Journal of Dental Research

The author pays tribute to the contributions of Michael J. Levine to the field of salivary biochemistry.

The periodontal diseases are infectious diseases caused by predominantly Gram-negative bacteria. However, as our understanding of the pathogenesis of the periodontal diseases grows, it is becoming clear that most of the tissue damage that characterizes periodontal disease is caused by the host response to infection, not by the infectious agent directly. Investigation into the mechanism of action of host-mediated tissue injury has revealed that the neutrophil plays an important role in destruction of host tissues. In this paper, we review the biochemical pathways and molecular mediators that are responsible for regulation of the inflammatory response in diseases such as periodontitis, with a focus on lipid mediators of inflammation. Pro-inflammatory mediators, such as prostaglandins and leukotrienes, are balanced by counter-regulatory signals provided by a class of molecules called lipoxins. The role of lipoxins in the control and resolution of inflammation is discussed, as is the possibility of the development of new therapeutic strategies for the control and prevention of neutrophil-mediated tissue injury in inflammatory diseases like periodontitis.

The c-Jun N-terminal kinase (JNK) pathway and heat-shock proteins (HSPs) are involved in stress-induced apoptosis. Here we examined the association of JNK, c-Jun, and anti-apoptotic HSPs with pulp apoptosis during wound healing. In normal pulp, c-Jun was activated only in a few pulp cells, but JNK was not. HSP70 was expressed in the cytoplasm of pulp cells. One day after injury, active JNK and c-Jun were detected in apoptotic pulp cells, whereas HSP70 was detected in non-apoptotic cells. We also found the translocation of HSP70 into nuclei of pulp cells, and co-localization with active JNK and c-Jun. Four days after injury, active JNK and c-Jun disappeared in pulp cells, and HSP70 was relocalized from nuclei to the cytoplasm. These results suggest that the JNK pathway may be one of the compartments inducing apoptosis in pulp cells, and that HSP70 may have an inhibitory role in the apoptosis of pulp cells during wound healing.

Class II major histocompatilibity complex (MHC)-expressing cells are usually distributed in dental pulp, and it was postulated that the colony-stimulating factor (CSF) derived from dental pulp fibroblasts contributes to the migration of class II MHC-expressing cells into pulp tissue. This study aimed to investigate the CSF production of human dental pulp fibroblasts. In pulp tissue sections, granulocyte (G)-CSF was detected from normal teeth, while G-CSF, macrophage (M)-CSF, and granulocyte-macrophage (GM)-CSF were detected from teeth with dentinal caries. In cultured dental pulp fibroblasts, G-CSF was detected by immunostaining, immunoprecipitation, and ELISA, and mRNAs of G-CSF, M-CSF, and GM-CSF were detected by RT-PCR. The dental pulp fibroblasts cultured with TNF-α were found to increase the G-CSF expression and to produce M-CSF and GM-CSF. These findings suggest that dental pulp fibroblasts usually produce G-CSF. In the presence of TNF-α, dental pulp fibroblast express M-CSF and GM-CSF.

The role of salivary glands in nitrate and nitrite metabolism is poorly understood. The aim of the present study was to investigate the effect of parotid gland ablation on dynamic metabolism of nitrate and nitrite in miniature pigs. The parotid glands of 5 healthy miniature pigs were bilaterally ablated by methyl violet. Concentrations of nitrate and nitrite of whole saliva, serum, and urine samples were analyzed by high-performance liquid chromatography. Results showed that bilateral ablation of the parotid glands led to a significant decrease of nitrate secretion from blood to saliva (P < 0.05) and thus low nitrite levels. Dysfunction of the parotid glands temporarily increased the systemic level of nitrate in miniature pigs after nitrate loading. This study suggests that the parotid glands play an important role in the balance of nitrate and nitrite levels in both whole saliva and the body.

The activity of the superior head of the human lateral pterygoid muscle (SHLP) is controversial. Given the non-parallel alignment of some SHLP fibers, the SHLP may be capable of differential activation. The aims were to clarify SHLP activity patterns in relation to location within SHLP. In 18 subjects, SHLP single motor units were intramuscularly recorded at computer-tomography-verified sites during horizontal (e.g., protrusion) and vertical (e.g., opening) jaw tasks (recorded by a jaw-tracking device) and at resting postural jaw position. None of 92 units was active at the resting postural position. Medially located units (21) showed activity during contralateral movement, protrusion, and opening; 5 were also active on jaw closing. There was a significant association between unit location and the number of units active during vertical tasks (i.e., jaw closing and clenching). Analysis of the data suggests differential activation within SHLP and raises the possibility of functional heterogeneity within SHLP.

Neither the etiology of muscle-related temporomandibular disorders (TMD) nor the reason for the disproportionate number of women suffering from these disorders is well-established. We tested the hypothesis that physiologically relevant exercise (i.e., chewing bubble gum for 6 min) increases masticatory muscle pain in patients, but not in asymptomatic control subjects, and that female patients experience a significantly greater increase than males. Chewing increased pain in both female and male patients and, unexpectedly, also in female control subjects. One hour after chewing, the pain remained above pre-test levels for female patients but not for the other groups. Thus, sex differences in chewing-induced pain were found in control subjects but not as hypothesized in patients. Because chewing-induced masticatory muscle pain was significantly greater in female control subjects than in males, and persisted longer in female patients than in males, these results suggest greater susceptibility in women.

Even though there is no “gold standard” for determining caries lesion activity, it is nonetheless possible to evaluate the validity of such diagnostic measures. The aim of this study was to estimate the construct and predictive validity of caries lesion activity assessments by means of their ability to reflect known effects of fluoride on caries. A three-year trial of the effect of daily supervised brushing with fluoride toothpaste was carried out among 273 12-year-old children. All children were examined clinically according to diagnostic criteria for activity assessment. The relative risk (fluoride vs. control) for caries lesion transitions among diagnostic categories was calculated. Fluoride inhibited progression of caries at all stages of lesion formation while at the same time enhancing lesion regression. The effects were most pronounced for active non-cavitated lesions. It is concluded that the clinical diagnostic criteria have construct and predictive validity for the assessment of caries lesion activity.

Dental status and its relationship to diet and nutritional status have been little explored. In this study of a representative sample of the US civilian, non-institutionalized population (NHANES III), we predicted that the intake of nutritious foods, dietary fiber, and levels of biochemical analytes would be lower, even after adjusting for potential social and behavioral factors, among those who were edentulous and wore complete dentures than for those who had all their natural teeth. Multivariate analyses indicated that intake of carrots and tossed salads among denture-wearers was, respectively, 2.1 and 1.5 times less than for the fully dentate (p < 0.0001), and dietary fiber intake was 1.2 times less (p < 0.05). Serum levels of beta carotene (9.8 μg/dL), folate (4.7 ng/dL), and vitamin C (0.87 mg/dL) were also lower among denture-wearers (p < 0.05). Intakes of some nutrient-rich foods and beta carotene, folate, and vitamin C serum levels were significantly lower in denture-wearers.

In humans, the inappropriate occlusal vertical dimension (OVD) causes several orofacial disorders, such as bruxism and pain in the masticatory muscles and temporomandibular joint. However, it is still unclear how strictly the OVD is adjusted. To answer this question, we studied the temporal change of the OVD in bite-raised young guinea pigs. The OVD was raised by fixation of a bite-raising appliance to the lower incisors, and increased by either 3 or 1.5 mm at the first molars. After the space produced between the upper and lower molars was filled within 10 days due to eruption of the molars, the appliance was removed. In the bite-raised animals, the raised OVD was reduced until it attained that observed in the control animals, after which the OVD increased according to cranial growth. These results show that the OVD is developmentally changed and strictly controlled.

Autologous bone is the most successful bone-grafting material; however, limited supply and donor site morbidity are problematic. Synthetic bone substitutes are effective, but healing is slow and unpredictable. Osseous wound healing may be enhanced if bone substitutes are combined with autologous bone marrow cells. To test this hypothesis, we created 40 calvarial defects in 20 12-week-old New Zealand White rabbits, divided into four groups: (1) unrepaired controls, (2) autologous bone grafts, (3) unseeded Caprotite^® (a polymer-ceramic composite) grafts, and (4) Caprotite^® grafts seeded with autologous bone marrow stromal cells. CT scans were obtained at 0, 6, and 12 weeks post-operatively, and defects were harvested for histology. Defects repaired with autologous bone had significantly (p < 0.05) more bone than the other three groups, although seeded Caprotite^® defects showed different wound-healing sequelae. Results suggest that seeded Caprotite^® scaffolds did not significantly enhance osseous defect healing compared with controls.

Resin-dentin bonds degrade over time. The objective of this study was to evaluate the influence of variables like hybridization effectiveness and diffusion/elution of interface components on degradation. Hypotheses tested were: (1) There is no difference in degradation over time between two- and three-step total-etch adhesives; and (2) a composite-enamel bond protects the adjacent composite-dentin bond against degradation. The micro-tensile bond strength (μTBS) to dentin of 2 three-step total-etch adhesives was compared with that of 2 two-step total-etch adhesives after 4 years of storage in water. Quantitative and qualitative failure analyses were conducted correlating Fe-SEM and TEM. Indirect exposure to water did not significantly reduce the μTBS of any adhesive, while direct exposure resulted in a significantly reduced μTBS of both two-step adhesives. It is concluded that resin bonded to enamel protected the resin-dentin bond against degradation, while direct exposure to water for 4 years affected bonds produced by two-step total-etch adhesives.

Although it is generally proposed that dentin bonding results from adhesive infiltration of superficially demineralized dentin, it is not clear how well the resin monomers seal the dentin collagen fibrils under wet bonding conditions. The aim of this study was to determine the quality and molecular structure of adhesive/dentin (a/d) interfaces formed with wet bonding as compared with adhesive-infiltrated demineralized dentin (AIDD) produced under controlled conditions (optimum hybrid). From each extracted, unerupted human 3rd molar, one fraction was demineralized, dehydrated, and infiltrated with Single Bond (SB) adhesive under optimum conditions; the remaining, adjacent fraction was treated with SB by wet bonding. AIDD and a/d interface sections were stained with Goldner’s trichrome; corresponding sections were analyzed with micro-Raman spectroscopy. The histomorphologic and spectroscopic results suggest that, under wet bonding, the a/d interface is a porous collagen web infiltrated primarily by the hydrolytically unstable HEMA.

The cariostatic effect of resin-modified glass ionomer (RMGI) on secondary root caries is well-documented. However, this beneficial effect may be dependent upon the mode of cavity surface treatment. To investigate this relationship, we studied 4 cavity surface treatments prior to the placement of RMGI: no treatment (None), polyacrylic acid (PAA), phosphoric acid (H₃PO₄), and Scotchbond Multi-Purpose adhesive (SMP) as a control. Specimens were aged for two weeks in synthetic saliva, thermocycled, and subjected to an artificial caries challenge (pH 4.4). Polarized light microscopy (PLM) and microradiography (MRG) showed significantly less demineralization with the H₃PO₄ cavity surface treatment as revealed by ANOVA and Tukey’s multiple comparisons (p ≤ 0.05). Dentin fluoride profiles determined by electron probe microanalysis (EPMA) supported PLM and MRG findings. It may be concluded that removal of the smear layer with phosphoric acid provides significantly enhanced resistance to secondary root caries formation adjacent to RMGI restorations.