Shipping amphorae and shipping sheep? Livestock mobility in the north-east Iberian peninsula during the Iron Age based on strontium isotopic analyses of sheep and goat tooth enamel.

Animal mobility is a common strategy to overcome scarcity of food and the related over-grazing of pastures. It is also essential to reduce the inbreeding rate of animal populations, which is known to have a negative impact on fertility and productivity. The present paper shows the geographic range of sheep provisioning in different phases of occupation at the Iron Age site of Turó de la Font de la Canya (7th to 3rd centuries BC). Strontium isotope ratios from 34 archaeological sheep and goat enamel, two archaeological bones and 14 modern tree leaves are presented. The isotopic results suggest that sheep and goats consumed at the site were reared locally (within a few kilometres radius) during the whole period of occupation. The paper discusses the isotopic results in light of the socio-political structure of this period, as complex, strongly territorial societies developed during the Iron Age in the north-east Iberian Peninsula.

Middle Stone Age human teeth from Magubike rockshelter, Iringa Region, Tanzania.

In 2006, six isolated hominin teeth were excavated from Middle Stone Age (MSA) deposits at the Magubike rockshelter in southern Tanzania. They comprise two central incisors, one lateral incisor, one canine, one third premolar, and one fourth premolar. All are fully developed and come from the maxilla. None of the teeth are duplicated, so they may represent a single individual. While there is some evidence of post-depositional alteration, the morphology of these teeth clearly shares features with anatomically modern Homo sapiens. Both metric and non-metric traits are compared to those from other African and non-African dental remains. The degree of biological relatedness between eastern and southern African Stone Age hunter-gatherers has long been a subject of interest, and several characteristics of the Magubike teeth resemble those of the San of southern Africa. Another notable feature is that the three incisors are marked on the labial crown by scratches that are much coarser than microwear striations. These non-masticatory scratches on the Magubike teeth suggest that the use of the front teeth as tools included regularly repeated activities undertaken throughout the life of the individual. The exact age of these teeth is not clear as ESR and radiocarbon dates on associated snail shells give varying results, but a conservative estimate of their minimum age is 45,000 years.

High-resolution isotopic evidence of specialised cattle herding in the European Neolithic.

Reconstructing stock herding strategies and land use is key to comprehending past human social organization and economy. We present laser-ablation strontium and carbon isotope data from 25 cattle (Bos taurus) to reconstruct mobility and infer herding management at the Swiss lakeside settlement of Arbon Bleiche 3, occupied for only 15 years (3384-3370 BC). Our results reveal three distinct isotopic patterns that likely reflect different herding strategies: 1) localized cattle herding, 2) seasonal movement, and 3) herding away from the site year-round. Different strategies of herding are not uniformly represented in various areas of the settlement, which indicates specialist modes of cattle management. The pressure on local fodder capacities and the need for alternative herding regimes must have involved diverse access to grazing resources. Consequently, the increasing importance of cattle in the local landscape was likely to have contributed to the progress of socio-economic differentiation in early agricultural societies in Europe.

Ph-activated nano-amorphous calcium phosphate-based cement to reduce dental enamel demineralization.

Enamel demineralization is destructive, esthetically compromised, and costly complications for orthodontic patients. Nano-sized amorphous calcium phosphate (NACP) has been explored to address this challenge. The 20% NACP-loaded ortho-cement notably exhibited favorable behavior on reducing demineralization of enamel around brackets in a caries model designed to simulate the carious attack. The 20% NACP-loaded ortho-cement markedly promotes higher calcium and phosphate release at a low pH, and the mineral loss was almost two fold lower and carious lesion depth decreased the by 1/3. This novel approach is promising co-adjuvant route for prevention of dental caries dissemination in millions of patients under orthodontic treatment.

Antibacterial effect of coffee: calcium concentration in a culture containing teeth/biofilm exposed to Coffea Canephora aqueous extract.

UNLABELLED: This study determined the changes of calcium concentration in a medium containing teeth/biofilm exposed to Coffea canephora extract (CCE). Enamel fragments were randomly fixed into two 24-well polystyrene plates containing BHI. Pooled human saliva was added to form biofilm on fragments. Specimens were divided into treatment groups (G, n = 8 per group) and treated with 50 µl daily for 1 min per week, as follows: G1, 20% CCE; G2, Milli-Q water (negative control); G3, antibiotic (positive control). Six fragments represented the blank control (G4). The calcium content was observed at baseline, 4 and 7 days of treatment by atomic-absorption spectrophotometry. Cross-sectional hardness of enamel was a demineralization indicator. Calcium increased in the medium after 4 and 7 days of treatment in G1 (3·80 ± 1·3 mg l(-1) and 4·93 ± 2·1 mg l(-1) , respectively) and G3 (4th day = 5·7 ± 1·8 mg l(-1) ; 7th day = 6·7 ± 3·5 mg l(-1) ) (P > 0·05). Calcium from G2 decreased after 7 days, which was different from G3 (P < 0·05). The lower calcium content, at the end of the experiment, was represented by G4, 2·16 ± 0·2 mg l(-1) . The increase in calcium after treatment with CCE is probably due to its antibacterial effect, which caused the bacterial lysis and consequent release of calcium in the medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed an inhibitory action of Coffea canephora against dental biofilm. This coffee species caused bacterial lysis and consequent release of calcium into the medium. Furthermore, the advantage of coffee as an antibacterial beverage is that it is consumed in a concentrated form (6-10%) as opposed to various medicinal infusions that have shown such effect in vitro and are usually consumed at 1-2%. Therefore, a light roasted C. canephora aqueous extract can be considered as a potential anticariogenic substance.

Topical laser application enhances enamel fluoride uptake and tribological properties.

Topical fluoride treatment prevents dental caries. However, the resulting calcium-fluoride-like deposits are soft and have poor wear resistance; therefore, frequent treatment is required. Lasers quickly heat surfaces and can be made portable and suitable for oral remedies. We examined the morphology, nanohardness, elastic modulus, nanowear, and fluoride uptake of fluoride-treated enamel followed by CO2 laser irradiation for 5 and 10 sec, respectively. We found that laser treatments significantly increased the mechanical properties of the calcium-fluoride-like deposits. The wear resistance of the calcium-fluoride-like deposits improved about 34% after laser irradiation for 5 sec and about 40% following irradiation for 10 sec. We also found that laser treatments increased fluoride uptake by at least 23%. Overall, laser treatment significantly improved fluoride incorporation into dental tissue and the wear resistance of the protective calcium-fluoride layer.

Fluoride plus functionalized ß-TCP: a promising combination for robust remineralization.

With more than 50 years of clinical success, fluoride serves as the gold standard agent for preventing tooth decay. In particular, the action of fluoride facilitates saliva-driven remineralization of demineralized enamel and alters solubility beneficially. Still, tooth decay remains problematic, and one way to address it may be through the development of new mineralizing agents. Laboratory and clinical studies have demonstrated that the combination of fluoride and functionalized ß-tricalcium phosphate (fTCP) produces stronger, more acid-resistant mineral relative to fluoride, native ß-TCP, or fTCP alone. In contrast to other calcium-based approaches that seem to rely on high levels of calcium and phosphate to drive remineralization, fTCP is a low-dose system designed to fit within existing topical fluoride preparations. The functionalization of ß-TCP with organic and/or inorganic molecules provides a barrier that prevents premature fluoride-calcium interactions and aids in mineralization when applied via common preparations and procedures. While additional clinical studies are warranted, supplementing with fTCP to enhance fluoride-based nucleation activity, with subsequent remineralization driven by dietary and salivary calcium and phosphate, appears to be a promising approach.

Mechanical and micromorphological evaluation of chlorhexidine-mediated dentin remineralization.

Chlorhexidine (CHX) has been reported to reduce self-degradation of collagen fibrils by inhibiting host-derived protease activity in demineralized dentin. Theoretically, if the collagen fibril scaffold of demineralized dentin maintains its original crosslinkage pattern on treatment with CHX and appropriate supplementation with necessary mineral sources, dentin remineralization may occur in demineralized lesions. In this study, we provide direct mechanical and micromorphological evidence for the ability of CHX to promote remineralization of demineralized dentin. Specifically, with respect to demineralized dentin blocks treated with different concentrations of CHX (0.02-2%) and stored in simulated body fluid, we have observed a significant increase in the elastic modulus of dentin treated with relatively high concentrations of CHX (0.2 and 2%) as storage time increased, whereas the elastic modulus of the non-CHX treated control group decreased. We have also observed a dense mineral deposition along collagen fibrils in the dentin group treated with 0.2 and 2% CHX via field emission scanning electron microscopy.

Effects of zinc and fluoride on the remineralisation of artificial carious lesions under simulated plaque fluid conditions.

The aim was to study the effects of zinc (Zn) and fluoride (F) on remineralisation at plaque fluid concentrations. Artificial carious lesions were created in 2 acid-gel demineralising systems (initially infinitely undersaturated and partially saturated with respect to enamel) giving lesions with different mineral distribution characteristics (high and low R values, respectively) but similar integrated mineral loss values. Lesions of both types were assigned to 1 of 4 groups and remineralised for 5 days at 37°C. Zn and F were added, based on plaque fluid concentrations 1 h after application, to give 4 treatments: 231 µmol/l Zn, 10.5 µmol/l F, Zn/F combined and an unmodified control solution (non-F/non-Zn). Subsequently remineralisation was measured using microradiography. High-R lesions were analysed for calcium, phosphorus, F and Zn using electron probe micro-analysis. All lesions underwent statistically significant remineralisation. For low-R lesions, remineralisation was in the order F(a) < non-F/non-Zn(a) < Zn(a, b) < Zn/F(b), and for high-R lesions F(a) < non-F/non-Zn(b) < Zn(b) < Zn/F(c) (treatments with the same superscript letter not significantly different, at p < 0.05). Qualitatively, remineralisation occurred throughout non-F/non-Zn and Zn groups, predominantly at the surface zone (F) and within the lesion body (Zn/F). Electron probe micro-analysis revealed Zn in relatively large amounts in the outer regions (Zn, Zn/F). F was abundant not only at the surface (F), but also in the lesion body (Zn/F). Calcium:phosphate ratios were similar to hydroxyapatite (all). To conclude, under static remineralising conditions simulating plaque fluid, Zn/F treatment gave significantly greater remineralisation than did F treatment, possibly because Zn in the Zn/F group maintained greater surface zone porosity compared with F, facilitating greater lesion body remineralisation.

Diode laser irradiation and fluoride uptake in human teeth.

AIM: To evaluate chemically the effects of diode laser on fluoride uptake before and after laser irradiation of enamel surfaces. METHODS: Crowns of 20 sound human teeth were halved and a 3 x 3 mm acid-resistant varnish uncovered window left for: A) no treatment; B) fluoride (Elmex gel); C) diode (fluoride + diode laser); D) diode (diode laser + fluoride). The dental surfaces were analysed using a fluoride ion-selective electrode, in order to evaluate the fluoride treatment in combination with a diode laser. Also, to investigate laser-induced compositional changes (contents in F(-)) in enamel before/after laser irradiation and topical fluoride application. RESULTS: The mean ± SD of fluoride uptake of teeth of group A was 1.55 ± 0.89 mg/l. Mean fluoride uptake increased sevenfold after fluoride gel treatment: 10.51 ± 3.38 mg/l for group B, up to 15 times after gel and laser treatment: 23.62 ± 3.58 mg/l for group C and was 22.7 ± 4.60 mg/l for group D (diode laser before fluoride application). The Kruskal Wallis test indicated a statistically significant effect of fluoride uptake for all three treatments (p<0.001). The Student-Newman-Keuls multiple comparison test indicated a statistically significant increase of fluoride uptake before and after all treatments, and also a statistically significant difference for laser treatment versus fluoride gel. However, there was no statistically significance difference between laser groups. CONCLUSIONS: There is an enhanced capability of lasers to increase fluoride uptake of enamel and providing protection to enamel surface from acid attack.

Comparison of effects of diode laser and CO2 laser on human teeth and their usefulness in topical fluoridation.

Various authors have reported more effective fluoridation from the use of lasers combined with topical fluoride than from conventional topical fluoridation. Besides the beneficial effect of lasers in reducing the acid solubility of an enamel surface, they can also increase the uptake of fluoride. The study objectives were to compare the action of CO(2) and GaAlAs diode lasers on dental enamel and their effects on pulp temperature and enamel fluoride uptake. Different groups of selected enamel surfaces were treated with amine fluoride and irradiated with CO(2) laser at an energy power of 1 or 2 W or with diode laser at 5 or 7 W for 15 s each and compared to enamel surfaces without treatment or topical fluoridated. Samples were examined by means of environmental scanning electron microscopy (ESEM). Surfaces of all enamel samples were then acid-etched, measuring the amount of fluoride deposited on the enamel by using a selective ion electrode. Other enamel surfaces selected under the same conditions were irradiated as described above, measuring the increase in pulp temperature with a thermocouple wire. Fluorination with CO(2) laser at 1 W and diode laser at 7 W produced a significantly greater fluoride uptake on enamel (89 ± 18 mg/l) and (77 ± 17 mg/l) versus topical fluoridation alone (58 ± 7 mg/l) and no treatment (20 ± 1 mg/l). Diode laser at 5 W produced a lesser alteration of the enamel surface compared to CO(2) laser at 1 W, but greater pulp safety was provided by CO(2) laser (ΔT° 1.60° ± 0.5) than by diode laser (ΔT° 3.16° ± 0.6). Diode laser at 7 W and CO(2) laser at 2 W both caused alterations on enamel surfaces, but great pulp safety was again obtained with CO(2) (ΔT° 4.44° ± 0.60) than with diode (ΔT° 5.25° ± 0.55). Our study demonstrates that CO(2) and diode laser irradiation of the enamel surface can both increase fluoride uptake; however, laser energy parameters must be carefully controlled in order to limit increases in pulpal temperature and alterations to the enamel surface.

Combination of high-fluoride toothpaste and no post-brushing water rinsing on enamel demineralization using an in-situ caries model with orthodontic bands.

OBJECTIVE: To compare the effects on enamel demineralization and fluoride (F) retention of two different brushing­rinsing regimens. MATERIAL AND METHODS: An in-situ caries model with orthodontic bands was used for 8­9 weeks. A total of 20 orthodontic patients participated. They were randomized into two groups: (1) a test group using 5000 ppm F (n = 10) with no post-brushing water rinsing; and (2) a control group using 1450 ppm F (n = 10) with three sessions of post-brushing water rinsing. Orthodontic stainless-steel bands were applied to the two upper first premolars, leaving 2­3 mm of space away from the exposed buccal surface in order to accumulate plaque and provoke initial caries development. The teeth were extracted after 8 and 9 weeks, then analysed using quantitative light-induced fluorescence (QLF). Additionally, oral F retention was compared for the two groups. RESULTS: In comparison to the control group, the test regimen resulted in a non-significant smaller QLF lesion area and a significantly lower average QLF loss of fluorescence (P < 0.05). The highest F retention concentration under the band was found in the test group (P < 0.001). CONCLUSIONS: The combination of using a 5000 ppm F toothpaste and no post-brushing water rinsing had a greater anti-caries potential and resulted in elevated oral F retention compared to a 1450 ppm F toothpaste with three sessions of post-brushing water rinsing.

Influence of chemical activation of a 35% hydrogen peroxide bleaching gel on its penetration and efficacy--in vitro study.

OBJECTIVES: The aim of this study was to evaluate the effects of chemical activation of hydrogen peroxide (HP) gel on colour changes and penetration through the tooth structure. METHODS: One hundred and four bovine incisors were used. One dentine (CD) disc and one enamel-dentine (ED) disc were prepared from each tooth. They were positioned over artificial pulpal chambers and the bleaching was performed with an experimental 35% HP gel. Two control and six experimental groups were prepared. In the positive control group (PC) no chemical activator was used. In the negative control group (NC) the specimens did not receive any bleaching. Each experimental group received a different chemical activator (manganese gluconate-MG; manganese chlorite-MC; ferrous sulphate-FS; ferrous chlorite-FC; and mulberries root extract-MRE). After the bleaching procedure a sample of solution was collected from the artificial pulpal chamber and the HP concentration was measured. The data were analysed using ANOVA, Tukey's, and Dunnett's tests. RESULTS: The groups MG and FS showed a significantly lower penetration of HP than the PC group. For the parameter Delta E, all the groups, with the exception of the group MRE, showed a significantly higher means in relation to the PC group in ED colour. For dentine colour, just the groups MG and FS had significant differences in relation to PC. CONCLUSIONS: The addition of MG and FS decreases the penetration of HP. The chemical activation using metal salts tested was effective in increasing the bleaching effect.

Chitosan effect on dental enamel de-remineralization: an in vitro evaluation.

OBJECTIVES: The aim of this work was to evaluate the in vitro effect of chitosan (concentration and time of action) treatment on enamel de-remineralization behavior upon a pH cycling assay. METHODS: Different group of human tooth samples were exposed to de-remineralizing solutions of controlled pH using a random experimental design. Microhardness and phosphorus chemical analysis were employed to evaluate the loss of phosphorus from the samples. Optical coherence tomography (OCT) images were obtained for selected specimens in order to evaluate the degree of penetration of chitosan into enamel. RESULTS: Vickers microhardness results were higher for samples treated with chitosan for concentration between 2.5mg/mL and 5.0mg/mL and time of action between 60s and 90 s. A maximum inhibition of mineral loss of 81% was obtained. Chemical analysis indicated lower net pohosphorus loss (net P loss) for samples treated with chitosan. Best results were obtained in the same conditions found out with microhardness measurements. Chitosan had little effect on the remineralization process. OCT results indicated a correlation of chitosan penetration with chitosan concentration. For chitosan concentrations of 2.5 g/mL and 5.0 g/mL the penetration was up to the dentin-enamel junction. CONCLUSIONS: Chitosan interferes with the process of demineralization of the tooth enamel inhibiting the release of phosphorus in this laboratory study. Demineralization is influenced by the concentration and exposure time of the biopolymer to the enamel. Microhardness measurements may be used as an indication of mineral loss from tooth enamel. Additionally, OCT images support the idea that chitosan may act as a barrier against acid penetration, contributing to its demineralization inhibition.

Surface dental enamel lead levels and antisocial behavior in Brazilian adolescents.

Lead poisoning has been reportedly linked to a high risk of learning disabilities, aggression and criminal offenses. To study the association between lead exposure and antisocial/delinquent behavior, a cross-sectional study was conducted with 173 Brazilian youths aged 14-18 and their parents (n=93), living in impoverished neighborhoods of Bauru-SP, with high criminality indices. Self-Reported Delinquency (SRD) and Child Behavior Checklist (CBCL) questionnaires were used to evaluate delinquent/antisocial behavior. Body lead burdens were evaluated in surface dental enamel acid microbiopsies. The dental enamel lead levels (DELL) were quantified by graphite furnace atomic absorption spectrometry (GFAAS) and phosphorus content was measured using inductively coupled plasma optical emission spectrometry (ICP-OES). Logistic regression was used to identify associations between DELL and each scale defined by CBCL and SRD scores. Odd ratios adjusted for familial and social covariates, considering a group of youths exposed to high lead levels (>or=75 percentile), indicated that high DELL is associated with increased risk of exceeding the clinical score for somatic complaints, social problems, rule-breaking behavior and externalizing problems (CI 95%). High DELL was not found to be associated with elevated SRD scores. In conclusion, our data support the hypothesis that high-level lead exposure can trigger antisocial behavior, which calls for public policies to prevent lead poisoning.

Effect of adhesive types on enamel discolouration around orthodontic brackets.

BACKGROUND: Unsightly colour changes may occur in the enamel around bonded orthodontic brackets. AIM: To determine the enamel colour changes around orthodontic brackets bonded with different adhesives. METHODS: One hundred extracted upper first premolars were divided into five equal groups. Fiducial holes were drilled in the buccal and palatal surfaces of the teeth and metal orthodontic brackets were bonded to the buccal surfaces with different combinations of adhesives and primers. The teeth were immersed in a tea-coffee solution for one week. The brackets were then debonded and the teeth polished. The colour of the enamel surface was measured around each fiducial hole before bonding (baseline), after debonding and after polishing. The colour differences (deltaE) were compared using one-way ANOVA and the paired t-test. RESULTS: There were no significant differences between the five adhesive groups in baseline-debonding deltaE and baseline-polishing deltaE, but statistically significant differences were found between the baseline-debonding deltaE on the buccal and palatal surfaces and the baseline-debonding deltaE versus baseline-polishing deltaE on the buccal surfaces. Although statistically significant, the colour changes would not be visible to many observers. Approximately 75 per cent of the staining was attributed to the enamel components. CONCLUSIONS: The type of adhesive and the methods of application had no effect on enamel colour change. Most of the colour change was attributed to uptake of stain by the components of the enamel, not the resin tags.

The susceptibility of bleached enamel to staining as measured by Quantitative Light-induced Fluorescence (QLF).

OBJECTIVES: To report the use of Quantitative Light-induced Fluorescence (QLF) to determine if there was a tendency for bleached enamel to take up extrinsic stains more than unbleached enamel. METHODS: Bovine teeth devoid of stains were selected, the roots removed and enamel gently pumiced. Each tooth was sectioned into two and each half randomly assigned to two groups (bleached or unbleached). Windows were created on each half using clear acid resistant varnish. 38% Hydrogen peroxide gel was applied to the exposed windows of the bleached group for 1 hour. The teeth were rinsed and dried. Bleached and unbleached halves of the same teeth were then mounted on glass rods attached to pot lids using green stick. QLF images were taken. The teeth were subjected to a cycle of artificial saliva, chlorhexidine and tea (2 minutes in each solution). This was repeated 5 times. QLF images were taken at the end of each cycle. RESULTS: The uptake and progression of stain was detected in all the sections by QLF. Using paired t- test (SPSS) there was no significant difference between the two groups for the change from baseline to the final stain cycle (p > 0.05), however there was variability in stain uptake within the groups as the cycles progressed. CONCLUSION: Bleaching of enamel in vitro does not appear to increase the susceptibility of enamel to extrinsic staining.

Effect of strontium in combination with fluoride on enamel remineralization in vitro.

Previous studies showed that strontium (Sr) as well as fluoride (F) can enhance enamel remineralization. The aim of this study was to evaluate the effects of Sr in combination with F on enamel remineralization in vitro. Sixty enamel specimens obtained from caries free human premolars were demineralised to produce caries-like lesions. Half of each lesion was covered with nail varnish as an untreated control. The specimens were then randomly divided into F and Sr+F treatment groups. The F group was exposed to remineralizing solutions (1.5mM CaCl(2), 0.9 mM KH(2)PO(4)) containing 1 ppm, 0.1 ppm or 0.05 ppm F. The Sr+F treatment group was exposed to the same solutions including 10 ppm Sr. After 2 weeks, lesion depth, mineral loss and percentage enamel remineralization were determined using transversal microradiography. There was a significant decrease in mineral loss in all groups (p<0.001). Lesion depth was significantly reduced for all groups (p<0.05) with the exception of group F. Remineralization was significantly affected by F concentration (p=0.000). The participation of Sr resulted in a significant enhancement of remineralization (p<0.001) with a synergistic effect of the Sr+F combination (p<0.01). It was concluded that while the remineralizing process was affected by the concentration of F, there was also an interaction between F and Sr when they were used in conjunction.

New insights into structural alteration of enamel apatite induced by citric acid and sodium fluoride solutions.

Attenuated total reflectance infrared spectroscopy and complementary scanning electron microscopy were applied to analyze the surface structure of enamel apatite exposed to citric acid and to investigate the protective potential of fluorine-containing reagents against citric acid-induced erosion. Enamel and, for comparison, geological hydroxylapatite samples were treated with aqueous solutions of citric acid and sodium fluoride of different concentrations, ranging from 0.01 to 0.5 mol/L for citric acid solutions and from 0.5 to 2.0% for fluoride solutions. The two solutions were applied either simultaneously or consecutively. The citric acid-induced structural modification of apatite increases with the increase in the citric acid concentration and the number of treatments. The application of sodium fluoride alone does not suppress the atomic level changes in apatite exposed to acidic agents. The addition of sodium fluoride to citric acid solutions leads to formation of surface CaF2 and considerably reduces the changes in the apatite P-O-Ca framework. However, the CaF2 globules deposited on the enamel surface seem to be insufficient to prevent the alteration of the apatite structure upon further exposure to acidic agents. No evidence for fluorine-induced recovery of the apatite structure was found.

The amyloid protein APin is highly expressed during enamel mineralization and maturation in rat incisors.

This study investigated the expression and localization of APin (which was previously identified and cloned from a rat odontoblast cDNA library), during ameloblast differentiation in rat incisors, by using in situ hybridization and immunohistochemistry. The subcellular localization of APin varied during ameloblast differentiation, but was stage-specific. APin mRNA was not expressed in pre-ameloblasts, was weakly expressed in secretory ameloblasts, and was strongly expressed in maturation-stage ameloblasts as well as in the junctional epithelium attached to the enamel of erupted molars. In the maturation-stage ameloblasts, APin protein was conspicuous in the supranuclear area (Golgi complex) of smooth-ended ameloblasts as well as in both the supranuclear area and the ruffle end of ruffle-ended ameloblasts. During ameloblast-lineage cell culture, APin was expressed at a low level in the early stages of culture, but at a high level in the late stage of culture, which was equivalent to the maturation stage. APin protein was efficiently secreted from transfected cells in culture. Furthermore, its overexpression and inactivation caused an increase and decrease in matrix metalloproteinase-20 (MMP-20) and tuftelin expression, respectively. These findings indicate a functional role for APin in the mineralization and maturation of enamel that is mediated by the expression of MMP-20 and tuftelin.