption of periapical bone.