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A confocal microscopic evaluation of resin-dentin interface using adhesive systems with three different solvents bonded to dry and moist dentinan in vitro study.

OBJECTIVE: Total dehydration of acid-etched dentin is known to cause the collapse of collagen fiber, which leads to poor hybridization. Dentin-bonding systems with water as a solvent are found to rehydrate the collapsed collagen. Acetone-based adhesives are found to compete with moisture, and the acetone carries the resin deep into the dentin. The question arises whether to dry the dentin and use a water-based adhesive, or to keep the dentin moist and use an acetone- or alcohol-based adhesive. The aim of this study was to compare different bonding systems and techniques to assess which is most successful. A confocal microscope was used to evaluate the amount of hybrid layer formation and the depth of resin tag formation. METHOD AND MATERIALS: Superficial occlusal dentin specimens from 120 noncarious, freshly extracted human premolars were used for the study. The dentin was etched using 36% phosphoric acid for 15 seconds and then rinsed. The specimens were then randomly divided into 4 groups for different drying procedures: group I: air-dried for 30 seconds; group II: air-dried for 3 seconds; group Ill: blotted dry; group IV: overwet. The specimens were further subdivided into 3 groups to be tested with different bonding systems: subgroup A: acetone-based adhesive (Prime & Bond NT); subgroup B: water-based adhesive (Syntac Single Component); subgroup C: water- and ethanol-based adhesive (Single Bond). The resulting resin-dentin interfaces were then examined and categorized via confocal microscopy, and relative values were assigned to each specimen. RESULTS: Group IV (overwet) showed the lowest values, and the highest values were obtained in group III. The highest values were seen in group III, subgroup A (blotted dry, acetone-based bonding agent, Prime & Bond NT). CONCLUSION: Under these conditions, using these three bonding systems, maximum hybridization and resin tag formation was achieved using acetone-based adhesive on etched dentin kept moist by blot drying.

Acetone↗

[Effects of soft laser irradiation on dentinal pain].

The use of soft lasers in dental therapy has increased dramatically in recent years. However, there is a dearth of reports in the literature of controlled in vivo and in vitro studies of soft laser effectiveness with respect to dentinal pain. The purpose of this study was to assess the analgesic effects of soft laser treatment by the following five studies: (1) Clinical evaluation of soft laser irradiation of hypersensitive dentin. (2) Morphological study of dentinal surfaces following soft laser irradiation. (3) Study of the effect of soft laser irradiation on dentinal fluid movement. (4) Study of the effect of soft laser irradiation on the jaw opening reflex (JOR) in rats. (5) Study of thermal changes on the internal wall of pulp chamber with soft laser irradiation. After soft laser treatment of 3 min to hypersensitive dentin, patients experiences diminished sensitivity in 46 of 55 teeth (83.6%). In model experiment with extracted human teeth, the energy levels of this laser were too low to produce any morphological changes on the surface of exposed dentin, and dentinal fluid movements induced by air blasts for 15 sec to exposed dentin were not affected by laser irradiation to the dentin for 3 min. Moreover, there was an immediate 0.7 degree C increase in the temperature of the internal wall of the pulp chamber by laser irradiation to the tooth surface. In experimental rats, the digastric EMG induced by 1.3 x T (T: threshold value producing the JOR) was reduced by 3 min soft laser irradiation to teeth or Ho-ku points, but not at 2.0 x T. These results suggest that soft lasers have an analgesic effect, but the latter is affected by the degree of stimuli-elicited pain, and may be due in addition to descending inhibition in the central nervous system.

Animals↗

The overwet phenomenon: an optical, micromorphological study of surface moisture in the acid-conditioned, resin-dentin interface.

PURPOSE: To evaluate the morphological effect of different degrees of surface wetness on "wet bonding" upon the acid-conditioned, resin-dentin interface using an acetone-containing adhesive system (All-Bond 2). MATERIALS AND METHODS: Dentin surfaces were conditioned with 10% phosphoric acid (20 seconds), washed and air dried (3 seconds) or blot dried. RESULTS: The surfaces were characterized by the formation of an intact layer of resin impregnated dentin. Dentin tubules were sealed with solid cores of primers commonly below which were intratubular primer globules. Excess moisture on the dentin surface resulted in the creation of blister-like spaces at the primer-dentin interface. A layer of resin-impregnated intertubular dentin was present and below such spaces were incompletely sealed tubules. Primer globules were also observed on the dentin surface. The presence of microscopic voids at the primer-dentin interface may offer a morphological explanation for the published decrease in bond strength when this system was applied in the presence of excessive surface moisture.

Acid Etching, Dental↗

The influence of the depth of substrate dentin surface and thickness of the current restorative samples on bond strength.

The effect of sample thickness and dentin depth on bond strength of composite, compomer and resin modified glass ionomer have been investigated. The occlusal surfaces of 84 non carious human third molars were used for bonding. 4 subgroups were tested, superficial dentin with sample thickness 1 and 2 mm and deep dentin with sample thickness 1 and 2 mm respectively SBMP + /Z 100 composite showed 26 +/- 3.2, 22.3 +/- 4.5, 17 +/- 3.2 and 21.8 +/- 4.2 MPa shear bond strength to S. dentin 1 mm, deep dentin 1 and 2 mm and S. dentin 2 mm respectively. Compoglass reported 10.4 +/- 1.57, 9.1 +/- 2.3, 5.0 +/- 0.6 and 9.24 +/- 3.1 MPa while, Vitremer achieved 4.7 +/- 0.49, 3.2 +/- 0.39, 3.0 +/- 0.81 and 3.2 +/- 0.53 MPa. It is concluded that the highest bond strength can be achieved to superficial dentin in thickness or increments not more than 1 mm. And that both dentin depth and sample thickness might influence the quality of the bond to dentin with an effect that varies from one material to another depending upon the mechanism of bond of each material and its chemical composition.

Composite Resins↗

Bond strength to crown and root dentin.

PURPOSE: To investigate (1) the tensile bond strengths of four commercial dentin bonding systems to bovine crown and root dentin and (2) the structure of the hybrid layers for each system bonded to the two dentin substrates. MATERIALS AND METHODS: Superficial surfaces were exposed in bovine crown and root dentin. The teeth were embedded in plaster and a 3 mm diameter bonding area was demarcated. The four bonding systems used were All-Bond 2, Super-Bond D-Liner Plus, Clearfil Liner Bond II, and ProBond. Bonding procedures followed the manufacturers' instructions with the exception of Super-Bond D-Liner Plus where the primer was left in situ for 60 seconds. Tensile bond strengths were tested after 24-hour storage in 37 degrees C deionized water. Specimens were also prepared for SEM observation of the hybrid layer, after treatment with 10% phosphoric acid, and 10% phosphoric acid and 5% sodium hypochlorite. RESULTS: Statistically lower bond strengths to crown dentin when compared with root dentin were observed for All-Bond 2, whereas Liner Bond II showed the opposite (P < 0.01). Both ProBond and Super-Bond D-Liner Plus showed no statistical differences between crown and root dentin (P > 0.05). Hybrid layers could be observed for All-Bond 2, Liner Bond II and Super-Bond D-Liner Plus, with no apparent differences between the hybrid layers of crown and root dentin. In the case of ProBond, where the smear layer was not removed during the priming stage, it appeared that the primer had infiltrated and caused hybridization of the smear layer. The differences in bond strength were thought to be related to the different bonding mechanisms of each material, as well as possible variations in the crown and root dentin substrates.

Acid Etching, Dental↗

2000 Judson C. Hickey Scientific Writing Award. Effect of impression materials on hybridized dentin.

STATEMENT OF PROBLEM: Tooth sensitivity after crown preparation is not an uncommon complication. For dentin bonding agents to be effective in preventing postoperative sensitivity, they must remain intact throughout the fixed prosthodontic procedures. PURPOSE: This study evaluated, by analyzing the change in dentin permeability, whether a dentin bonding agent was removed from the surface of prepared teeth in the process of making an impression. MATERIAL AND METHODS: Eighty extracted human molars were cut horizontally apical to the cemento-enamel junction and horizontally through the middle of the clinical crown. The specimens were prepared to measure dentin permeability (fluid flow rate, Lp). The specimens (n = 40) were randomly assigned to either the primer-only group (PO) or the primer + adhesive group (PA). Five different impression materials were tested: vinyl polysiloxane, polyether, polysulfide, irreversible hydrocolloid, and reversible hydrocolloid. Dentinal permeability was measured after smear layer removal (control, etched Lp = ELp), after coating the dentin with either PO or PA (coated Lp = CLp), and again after making an impression (impression Lp = ILp). A 1-way ANOVA of differences, followed by a Student-Newman-Keuls test (alpha=.05), was performed to compare the ELp, CLp, and ILp values. RESULTS: A statistically significant difference was found between the CLp and ELp for all 5 impression materials in both groups (PO and PA). No difference was found between the CLp and ILp values for any of the impression materials. No significant differences were found between any of the 5 impression materials or either of the dentin surface treatments (PO or PA) when measuring the CLp and ILp. CONCLUSION: The dentin bonding agent had a significant effect in decreasing the Lp of the treated specimens. None of the impression materials significantly affected the measured CLp. There was no significant difference between the PO or PA (CLp) values.

Acid Etching, Dental↗

Fluid movement across the resin-dentin interface during and after bonding.

This study evaluated the extent of water penetration through resin-dentin interfaces before and after being sealed with adhesives. Four adhesive resin systems (2 total-etch adhesives and 2 self-etching primer adhesives) were used in this study. Dentin disks were placed in a split-chamber device, and in situ fluid movement across dentin was measured, with and without physiological pressure, during bonding procedures or 24 hrs after bonding. The fluid movement across dentin occurs via dentin tubules after acid-etching. Large outward or inward fluid shifts across dentin were observed during air-drying and light-curing for resin application. The amount of fluid movement across resin-bonded dentin when total-etch adhesives were used was significantly greater than that with self-etching adhesives. The milder acid-etching effects of self-etching primers may retain hybridized smear plugs within the tubules that reduce outward fluid flow, resulting in superior dentin sealing.

Acid Etching, Dental↗

Bonding durability of a self-etching primer system to normal and caries-affected dentin under hydrostatic pulpal pressure in vitro.

PURPOSE: To evaluate the bonding durability of a self-etching primer system to normal and caries-affected dentin under hydrostatic pulpal pressure. METHODS: 18 extracted human molars with occlusal caries were used. Their occlusal dentin surfaces were ground flat to expose normal and caries-affected dentin using #600 SiC paper under running water. Clearfil SE Bond was placed on the dentin surface including the caries-affected dentin according to the manufacturer's instructions and then the crowns were built up with resin composite (Clearfil AP-X) under either a pulpal pressure of 15 cm H2O or none (control). The bonded specimens were stored in 100% humidity for 1 day (control) or for 1 week and 1 month with hydrostatic pulpal pressure. After storage, the specimens were serially sectioned into 0.7 mm-thick slabs and trimmed to an hour-glass shape with a 1 mm2 cross-section, isolated by normal or caries-affected dentin, and then subjected to the micro-tensile bond test. Data were analyzed by two-way ANOVA and Tukey's test (P< 0.05). RESULTS: Hydrostatic pulpal pressure significantly reduced the bond strength to normal dentin after 1-month storage (P< 0.05), but did not affect the bond strength to caries-affected dentin.

Analysis of Variance↗

Effects of N-methacryloyl-omega-amino acid primer pretreatment on the bond strength of the resin to acid-etched dentin.

To ascertain the adhesion mechanism of resins to etched dentin treated with hydrophilic primers such as N-methacryloyl-omega-amino acids (NM omega A), the effect of the application of NM omega A primers on the bond strength of the resin and also the characteristics of the "hybrid layer" were investigated. Here, the concept of "hybrid layer" has been proposed previously by Nakabayashi et al. When the demineralized dentin was treated with the NM omega A solution, the bond strength increased remarkably, thus indicating the formation of a hybrid layer. It can be construed that NM omega A primers allowed for diffusion of the bonding agent to the dentinal collageous layer that was exposed by acid etching, and thereby the creation of a hybrid layer. To obtain an understanding of how NM omega A primers improved bond strength at the interface between the resin and dentinal collagen, the 13C-NMR spectra of NM alpha A were observed in the absence and presence of demineralized dentin. The 13C peak intensities of all of the carbons of the NM alpha A species were dramatically reduced in the presence of the dentin. Specifically, the reduction of the carbon peak intensity of carboxylic acid in the NM alpha A species was reduced by 30%. This indicated that the unionized carboxylic acid in the NM alpha A primer interacted with the dentinal collagen. Thus, the composite resin can be considered to adhere to the dentinal collagen through the unionized NM alpha A that interacts with the dentinal collagen.

Amino Acids↗

Effect of acid etching and collagen removal on dentin wettability and roughness.

It was recently reported that removal of the collagen network from etched dentin does not affect dentin bond strengths. The aim of this study was to determine if the removal of the collagen fibers results in changes in dentin roughness and wettability. Twenty cary-free extracted human third molars were sectioned parallel to the occlusal surface to expose either superficial or deep dentin. Dentin was ground flat through 600-grit SiC abrasive paper under water to provide uniform surfaces. Observed contact angle measurements were performed to assess wettability by using the axisymmetric drop shape analysis technique using water and a water-based primer. Average roughness was determined with a profilometer. The specimens were analyzed just after being ground, after etching with 35% H(3)PO(4) gel for 15 s, and after etching and deproteinization with 5% NaOCl for 2 min. Data were analyzed with two-way ANOVA and Newman-Keuls multiple comparison t test procedure. Etching resulted in an increase in surface roughness and dentin wettability. For deep dentin, collagen removal did not influence the average roughness, but it resulted in a significantly greater degree of wettability. The degree of wettability for deep dentin was greater than for superficial dentin.

Acid Etching, Dental↗

Dentin resorption mediated by odontoclasts in physiological root resorption of human deciduous teeth.

Extracted human deciduous teeth undergoing physiological root resorption were fixed with a mixture of formaldehyde and glutaraldehyde and processed for scanning (SEM) and analytical transmission electron microscopy, as well as acid trimetaphosphatase cytochemistry. The granulated tissues, which are rich in multinucleated odontoclasts and capillary vessels, formed various resorption lacunae on the resorbing dentin surfaces. SEM observations of dentin surfaces treated with sodium hypochlorite revealed two types of resorption lacunae: deep, round lacunae in which the peritubular matrix of dentinal tubules was strongly dissolved; and shallow, irregular lacunae with intact peritubular matrix. In trypsin-treated materials, the resorption surfaces were characterized by the presence of numerous collagen fibers in both the peritubular and intertubular matrices, suggesting demineralization of the surface dentin. Odontoclasts were characterized by the presence of abundant mitochondria, perinuclear stacks of Golgi membranes, various lysosomes, numerous endocytotic vacuoles, and a well-developed ruffled border against the resorption lacunae. Most endocytotic vacuoles were distributed in the cytoplasm between the ruffled border and the nuclei. In undemineralized ultrathin sections, the surface dentin of resorption lacunae consisted of collagen fibers and apatite crystals and had a lower packing density than those in unresorbed, deeper dentin. Many apatite crystals were demonstrated to be present in the extracellular channels of the ruffled border and in adjacent endocytotic vacuoles derived from it. Lysosomes located in the perinuclear cytoplasm of odontoclasts contained amorphous dense material and/or a small amount of crystals. An energy-dispersive x-ray microanalysis of apatite crystals in undemineralized sections indicated that the energy spectrum peaks of Ca and P detected from crystals in resorbing dentin were much lower than those in unresorbed dentin. Similarly, lower spectrum peaks of Ca and P were obtained from crystals found in the ruffled border and endocytotic vacuoles of odontoclasts. A slight trace Ca peak also was detected in the amorphous dense material in lysosomes of odontoclasts. The enzyme cytochemistry of lysosomal acid trimetaphosphatase indicated that odontoclasts had intense enzymatic activity in the Golgi membranes, endoplasmic reticulum cisternae, lysosomes, and endocytotic vacuoles. Dense reaction precipitates of enzymatic activity also were found along the dentin surfaces of resorption lacunae occupied by odontoclast ruffled borders.(ABSTRACT TRUNCATED AT 400 WORDS)

Acid Anhydride Hydrolases↗

Improved wet bonding of methyl methacrylate-tri-n-butylborane resin to dentin etched with ten percent phosphoric acid in the presence of ferric ions.

The objective of this study was to determine the influence of dissolved dentinal substances in demineralized dentin on the hybridization of resin for bonding to dentin. It was hypothesized that these substances, including polyelectrolytes, significantly change the substrates, which could then be assessed by the addition of Na(+), Ca(2+), or Fe(3+) in 10% phosphoric acid. Bovine dentin specimens were etched for 10 s with a solution of 10% phosphoric acid (control) or of 22.0 mM dissolved sodium chloride (10P-Na), calcium chloride (10P-Ca), or ferric chloride (10P-Fe). The specimens were then rinsed, blot-dried, and primed three times with 5% 4-methacryloyloxyethyl trimellitate anhydride in acetone for 60 s. Methyl methacrylate-tri-n-butylborane resin was then applied. The tensile bond strength of each of the dumbbell-shaped specimens was then measured. The fractured surfaces and modified cross-sections were examined by scanning electron microscopy. The cross-sections were soaked in 6N HCl for 10 s and then in 1% sodium hypochlorite for 30 min to determine the resin content in the hybridized specimens. Shrinkage of the demineralized dentins upon drying was assessed by atomic force microscopy. The tensile bond strengths were 10.8 +/- 4.5 (control), 15.0 +/- 7.0 (10P-Na), 19.3 +/- 5.5 (10P-Ca), and 27.8 +/- 8.1 (10P-Fe) MPa. The atomic force microscopy studies showed that Fe(3+) minimized the shrinkage by drying for 10 s but Ca(2+) and Na(+) did not decrease the shrinkage the same as the control. The results support the hypothesis that the monomer permeability of wet demineralized dentin is effectively improved by dissolving ferric ions in the phosphoric acid, resulting in a greater bond strength and higher resin content in the hybridized dentin. The dissolved dentinal substances, including the polyelectrolytes, had a significant influence on the characteristics of the demineralized dentin, changing the degree of hybridization and bonding.

Animals↗

Local properties of a functionally graded interphase between cementum and dentin.

The study of natural interfaces may provide information necessary to engineer functionally graded biomaterials for bioengineering applications. In this study, the mechanical, structural, and chemical composition variations associated with a region between cementum and dentin were studied with the use of nanoindentation, microindentation, optical microscopy, and Raman microspectroscopy techniques. Three-millimeter-thick transverse sections (N = 5) were obtained from the apical one-third of the roots of sterilized human molars. The samples were ultrasectioned at room temperature with the use of a diamond knife and an ultramicrotome. Longitudinal ground sections of 100 microm thickness were prepared and stained with von Kossa stain to determine the mineralized regions within the molar roots. Raman microspectroscopy was used to determine the relative inorganic content, mainly apatite (PO4(3-)nu1 mode at 960 cm(-1)) and organic content, mainly collagen (C--H stretch at 2940 cm(-1)) between cementum and dentin bulk tissues. The microindentation and nanoindentation results indicated a gradual transition in hardness from cementum to dentin over a width ranging from 100 to 200 microm. However, the variation in hardness data for cementum and dentin by nanoindentation was larger (0.62 +/- 0.21, 0.77 +/- 0.14 GPa) than from microindentation (0.49 +/- 0.03, 0.69 +/- 0.07 GPa). Within the 100 to 200 microm region there was a 10 to 50 microm fibrillar hydrophilic cementum-dentin junction (CDJ) with mechanical properties significantly lower than either the cementum or the dentin side of CDJ. Light microscopy revealed a 100 to 200 microm translucent region between cementum and dentin. Raman microspectroscopy results showed a variation in organic and inorganic composition 80 to 140 microm wide. It was concluded that a morphologically and biomechanically different CDJ lies within a wider cementum-dentin interphase. Hence, cementum, dentin, and the interphase can be classified as a functionally graded dental tissue within the root of a tooth.

Biomechanical Phenomena↗

Bactericidal effects of diode laser on Streptococcus mutans after irradiation through different thickness of dentin.

BACKGROUND AND OBJECTIVES: A reliable method to eradicate the bacteria of residual carious dentin has not yet been developed. The aim of this study was to evaluate the antibacterial effect of a diode laser on Streptococcus mutans through different thickness (500, 1,000, and 2,000 microm) of human dentin. The thermal effect of laser irradiation was also investigated. STUDY DESIGN/MATERIALS AND METHODS: Dentin specimens were inoculated with 2 microl of S. mutans on one side and irradiated by a diode laser on the other side with a power output ranging from 0.5 to 7 W. The laser tip was swept with the whole irradiation area of 7 mm x 3 mm at a speed of about 10 mm/second with a total irradiation time of 30 seconds. Cooling with distilled water (30 ml/minute) was applied simultaneously during laser irradiation. After laser irradiation, the bacteria was removed from the dentin surfaces and cultured for 48 hours at 37 degrees C anaerobically to assess the colony forming units (CFU) per ml. The morphology of the lased bacteria and the temperature rise during laser irradiation were observed by scanning electron microscope (SEM) and measured by thermocouple, respectively. RESULTS: The results revealed that 7 W of laser power could kill 97.7% of CFU through 500 microm thickness of dentin. However, the bactericidal efficiency was significantly reduced as the dentin thickness was increased. The morphological changes of lased bacteria ranged from less affected such as loss of their wall bands and existence of minicells to more severely degenerated, such as disintegration and fusion of cells with pores on the cell wall. Only the dentin specimens with a thickness of 500 microm exhibited a temperature rise greater than 5.5 degrees C after receiving 5 or 7 W of laser irradiation. CONCLUSIONS: A diode laser can eliminate the Streptococcus mutans of the residual carious dentin without inducing high pulpal temperature rise when the remaining dentin thickness is greater than 1 mm.

Dental Caries↗

1-Hydroxyethylidene-1,1-bisphosphonate (HEBP) simultaneously induces two distinct types of hypomineralization in the rat incisor dentine.

Effects of 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) on the dentinogenesis of the rat maxillary incisor were microradiographically examined 96 hours after a single subcutaneous injection. HEBP produced hypomineralized incremental bands in the enamel (HPOe) and in the dentine (outer HPO1 and inner HPO2) in a dose-dependent manner. In the labial dentine, both HPO1 and HPO2 did not extend apically beyond the apical limit of the HPOe and were thus located in the circumpulpal dentine. The two lesions in the labial dentine fully developed with a dosage of 10 mg/ P/kg or larger, and with 15 mg P/kg or larger, gross hypoplastic lesions developed. Therefore, histological analysis at shorter postinjection intervals was carried out on the labial dentine with 10 mg P/kg of HEBP. The HPO1 appeared along the dentine-predentine junction by 24 hours and was characterized by reduced numbers of inorganic crystals and 10 nm particles which invested the collagen fibrils. Therefore, the HPO1 was judged to be produced by the disturbance of transformation of the predentine to dentine. The precursor lesion of HPO2 appeared in the proximal predentine by 4 hours, and was characterized by reduced number of collagen fibrils and unusually coarse interfibrillar stippled material. This lesion migrated to the dentine by 96 hours and became the HPO2, which was characterized by reduced number of collagen fibrils. Individual collagen fibrils in the HPO2 were invested by as many inorganic crystals and 10 nm particles as in the normal dentine. The HPO2 is considered to be produced by the disturbance of organic matrix formation.

Animals↗

Ameloblastin fusion protein enhances pulpal healing and dentin formation in porcine teeth.

Ameloblastin (Ambn, also named "amelin" or "sheathlin") is a protein participating in enamel formation and mesenchymal-ectodermal interaction during early dentin formation in developing teeth. Experiments have demonstrated an association between Ambn expression and healing of acute pulp wounds. The purpose of this study was to investigate if local application of recombinant fusion Ambn (rAmbn) could influence reparative dentin formation in pulpotomized teeth. In this randomized, double-blinded study, pulpotomy was performed in 28 lower central incisors in 17 adult miniature pigs. Following the surgical procedure, the exposed pulp tissue was covered either with rAmbn or with calcium hydroxide. After 2, 4, or 8 weeks, the teeth were extracted and examined by histomorphometry and immunohistochemistry using antibodies against porcine ameloblastin, collagen type I, and dentin sialoprotein (DSP). In rAmbn-treated teeth, a substantial amount of newly formed reparative dentin was observed at the application site, completely bridging the pulpal wound. Dentin formation was also observed in calcium hydroxide-treated teeth; however, the amount of reparative dentin was significantly smaller (P < 0.001) than after rAmbn treatment. Immunohistochemistry confirmed that the new hard tissue formed was similar to dentin. This is the first time a direct link between ameloblastin and dentin formation has been made in vivo. The results suggest potential for rAmbn as a biologically active pulp-dressing agent for enhanced pulpal wound healing and reparative dentin formation after pulpotomy procedures.

Animals↗

Odontoblast processes in human dentin revealed by fluorescence labeling and transmission electron microscopy.

In the present undertaking, the distribution of odontoblast processes in human dentin was determined through the DiI carbocyanine dye fluorescent staining of the cell membrane, while F-actin was identified by rhodamine-phalloidin. Confocal laser scanning microscopy revealed intense labeling for both agents in inner dentin, while transmission electron microscopy (TEM) identified dentinal tubules including odontoblast processes in this area, each process being surrounded by a cell membrane and containing an abundance of filamentous structures. Electron-dense "lamina limitans" lined the dentinal tubules. Individual cell processes became narrower toward the middle area, and their overall numbers decreased as well under TEM. Labeling for F-actin was absent in both middle and outer dentin, while faint labeling for DiI was visible along the dentinal tubules as far as the dentino-enamel junction (DEJ), where it was also recognized within the tubules themselves. Under TEM, the dentinal tubules lined with electron-dense structures were, in fact, empty in the middle and outer dentin. Immediately below the DEJ, however, the tubules manifested dense concentrations of fine granular material. Our study, therefore, appears to suggest that odontoblast processes do not extend beyond the inner dentin of fully erupted human premolars.

Actins↗

High-resolution electron-microscopic study of the relationship between human enamel and dentin crystals at the dentinoenamel junction.

The development of dentin and of enamel share a common starting locus: the dentinoenamel junction (DEJ). In this study the relationship between enamel and dentin crystals has been investigated in order to highlight the guiding or modulating role of the previously mineralized dentin layer during enamel formation. Observations were made with a high-resolution electron microscope and, after digitalization, image-analysis software was used to obtain digital diffractograms of individual crystals. In general no direct epitaxial growth of enamel crystals onto dentin crystals could be demonstrated. The absence of direct contact between the two kinds of crystals and the presence of amorphous areas within enamel particles at the junction with dentin crystals were always noted. Only in a few cases was the relationship between enamel and dentin crystals observed, which suggested a preorganization of the enamel matrix influenced by the dentin surface structure. This could be explained either by the existence of a proteinaceous continuum between enamel and dentin or by the orientation of enamel proteins by dentin crystals.

Crystallization↗