Journal of Dental Research

The word "serendipity" comes from the tale "The Three Princes from Serendip" by Horace Walpole. It is defined as "an aptitude for making fortunate discoveries accidentally". The demonstration of crevicular fluid was truly an accidental discovery. Niels Brill, a successful private practitioner in Copenhagen with prosthodontics as a specialty, and Bo Krasse, associate professor of cariology, were examining the microflora of gingival pockets in a dog. Before the samples were taken, gingiva and teeth were disinfected with an iodine solution. The solution disappeared from different pockets at different speeds. After intravenous injection of a fluorescein solution, the occurrence of fluorescein could be recorded on filter paper strips. Thus, after stumbling on the disappearance of iodine from the cervical areas of the teeth, two nonperiodontologists managed to demonstrate why it disappeared. This article describes the details of the discovery and subsequent studies, and offers some reflections by one of the original authors 37 years later.

A few recent studies have shown associations between poor oral health and coronary heart disease (CHD). The objective of this study was to examine the incidence of CHD in relation to number of teeth present and periodontal disease, and to explore potential mediators of this association, in a prospective cohort study. This study is a part of the ongoing Health Professionals Follow-Up Study (HPFS). Participants included a US national sample of 44,119 male health professionals (58% of whom were dentists), from 40 to 75 years of age, who reported no diagnosed CHD, cancer, or diabetes at baseline. We recorded 757 incident cases of CHD, including fatal and non-fatal mvocardial infarction and sudden death, in six years of follow-up. Among men who reported pre-existing periodontal disease, those with 10 or fewer teeth were at increased risk of CHD compared with men with 25 or more teeth (relative risk = 1.67; 95% confidence interval, 1.03 to 2.71), after adjustment for standard CHD risk factors. Among men without pre-existing periodontal disease, no relationship was found (relative risk = 1.11; 95% confidence interval, 0.74 to 1.68). The associations were only slightly attenuated after we controlled for dietary factors. No overall associations were found between periodontal disease and coronary heart disease. Tooth loss may be associated with increased risk of CHD, primarily among those with a positive periodontal disease history; diet was only a small mediator of this association.

In a previous study in cats, we found that electrical stimulation of the tooth pulp caused blood flow increases at various sites in the ipsilateral oral mucosa (upper and lower gingivae, lower lip, buccal mucosa and tongue, and, notably, bilaterally in the palatal mucosa). Tooth-pulp stimulation is well-known to induce severe pain and to evoke autonomic reflex responses in other organs and tissues. The purpose of this study was: to confirm that tooth-pulp stimulation may indeed induce autonomically mediated vasodilator responses in the feline oral mucosa away from the stimulated tooth, and to test our hypothesis that the reflex pathway involves parasympathetic vasodilator fibers as efferents. Dynamic changes in palatal mucosal blood flow (PMBF), with lower lip blood flow (LBF) as a control, were investigated in anesthetized, cervically sympathectomized cats by means of Laser Doppler Flowmetry. Unilateral electrical stimulation of the maxillarv canine tooth pulp produced hexamethonium-sensitive bilateral increases in PMBF in a stimulus-intensity-dependent manner, without an increase in systemic blood pressure; LBF increased only ipsilaterally. Bilateral section of the glossopharyngeal nerve roots had no effect on the vasodilator responses, while unilateral section of the facial nerve root or lesion of the pterygopalatine ganglion (PPG) abolished the response on that side. Intracranial electrical stimulation of the peripheral cut ends of the facial or glossopharyngeal nerve roots caused an increase in ipsilateral PMBF. These results indicate that unilateral tooth-pulp stimulation induces a bilateral reflex vasodilator response in the palatal mucosa mediated viaparasympathetic vasodilator fibers that emerge from the brain stem with the facial nerve and reach the blood vessels viaPPG. Although there is a dual innervation of the cat palatal mucosa by parasympathetic vasodilator fibers running viathe facial and glossopharyngeal nerve roots, the latter are not involved in the bilateral reflex responses.

Root resorption associated with tooth movement is an unsolved problem in orthodontics. If such root resorption could be prevented, it would be an important contribution toward reducing risk factors in orthodontic treatment. The purpose of this study was to examine the effects of the topical administration of a bisphosphonate, risedronate, which is known to be a potent blocker of bone resorption, on root resorption during tooth movement and on the repair of the resorbed root surface after tooth movement in rats. In the first experiment, both the right and left upper first molars were moved buccally with a standardized expansion spring under administration of risedronate. After day 7, extensive root resorption had occurred on the control side, and the area of root resorption reached a maximum on day 14. The topical administration of risedronate caused a significant and dose-dependent inhibition of root resorption after the orthodontic force was applied. In the second experiment, the right and left upper molars were first moved buccally for 3 weeks. Risedronate treatment began on the day the spring was removed. After the force was withdrawn, the resorbed root surfaces on both the control and risedronate-treated sides were gradually restored by apposition of repair cementum (cementoid). The topical administration of risedronate did not appear to inhibit the repair process of root resorption. These results suggest that the topical administration of risedronate may be useful in preventing root resorption of teeth during orthodontic treatment.

Tooth resorption, a common feline dental problem, is often initiated at the cemento-enamel junction and hence is called cat 'neck' lesion. Studies have demonstrated that osteoclasts/odontoclasts are increased and activated at resorption sites, and that areas of resorption are partly repaired by formation of tissues resembling bone, cementum, and possibly dentin. However, the cellular/molecular mechanisms/factors involved in resorption and repair are unknown. In this study of tissues from cats with 'neck' lesions, we used specific antibodies and immunohistochemical analyses to examine adhesion molecules associated with mineralized tissues, bone sialoprotein (BSP) and osteopontin (OPN), and a cell-surface receptor linked with these molecules, α _vβ₃, for their localization in these lesions. In addition, to determine general cellular activity during repair, we performed in situ hybridization using a type I collagen riboprobe.

Results showed OPN localized to resorption fronts and reversal lines, while BSP was localized to reversal lines. However, some osteoclasts and odontoblasts "sat" on mineralized surfaces not associated with OPN. The cell-surface receptor, α_vβ ₃, was localized to surfaces of osteoclasts/odontoclasts. Type I collagen mRNA was expressed where osteoblasts attempted to repair mineralized tissue. In contrast, odontoblasts did not express mRNA for type I collagen. This study suggests that osteoclastic resorption is the predominant activity in 'neck' lesions and that this activity was accompanied, at least in part, by increased concentrations of OPN and an associated integrin, α _vβ₃, at resorption sites. Lack of collagen expression by odontoblasts indicates that odontoblasts do not play an active role in attempts at repair.

The epithelial proliferation associated with inflammatory periapical lesions and with periapical cyst formation represents an interesting but poorly understood pathological change. Keratinocyte growth factor (KGF) is a recently identified growth factor that is produced by stromal fibroblasts and acts specifically to stimulate epithelial growth and differentiation. To investigate its possible role in the activation of the normally quiescent rests of Malassez, we examined the expression of KGF by in situ hybridization of sections of normal periodontal ligament (PDL) and of 12 periapical granulomas or cysts. Normal PDL and periapical granulomas with scant inflammatory infiltration showed few cells expressing message for KGF. However, KGFexpressing cells were found in the connective tissue stroma close to dense foci of inflammatory cells and to proliferating epithelial elements and cystic epithelial linings. Examination of tissues by the reverse-transcription polymerase chain reaction (RT-PCR) showed KGF expression in 4 specimens of periapical lesions but low or undetectable levels in normal PDL. These observations suggest that the induction of KGF expression in the stromal cells of periapical lesions may play an important role in stimulating the epithelial proliferation associated with cyst formation.

Because diabetic patients are easily led to manifest severe periodontitis, we wanted to determine whether various glucose levels interfere with normal cellular function. Human periodontal ligament (PDL) cells were cultured in glucose-free medium, or in medium containing either 1100 mg/L of glucose (normal-glucose medium) or 4500 mg/L of glucose (high-glucose medium). Cells cultured in glucose-free medium changed their morphology from spindle-shaped to round, and incorporated trypan blue in a time-dependent manner. The incorporation rate was much faster in cells with shorter cell cycles than in those with longer cycles, suggesting the involvement of cell-cycle progression in cell death. However, fragmented DNA, which suggests apoptotic cell death, was not observed in these cells. We reasoned that initial cell rounding and detachment from the culture plate might be due to the conformational changes in cell-surface receptors to fibronectin, a major extracellular matrix for fibroblasts. Western blot analysis revealed that cells cultured in glucose-free medium lost their fibronectin receptor in a time-dependent manner. In addition, fibronectin receptor expression was much higher in cells cultured in high-glucose medium than in cells cultured in normal-glucose medium. Furthermore, the over-expression of the fibronectin receptor resulted in a suppressed chemotactic response of these cells to platelet-derived growth factor. On the basis of these data, it was hypothesized that a high glucose level induced over-expression of these receptors. This might be the mechanism by which a high glucose level compromises wound healing in diabetic patients and, at least in part, might be the reason diabetic patients are subject to severe periodontal destruction.

Previous reports have linked the prevalence of tooth abnormalities with high caries experience in the different types of epidermolysis bullosa (EB). However, it is not known to what extent the apparent susceptibility to enamel caries is due to disease-related altered enamel chemistry in these cases. The aim of this study was to characterize the enamel of teeth from patients suffering from recessive epidermolysis bullosa dystrophica (rEBD) in terms of its mineral content, carbonate content, protein content, and amino acid composition. The results showed that dental enamel from these patients was essentially normal in terms of its chemistry. It is therefore concluded that the high caries experience in recessive dystrophic epidermolysis bullosa patients is probably related to other factors, such as compromised oral hygiene and prolonged oral clearance due to extensive oral soft tissue damage and a cariogenic diet.

Recent studies show that methacrylate-based composites with amorphous calcium phosphate (ACP) as a filler can release supersaturating levels of calcium and phosphate ions in proportions favorable for apatite formation. These findings suggest that such composites could be effectively used as coatings for remineralizing teeth damaged by tooth decay. To examine this hypothesis, we tested composites in vitro for their efficacy to remineralize artificially formed caries-like lesions in extracted bovine incisors. Single 120-μm-thick sagittal tooth sections were placed in holders that exposed only the carious enamel surface. The exposed surfaces were coated with a 1-mm- to 1.5-mm-thick layer of the composite containing, by mass, 40% apatite, silica, or P₂O₇ ^4--stabilized ACP and 60% photoactivated resin comprised of Bis-GMA, TEGDMA, HEMA, and ZrM. The photocured composite-coated sections were immersed either in a remineralizing solution for 4 weeks at 37°C (static model) or cyclically immersed in demineralizing (0.5 h) and remineralizing solutions (11.5 h) for 2 weeks (dynamic model). Quantitative digital image analysis of matched 102 μm x 220 um areas from contact microradiographs taken of the sections before and after immersion showed that lesions coated with ACP-filled composites fractionally recovered 71% ± 33% of their lost mineral compared with 14% ± 13% for apatite controls in the static model and 38% ± 16% compared with -6% ± 24% in the dynamic model. The results suggest that sealants based on ACP-filled methacrylate composites have the potential to remineralize carious enamel lesions.

Recently, we introduced an intra-oral dentin demineralization model to evaluate the efficacy of a fluoridated toothpaste at plaque-retention sites with limited access to salivary components and fluoride. Usage of the fluoridated toothpaste reduced mineral loss from grooves in sound dentin by 15% compared with a non-fluoridated toothpaste (Lagerweij et al., 1997). In the current study, preformed dentinal lesions were subjected to similar toothpaste treatments. To improve the power of the model in discriminating between toothpaste groups, we changed the design from monadic to crossover to reduce the influence of variation between the participants. Two sets of specimens were placed in the partial prostheses of 29 participants who used a toothpaste containing either 1000 ppm fluoride or no fluoride during two consecutive six-week periods. Changes in mineral content were analyzed by transverse microradiography. The average mineral loss after the non-fluoride period was 1785 vol% x μm (SD 2399), and 1335 vol% x μm (SD 2039) after the fluoride period, a 25% inhibition in demineralization. Analysis of variance showed that the demineralization was significantly influenced not only by the type of treatment (p < 0.001 level), but also by the participant (p < 0.001), the width of the groove (p < 0.001), and the depth into the groove where changes in mineral content were measured (p < 0.017). Twenty-one percent of all the lesions showed remineralization. No significant difference in the extent or the occurrence of remineralization was found between the treatments.

Though dental composite materials leach filler elements when stored in distilled water, it is not known whether similar leaching occurs in saliva. The hypothesis to be tested was that due to ion exchange occurring at the filler surfaces, more filler elements leach from composites stored in a salt solution simulating saliva than from composites stored in distilled water. Another aim was to determine how matrix selection, filler composition, and filler silanization affect filler leachability of composites after storage in the simulated saliva and water media. We made 128 batches of experimental composites. Half of these used a bis-GMA/TEGDMA matrix and the other a UEDMA/TEGDMA matrix. Either silica or barium glass filler particles were incorporated into these matrices. Filler silanization was followed by a filler drying at 60°C for 24 h. Half of the silanized particles received an additional heat treatment for 1 h at 110°C in vacuum. One specimen per batch was stored in distilled water and the other in artificial saliva at 37°C. After each 30-day interval for one year, the specimens were transferred to either freshly distilled water or newly mixed artificial saliva. The "old" solutions were analyzed by ICP for determination of the Si, Ba, and Al concentrations. Analysis of variance revealed that storage solution, filler composition, and total time in the storage solution had strong effects on the leachability (p < 0.0001 in all cases). The average monthly leakage of Si for quartz-filled composites was 0.22 ± 0.20 μg/mL (distilled water) and 2.80 ± 1.20 μg/mL (artificial saliva). For barium-glass-filled composites, the corresponding Si leaching values were 0.73 ± 0.48 μg/mL and 5.00 ± 2.20 ug/mL. The monthly means of the barium leaching values were 2.00 ± 1.00 ug/mL (distilled water) and 3.10 ± 1.80 ug/mL (artificial saliva). The large difference between leaching in artificial saliva and in distilled water, as well as the interaction between storage medium and filler, cast doubt on the clinical relevance of in vitro studies using distilled water.

Clinical studies measuring the lifetime of dental prostheses produce censored data when not all specimens have failed during the course of the study. Such clinical data can be analyzed by the Weibull probability distribution function. Algorithms are presented that provide the maximum likelihood estimates of the distribution's parameters. These parameters are the characteristic lifetime (time to failure for 63% of the specimens of the total sample) and the Weibull or shape parameter. Two iterative methods for solving the maximum likelihood equations are given. These mathematical methods have been applied to the results from a retrospective clinical investigation into the lifetime assessment of resin-bonded prostheses. This study evaluated 164 resin-bonded prostheses (for 146 patients) placed between January, 1980, and May, 1985. To date (April, 1995), 47 prostheses (29%) have failed with a median time in service of 74 months (6.2 yr). For the surviving prostheses, the median time in service is 123 months (10.3 yr) and still increasing. The maximum likelihood estimate of the characteristic lifetime for these restorations is 255 months (21.3 yr). Differences in the characteristic lifetime were observed between prostheses placed anteriorly, 338 months (28 yr), and posteriorly, 207 months (17 yr). Since there are no rigorous confidence intervals for deeply censored samples, only provisional confidence bounds could be determined, which substantiated the observed differences. The Weibull modulus value of 1.5 indicates that the probability of failure for resin-bonded prostheses begins to decrease after 10 years in service.

It has been reported that the presence of a smear layer on dentinal substrates can compromise bonding. Typically, smear layers are removed by acidic agents that selectively extract calcium salts from dentin surfaces to leave a collagen-rich substrate. Acid-conditioned dentin (i.e., demineralized) is then primed and an adhesive agent applied. In the present study, we removed smear layers by "polishing" dentin specimens with a hydroxyapatite paste and ultrasonication. Bonding procedures were carried out by means of an aqueous solution of 20% 2-methacryloyloxyethyl phenyl phosphoric acid (phenyl-P) and 30% 2-hydroxyethyl methacrylate, referred to as 20P-30H, a "self-etching primer". The 20P-30H solution was applied to "intact" dentin (i.e., non-demineralized) for either 30 or 60 s. Control samples received no application (0 s) of the self-etching primer. Mean tensile bond strengths (10 MPa) were similar in both the 30-second- and 60-second-primed groups. The widths of formed hybrid layers varied from 0.3 ± 0.2 pm at 0 s application (control) to 2.1 ± 0.3 μm for the 30-second group and 4.1 ± 0.2 pm for the 60-second group. SEM and TEM observations revealed that the 20P-30H self-etching primer created diffusion channels into "intact", calcium-rich dentin which permitted monomer to infiltrate dentin substrates. Hybrid layers identified under microscopic examination demonstrated resistance to both HCl and NaOCI treatments, suggesting that the hybrid layer was not defective, and that bonding was stable.