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Mineral content of the dentine remaining after chemomechanical caries removal.

Although the dentine remaining after chemomechanical caries removal appears sound by normal clinical criteria, no definitive evidence has yet been obtained to confirm that the dentine surface is in fact mineralised. The aim of this study was to use backscattered electron (BSE) imaging and electron probe micro-analysis (EPMA) to ascertain the level of mineralisation of the dentine remaining in cavities prepared by this technique. Carious dentine was removed from carious lesions by means of N-monochloro-DL-2-aminobutyric acid (NMAB) or NMAB containing 2 mol/l urea. Sections of teeth in which caries removal was complete by normal clinical criteria were examined by EPMA and BSE. Dentine adjacent to the pulp was found to be less mineralised than the surrounding dentine. Although the superficial layer of dentine remaining on the cavity floors frequently appeared to have a slightly reduced mineral content, the results clearly indicated that there was no significant difference between this dentine and the underlying sound dentine.

Aminobutyrates↗

Bacterial colonization of mineralized and completely demineralized dentine in situ.

The changing environment in a developing root lesion may result in a succession of the microbial flora in the dentine. As demineralization proceeds, the collagenous matrix is exposed, which could be conducive to the growth of specific microorganisms. In this study both sound and completely demineralized dentine were placed together in the partial prothesis of 8 individuals to test whether the type of substrate influenced the composition of the bacterial flora. After 6 weeks the degradation of the collagenous matrix, the demineralization of the dentine and the microbial composition were assessed. The collagen loss varied between 0 and 69 wt%. Mineral loss from the originally sound dentine specimens ranged from virtually none to complete demineralization. Percentages of total streptococci, mutans streptococci, Actinomyces and lactobacilli isolated from both dentinal substrates did not differ significantly. The percentage of lactobacilli in the dentine specimens was positively correlated to the lesion depth. The percentage of Actinomyces species was significantly higher in both the dentine specimens that had been demineralized in vitro and those that were found to be completely demineralized in situ compared to the partially demineralized dentine specimens. In vitro, no collagenolytic activity of the predominant flora isolated from both dentinal substrates could be shown.

Actinomyces↗

Structural events in the caries process in enamel, cementum, and dentin.

The structural events observed in enamel, cementum, and dentin during the caries process have been reviewed. In incipient enamel lesions, the prevailing concept of an almost intact surface layer has been seriously challenged by SEM and TEM observations demonstrating structural pathways (such as enlarged prism junctions or sheaths) from the enamel surface to the sub-surface lesion. The destruction in this latter location consisted of (1) enlarged prism junctions, (2) diffuse mineral destruction in the prism cores, and (3) destruction of the interprismatic substance. In root caries, the destruction of cementum started along junctions between calcified layers of extrinsic (Sharpey) and intrinsic collagen fibers as well as along incremental lines. Invasion of Gram-positive micro-organisms followed these enlarged junctions. Dentin caries was similar in coronal and root caries. It consisted of sclerosis of the lumens of the dentinal tubules, followed by an important gradient of demineralization of intertubular dentin and destruction of occluded tubular lumens and peritubular dentin. Bacterial penetration occurred initially in the dentinal tubules and was followed by bacterial invasion and destruction of the intertubular dentin. Various phenomena of crystalline remineralization were described in enamel and dentin. Whereas in enamel and dentin caries, an important gradient of demineralization was observed before bacterial invasion, a simultaneous destruction of the mineral and organic components seemed to occur in cementum.

Dental Caries↗

The importance of intrafibrillar mineralization of collagen on the mechanical properties of dentin.

It is widely held that the hardness and modulus of dentin increase in proportion to the mineral concentration. To test this belief, we measured hardness and modulus of normal dentin and an altered form of dentin without gap-zone mineralization in wet and dry conditions by AFM nanoindentation to determine if the modulus and hardness scale linearly with mineral concentration. Mineral concentrations in the mid-coronal location of the normal and altered dentins were 44.4 vol% and 30.9 vol%, respectively. Surrounding the pulp of the altered dentin was a region of higher mineralization, 40.5 vol%. The indentation modulus of normal dentin was 23.9 (SD = 1.1) GPa dry and 20.0 (SD = 1.0) GPa wet. In mid-coronal regions of the altered dentin, the indentation modulus was 13.8 (SD = 2.0) GPa dry and 5.7 (SD = 1.4) GPa wet. In the more mineralized regions of the altered dentin, the modulus was 20.4 (SD = 1.8) GPa dry and 5.3 (SD = 0.8) GPa wet; the properties of the altered wet dentin did not correlate with mineral concentration. The results of this study raise doubt as to whether mineral concentration alone is a sufficient endpoint for assessing the success or failure of remineralization approaches in restorative dentistry.

Adolescent↗

Immunohistochemical distribution of collagens types IV, V, and VI and of pro-collagens types I and III in human alveolar bone and dentine.

The aim of the present study was to characterize the composition of the organic matrix in alveolar jaw bone and dentine using antibodies against pro-collagens Types I and III and collagens Types IV, V, and VI. After demineralization of oral hard tissues in 0.2 N HCl, antigenicity was well preserved and the distribution of the pro-collagens and collagens could be demonstrated. Staining for pro-collagen Type I was prominent around osteoblasts and in pre-dentine, indicating active de novo synthesis of Type I pro-collagen. Pro-collagen Type I was ubiquitous but was less abundant in bone and dentine, whereas pro-collagen Type III was seen only in areas of bone remodeling, in peritubular spaces, and in pre-dentine. Type IV collagen was limited to the basement membranes of vessels in osteons and bone marrow. Type V collagen was detected neither in pre-dentine nor in bone. In contrast, Type VI collagen was found in dentine and bone, showing a faint but homogeneous staining which, similarly to pro-collagen Type III, was pronounced around osteoblasts and in pre-dentine, areas of active bone and dentine formation. This study showed that the organic matrix of dentine and bone contains Type VI as well as Type I collagen. Pro-collagen Type III (and to a lesser extent collagen Type VI) is transiently produced during new formation and remodeling of oral hard tissues, and disappears once the matrix calcifies. Type I pro-collagen qualifies as a general marker protein for increased osteoblastic activity. We conclude that immunostaining for the different collagen/pro-collagen types can be used to assess normal or abnormal stages of bone/dentine formation.

Adolescent↗

Intermittent inhibition of dentin mineralization of rat incisors under continual infusion of 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP) using a subcutaneous mini osmotic pump.

The inhibitory effect of the continual administration of 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP) (8 mgP/kg/day) through a mini osmotic pump on dentin mineralization was examined in relation to the diurnal rhythm of the rat and compared with that of daily injections of same amounts of HEBP known to inhibit dentin mineralization. After daily injections of HEBP, a series of alternating rows of mineralized and non-mineralized dentin islands appeared in the newly formed portion of the crown-analogue of rat incisors. A similar phenomenon occurred under the continual administration of HEBP in rats raised either under regular environmental photofraction or constant lighting conditions. The average distance between the adjacent mineralized dentin islands was 521.0 +/- 51.3 microns in the injected rats. After continual HEBP administration, this was 426.0 +/- 13.2 microns and 416.5 +/- 19.4 microns under ordinary photofraction and constant light, respectively. Although the pattern of individual mineralized dentin islands tended to become irregular in nocturnal rats, no statistical difference was noted between the two values. Rows of mineralized and non-mineralized dentin islands also appeared in the root analogue dentin. No sign of the intermittent inhibition of mineralization was recognized in mesodermal hard tissues other than dentin in the HEBP-affected animals. These data implicate the presence of intrinsic cycles in dentin mineralization at the growing end of rat incisors independent of environmental photofraction as well as the ameloblast function.

Animals↗

Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts.

BACKGROUND: Implant surface roughness and chemical composition, as well as other factors, affect the ability of osteogenic cells to form bone adjacent to an implant. The same principles may also apply to the tooth root and some reports have shown that surface modification of the root may lead to improved restoration of the periodontal apparatus. The most common of these surface modification techniques involves demineralization with citric acid or treatment with tetracycline to expose collagen fibrils. In addition, during normal bone remodeling, osteoclasts demineralize the extracellular matrix, leaving resorption pits and exposed collagen fibrils. In this study, the effect of different dentin surface-preparation techniques on osteoblasts were compared. METHODS: Slices of sperm whale dentin were mechanically polished and surfaces were treated with tetracycline-HCl (TCN) or were cultured with mouse bone marrow cells to create a surface with osteoclast (OC) resorption pits or left untreated. Profilometry, x-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM) were used to evaluate the 3 different dentin surfaces. MG63 osteoblast-like cells were cultured on the 3 different surfaces and the effect of dentin surface preparation technique on MG63 cell proliferation (cell number), differentiaton (alkaline phosphatase specific activity of isolated cells and cell layer lysates; osteocalcin production), and local factor production (transforming growth factor (TGF)-beta1 and prostaglandin E2 (PGE2) compared. RESULTS: Profilometry showed the polished and TCN surfaces were smooth with comparable Ra values, whereas the OC surfaces were slightly rougher due to resorption pits which covered 3.7% of the surface. XPS measurements showed that TCN treatment reduced the Ca and P content of the surface, indicating that it had dissolved the mineral. Osteoclast-resorption also reduced the Ca and P content, but to a lesser extent. MG63 cell proliferation on polished dentin and tissue culture polystyrene was equivalent. In contrast, cells grown on the TCN- and OC-treated surfaces exhibited increased proliferation. No effect of surface treatment on cell alkaline phosphatase activity was observed, but activity in the cell layer lysates was increased on the TCN- and OC-treated surfaces. Osteocalcin production was reduced on all dentin surfaces, but the greatest reduction was found on the TCN-treated surface. Production of both TGF-beta1 and PGE2 was increased on the treated surfaces. All effects were greatest in cultures grown on the TCN-treated dentin. CONCLUSIONS: These data indicate that demineralization of the dentin surface promotes proliferation of osteoblasts and early differentiation events like production of alkaline phosphatase and autocrine mediators such as PGE2 and TGF-beta1. However, later differentiation events like osteocalcin production are decreased. Osteoclast-mediated bone resorption elicits similar responses; less than 4% of the dentin surface resulted in approximately 75% of the response caused by TCN treatment. These observations suggest that greater attention should be paid to the effects of osteoclastic resorption in designing methods for enhancing bone and cementum formation adjacent to root surfaces.

Alkaline Phosphatase↗

The effect of phenytoin medication on dentin apposition, root length, and caries progression in rat molars.

To determine the effect of phenytoin (PHT) on dentin apposition and the rate of caries progression in dentin, 40 Wistar rats were treated daily for 35 days with intraperitoneal injections of 5,5-diphenylhydantoin. Twenty-eight controls received saline with pH adjusted to that of PHT. One PHT and one control group were fed a 43% sucrose diet, and the others a non-cariogenic pellet diet. Streptococcus sobrinus was inoculated to induce caries. Schiff staining was used to determine caries. The areas of dentin apposition and dentinal caries lesions were quantified after tetracycline staining. The root lengths were measured. PHT reduced slightly the dentin apposition and activated significantly the progression of the dentinal carious lesions. No difference in caries initiation was found. The high-sucrose diet reduced dentin apposition and increased the rate of progression of existing caries greatly. Our study suggests that both the high-sucrose diet and PHT have an effect on secreting odontoblasts, which can be seen as an alteration in dentin apposition and caries progression rates in dentin.

Animals↗

Identification of dentin phosphophoryn localization by histochemical stainings.

Phosphophoryn, the most abundant of the dentin non-collagenous proteins, has been considered to be related in function to the mineralization process. In the present study, identification of dentin phosphophoryn localization was attempted using newly developed, precautionary histological methods by which phosphophoryn was retained in the sections during the specimen preparation and stained selectively in situ. Phosphophoryn was found to be present widely in all of the calcified dentin except the mantle dentin, the external, first-formed portion of dentin, but was not found in the predentin, the inner, uncalcified layer of dentin. These results indicate that phosphophoryn is apparently related to the mineral phase of calcified dentin and that the mineralization process of mantle dentin, which is formed before the odontoblasts are fully differentiated, may be different from that of circumpulpal dentin.

Acetates↗

Thermal stability of mineralized and demineralized dentin: a differential scanning calorimetric study.

The purpose of this study was to determine the difference, if any, in the thermal stability of collagen in mineralized and demineralized dentine compared to that in unmineralized tissues, using differential scanning calorimetry, DSC. Human tooth dentin blocks, about 1 x 1 x 2 mm in size, were used in this study. Some dentin blocks were demineralized using a Plank Rychlo solution; others, using EDTA solution. The mineralized dentin showed an exothermic peak at about 310 degrees C and the combustion of organic materials was completed at about 450 degrees C. For the demineralized dentin, the combustion was completed at higher temperature range and showed a strong exothermic peak at about 470 degrees C. An exotherm at the temperature between 450 degrees C and 470 degrees C was also observed in DSC pattern of native type I collagen from calf skin and rat tail tendon. DSC pattern of rat tail collagen showed a close similarity to that of the demineralized dentin. Statistically, the same heat flow value was obtained both from the mineralized dentin and the demineralized dentin and from the native type I collagen. These findings indicated that the thermal stability of collagen in dentin is lower than collagen in uncalcified connective tissue. It is suggested that in calcified collagen, the apatite crystallites may have intruded into spaces of the crosslinks of intra- and inter-fibrils, and in so doing, destroyed the crosslinks.

Animals↗

Effect of depth and tubule direction on ultimate tensile strength of human coronal dentin.

The purpose of this study was to evaluate the effect of dentin depth and tubule direction on the ultimate tensile strength (UTS) of human dentin. Dentin slabs of 0.5-mm thickness were trimmed either from the mesial and distal (for specimens with the tubules parallel to the tensile force; parallel group) or from the occlusal and pulpal surfaces (perpendicular group) to reduce the cross-sectional area of the superficial, middle, and deep regions to 0.25 mm2, and subjected to microtensile testing. From SEM photomicrographs of the fractured specimens of the parallel group, the tubule density was investigated. For both parallel and perpendicular groups, superficial dentin showed a significantly higher UTS than deep dentin. The tubule density of superficial dentin was significantly lower than that of middle and deep dentin. When performing the microtensile bond test to deep dentin, it is possible that cohesive failure of dentin can occur at relatively low tensile stresses.

Dental Restoration Failure↗

Evaluation of Er:YAG laser and EDTAC on dentin adhesion of six endodontic sealers.

The effect of Er:YAG laser application and EDTAC on the adhesion of epoxy resin-based endodontic sealers to human dentin was evaluated in vitro. A total of 99 extracted human maxillary molars with their crowns worn flat were used. The teeth were divided into 3 groups: group 1, the dentin surface received no treatment; group 2, EDTAC was applied to the dentin surface for 5 min; group 3, the dentin surface received Er:YAG laser application (2.25 W potency; 11 mm focal distance; 4 Hz frequency; 200 mJ energy; 62 J total energy; 313 mean impulse). Three teeth from each group were analyzed by scanning electron microscopy for changes in dentin surface. The epoxy resin root canal sealers used were: AH Plus, Topseal, Sealer 26, AH 26, and Sealer Plus. The zinc oxide eugenol-based sealer Fillcanal was used as control. Adhesion was measured with a Universal testing machine. The results showed a statistically significant difference at the level of 1% among the dentin treatments. The dentin treated with Er:YAG laser showed greater adhesion with the sealers than dentin treated with EDTAC which was greater than dentin that received no treatment. The Tukey test showed the formation of 5 groups in decreasing order of adhesion: AH Plus, Topseal and Sealer 26, AH 26, Sealer Plus, and Fillcanal (Grossman cement).

Adhesives↗

Effects of Nd: YAG laser, air-abrasion and acid-etching on human enamel and dentin.

The effects of the Nd:YAG laser, air-abrasion and acid-etching systems on mineral content and surface morphology of cut dentin and enamel were examined in 10 extracted human teeth. Enamel specimens were lased for two seconds at a fluence of 0.75 J and a frequency of 15 Hz, air-abraded for two seconds with 50 micron Al-oxide and etched for 60 seconds with 37% ortho-phosphoric acid. Dentinal specimens were subjected to the same procedure for half the time. Untreated areas of the same specimens served as the control. Morphologically, the lased dentin showed an apparently melted surface with partial obstruction of the dentin tubules, as well as cracks along the lased surface. Air-abrasion created very irregular surfaces on enamel and dentin. Dentin tubules were observed on the acid-etched dentin samples but not the air-abraded surfaces. The Nd:YAG laser created the most surface irregularity on both enamel and dentin. Laser treatment appeared to alter the chemical structure and surface morphology of the dentin and enamel.

Acid Etching, Dental↗

Evaluation of active and arrested carious dentin using a pH-imaging microscope and an X-ray analytical microscope.

A pH-imaging microscope was tested on carious teeth to examine the acid-base characteristics of active and arrested carious dentinal lesions and compared with an assessment of mineral loss in carious affected lesions using an x-ray analytical microscope. Extracted human molars, 22 active and 83 arrested carious teeth, were sliced vertically to maximize the presence of visible carious lesions. The pH change on the dentin surface was measured with a pH-imaging microscope, and the mineral loss was measured with an x-ray analytical microscope. The pH value of intact dentin and active and arrested carious dentinal lesions showed 7.0 (n=105), 5.8 (n=22) and 6.6 (n=83), respectively (analyzed by Kruskal-Wallis). The mineral loss was distinguishable in active dentinal lesions (4.92 times as much as intact dentin), while arrested lesions showed a slight mineral loss (2.19 times as much as intact dentin). The changes in pH value and mineral loss were significantly correlated (Pearson's correlation coefficients r=-0.8024) in active carious dentin lesions and weakly correlated (r=-0.1480) in arrested carious dentin lesions. Changes in tooth substance with the progression of the active carious process were reflected in a reduction in pH value and an increase in the amount of mineral loss.

Adolescent↗

Clinical, histological and microbiological study of hand-excavated carious dentine in extracted permanent teeth.

Changes in cultivable flora in dentine samples collected before and after hand excavation were examined in association with clinical status of the cavity surface, light microscopy and scanning electron microscopy (SEM). Thirty-five extracted permanent molar teeth with an occlusal caries lesion were excavated with hand instruments according to the atraumatic restorative treatment (ART) approach. Excavation pressure, dentine colour and consistency were recorded at the dentine-enamel junction (DEJ) prior to carious dentine removal and at the cavity floor after the final excavation; a microbiological sample of dentine was taken at both stages. Twelve restored teeth; six with positive and six with negative bacterial growth on the second sample, were selected for light microscopy and SEM. The hand-excavation removed tooth structure was soft, irreversibly damaged, dark and highly infected. Hand excavation reached dentine of increased hardness with a more normal colour to provide a sound structural base for restoration. Light and SEM examination of the cavity floor showed infected dentinal tubules in all 12 teeth examined. Linear logistic analysis showed a statistical association between light-yellow dentine on the cavity floor and an absence of bacterial growth (P = 0.006). This short-term in vitro study showed that caries-producing bacteria remained in dentine close to the cavity floor in 26/35 teeth despite clinical observations that indicated a suitably prepared cavity floor.

Bacteria↗

Effect of Nd:YAG laser irradiation on shear bond strength of glass-lonomer luting cement to dentin surface.

PURPOSE: The purposes of this study were to evaluate the shear bond strength of luting glass-ionomer cement to a dentin surface treated by pulsed Nd:YAG laser irradiation, and to prove the hypothesis that the bond strength of glass-ionomer luting cement to dentin is favorably altered after Nd:YAG laser irradiation. MATERIALS AND METHODS: Sixty-four extracted human molars with an exposed flat dentin surface were divided into four groups (n = 16). After painting black ink on their surfaces, the teeth of groups 1, 2, and 3 were irradiated by an Nd:YAG laser at 1.064-microm wavelength at 1, 2, and 3 W, respectively; group 4 was untreated and served as a control. Some specimens of each group were used for morphologic and atomic analytic study, and the others were used for shear bond testing. The shear bond test was performed after cylindric titanium specimens were cemented with glass-ionomer cement on a circular dentin area and specimens were immersed in distilled water. The tested specimens were also morphologically investigated. RESULTS: Shear bond strength of group 3 was significantly higher than that of the control group. Morphologic observation of the cement-dentin interface showed good adaptation of the luting cement to laser-treated dentin. Analysis of atomic contents on the dentin surface showed a significant increase of calcium:phosphorus ratio after laser irradiation. CONCLUSION: The dentin surface was modified morphologically and chemically, and the shear bond strength of glass-ionomer luting cement to dentin was increased by Nd:YAG laser irradiation at 3 W.

Aluminum Silicates↗

In vitro effect of cavity disinfectants on the bond strength of dentin bonding systems.

OBJECTIVE: To evaluate the effect of two cavity disinfectants, a 2% chlorhexidine and a 1% benzalkonium chloride solution, on the shear and tensile bond strengths of dentin bonding systems to dentin. METHOD AND MATERIALS: Superficial dentin was exposed from 84 freshly extracted human third molars. The teeth were randomly assigned to two main groups according to the bonding agent used, either One Step or Optibond Solo. Each dentin bonding system had six test groups (three for shear, three for tensile testing), and each test group had a control and two cavity disinfectant groups. In the control groups, dentin bonding systems were applied after etching the dentin, whereas in the cavity disinfectant groups, dentin was conditioned and treated for 20 seconds with the disinfectants before applying the dentin bonding systems. A hybrid resin composite then was applied to all treated samples. After storage in distilled water at 37 degrees C for 24 hours, shear and tensile tests were performed. Data were analyzed using Kruskall-Wallis and Mann-Whitney U tests. RESULTS: The mean shear and tensile bond strengths of One Step and Optibond Solo were not significantly different from each other, and the cavity disinfectants also had no significant effects on shear and tensile bond strength values versus the controls. CONCLUSION: The results indicate that the use of 2% chlorhexidine and 1% benzalkonium chloride solutions as cavity disinfectants after etching the dentin did not affect the shear and tensile bond strengths of One Step and Optibond Solo.

Anti-Infective Agents, Local↗

The relationship between the color of carious dentin stained with a caries detector dye and bacterial infection.

This in vitro study aimed to design a method for the objective evaluation of carious dentin using numerical values. This study also investigated the relationship between the color of carious dentin stained with a caries detector dye using this objective method and the rate of bacterial detection as detected by a polymerase chain reaction (PCR). In 15 molars with occlusal dentin caries and three extracted sound molars, dentin was removed in multiple steps with 300 microm removed each step. Before and after every removal, images of a color-matching sticker and carious surfaces stained with a caries detector dye were acquired simultaneously using a CCD camera and dentinal tissue samples were removed with a round bur. Next, corrected L*, a* and b* values of the carious surfaces (CIE 1976 L*a*b* color system) were calculated from the color changes of the stickers in the images. In addition, bacterial DNA in the dentinal tissue was detected by PCR. From evaluations of the receiver operating characteristic curves for the L*, a* and b* values, the L* value was determined to be a more useful parameter than a* or b* for detecting bacterial infection using the caries detector dye. The bacterial detection rates of carious dentin decreased as the L* values of carious dentin stained with the dye increased. When the L* values were more than 60, the dentin had no bacterial infection. This study clarified the relationship between the colors of lesions stained with a caries detector dye and the rates of bacterial detection.

Bacterial Infections↗