MEMO1 Is Required for Ameloblast Maturation and Functional Enamel Formation.

Coordinated mineralization of soft tissue is central to organismal form and function, while dysregulated mineralization underlies several human pathologies. Oral epithelial-derived ameloblasts are polarized, secretory cells responsible for generating enamel, the most mineralized substance in the human body. Defects in ameloblast development result in enamel anomalies, including amelogenesis imperfecta. Identifying proteins critical in ameloblast development can provide insight into specific pathologies associated with enamel-related disorders or, more broadly, mechanisms of mineralization. Previous studies identified a role for MEMO1 in bone mineralization; however, whether MEMO1 functions in the generation of additional mineralized structures remains unknown. Here, we identify a critical role for MEMO1 in enamel mineralization. First, we show that Memo1 is expressed in ameloblasts and, second, that its conditional deletion from ameloblasts results in enamel defects, characterized by a decline in mineral density and tooth integrity. Histology revealed that the mineralization defects in Memo1 mutant ameloblasts correlated with a disruption in ameloblast morphology. Finally, molecular profiling of ameloblasts and their progenitors in Memo1 oral epithelial mutants revealed a disruption to cytoskeletal-associated genes and a reduction in late-stage ameloblast markers, relative to controls. Collectively, our findings integrate MEMO1 into an emerging network of molecules important for ameloblast development and provide a system to further interrogate the relationship of cytoskeletal and amelogenesis-related defects.

Method for the analysis of total fluoride in fluoride-releasing dental varnishes.

Today's fluoride-releasing varnishes (F-varnish) contain a wide variety of ingredients which present analytical challenges for measuring their total fluoride content. This study reports improved methods to measure fluoride content in F-varnishes. Six different commercially available F-varnishes that contain difluorosilane (0.1% F) or NaF (2.26% F) alone or in combination with calcium-phosphates were analyzed. In a vial, 1-3 drops (0.05-0.15 g) of varnish product was dispensed, dissolved in chloroform, equilibrated in TISAB and analyzed via fluoride ion-selective electrode. The average weight percentage of fluoride for all F-varnishes containing NaF ranged from 2.03 to 2.24% F, which is within 90% of the declared label concentration of 2.26% F. Analysis of the difluorosilane-containing product required an additional hydrolysis step. ANOVA found no significant difference between the 5% NaF varnishes at p < 0.05. This method for fluoride analysis yields reliable and reproducible results and can be used for a wide variety of F-varnishes. The standard uncertainty for this method is ±4%. This method may become the basis for national and international standards that ensure the F-varnish products used in clinical practice have the fluoride content declared in the product literature.

Microbial biogeochemistry of Boiling Springs Lake: a physically dynamic, oligotrophic, low-pH geothermal ecosystem.

Boiling Springs Lake (BSL) in Lassen Volcanic National Park, California, is North America's largest hot spring, but little is known about the physical, chemical, and biological features of the system. Using a remotely operated vessel, we characterized the bathymetry and near-surface temperatures at sub-meter resolution. The majority of the 1.2 ha, pH 2.2 lake is 10 m deep and 50-52 °C, but temperatures reach 93 °C locally. We extracted DNA from water and sediments collected from warm (52 °C) and hot (73-83 °C) sites separated by 180 m. Gene clone libraries and functional gene microarray (GeoChip 3.0) were used to investigate the BSL community, and uptake of radiolabeled carbon sources was used to assess the relative importance of heterotrophic vs. autotrophic production. Microbial assemblages are similar in both sites despite the strong temperature differential, supporting observations of a dynamic, convectively mixed system. Bacteria in the Actinobacteria and Aquificales phyla are abundant in the water column, and Archaea distantly related to known taxa are abundant in sediments. The functional potential appears similar across a 5-year time span, indicating a stable community with little inter-annual variation, despite the documented seasonal temperature cycle. BSL water-derived DNA contains genes for complete C, N, and S cycles, and low hybridization to probes for N and S oxidation suggests that reductive processes dominate. Many of the detected genes for these processes were from uncultivated bacteria, suggesting novel organisms are responsible for key ecosystem services. Selection imposed by low nutrients, low pH, and high temperature appear to result in low diversity and evenness of genes for key functions involved in C, N, and S cycling. Conversely, organic degradation genes appear to be functionally redundant, and the rapid assimilation of radiolabeled organic carbon into BSL cells suggests the importance of allochthonous C fueling heterotrophic production in the BSL C cycle.

Improved Contact X-Ray Microradiographic Method to Measure Mineral Density of Hard Dental Tissues.

Contact X-ray microradiography is the current gold standard for measuring mineral densities of partially demineralized tooth specimens. The X-ray sensitive film specified in the last J Res NIST publication on the subject is no longer commercially available. OBJECTIVES: Develop a new microradiographic method by identifying a commercially available film with greater than 3000 lines per millimeter resolution, which is sensitive to X rays, and develop correct film processing for X-ray microradiographic application. METHODS: A holographic film was identified as a potential replacement film. Proper exposure was determined utilizing a thick nickel plate to create test-strips. Film development was bracketed around manufacturer suggestions. Film linearity was determined with aluminum step-wedges. Microradiographs of 100 µm thick tooth sections, before and after acidic challenges, were a final test for film. Magnified images were captured with a digital microscope camera with 0.305 micrometers per pixel resolution. RESULTS: The appropriate film exposure was 30 minutes at 80 kV(p) and 3 mA with a development time of 2 minutes. Step-wedge experiments show the system to be linear in terms of pixel intensities with respect to x-ray attenuation for normalized pixel intensity values that are 10% to 90% of full scale (r(2) = 0.997) which encompasses the full exposure region of tooth tissue. Enamel sections were analyzed and show distinctive differences between erosion and demineralization. The image capture device resolution of 0.305 micrometers per pixel limits the system resolution. CONCLUSION: Use of the identified holographic film when combined with the described processing modifications has resulted in an improved X-ray microradiographic method for the measurement of mineral density of dental hard tissues. The method described can be further improved by using a higher resolution digitization system. The method is appropriate for quantitatively measuring changes in mineral density and erosion.

Calcium pre-rinse greatly increases overnight salivary fluoride after a 228 ppm fluoride rinse.

BACKGROUND: Large increases in salivary fluoride were reported 1 h after a calcium pre-rinse/NaF rinse. AIMS: This study examined the persistence of these increases. METHODS: 12 subjects rinsed in the evening with water, with a 228 microg/g (ppm) F rinse or with 150 mmol/l calcium lactate followed by a 228 microg/g F rinse. In a second experiment these same patients rinsed with a 912 microg/g F rinse. Saliva samples were obtained the morning after rinsing, centrifuged and the supernatants analyzed. RESULTS: The Ca pre-rinse/228 microg/g F rinse induced an increase in overnight salivary F over the 912 microg/g F rinse (approximately 2.5 times) and a statistically significant increase over the 228 mug/g F rinse (approximately 5.5 times). CONCLUSIONS: The results suggest that a Ca pretreatment may increase the cariostatic effect of topical F agents.

Ca pre-rinse greatly increases plaque and plaque fluid F.

Previous studies demonstrated that a Ca pre-treatment greatly increases salivary F from a subsequent NaF rinse. This study examines if these increases are found in plaque and plaque fluid F. Thirteen individuals accumulated plaque before rinsing with: (1) 12 mmol/L NaF (228 microg/g F), (2) 150 mmol/L Ca rinse, or (3) the Ca rinse followed by the F rinse. One hr later, plaque samples were collected, the plaque fluid was recovered, and the plaque residues were extracted 5 times with pH 6.8 or pH 4.8 buffers, and then by acid. The F in each extract after the Ca rinse/F rinse greatly exceeded the corresponding F from the NaF rinse. Consequently, the Ca rinse/F rinse increased the total plaque F and the plaque fluid F by 12x and 5x, compared with the NaF rinse alone. These and the previous salivary results suggest that a Ca pre-treatment may increase the cariostatic effects of topical F agents.

Salivary fluoride from fluoride dentifrices or rinses after use of a calcium pre-rinse or calcium dentifrice.

The low concentration of available calcium (Ca) in oral fluids limits the formation of Ca-mediated fluoride deposits that maintain oral fluoride (F) after a topical F treatment. The purpose of this study was to examine if a high concentration of Ca would increase salivary F when used before a F rinse or dentifrice. We found that a Ca pre-rinse (150 mmol/l Ca lactate) or Ca dentifrice (0.084 g Ca glycerolphosphate per gram dentifrice) used immediately before a 60 s 228-ppm F rinse (12 mmol/l NaF) produced a 4.6x or 3.6x increase (p < 0.05) respectively in the 1 h salivary F concentrations over the F rinse alone. Reducing the post-Ca F rinse to 10 s still produced a significant 2.2x increase in salivary F compared to the 60 s F rinse alone. Used with a conventional 1,100 ppm F (i.e. 1,100 microg F per gram) NaF dentifrice (Crest), the above Ca pre-rinse increased 1 h salivary F levels by 2.3x over the F dentifrice alone. However, a F rinse given before a Ca rinse produced no increase in 1 h salivary F concentrations. Although the persistence of these increases requires further study, these results suggest that a moderately high concentration of Ca given shortly before a F rinse or F dentifrice may increase the cariostatic effect of the F product.

Effect of a calcium prerinse on salivary fluoride after a 228-ppm fluoride rinse.

The objective of this study was to determine if a concentrated calcium prerinse given before a fluoride rinse would cause an increase in the post rinse sali vary fluoride (F). A panel of 5 subjects used a 30, 150 or 300 mmol/l calcium lactate prerinse followed by a 1-min NaF rinse. All calcium prerinses significantly increased the 1-hour saliva F relative to the NaF control without a prerinse. The maximum increase was produced by the 150 mmol/l calcium lactate prerinse and was about ninefold higher than the NaF control.

Biology, persistence and detection of Cryptosporidium parvum and Cryptosporidium hominis oocyst.

Cryptosporidium parvum and Cryptosporidium hominis are obligate enteric protozoan parasites which infect the gastrointestinal tract of animals and humans. The mechanism(s) by which these parasites cause gastrointestinal distress in their hosts is not well understood. The risk of waterborne transmission of Cryptosporidium is a serious global issue in drinking water safety. Oocysts from these organisms are extremely robust, prevalent in source water supplies and capable of surviving in the environment for extended periods of time. Resistance to conventional water treatment by chlorination, lack of correlation with biological indicator microorganisms and the absence of adequate methods to detect the presence of infectious oocysts necessitates the development of consistent and effective means of parasite removal from the water supply. Additional research into improving water treatment and sewage treatment practices is needed, particularly in testing the efficiency of ozone in oocyst inactivation. Timely and efficient detection of infectious C. parvum and C. hominis oocysts in environmental samples requires the development of rapid and sensitive techniques for the concentration, purification and detection of these parasites. A major factor confounding proper detection remains the inability to adequately and efficiently concentrate oocysts from environmental samples, while limiting the presence of extraneous materials. Molecular-based techniques are the most promising methods for the sensitive and accurate detection of C. parvum and C. hominis. With the availability of numerous target sequences, RT-PCR will likely emerge as an important method to assess oocyst viability. In addition, a multiplex PCR for the simultaneous detection of C. parvum, C. hominis and other waterborne pathogens such as Giardia lamblia would greatly benefit the water industry and protect human health.

Hypertriglyceridaemia in Alström's syndrome: causes and associations in 37 cases.

OBJECTIVE: To document frequency of severe hypertriglyceridaemia in Alström's syndrome (AS) and its relationship to hepatic and renal function, glycaemia and insulin resistance. PATIENTS AND METHODS: Thirty-seven subjects with AS aged 5-35 years, 51% male, were assessed at multidisciplinary clinics in Canada, UK and Italy. Diagnostic criteria were: severe cone/rod dystrophy leading to severe visual impairment in early childhood, sensorineural deafness, moderate overall obesity and normal intelligence. Three patients were treated with thyroxine for primary hypothyroidism and one female patient for secondary amenorrhoea with 20 micro g ethinyloestradial combined oral contraceptive. Two male patients were receiving monthly intramuscular testosterone enanthate for secondary hypogonadism. Fasting bloods were taken for serum insulin, serum glucose, serum triglycerides, hepatic and renal function and glycosylated Hb. Triglyceride levels > 8 mmol/l and fasting serum insulin levels > 16 microunits/ml were considered to represent severe hypertriglyceridaemia and severe insulin resistance, respectively. All subjects with (23) hypertriglyceridaemia also had high insulin resistance, as measured by HOMA modelling. However, there was no significant correlation between log tyriglyceride and log serum insulin or HOMA in the whole group (P = 0.2 and 0.14, respectively). There was no clear relationship between serum triglyceride levels and age, body mass index (BMI), hepatic or renal impairment or glycaemia. CONCLUSION: The first overview of serum triglyceride levels in a significant number of reported cases of Alström Syndrome shows an overlap between severe hypertriglyceridaemia and severe hyperinsulinism, but not a direct correlation between the two nor with insulin resistance measured by HOMA. Triglyceride levels were not related to glycaemia, hepatic or renal dysfunction.

Fluoride release from a resin-modified glass-ionomer cement in a continuous-flow system. Effect of pH.

Fluoride is added to many dental restorative materials, including glass-ionomer cements, for the specific purpose of leaching fluoride into the surrounding tissues to provide secondary caries inhibition. During the caries process, an acidic environment attacks the dental tissues as well as the glass-ionomer cement. We hypothesized that pH significantly affects the rate of release of fluoride from the glass-ionomer cement. A continuous-flow fluoride-measuring system that monitors the amount of fluoride released over time was used to determine the release of fluoride from a resin-modified glass-ionomer cement (KetacFil). The results show that the release rate began with a fast burst of fluoride which quickly diminished to low levels in 3 days. Under neutral pH conditions, the rate of fluoride release at 72 hrs was significantly slower than at pH 4.

Effect in vitro acidification on plaque fluid composition with and without a NaF or a controlled-release fluoride rinse.

Plaque fluid ion concentration changes, especially fluoride, in response to the pH decrease associated with a cariogenic episode are important components of the caries process. A "controlled-release" (CR) fluoride rinse, based on the controlled release of fluoride in the presence of calcium, has been shown to form large fluoride reservoirs in resting plaque. In this study, the in vitro acid-induced release of fluoride, and other ions, was examined in 48-hour-fasted plaque fluid from subjects (n = 11) who received no rinse, or who used a 228-ppm CR or NaF fluoride rinse 1 hr before being sampled. After collection, the plaque was centrifuged to yield plaque fluid, acidified (0.1 microL of 0.5 mol/L HCl per milligram plaque), and then re-centrifuged before a second sample was obtained. Although previous studies indicated a higher plaque fluid fluoride after the new rinse relative to NaF, no statistically significant difference was observed here. Average fluoride release after acidification (average pH, 5.2) was statistically greater following the use of the CR rinse (153 micromol/L) compared with the NaF rinse (17 micromol/L). No fluoride release was seen in the no-rinse samples. The pH, free calcium, phosphate, acetate, propionate, and buffer capacity were not affected by the different amounts of fluoride deposited in the plaque. However, following acid addition, an increase in free calcium and phosphate was observed, which was also independent of the rinse. The large release of fluoride following acidification suggests that the new rinse may provide an improved cariostatic effect.

Remineralization effects of a two-solution fluoride mouthrinse: an in situ study.

Results from previous studies show that a two-solution fluoride (F) rinse is significantly more effective than a NaF rinse of the same F content of 250 microg/g (ppm) in remineralizing enamel and root lesions in an in vitro cyclic de- and remineralization model. In the present study, the two-solution rinse and two NaF rinses with F contents of 250 ppm and 1000 ppm were evaluated in an intra-oral remineralization model. Caries-like lesions were formed in the enamel of extracted human molars with the use of a pH 4 demineralizing solution. Thin sections of the enamel (approximately 120 microm) containing lesions were prepared, and the mineral contents of the lesions were assessed by quantitative microradiography. With the cut surfaces protected by nail varnish, 3 enamel specimens were mounted with wax in the lingual areas of a removable mandibular appliance. The study used a randomized, crossover design with seven subjects. In each of the 3 legs of the study, subjects wore the appliances continuously except when eating, drinking, and brushing their teeth. Twice daily (after breakfast and before bedtime), the subjects received a one-minute rinse with 20 mL of (1) 250-ppm-F NaF rinse, (2) 1000-ppm-F NaF rinse, or (3) 228-ppm-F two-solution F rinse. At the end of the 14-day experimental period, the sections were retrieved, and the mineral contents of the lesions were again assessed quantitatively. The results show that both the 1000-ppm-F NaF and 228-ppm-F two-solution rinses produced a greater (p < 0.05) remineralization than did the 250-ppm-F NaF rinse. The remineralization produced by the two-solution rinse was not statistically different (p > 0.05) from that produced by the NaF rinse with 4x the F content (1000 ppm F).

Composition of plaque and saliva following use of an alpha-tricalcium-phosphate-containing chewing gum and a subsequent sucrose challenge.

Previous studies demonstrated that the chewing of a 2.5% (mass fraction) alpha-tricalcium-phosphate-fortified (alpha-TCP) experimental chewing gum released sufficient calcium and phosphate to eliminate any fall in the tooth mineral saturation of plaque fluid after a sucrose rinse (Vogel et al., 1998). In contrast, the chewing of a conventional sugar-free gum did not eliminate this decrease in saturation. The purpose of this study was to examine if the release of ions from plaque calcium-phosphate pools induced by this gum could provide protection during subsequent exposure to cariogenic conditions. Fourteen subjects accumulated plaque for 48 hrs, fasted overnight, chewed a control or experimental gum for 15 min, and subsequently rinsed 1 min with a mass fraction 10% sucrose solution. Before gum chewing, and at 7 min and 15 min afterward, whole plaque, plaque fluid, and salivary samples were obtained and analyzed by micro-analytical techniques. Additional samples were collected and analyzed at 25 min (7 min after the sucrose rinse). Although the results confirmed the deposition of large amounts of calcium and phosphates in plaque seen in the previous study, only a small increase was seen in plaque-fluid-free calcium and phosphate before sucrose administration. This suggests that few of the mineral ions were mobilized under non-cariogenic conditions. However, 7 min after the sucrose rinsing, an increase in these concentrations was seen which, based on hydroxyapatite ion activity product calculations, indicated a decrease in the driving force for demineralization compared with that seen with the control gum. These results suggest that the chewing of the experimental gum deposits a labile mineral reservoir in plaque that can resist a subsequent cariogenic challenge.

Measurement of Calcium Activity in Oral Fluids by Ion Selective Electrode: Method Evaluation and Simplified Calculation of Ion Activity Products.

The activity of calcium in plaque fluid is needed to calculate the saturation level of that fluid relative to the tooth mineral. One method to determine the calcium activity in very small plaque fluid samples is by micro ion-selective electrode (ISE). Two commercially available calcium ionophores, a neutral-carrier and a charged-carrier, were evaluated in micro ISEs and compared to a commercially available macro ISE using saliva as a model for plaque fluid. The neutral-carrier containing ISEs gave results consistent with those of the macro ISE. Calcium activity measurements made with micro ISEs that contained the neutral ion-carrier of whole plaque samples and plaque fluid samples obtained by centrifugation of whole plaque showed that the activities did not change due to centrifugation. Estimates of the saturation with respect to hydroxyapatite were made from these measurements. A simplified calculation method is presented to estimate the ion activity product (IAP) of the calcium-phosphate minerals. The method is based on the relative abundance of some of the possible calcium-binding species and a fixed ionic strength for plaque fluid. Calculations show that within a normal pH range for plaque fluid (5.0 to 7.5) the differences in the IAP calculations for hydroxyapatite using the simplified method are less than those estimated from propagation of uncertainty calculations.

Composition of plaque and saliva following a sucrose challenge and use of an alpha-tricalcium-phosphate-containing chewing gum.

Calcium phosphate concentrations in plaque, plaque fluid, and saliva play an important role in caries prevention. In this study, we used a microanalytical technique to examine the anticaries potential of a 2.5% (mass fraction) alpha-tricalcium-phosphate-fortified experimental gum by measuring the pH, free and total calcium, and total phosphate in plaque fluid, whole plaque, and saliva, and centrifuged saliva from 14 subjects who (1) accumulated plaque for 48 hours, (2) fasted overnight, (3) rinsed for 1 min with sucrose, and (4) chewed a control or experimental gum for 15 min. From these data, the hydroxyapatite (HAp) ion activity products (IAP[HAp]) of saliva and plaque fluid were calculated as a measure of tooth mineral saturation. Results, compared with those of the control gum, show significant increases in pH and in free calcium and phosphate concentrations in plaque fluid and saliva when the experimental gum was chewed following sucrose ingestion. These increases result in a rise in fluid saturation with respect to tooth mineral that, for plaque fluid, nearly cancels the decrease seen with the control gum after the sucrose rinse. This suggests that the experimental gum may be more effective than a conventional gum in ameliorating the cariogenic effects of sucrose. Similar statistically significant increases were also seen in the total calcium content of the plaque fluid, centrifuged saliva, whole saliva, and whole plaque, and in the total phosphate of whole plaque and whole saliva. These results suggest that the deposition of a mineral reservoir in plaque and saliva by the experimental gum may help resist future cariogenic challenges.

Choroid plexus papilloma and cysts in the Aicardi syndrome: case reports.

Two female infants with callosal agenesis, infantile spasms, chorioretinal lacunae, optic disc colobomas and cortical heterotopias were diagnosed with Aicardi syndrome. A choroid plexus papilloma was found in one patient, and choroid plexus cysts were found in the other. Choroid plexus lesions are common findings in the Aicardi syndrome and are discussed in this paper.

Increased overnight fluoride concentrations in saliva, plaque, and plaque fluid after a novel two-solution rinse.

Recent studies showed that salivary, plaque-fluid, and whole-plaque fluoride were significantly higher 120 min after subjects rinsed with a novel two-solution rinse than after they rinsed with a NaF rinse of the same fluoride concentration. In this study, the persistence of these increases was investigated overnight, a period of time that is more clinically relevant. Improved analytical techniques for the ultramicro determination of whole-plaque and plaque-fluid fluoride from the same sample are also described. Thirteen subjects abstained from toothbrushing for 48 hrs and rinsed for 1 min with a 12 mmol/L (228 ppm) NaF or the two-solution rinse before bedtime. Samples were then collected the following morning before breakfast: (1) Saliva samples were either clarified by centrifugation or acid-extracted with 1 mol/L HClO4; and (2) single-site molar plaque samples were centrifuged to obtain plaque fluid and/or extracted with 1 mol/L HClO4. Results showed that, compared with NaF, the two-solution rinse produced significantly higher fluoride concentrations in all samples: The concentration of fluoride in whole plaque and whole saliva following the new rinse exceeded concentrations found after the NaF rinse by factors of three and four, respectively, while in plaque fluid, the two-solution rinse produced about a two-fold increase over NaF values, which were near baseline levels. This increase, however, was only about 20% in centrifuged saliva. The increases in saliva and especially in plaque-fluid fluoride after the two-solution rinse indicate a greater remineralization potential, while the enhanced fluoride reservoirs found in plaque overnight after this rinse constitute a reserve that may release fluoride into the plaque fluid over an extended period of time.

Changes in the permselectivity of human teeth during caries attack.

Previous studies have shown that enamel permselectivity can influence fluid composition within caries lesions during de- and remineralization. The permselectivity of human enamel, cementum, and dentin sections was examined, in a microwell model, by measurement of the membrane potential developed by KCl diffusion while the sections were immersed in solutions simulating resting (pH = 5.6) and cariogenic plaque fluid (pH = 4.8). In a second experiment, the effects of charged compounds (phytate and Zonyl-FSC) on the tooth permselectivity were examined. The average membrane potentials (+/- SD) in "resting plaque" solution were: sound enamel, 18.9 +/- 3.2 mV, n = 66; dentin, 0.9 +/- 9.2 mV, n = 59; and cementum, -0.8 +/- 8.2 mV, n = 42, with a positive sing indicating cation selectivity. The average membrane potentials became more negative in "cariogenic plaque" solution for all types of sections: sound enamel, 5.2 +/- 2.1 mV, n = 46; dentin, -8.1 +/- 7.4 mV, n = 45; and cementum, -14.3 +/- 8.0 mV, n = 34. In lesion enamel sections, the membrane potential was reduced from the non-lesion wells in both types of test solutions, while phytate treatment caused an increase of approximately 10 mV in potential (increased cation selectivity) in every enamel well in either "resting" or "cariogenic" solution. Treatment of enamel sections with Zonyl-FSC caused the membrane potential to become more negative in both test solutions, with many of the wells showing anion selectivity in the cariogenic "plaque-like" solution. However, the changes in enamel membrane potentials induced by Zonyl-FSC slowly increased toward the initial values after treatment, while the effects of the phytate pre-treatment persisted. Most dentin sections treated with phytate also showed an increase in potential after phytate treatment; however, Zonyl-FSC seemed to have little effect on the membrane potential of dentin. The results of this study suggest that modification of tooth permselectivity by surface-active agents may be a viable method of decreasing the rate of caries progression.

An in vitro diffusion model for the study of calcification of bovine pericardium tissue.

Bovine pericardium (BP) is extensively used for the production of heart valve bioprostheses. BP has excellent mechanical properties but a limited lifespan because of intrinsic subsurface calcification in vivo. In this study, the in vitro mineralization of BP was investigated by a novel diffusion cell model. In two sets of experiments, glutaraldehyde-treated BP membranes were placed between two compartments, both of which contained calcium phosphate solutions made by equilibration of octacalcium phosphate (Exp I) or dicalcium phosphate dihydrate (Exp II) in phosphoric acid. The movement of calcium (Ca), phosphate (P), and protons through the BP membrane was followed throughout the diffusion process. Histology, scanning electron microscopy, wet chemical analysis, and energy dispersive X-ray analyses provided good evidence of subsurface mineralization of BP that resembled in vivo mineral deposition. Energy dispersive x-ray microanalyses found a Ca/P heterogeneity of the early subsurface mineral that formed in the membrane. The use of a diffusion cell to model BP calcification under well-characterized conditions has led to in vitro mineralization that more closely matches that observed in vivo. The results suggest that this in vitro diffusion model can be used to study the mechanism of pathological mineralization. This model has the potential to provide rapid, inexpensive, basic information about the mineralization process.