Dental Calculus as a Potential Biosource for Human Papillomavirus Detection in Oral Squamous Cell Carcinoma.

OBJECTIVE: The infection of human papillomaviruses (HPVs) plays a role in the development of oral squamous cell carcinoma (OSCC). A poor oral hygiene and dental calculus may cause the infection to persist. Therefore, this study aimed to assess whether this dental calculus could serve as a potential biosource in early detection of HPVs in patients with OSCC. METHODS: DNA was isolated from the dental calculus of people diagnosed with OSCC, and MY09/11 primer set was used to detect the presence of HPV. The positive samples were further sequenced and aligned using megablast NCBI BLAST tool to identify the HPV genotype. RESULTS: Electrophoresis examination showed that 4 of 14 samples collected (29%) had a clear single band, of which three had 97% to 99% similarity to a high-risk genotype HPV-58. Meanwhile, the other sample had 99% similarity to an unclassified papillomaviridae. CONCLUSION: Dental calculus is a promising source of HPV in oral cavity and could be used as a biomarker for early detection.

Ancient DNA analysis of food remains in human dental calculus from the Edo period, Japan.

Although there are many methods for reconstructing diets of the past, detailed taxon identification is still challenging, and most plants hardly remain at a site. In this study, we applied DNA metabarcoding to dental calculus of premodern Japan for the taxonomic identification of food items. DNA was extracted from 13 human dental calculi from the Unko-in site (18th-19th century) of the Edo period, Japan. Polymerase chain reaction (PCR) and sequencing were performed using a primer set specific to the genus Oryza because rice (Oryza sativa) was a staple food and this was the only member of this genus present in Japan at that time. DNA metabarcoding targeting plants, animals (meat and fish), and fungi were also carried out to investigate dietary diversity. We detected amplified products of the genus Oryza from more than half of the samples using PCR and Sanger sequencing. DNA metabarcoding enabled us to identify taxa of plants and fungi, although taxa of animals were not detected, except human. Most of the plant taxonomic groups (family/genus level) are present in Japan and include candidate species consumed as food at that time, as confirmed by historical literature. The other groups featured in the lifestyle of Edo people, such as for medicinal purposes and tobacco. The results indicate that plant DNA analysis from calculus provides information about food diversity and lifestyle habits from the past and can complement other analytical methods such as microparticle analysis and stable isotope analysis.

Lifestyle of a Roman Imperial community: ethnobotanical evidence from dental calculus of the Ager Curensis inhabitants.

BACKGROUND: The analysis of ancient calcified dental plaque is a powerful archaeobotanical method to elucidate the key role of the plants in human history. METHODS: In this research, by applying both optic microscopy and gas chromatography mass spectrometry on this matrix, a detailed qualitative investigation for reconstructing the lifestyle of a Roman imperial community of the Ager Curensis (Sabina Tiberina, Central Italy) was performed. RESULTS: The detection of animal micro-remains and molecules (e.g., hairs, feather barbules, markers of dairy products), starch granules of several cereals and legumes, pollen (e.g., Juglans regia L., Hedera sp. L.) and other plant micro-debris (e.g., trichome of Olea sp., hemp fibers), and phytochemicals (e.g., Brassicaceae, Lamiaceae herbs, Ferula sp., Trigonella foenum-graecum L., wine, and Humulus lupulus L.) in the dental calculus sample demonstrated that plant-derived foods were regularly consumed together with animal resources. CONCLUSIONS: This nutritional plan, consistent with the information reported in ancient written texts, suggested that the studied population based its own subsistence on both agriculture and husbandry, probably also including beekeeping and hunting activities. All together, these results represent proofs for the comprehension of food habits, phytotherapeutic practices, and cultural traditions of one of the first Roman settlements in the Sabina Tiberina area.

Evaluation of Cadmium Levels in Dental Calculus of Male Oral SCC Patients with Betel-Quid Chewing in Hunan Province of China.

Cadmium (Cd) is a widespread trace toxic heavy metal with long biological half-life and may induce higher risk of cancer on multiple organs of human body. Recent studies have confirmed that dental calculus has enormous potential for investigation of exposure to Cd in the human mouth by acting as a time capsule. We aimed to examine relationship between Cd levels in dental calculus due to betel-quid chewing and risk of oral cancer. This study included 85 male oral squamous cell carcinoma (SCC) cases with betel-quid chewing and smoking as observation subjects (group A) and 67 healthy people with smoking but without betel-quid chewing as control subjects (group B) in Hunan province of Mainland China. Cd levels in calcified dental calculus samples from all participants were measured by inductively coupled plasma mass spectrometry (ICP-MS). The results of this study indicated that cadmium levels in dental calculus were significantly higher in male oral SCC patients with betel-quid chewing and smoking than that in healthy individuals without habit of betel-quid chewing and with smoking (p < 0.0001). This study gives some evidence to support that there may be a positive relationship between cadmium in dental calculus due to betel-quid chewing and risk of oral SCC.

Cadmium Profiles in Dental Calculus: a Cross-Sectional Population-Based Study in Hunan Province of China.

We aimed to investigate whether the cadmium concentrations differ in human dental calculus obtained from the residents with no smoking living in the contaminated area and those with no smoking living in noncontaminated area. In total, there were 260 samples of dental calculus from the adults (n = 50) with no smoking living in contaminated area, the adults (n = 60) with no smoking living in mountainous area, and the adults (n = 150) with no smoking living in low altitude area in Hunan province of China. All samples were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) for cadmium levels. The cadmium levels in dental calculus were significantly higher in the adults with no smoking living in contaminated area than those living in mountainous area and in low altitude area (p < 0.01). The cadmium levels in dental calculus were also higher in the adults with no smoking living in low altitude area than those living in mountainous region (p < 0.01). The results suggested that measuring cadmium levels in dental calculus may be a useful noninvasive method for analysis of environmental exposure to cadmium in the human oral cavity. The low altitude region may have an area contaminated with cadmium in Hunan province of China.

Crystalline structure of pulverized dental calculus induces cell death in oral epithelial cells.

BACKGROUND AND OBJECTIVE: Dental calculus is a mineralized deposit attached to the tooth surface. We have shown that cellular uptake of dental calculus triggers nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome activation, leading to the processing of the interleukin-1ß precursor into its mature form in mouse and human phagocytes. The activation of the NLRP3 inflammasome also induced a lytic form of programmed cell death, pyroptosis, in these cells. However, the effects of dental calculus on other cell types in periodontal tissue have not been investigated. The aim of this study was to determine whether dental calculus can induce cell death in oral epithelial cells. MATERIAL AND METHODS: HSC-2 human oral squamous carcinoma cells, HOMK107 human primary oral epithelial cells and immortalized mouse macrophages were exposed to dental calculus or 1 of its components, hydroxyapatite crystals. For inhibition assays, the cells were exposed to dental calculus in the presence or absence of cytochalasin D (endocytosis inhibitor), z-YVAD-fmk (caspase-1 inhibitor) or glyburide (NLRP3 inflammasome inhibitor). Cytotoxicity was determined by measuring lactate dehydrogenase (LDH) release and staining with propidium iodide. Tumor necrosis factor-α production was quantified by enzyme-linked immunosorbent assay. Oral epithelial barrier function was examined by permeability assay. RESULTS: Dental calculus induced cell death in HSC-2 cells, as judged by LDH release and propidium iodide staining. Dental calculus also induced LDH release from HOMK107 cells. Following heat treatment, dental calculus lost its capacity to induce tumor necrosis factor-α in mouse macrophages, but could induce LDH release in HSC-2 cells, indicating a major role of inorganic components in cell death. Hydroxyapatite crystals also induced cell death in both HSC-2 and HOMK107 cells, as judged by LDH release, indicating the capacity of crystal particles to induce cell death. Cell death induced by dental calculus was significantly inhibited by cytochalasin D, z-YVAD-fmk and glyburide, indicating NLRP3 inflammasome involvement. In permeability assays, dental calculus attenuated the barrier function of HSC-2 cell monolayers. CONCLUSION: Dental calculus induces pyroptotic cell death in human oral epithelial cells and the crystalline structure plays a major role in this process. Oral epithelial cell death induced by dental calculus might be important for the etiology of periodontitis.

High levels of heavy metal accumulation in dental calculus of smokers: a pilot inductively coupled plasma mass spectrometry study.

BACKGROUND AND OBJECTIVE: Various trace elements, including toxic heavy metals, may exist in dental calculus. However, the effect of environmental factors on heavy metal composition of dental calculus is unknown. Smoking is a major environmental source for chronic toxic heavy metal exposition. The aim of this study is to compare toxic heavy metal accumulation levels in supragingival dental calculus of smokers and non-smokers. MATERIAL AND METHODS: A total of 29 supragingival dental calculus samples were obtained from non-smoker (n = 14) and smoker (n = 15) individuals. Subjects with a probability of occupational exposure were excluded from the study. Samples were analyzed by inductively coupled plasma mass spectrometry in terms of 26 metals and metalloids, including toxic heavy metals. RESULTS: Toxic heavy metals, arsenic (p < 0.05), cadmium (p < 0.05), lead (p < 0.01), manganese (p < 0.01) and vanadium (p < 0.01) levels were significantly higher in smokers than non-smokers. The levels of other examined elements were similar in both groups (p > 0.05). CONCLUSION: Within the limitations of this study, it can be concluded that the elementary composition of dental calculus may be affected by environmental factors such as tobacco smoke. Therefore, dental calculus may be utilized as a non-invasive diagnostic biological material for monitoring chronic oral heavy metal exposition. However, further studies are required to evaluate its diagnostic potential.

Patients with dental calculus have increased saliva and gingival crevicular fluid fetuin-A levels but no association with fetuin-A polymorphisms

ABSTRACT: Fetuin-A is a potent inhibitor of calcium-phosphate precipitation and of the calcification process, therefore it can also be related with dental calculus. Thus, we aimed to investigate a possible relationship between fetuin-A gene polymorphism and the presence of dental calculus. A possible relationship between serum, saliva and gingival crevicular fluid (GCF) levels of fetuin-A was also investigated. Fetuin-A c.742C > T and c.766C > G polymorphisms were investigated in 103 patients with or without dental calculus. Additionally, serum, saliva and GCF fetuin-A levels of patients were compared according to dental calculus presence. A significant difference was not observed in the distribution of the fetuin-A c.742C > T and c.766C > G polymorphisms between patients with or without dental calculus. Saliva and GCF fetuin-A concentrations of patients with dental calculus were statistically higher than those without dental calculus (P=0.001, P=0.036 respectively). According to our results, fetuin-A c.742C > T and c.766C > G polymorphisms were not associated with presence of dental calculus. However, higher GCF and saliva fetuin-A levels were detected in patients with dental calculus than in patients without dental calculus, which may result from an adaptive mechanism to inhibit mineral precipitation and eventually calculus formation.

Chemical salivary composition and its relationship with periodontal disease and dental calculus

Aim: To determine the relationship between the chemical composition of saliva, periodontal disease and dental calculus. Methods: An observational analytical cross-sectional study was conducted with patients over 55 years of age. Ethical principles of autonomy and risk protection were applied according to the international standards. Sociodemographic and diagnosis variables (presence of dental calculus and periodontal status) were considered to measure salivary concentrations of glucose (by the glucose oxidase/peroxidase method, amylase (by the colorimetric test), urea (by the amount of indophenol), total protein (by the Bradford method) and albumin (by the nephelometric method). Patients chewed a sterile rubber band and 3 mL of stimulated saliva were collected. The samples were stored at -5 °C, centrifuged at 2,800 rpm for 10 min, and the supernatant was removed and stored at -20 °C. Data were presented as frequencies and proportions for qualitative variables and measures of central tendency and dispersion for quantitative variables. Data were analyzed by either analysis of variance or Kruskal Wallis test . A p value <0.05 was considered statistically significant. Results: Significant relationships were observed between the concentration of salivary urea and periodontal status (p = 0.03) and the presence of dental calculus and urea (p = 0.04) was demonstrated. Conclusions: A relationship between the salivary urea concentration and the presence of periodontal disease and dental calculus is suggested.

Mineral density volume gradients in normal and diseased human tissues.

Clinical computed tomography provides a single mineral density (MD) value for heterogeneous calcified tissues containing early and late stage pathologic formations. The novel aspect of this study is that, it extends current quantitative methods of mapping mineral density gradients to three dimensions, discretizes early and late mineralized stages, identifies elemental distribution in discretized volumes, and correlates measured MD with respective calcium (Ca) to phosphorus (P) and Ca to zinc (Zn) elemental ratios. To accomplish this, MD variations identified using polychromatic radiation from a high resolution micro-computed tomography (micro-CT) benchtop unit were correlated with elemental mapping obtained from a microprobe X-ray fluorescence (XRF) using synchrotron monochromatic radiation. Digital segmentation of tomograms from normal and diseased tissues (N=5 per group; 40-60 year old males) contained significant mineral density variations (enamel: 2820-3095 mg/cc, bone: 570-1415 mg/cc, cementum: 1240-1340 mg/cc, dentin: 1480-1590 mg/cc, cementum affected by periodontitis: 1100-1220 mg/cc, hypomineralized carious dentin: 345-1450 mg/cc, hypermineralized carious dentin: 1815-2740 mg/cc, and dental calculus: 1290-1770 mg/cc). A plausible linear correlation between segmented MD volumes and elemental ratios within these volumes was established, and Ca/P ratios for dentin (1.49), hypomineralized dentin (0.32-0.46), cementum (1.51), and bone (1.68) were observed. Furthermore, varying Ca/Zn ratios were distinguished in adapted compared to normal tissues, such as in bone (855-2765) and in cementum (595-990), highlighting Zn as an influential element in prompting observed adaptive properties. Hence, results provide insights on mineral density gradients with elemental concentrations and elemental footprints that in turn could aid in elucidating mechanistic processes for pathologic formations.

Calcium phosphate particles induce interleukin-8 expression in a human gingival epithelial cell line via the nuclear factor-κB signaling pathway.

BACKGROUND: Dental calculus is calcified plaque composed primarily of calcium phosphate mineral salts, and there is a clear association between the presence of calculus and the initiation/progression of periodontitis. However, it is still inconclusive whether dental calculus can be a direct causative factor. The authors examined the effect of nano/microsized calcium phosphate particles, which may be generated in the process of early precipitation and/or dissolution of calcium phosphate mineral, on the expression of interleukin (IL)-8 in human gingival epithelial cells. METHODS: Primary human gingival epithelial cells and/or a human gingival carcinoma cell line (Ca9-22) were stimulated with calcium phosphate particles. Gene and protein levels were assessed by real-time polymerase chain reaction analysis and enzyme-linked immunosorbent assay, respectively. The activity of nuclear factor (NF)-κB signaling was measured by an immunofluorescence assay to evaluate NF-κB p65 nuclear translocation. RESULTS: The results show that nano/microsized particles stimulate IL-8 expression in human gingival epithelial cells at gene and protein levels. The activity to induce IL-8 expression depends on the particle size: particles with a diameter of 200 nm are more effective than those of 40-nm and 5-µm diameters. Calcium phosphate particles (diameter 200 nm) stimulated NF-κB activity. Pretreatment with BMS-345541, an NF-κB signaling inhibitor, inhibited the particle-mediated IL-8 gene induction, suggesting a requirement for the NF-κB signaling pathway. CONCLUSION: These findings suggest that calcium phosphate particles, which may be related to calculus development, may act as a direct causative factor in the pathogenesis of gingival epithelium.

Staining and calculus formation after 0.12 percent chlorhexidine rinses in plaque-free and plaque covered surfaces: a randomized trial

OBJECTIVES: Studies concerning side effects of chlorhexidine as related to the presence of plaque are scarce. The purpose of this study was to compare the side effects of 0.12 percent chlorhexidine gluconate (CHX) on previously plaque-free (control group) and plaque-covered surfaces (test group). METHODS: This study had a single-blind, randomized, split-mouth, 21 days-experimental gingivitis design, including 20 individuals who abandoned all mechanical plaque control methods during 25 days. After 4 days of plaque accumulation, the individuals had 2 randomized quadrants cleaned, remaining 2 quadrants with plaque-covered dental surfaces. On the fourth day, the individuals started with 0.12 percent CHX rinsing lasting for 21 days. Stain index intensity and extent as well as calculus formation were evaluated during the experimental period. RESULTS: Intergroup comparisons showed statistically higher (p<0.05) stain intensity and extent index as well as calculus formation over the study in test surfaces as compared to control surfaces. Thus, 26.19 percent of test surfaces presented calculus, whereas calculus was observed in 4.52 percent in control surfaces. CONCLUSIONS: The presence of plaque increased 0.12 percent CHX side effects. These results strengthen the necessity of biofilm disruption prior to the start of CHX mouthrinses in order to reduce side effects.

Evidence for calcifying nanoparticles in gingival crevicular fluid and dental calculus in periodontitis.

BACKGROUND: Calcifying nanoparticles (CNPs), also known as nanobacteria, can produce carbonate apatite on their cell walls and initiate pathologic calcification. The objective of this study was to determine whether CNPs are present in the gingival crevicular fluid (GCF) from subjects with periodontal disease and whether they can induce the pathologic calcification of primary cultured human gingival epithelial cells. METHODS: GCF and dental calculus samples were collected from 10 subjects with gingivitis and 10 subjects with chronic periodontitis. CNPs in GCF and calculus filtrates were detected with nanocapture enzyme-linked immunosorbent assay kits. The CNPs in cultures of dental calculus filtrates were also identified using immunofluorescence staining, transmission electron microscopy (TEM), and chemical analysis. Pathologic changes in the CNP-treated gingival epithelial cells were observed with TEM, alizarin red staining, and disk-scanning confocal microscopy. RESULTS: CNPs were found in GCF samples from two subjects with chronic periodontitis. Based on chemical analysis, the surface-associated material from CNPs isolated and cultured from calculus has a composition similar to dental calculus. The pathologic calcification of CNP-treated gingival epithelial cells was also observed. CONCLUSIONS: Self-replicating calcifying nanoparticles can be cultured and identified from dental calculus. This raises the issue of whether CNPs contribute to the pathogenesis of periodontitis.

[Preliminary study of homogeneous phase redeposition of dissolved dental subgingival calculus].

Fresh dental calculus were scratched and rinsed with distilled water, and then dissolved by HNO3. Simulated body fluid was used as control. Aqueous ammonia was added to step up the pH. FSEM and FI-IR were used to analyze the crystal character of deposition. Turbid occurred when pH = 5.4 and deposition occurred when pH = 5.6. Ribbon-like crystal, which was the same as the crystal in natural dental calculus was observed in experimental group and was evidenced to be hydroxyapatite (HAP) by FT-IR. HAP formation through homogeneous phase redeposition of dissolved dental subgingival calculus may be related with the existing template molecules in dental subgingival calculus resolution, which induce the biomineralization of HAP formation.

Dental calculus composition following use of essential-oil/ZnCl2 mouthrinse.

PURPOSE: To test the hypothesis that anticalculus agents cannot completely inhibit calculus formation but can influence the types of calcium phosphate which form, i.e., they can influence the composition of the inorganic component of human dental calculus (HDC). MATERIALS AND METHODS: The composition of HDC specimens obtained from a 16-week multi-center clinical study using three regimens were analyzed, investigators blinded. The treatment regimens were: (a) standard dentifrice (SD), (b) pyrophosphate antitartar dentifrice, and (c) SD with Tartar Control Listerine Antiseptic mouthrinse (containing essential oils and 0.09% zinc chloride). 25 individual samples and eight pooled samples from each group were analyzed using X-ray diffraction, infrared spectroscopy, and scanning electron microscopy. RESULTS: (1) relative frequency of occurrence for: (a) bacteria: Group A = 100%, Group B = 60%, and Group C = 25%; (b) Carbonate hydroxyapatite (CHA): Groups A, B, and C = 100%; (c) dicalcium phosphate dihydrate (DCPD): Group A = 55%; Group B = 45%; Group C = 80%; (2) The relative amount of DCPD is inversely proportional to that of CHA in HDC: the higher the amount of DCPD, the lower the amount of CHA. Group C regimen with essential oil/ZnCl2 mouthrinse and standard dentifrice showed a significant anti-microbial effect and favored the formation of DCPD, the most soluble Ca-P.

Subgingival calculus detection with fluorescence induced by 655 nm InGaAsP diode laser radiation.

BACKGROUND: It has been suggested that the 655 nm InGaAsP diode laser radiation can induce considerably higher amounts of fluorescence in subgingival calculus than in cementum. This in vitro study evaluated the use of fluorescence induced by the diode laser radiation for the detection of subgingival calculus. METHODS: The study sample consisted of 30 extracted molars, premolars, canines, and incisors. The root surface of each sample was partially covered with subgingival calculus. The source of laser radiation was an InGaAsP diode laser emitting visible radiation at a wavelength of 655 nm. The fluorescent radiation reflected from the cementum and calculus was detected by a photoelectric cell integrated within the diagnostic system. The samples were randomly assigned to 3 experimental groups of 10 teeth each for detection of calculus in various media. Specifically, the fluorescence was determined in air (group A), in electrolytic salt solution (group B), and in blood (group C). For each sample, the intensity of the fluorescent light was evaluated separately for cementum and calculus in triplicate measurements. The statistical analysis was performed using a t test at a level of significance of 5% (P<0.05). RESULTS: The intensities of fluorescence radiation emitted by cementum were 0.4 (+/-0.51) in air, 0.4 (+/-0.51) in electrolytic solution, and 2.13 (+/-0.59) in blood. For calculus, the relative intensities of fluorescence radiation were 54.1 (+/-29.09) in air, 60.77 (+/-28.35) in electrolytic salt solution, and 39.63 (+/-29.79) in blood. The differences in fluorescence between cementum and calculus within each experimental unit were significant (group A: P= 0.0002, group B: P= 0.0001, group C: P= 0.031). The intensity of fluorescence in calculus was also significantly different between the 3 experimental groups. CONCLUSION: The present study indicates that subgingival calculus can be reliably detected on extracted teeth using laser fluorescence induced by the 655 nm diode-laser radiation. In vivo studies are necessary to evaluate the clinical utility of this procedure.

Spectrochemical analysis of dental calculus by synchrotron radiation X-ray fluorescence.

This work presents elemental composition studies of human dental calculus by X-ray fluorescence analysis using synchrotron radiation. The intrinsic characteristics of synchrotron light allow for a quantitative analysis of major, minor, and trace elements of very small samples in a single measurement. At present, several theories of calculus formation exist, but none of them can describe completely all the complicated mechanisms involved in the mineralization processes. For this reason, and taking into account that experimental data are long overdue, several calculi with certain degrees of formation were collected from adult patients for analysis. The ratio of calcium/phosphorus was used as an indicator of the major crystalline structure and the state of formation of the calculus. The results demonstrate a clear correlation between the concentrations of certain elements and the type of calculus (supra- or sub-gingival). In addition, the possible correlation between the elements was analyzed.

A cariogenicidade da rapadura in situ em humanos / The cariogenicity od rapadura in situ in humans

A cariogenicidade da rapadura foi avaliada, in situ, em oito voluntários por meio de microdureza superficial do esmalte e pH do biofilme dental. Dois blocos de esmalte bovino, avaliados por microdureza, foram colocados em aparelhos palatinos e usados durante uma semana. Os blocos de esmalte foram supersaturados (4X/dia) com 10 gotas de uma soluçäo de rapadura (1: 3 em água deionizada, teste). Em outra semana, novos blocos foram supersaturados (4X/dia), com uma soluçäo de 10 por cento de sorbitol (controle positivo), e em uma terceira semana com uma soluçäo de 10 por cento de sorbitol (controle negativo). Os resultados mostraram que a rapadura é täo cariogênica quanto a sacarose

New in vitro model for the acquired enamel pellicle: pellicles formed from whole saliva show inter-subject consistency in protein composition and proteolytic fragmentation patterns.

The present investigation was undertaken to investigate the variability of proteins in whole saliva which adsorb to hydroxyapatite and are thus likely to be precursors of the acquired enamel pellicle. Whole-saliva proteins from 18 subjects were absorbed to hydroxyapatite, and the gel filtration patterns of released proteins revealed four major peaks and three minor peaks eluting between the major peaks. Amino acid analysis indicated that minor peaks contained fragments of proteins in major peaks, and this was confirmed by sequence analysis. Major peaks comprised 95% and minor peaks comprised 5% of protein absorbed to hydroxyapatite, suggesting a limited proteolytic capacity of whole saliva. HPLC elution patterns of components in minor peaks suggested that proteolysis is not totally random but is an orderly and consistent process. These studies suggest that whole saliva may be suitable as a model system for the investigation of post-secretory modifications of salivary proteins important for the formation of the acquired enamel pellicle.

Morphology and elemental composition of subgingival calculus in two ethnic groups.

BACKGROUND: The aim of the present study was to compare the morphology and elemental composition of subgingival calculus between Indo-Pakistani and Caucasian patient groups. METHODS: Extracted teeth from 14 Indo-Pakistani and 19 Caucasian subjects were collected. Of these, 12 Indo-Pakistani and 10 Caucasian teeth had sufficient subgingival calculus for analysis. Subgingival calculus present on the 22 teeth was classified into six morphological types: 1) crusty/spiny/nodular; 2) ledge/ ring; 3) thin, smooth veneers; 4) finger/fern-like; 5) individual islands/spots; or 6) supramarginal on submarginal. Subgingival calculus was zoned: coronal, mid, and apical. A sample obtained from each zone was subdivided to allow 3 separate analyses: transmission electron microscopic (TEM) x-ray microanalysis for elemental composition, fluoride analysis, and carbonate analysis. RESULTS: Crusty/spiny/nodular, ledge/ring, and thin, smooth veneers were more commonly found in the Indo-Pakistani group; individual islands were more prevalent in the Caucasian subjects. Supramarginal on submarginal calculus was found only in the Indo-Pakistani group. No finger/fern-like deposits were found. No differences within or between the two ethnic groups were found with regard to calcium:phosphate ratios, fluoride, or carbonate content. However, the Indo-Pakistani group showed significantly lower levels of sodium in apical samples than in coronal samples (ANOVA, F1,16 = 5.98, P= 0.03), and significantly lower levels of sodium (ANOVA, F1,12 = 4.75, P= 0.05) and magnesium (ANOVA, F1,12 = 5.16, P= 0.04) in apical samples than in those from Caucasians. After adjusting for smoking, the magnesium results remained significant (ANOVA, F2,11 = 4.64, P= 0.05). CONCLUSIONS: Subgingival calculus demonstrated differences in morphology between these two ethnic groups and differences in elemental composition, which may influence its overall solubility and contribute to its greater accretion in the Indo-Pakistani subjects.