Catalase activity in healthy and inflamed pulp tissues of permanent teeth in young people.

AIM: To evaluate catalase (CAT, EC 1.11.1.6) activity in healthy and inflamed dental pulp of young patient's teeth and to investigate if an active defense system oxidizing agents is present as a response to bacterial invasion. MATERIALS AND METHODS: Twenty young patients between 15 and 25 ages, who were diagnosed to be healthy, were the source of the pulp tissue. The situation of the dental pulps was evaluated using clinical and radiographic assessments. The patients were divided two groups from healthy, and inflamed pulp tissues were obtained; each participant provided one pulp tissue specimens. The specimens were collected during endodontic treatment or by longitudinally grooving and splitting the teeth (if extracted). Catalase activity was determined through spectrophotometric methods and an independent sample t-test assessed the significance of differences between the groups. RESULTS: There was statistically a difference between healthy pulp tissue and inflamed pulp tissue (P < 0.005, independent sample t-test). The catalase activity of healthy group was significantly lower than inflamed pulp groups. CONCLUSION: The present study has shown that a significant increase in catalase activity is determined in inflamed dental pulps, which is due to pulpitis in comparison to healthy dental pulp.

Age-related Changes in the Alkaline Phosphatase Activity of Healthy and Inflamed Human Dental Pulp.

INTRODUCTION: Alkaline phosphatase (ALP) plays an important role in inducing mineralization events in the dental pulp. This study investigated and compared the ALP levels in healthy and inflamed pulp in young and old human pulp. METHODS: Tissue samples were collected from young (<30 years) and old (>60 years) donors. In both age groups, healthy human pulp (n = 18) were collected from extracted wisdom teeth. For reversible and irreversible pulpitis, pulp samples (n = 18 each) were obtained during endodontic treatment. ALP activity was assessed by spectrophotometry and immunhistochemistry. RESULTS: Regardless of age, reversible pulpitis group samples showed a slight elevation in ALP activity compared with normal healthy pulp. In elderly patients, ALP expression with irreversible pulpitis was significantly higher than those with a healthy pulp (P < .05). CONCLUSIONS: In the hyperemic state, both the young and old pulp shows a slight increase in ALP activity, whereas in irreversible pulpitis, only the old pulp shows significantly elevated ALP levels. Such an increase may trigger calcification events, which may eventually cause difficulties in endodontic treatment procedures in elderly individuals.

Immune Cells and Molecular Networks in Experimentally Induced Pulpitis.

Dental pulp is a dynamic tissue able to resist external irritation during tooth decay by using immunocompetent cells involved in innate and adaptive responses. To better understand the immune response of pulp toward gram-negative bacteria, we analyzed biological mediators and immunocompetent cells in rat incisor pulp experimentally inflamed by either lipopolysaccharide (LPS) or saline solution (phosphate-buffered saline [PBS]). Untreated teeth were used as control. Expression of pro- and anti-inflammatory cytokines, chemokine ligands, growth factors, and enzymes were evaluated at the transcript level, and the recruitment of the different leukocytes in pulp was measured by fluorescence-activated cell-sorting analysis after 3 h, 9 h, and 3 d post-PBS or post-LPS treatment. After 3 d, injured rat incisors showed pulp wound healing and production of reparative dentin in both LPS and PBS conditions, testifying to the reversible pulpitis status of this model. IL6, IL1-ß, TNF-α, CCL2, CXCL1, CXCL2, MMP9, and iNOS gene expression were significantly upregulated after 3 h of LPS stimulation as compared with PBS. The immunoregulatory cytokine IL10 was also upregulated after 3 h, suggesting that LPS stimulates not only inflammation but also immunoregulation. Fluorescence-activated cell-sorting analysis revealed a significant, rapid, and transient increase in leukocyte levels 9 h after PBS and LPS stimulation. The quantity of dendritic cells was significantly upregulated with LPS versus PBS. Interestingly, we identified a myeloid-derived suppressor cell-enriched cell population in noninjured rodent incisor dental pulp. The percentage of this population, known to regulate immune response, was higher 9 h after inflammation triggered with PBS and LPS as compared with the control. Taken together, these data offer a better understanding of the mechanisms involved in the regulation of dental pulp immunity that may be elicited by gram-negative bacteria.

Investigation of functional activity human dental pulp stem cells at acute and chronic pulpitis.

UNLABELLED: It is already recognized that together with the other connective tissues organ-specific progenic stem cells are also found in postnatal dental pulp. This group of undifferentiated cells is only 1% of total cell population of the pulp. The aim of the study was the identification of stem cells in human dental pulp, detection of their localization and assessment of functional activity during inflammation process and/or at norm. The obtained results showed that at acute pulpitis the pulp stroma is hypocellular in comparison with the norm but cells proliferative activity is low. CD 133 and NCAM (CD 56) positive stem cells were found in perivascularl space of the pulp stroma and in Hohle layer. At process prolongation and transition to the chronic phase pulp stroma is hypercellular, the cells with large, rounded or oval-shaped nuclei with clear chromatin appear together with fibroblasts. They are distributed as about entire thickness of the stroma as especially Hohle layer. In such cells higher proliferative activity (Ki67 expression) was observed. The cells in the mentioned proliferation phase are intensively marked by CD133, the rate of which is high in Hohle layer and along it. A large number of NCAM (CD 56) positive cells appear in pulp stroma. CONCLUSIONS: During pulpitis an involvement of stem cells into the process of reparative dentinogenesis should be conducted stepwise. In acute cases of the disease, stem cell perivascularl mobilization and proliferation and its migration to Hohle layer occur in response to irritation /stimulation. Chronification of the process leads not only to the migration of stem cells to the periphery of the pulp but also s their В«maturationВ¼ (increase of NCAM expression in the stem cells), which causes an increase the number of dentin producing active odontoblasts and initiation of reparative dentinogenesis.

Evaluation of gelatinases, tissue inhibitor of matrix metalloproteinase-2, and myeloperoxidase protein in healthy and inflamed human dental pulp tissue.

INTRODUCTION: The aim of this study was to compare the gelatinolytic activity of matrix metalloproteinase (MMP)-2 and MMP-9 and the expression of tissue inhibitor of matrix metalloproteinase (TIMP)-2 and myeloperoxidase protein (MPO) in clinically healthy human pulp and inflamed pulp tissue specimens. METHODS: Twenty dental pulps clinically diagnosed as inflammatory tissues and 20 healthy pulp tissues from enclosed third molars were harvested and evaluated. The gelatinolytic activity for MMP-2 and MMP-9 was assessed by using the zymography technique, TIMP-2 gene expression was evaluated using the enzyme-linked immunosorbent assay, and MPO was determined using the MPO assay. RESULTS: Data showed increased levels of MMP-9, active MMP-2, TIMP-2, and MPO in inflammatory pulp tissues compared with healthy tissues (P < .05). No statistical difference could be observed for pro-MMP-2 (P > .05). CONCLUSIONS: Although all samples were associated with MMP-2 expression, the active form of this MMP was observed only in inflamed pulps. Inflamed pulps showed an up-regulation of MMP-9, TIMP-2, and MPO.

Concentration of collagenases (MMP-1, -8, -13) in patients with chronically inflamed dental pulp tissue.

Matrix metalloproteinases (MMPs) form an enzyme family capable of degrading almost all extracellular matrix (ECM) and basement membrane (BM) components. They play an important role in normal tissue remodelling and growth, as well as in many destructive pathological conditions such as inflammation, tumour growth and metastasis. The role of MMPs in the breakdown of pulp tissue of teeth with pulpitis has not yet been directly elucidated. The purpose of this study was to evaluate the tissue levels of collagenases (MMP-1, -8, -13) and their distributions in the clinically healthy and chronically inflamed human dental pulps of 30 patients, aged 15-70 years. Twenty pulps were collected from subjects diagnosed with chronic pulpitis, and 10 control pulps were obtained from 10 subjects following molar extraction for orthodontic reasons. The levels of collagenases were determined with an enzyme-linked immunosorbent assay (ELISA). Results reveal that levels of collagenases were significantly higher in chronically inflamed vs. clinically normal pulps. Overall, these results show that MMPs play an important role in ECM destruction during the inflammatory processes of pulpitis, as well as reflecting the special characteristics of them. This investigation opens a new opportunity for one contemporary method for the diagnosis of pulp inflammations and monitoring of the inflammatory processes.

The anti-inflammatory effects of matrix metalloproteinase-3 on irreversible pulpitis of mature erupted teeth.

Matrix metalloproteinases (MMPs) are involved in extracellular matrix degradation and the modulation of cell behavior. These proteinases have also been implicated in tissue repair and regeneration. Our previous studies have demonstrated that MMP-3 elicits stimulatory effects on the proliferation and the migration of endothelial cells as well as anti-apoptotic effects on these cells in vitro. In addition, we found that MMP-3 enhanced the regeneration of lost pulp tissue in a rat incisor pulp injury model. However, continuously erupting rodent incisors exhibit significantly different pulp organization compared with mature erupted teeth. Therefore, we have further extended these studies using a canine irreversible pulpitis model to investigate the effects of MMP-3. In this study, the crowns of the canine mature premolars were removed and the pulp tissues were amputated. The amputated pulp tissues remained exposed for 24 or 72 hours to induce mild or severe irreversible pulpitis, respectively, followed by sealing of the cavities. In both models, the whole pulp tissues became necrotic by day 14. In this mild pulpitis model, the regeneration of pulp tissue with vasculature and nerves was observed until 14 days after sealing with MMP-3, followed by extracellular matrix formation in the regenerated pulp tissues until day 28. The treatment with MMP-3 resulted in a decrease in the number of macrophage and antigen-presenting cells and a significant inhibition of IL-6 expression on day 3. The inhibition of MMP-3 activity abolished these anti-inflammatory effects. Immunofluorescence staining demonstrated that MMP-3 was involved in the modification of serum-derived hyaluronan-associated proteins and hyaluronan (SHAP-HA) complexes possibly through the degradation of versican. These results demonstrate that MMP-3 can act as an anti-inflammatory agent and suggest that MMP-3 might represent a useful therapy for the treatment of mild irreversible pulpitis.

Irreversible inflammation is associated with decreased levels of the alpha1-, beta1-, and alpha2-subunits of sGC in human odontoblasts.

The nitric oxide (NO) receptor enzyme soluble guanylate cyclase (sGC) contains one prosthetic heme group as an αß heterodimer, and two heterodimer isoforms (α(1)ß(1), α(2)ß(1)) were characterized to have enzyme activity. To test the irreversible inflammation-dependent regulation of sGC in odontoblasts, we incubated decalcified frozen sections of healthy and inflamed human third molars with antibodies against ß-actin, nitrotyrosine, inducible nitric oxide synthase (iNOS), α(1)-, ß(1)-, and α(2)-subunits of sGC and analyzed them at protein levels by quantitative immunohistochemistry. The irreversible inflammation induced an increase in the signal intensities for nitrotyrosine and iNOS and a decrease for the α(1)-, ß(1)-, and α(2)-subunits of sGC in odontoblasts. Inflammatory mediators, reactive oxygen, and nitrogen species may impair the expression of the α(1)-, ß(1)-, and α(2)-subunits in odontoblasts. The decrease of sGC at the protein level in inflamed odontoblasts is compatible with a critical role for sGC to mediate biological effects of NO in health.

A first study on the usefulness of matrix metalloproteinase 9 from dentinal fluid to indicate pulp inflammation.

INTRODUCTION: Pulpal diagnostics might be improved by using molecular markers found in dentinal fluid. In the current pilot study, we tested whether matrix metallopeptidase 9 (MMP-9) levels in dentinal fluid were detectable and differed between pulps from symptomatic teeth diagnosed with irreversible pulpitis and healthy counterparts. METHODS: Thirty-one patients participated; 19 were diagnosed with irreversible pulpitis, and 12 were in need of replacement of a filling close to the pulp space in a clinically healthy tooth. Dentinal fluid was collected during a period of 2 minutes from dentin cavities by using folded polyvinylidene difluoride (PVDF) membranes, which were then transferred to microcentrifugation tubes containing physiologic saline solution. Total MMP-9 levels in these solutions were assessed by using a human MMP-9 fluorescent assay, detection limit 0.25 ng/mL. MMP-9 levels between groups were compared by using Mann-Whitney U test (alpha <0.05). RESULTS: Three specimens from the symptomatic teeth were not included because coronal pulps proved to be necrotic on access. Dentinal fluid samples from symptomatic teeth had significantly higher MMP-9 levels than those from clinically healthy counterparts (P < .05). However, merely 7 of the 16 pulpitis samples contained detectable levels of MMP-9. None of the clinically healthy control specimens contained any detectable amounts of MMP-9. CONCLUSIONS: With a sensitive assay, it was possible to detect an enzyme linked with pulp tissue destruction (MMP-9) in dentinal fluid. However, the collection method needs to be improved to provide predictable fluid yields. Longitudinal studies should be performed to assess the predictive value of molecular markers in dentinal fluid on pulpal pathosis.

Differential cholinoceptor modulation of nitric oxide isoforms in experimentally-induced inflammation of dental pulp tissue.

AIM: The aim of the study was to investigate the role of muscarinic acetylcholine receptor (mAChR) activity in the regulation of endothelial (e), neuronal (n) and inducible (i) nitric oxide synthase (NOS) activity and expression in experimentally induced inflammation of rat dental pulp tissue. METHODOLOGY: Inflammation was induced by application of bacterial lipopolysaccharide (LPS) to the pulp. Extirpated pulp-tissue samples were incubated in saline solution until the various experiments were performed. Saline-treated pulp and healthy pulp tissues were used as controls. NOS activity was measured by the production of [U-(14)C]-citrulline from [U-(14)C]-arginine. Nitrite/nitrate assay was evaluated by the conversion of nitrate to nitrite in the presence of nicotinamide adenine dinucleotide phosphate. i-nos, e-nos and n-nos mRNA levels were measured using reverse-transcriptase polymerase chain reaction by co-amplification of target cDNA with a single set of primers. RESULTS: Application of LPS to the pulp increased NOS activity and nitrate production (P < 0.001), generated by iNOS over-activity and expression. Pilocarpine acting on mAChRs triggered a biphasic action on NOS activity and NO accumulation. At low concentrations, pilocarpine induced a negative effect associated with a decrease in i-nos mRNA level, whilst at high concentration, it produced a positive effect associated with increased e-nos and n-nos mRNA levels. In control pulp tissue, only the positive effect of pilocarpine was observed. CONCLUSIONS: Irreversible pulpitis changes mAChR conformation increasing its efficiency of coupling to transducing molecules that in turn induce activate iNOS. The capacity of pilocarpine to prevent NO accumulation and iNOS activity, by acting on mAChR mutation induced by pulpitis, might be useful therapeutically as a local treatment.

Cholinoceptor modulation on nitric oxide regulates prostaglandin E(2) and metalloproteinase-3 production in experimentally induced inflammation of rat dental pulp.

The purpose of this study was to investigate the role of muscarinic acetylcholine receptor (mAChR) activity in the regulation of inducible nitric oxide synthase (iNOS) activity, prostaglandin E(2) (PGE(2)), and metalloproteinase-3 (MMP-3) in experimentally induced inflammation of rat incisors dental pulp. Inflammation was induced by application of bacterial lipopolysaccharide (LPS) to the pulp. Extirpated pulp tissue samples were incubated in saline solution until the various experiments were performed. Saline-treated pulp and healthy pulp were used as controls. NOS activity was measured by the production of [U-(14)C]-citrulline from [U-(14)C]-arginine. PGE(2) and MMP-3 production were evaluated by an enzyme-linked immunosorbent assay (ELISA) and cyclooxygenase (cox-1 and cox-2) messenger RNA levels were measured using a reverse-transcriptase polymerase chain reaction by coamplification of target complementary DNA with a single set of primers. The application of LPS to the pulp increased NOS activity, PGE(2), and MMP-3 production associated with iNOS overactivity. Moreover, PGE(2) and MMP-3 production were the result of cox-2 expression. Pilocarpine (5 x 10(-11) mol/L to 5 x 10(-9) mol/L), acting on mAChRs, triggered a negative effect on NOS activity, PGE(2), and MMP-3 production. In control pulp, no action of pilocarpine was observed. Pulpitis changed mAChR conformation, increasing its coupling efficiency to transducing molecules that in turn activate iNOS. The capacity of pilocarpine to prevent iNOS activity, PGE(2), and MMP-3 by acting on mAChR mutation induced by pulpitis might be useful therapeutically as a local treatment.

Extracellular proteolytic activities expressed by Bacillus pumilus isolated from endodontic and periodontal lesions.

The purpose of the present study was to identify 12 Bacillus isolates that had been obtained from root canals of teeth requiring endodontic therapy and from periodontal pockets in severe marginal periodontitis, and to determine whether these isolates exhibited extracellular proteolytic activity and, using in vitro assays, whether any such activity could degrade substrates that would be pathophysiologically relevant with regard to the production of endodontic and periodontal lesions. Biochemical and carbohydrate fermentation patterns were used in the identification of all strains, which was confirmed by determination of the16S rRNA gene sequence for strain BJ0055. Screening for production of extracellular proteolytic activity by all strains was done with a general proteinase substrate. All isolates were identified as representing Bacillus pumilus and all exhibited extracellular proteolytic activity. The putative pathophysiological relevance of extracellular proteinase production in strain BJ0055 was assessed using fluorophore-labelled elastin and collagen and several chromogenic peptides. Probable classes of proteinases acting on each substrate were investigated using class-specific inhibitors. Activity-pH profiles were determined in buffers at different pH values. Extracellular activities that were caseinolytic, elastinolytic, collagenolytic, glutamyl endopeptidase-like, and alanyl tripeptidyl peptidase-like were observed. No trypsin-like activities were detected. Serine- and chymotrypsin-like serine proteinase activities were detected, with activity observed at neutral and alkaline, but not acidic, pH. B. pumilus strains isolated from endodontic and periodontal lesions exhibited extracellular activities that degrade elastin, collagen and other substrates. These activities may be virulence factors that contribute to tissue damage in apical periodontitis and severe marginal periodontitis.

Quantification of neural protein in extirpated tooth pulp.

Because the pulp tissue extirpated during root canal procedures might serve as a valuable resource with which to assess underlying mechanisms of persistent pain, we sought to determine whether standard Western blotting techniques could be used to quantify neural proteins in pulp extirpated from teeth with irreversible pulpitis. Pulp harvested from healthy intact teeth extracted for orthodontic reasons was used for comparison. The neural marker PGP9.5 was detectable in all samples tested. A membrane enrichment protocol enabled detection of even low abundance, high molecular weight proteins such as the sodium channel alpha-subunit NaV1.8. Importantly, it was possible to quantify a approximately 6-fold increase in the relative density of NaV1.8 in inflamed pulp compared with control pulp. Our results suggest that it should be possible to use extirpated tooth pulp to validate mechanisms of persistent pain implicated in preclinical studies as well as evaluate the therapeutic efficacy of novel antinociceptive interventions.

Elevated expression of Cu, Zn-SOD and Mn-SOD mRNA in inflamed dental pulp tissue.

AIM: To determine the mRNA expression levels of copper-zinc superoxide dismutase (Cu, Zn-SOD) and manganese SOD (Mn-SOD) in healthy and inflamed human dental pulp tissue. METHODOLOGY: Sixteen patients with symptomatic irreversible pulpitis (eight females and eight males) were selected for study. Normal healthy pulps were removed from extracted mandibular third molar teeth from 10 systemically healthy individuals (six females and four males). QRT-PCR analysis of Cu, Zn-SOD and Mn-SOD mRNA expression was carried out in 16 cases of irreversible pulpitis and in 10 cases of systemically healthy donors. The Shapiro-Wilk's test was used to test the normality of data, whereas the Mann-Whitney U-test was used to evaluate the significance of the differences between groups. Differences in the expression levels were considered to be statistically significant for P-values <0.05. RESULTS: A significant increase (P < 0.05) occurred in both Cu, Zn-SOD and Mn-SOD mRNA expression in cases of irreversible pulpitis. The increase in Mn-SOD level was significantly higher (P < 0.05) than the change observed for Cu, Zn-SOD. CONCLUSIONS: The development of pulpitis is associated with elevated transcription of both Cu, Zn-SOD and Mn-SOD; pulp tissue inflammation generated higher Mn-SOD transcription compared with Cu, Zn-SOD.

A real time quantitative PCR analysis and correlation of COX-1 and COX-2 enzymes in inflamed dental pulps following administration of three different NSAIDs.

Dental pain is encountered daily by clinicians. Nonsteroidal anti-inflammatory drugs (NSAIDs) commonly used for pain management are traditionally cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) inhibitors, and more recently selective COX-2 inhibitors. This study was designed to identify and quantify COX-1 and COX-2 gene expression level in inflamed rat molar pulps after administration of three NSAIDs: Celebrex, Vioxx, and Advil. Fifty male Wistar rats had their first and second molar pulps exposed and sealed with Cavit for 4 days. Rats were randomly divided into the three drug groups and two control groups. RNA was isolated from the rat pulps. Real Time Quantitative Reverse Transcriptase-Polymerase Chain Reaction assay, a relatively new PCR technique, was used to quantify COX-1 and COX-2 mRNA. Statistical analysis demonstrated no significant differences in COX-1 and COX-2 levels among the drug groups. However, Vioxx and Advil significantly reduced COX-2 expression levels compared to inflamed (positive control) pulps (p < 0.05).

The role of endothelial nitric oxide in the Substance P induced vasodilation in bovine dental pulp.

Vasodilation, an important response in neurogenic inflammation, involves release of Substance P (SP) from the sensory nerve endings. It is now well known that SP causes edema formation and vascular relaxation in nondental tissues, however, the SP vasodilatory mechanism in the dental pulp is not completely understood. Endothelium-dependent relaxation is mediated by nitric oxide (NO) release with consecutive intracellular cyclic-GMP elevation in many vascular preparations. Recently, it has been shown in different vascular systems that SP-induced vasodilation is mediated by cyclic-GMP production through different pathways involving endothelial NO or direct endothelial-independent pathways. In the present study, the role of endothelial NO in SP induced vasodilation in the dental pulp was investigated to better understand the inflammatory mechanisms. Freshly extracted bovine dental pulp was used to measure NO production. Sodium nitroprusside (SNP), L-NAME and SP were utilized to induce and to inhibit NO production in endothelial cells. Released NO byproducts were measured with chemiluminescence assay technique. The present data demonstrate that SP induces NO production by activating NOsynthase (NOS) in endothelial cells. The NOS inhibitor L-NAME blocks NO production completely. In conclusion, in the bovine dental pulp, SP-induced vascular relaxation can be mediated by inducing NOS, and subsequently NO production in endothelial cells.

Effect of NOS inhibitor on cytokine and COX2 expression in rat pulpitis.

Various kinds of chemical mediators are synthesized in the course of pulpitis; thus, control of their production would assist in inducing a reduction in pulpal inflammation. We hypothesized that nitric oxide (NO) would be an important mediator of pulpal inflammation. Pulpal inflammation was induced by the application of LPS in rat incisor pulp, and inducible nitric oxide synthase (iNOS) expression was evaluated by reverse-transcription/polymerase chain-reaction and immunohistochemical staining. After LPS application, iNOS mRNA was first detected after 3 hrs, peaked at 6 hrs, and decreased thereafter. iNOS-positive cells were macrophages and neutrophils. An NOS inhibitor caused drastic decreases in the expression of pro-inflammatory cytokines and COX2 mRNA, which was highly induced in the LPS-induced pulpitis. These results indicate that NO synthesis is related to the initiation of mediator production, and that its down-regulation should contribute to the prevention of pro-inflammatory mediator synthesis.

Upregulation of tissue-type plasminogen activator in inflamed human dental pulps.

AIM: To compare tissue-type plasminogen activator (t-PA) expression in normal human pulp and inflamed human pulp tissue specimens. METHODOLOGY: Thirty pulpal tissue specimens (13 normal and 17 inflamed pulps) were obtained from extracted third molars. The levels of t-PA between normal pulp and inflamed pulp tissues were compared using the quantitative reverse-transcriptase polymerase chain reaction analysis. In addition, immunohistochemistry was used to identify the in situ localization of t-PA expression in pulp specimens. Wilcoxon-Mann-Whitney rank sum test was applied for the statistical analysis of the results. RESULTS: t-PA mRNA gene was found more in inflamed pulps when compared with normal pulp tissue (P<0.05). The results from immunohistochemistry demonstrated that t-PA expression was significantly higher in the inflamed pulp (P=0.025). t-PA stain was detected in the fibroblasts, inflammatory infiltrates and endothelial cells. CONCLUSIONS: t-PA expression was significantly higher in inflamed pulp tissue. t-PA may play an important role in the pathogenesis of pulpal inflammation.

Copper-zinc superoxide dismutase activity in healthy and inflamed human dental pulp.

AIM: To examine copper-zinc superoxide dismutase (Cu, Zn-SOD) activity in clinically healthy and symptomatic human dental pulps. METHODOLOGY: Twenty-five systemically healthy patients, 14 females and 11 males (age: 13.1-34.6 years; mean: 21.7 +/- 6.3), were the source of the pulp tissue. The condition of the pulps was assessed using clinical and radiographic evaluation. The pulp tissue was collected by longitudinally grooving and splitting the teeth (if extracted) or during endodontic treatment, and were age- and sex-matched between the healthy and the irreversible symptomatic pulpitis tissue groups. Cu, Zn-SOD activity was determined through spectrophotometric methods and a Mann-Whitney test assessed the significance of differences between the groups. RESULTS: The enzyme activities were 144.8 +/- 42.2 and 68.1 +/- 25.0 U mg(-1) in the healthy and irreversible symptomatic pulp tissue, respectively. The difference between the groups was statistically significant (P < 0.001). CONCLUSIONS: These results demonstrate a potential role for Cu, Zn-SOD during dental pulp inflammation in humans.

Nitric oxide synthase in healthy and inflamed human dental pulp.

Nitric oxide synthase (NOS) plays a significant role in the pathogenesis of pulpitis. In this study, we hypothesized the existence of endothelial (eNOS) and inducible (iNOS) enzyme isoforms in human dental pulp. Extracted third molar pulps were divided into groups based on clinical diagnosis: healthy, hyperemic, and irreversible pulpitis. We have localized the eNOS and iNOS by immunohistochemistry and have tested their mRNA expression by RT-PCR and protein levels by Western blots. eNOS is present in the endothelial cells and odontoblasts of the healthy pulp, but an elevation of eNOS mRNA and protein levels with a concomitant dilation of vessels was characteristic under pathological conditions. Healthy pulp tissue failed to exhibit any iNOS; however, acute inflammation enhanced the mRNA and protein levels of iNOS, mainly in the leukocytes. There are differences in localization and expression between eNOS and iNOS in healthy and inflamed dental pulp.