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PubMedDate not supplied

The effect of antimicrobial pre-treatments on the performance of resin composite restorations.

This study evaluated the effects of four cavity pre-treatments on the microleakage of a fifth generation adhesive system and accompanying resin composite. Sixty human molars were randomly selected for cavity pre-treatment for the following material groups: Ultra-Etch 35% phosphoric acid gel, Ultra-Etch AB 35% phosphoric acid gel with cetylpyridinium chloride antimicrobial agent, Ultra-Etch 35% phosphoric acid gel followed by Concepsis chlorhexidine gluconate antimicrobial solution, Ultra-Etch 35% phosphoric acid gel followed by Consepsis Scrub chlorhexidine gluconate antimicrobial slurry and no etchant and/or cavity pre-treatment. Circular preparations were cut on the facial or lingual surface of each tooth and treated with the appropriate etchant/cavity disinfectant. PQ1 self-priming adhesive was placed in the preparations followed by Vitalescence resin composite. The molars were thermocycled, sealed with nail varnish, placed in 5% methylene blue dye and sectioned. Leakage was assessed by dye penetration at the occlusal and gingival surface positions of the specimen blocks using a binocular microscope. The Consepsis Scrub and no pre-treatment groups revealed significantly greater leakage than the other groups treated with antimicrobial disinfectants or etchants. No significant differ. ences were shown in the two etchant groups that were compared, Ultra-Etch and Ultra-Etch AB. Significantly greater leakage was not exhibited by the same materials at the occlusal vs gingival surface positions.

Acid Etching, Dental↗
PubMed1990

[Penetration of calcium hydroxide based cements by dental enamel conditioning acids].

It was verified the penetration of phosphoric acid into 3 commercial calcium hydroxide-based cements (Life, Renew and Prisma VLC Dycal). The colorimetric method employed permitted the identidication of phosphorus amount in representative samples of 6 successive layers 0.1 mm thick of each material. The acid etching used were the commercial products Scotchbond Etching Gel--3M at 36.114% by weight and Solução Condicionadora--Johnson & Johnson at 36.054% by weight. The contact time was 60 seconds. The result showed that layers 0.1 mm tick for Life and Prisma VLC Dycal and 0.2 mm thick for Renew were able to block the penetration of phosphoric acid solution whereas layers 0.1 mm thick for the 3 cements were able to block the penetration of phosphoric acid gel.

Acid Etching, Dental↗
PubMed2004

An in vitro investigation of penetration depth of dentine bonding agents into carious dentine.

The aim of the present in vitro study was to determine the penetration depth of dentine bonding agents into carious dentine as a possible strategy in caries treatment of deep lesions. Forty-eight extracted carious teeth were used for the experiments and divided in four groups. In a split-tooth design, one half of each tooth was treated without caries excavation, and the second half after caries excavation. In group 1 the teeth were treated with Prime & Bond 2.0 without phosphoric acid etching, and in group 2 with phosphoric acid etching. In group 3, the teeth were treated with All-Bond 2 without phosphoric acid etching, and in group 4 with phosphoric acid etching. Bonding agents were marked with an orange dye prior to application. The composition of the bonding agents had no significant influence on penetration depths. The mean thickness of the outer, necrotic caries zone in unexcavated specimens was 117 microm (+/-126). Bonding agents penetrated significantly deeper into unexcavated carious dentine (mean 124 +/- 76 microm) compared with excavated dentine (11 +/- 14 microm) and sound dentine (3 +/- 4 microm). Acid etching significantly reduced the thickness of the outer caries zone, and the penetration depth of the bonding agents into unexcavated carious specimens by approximately 50%, and increased it into sound dentine. The results indicate that dentine bonding agents could completely penetrate the unexcavated outer carious dentine in 90% of specimens. Further studies are required to determine whether further caries activity is arrested after treatment with dentine bonding procedures.

Dental Caries↗
PubMed1981

Phosphoric acid-its use in the extraction of RNA: staining of DNA in mammalian tissue sections.

This communication presents highly satisfactory methods for the demonstration of DNA in animal materials. The method involves selective extraction of RNA with concentrated, 90% or 75% phosphoric acid of 5 degrees C for 20, 40 and 120 min, respectively, followed by staining with aqueous solutions of basic dyes, such as setoglaucine, setocyanine, pinakryptol green 2) and alcoholic aniline blue without SO2. Perfect blue nuclei were seen when staining was performed with aqueous solutions of setoglaucine and setocyanine at pHs 3.5 and 4.0 to 4.5, respectively. Sections of tissues from which RNA has been extracted and then hydrolysed in 6 N HCl at 28 degrees C for 15 min followed by staining with these dyes also revealed perfect colouration of the nuclei. Acid-hydrolysed sections when stained with alcoholic aniline blue-SW2 prepared with N HCl and sodium thiosulphate revealed nuclei of magenta colour, and sections from whcih RNA has been extracted and then hydrolysed in hydrochloric acid and stained with this dye-reagent revealed nuclei of purplish colour. Sections of tissues fixed in Carnoy, 10% buffered neutral formalin as well as paraformaldehyde were found to be most suitable for staining with these dyes. The in situ absorption spectra of nuclei stained with aqueous solutions of setoglaucine, setocyanine and alcoholic aniline blue without SO2, after extraction of RNA as well as those of nuclei in tissue sections from which RNA has been extracted and then acid-hydrolysed and stained with alcoholic aniline blue-SO2 have been presented. Also presented herein are absorption data of nuclei in tissue sections which wee hydrolysed in hydrochloric acid and then stained with alcoholic aniline blue-SO2. Some implications of these findings have been discussed.

Aniline Compounds↗
PubMedDate not supplied

A comparison of six enamel treatment procedures for sealant bonding.

This in-vitro study evaluated the effectiveness of six different enamel treatment procedures for bonding a dental sealant. Sixty extracted human molar teeth were separated into the following enamel treatment groups (10 teeth each): group 1 (control)-etched with 37% phosphoric acid; group 2-air polished (air abraded) with 45 microns particles of sodium bicarbonate; group 3-air abraded with 50 microns aluminum oxide particles; group 4-etched with 2.5% nitric acid; group 5-air abraded with sodium bicarbonate particles and etched with 37% phosphoric acid; and group 6-air abraded with 50-microns aluminum oxide particles and etched with 37% phosphoric acid. The enamel treatment procedures were accomplished on intact mesial or distal surfaces. Following the enamel treatment, a sealant was bonded to the surfaces using a plastic matrix technique. After 24 hr of water storage at 37 degrees C, the specimens were debonded using an Instron machine. The mean shear bond strengths (MPa) were as follows: group 1-9.19 +/- 1.34 MPa; group 2-2.03 +/- 1.67 MPa; group 3-1.50 +/- 0.93 MPa; group 4-4.99 +/- 1.26 MPa; group 5-11.61 +/- 4.51; and group 6-11.14 +/- 1.70 MPa. Statistical analysis using a one-way ANOVA and Scheffe F-test revealed no significant difference (P > 0.05) among groups 1,5, and 6. However, there was a significant difference (P < 0.05) between groups 1,5, and 6 and the other three groups (2,3, and 4). In conclusion, 37% phosphoric acid treatment of intact enamel, or a combination of air abrasion with sodium bicarbonate or aluminum oxide followed by phosphoric acid, provides significantly higher bond strengths of a sealant material than enamel conditioning with 2.5% nitric acid or air abrasion with sodium bicarbonate or aluminum oxide.

Acid Etching, Dental↗
PubMed2007

Optimization of phosphopeptide elution conditions in immobilized Fe(III) affinity chromatography.

While immobilized metal affinity chromatography (IMAC) has been widely used for affinity purification of phosphopeptides, the technique suffers from insufficient specificity. Therefore, there is an urgent need for IMAC optimization to yield the selectivity and sensitivity that is required for more challenging analyses. Recently, 2,5-dihydroxybenzoic acid (DHB) and phosphoric acid mixture has been reported as an efficient IMAC eluant. The disadvantage of DHB is that is not suitable for electrospray ionization-mass spectrometry. While further developing the IMAC elution protocol to overcome this problem, we noticed that DHB is not necessary and found a novel combination of phosphoric acid and acetonitrile to be more efficient. The purification efficacy of the novel protocol is superior to all previously described methods, while still being compatible with the most commonly used mass-spectrometric techniques in phosphoproteomics.

Chromatography, Affinity↗
PubMed2002

Gas chromatography-mass spectrometry assay of the (18)o enrichment of water as trimethyl phosphate.

We have developed an assay for determining the 18O enrichment of water in biological fluids. Urine, plasma, or whole blood is reacted with phosphorous pentachloride to yield phosphoric acid. Derivatization of phosphoric acid with diazomethane generates trimethyl phosphate. The enrichment of trimethyl phosphate is nearly four times that of water and is assayed using gas chromatography-mass spectrometry (electron impact ionization). Yang et al. (1998, Anal. Biochem. 258, 315-321) assayed the 2H enrichment of body water after exchange with acetone, by gas chromatography-mass spectrometry. The combination of our 18O method and the 2H method of Yang et al. allows one to measure energy expenditure via "doubly labeled" water (2H(2)O + H(2)18O), using small samples of body fluids. These techniques were used to measure energy expenditure in mice, in which the 18O enrichment of body water can be monitored down to 0.025%.

Animals↗
PubMed1989

[Application of succinoxy methacrylates for direct bonding system].

Three types of succinoxy methacrylates [2-succinoxyethyl methacrylate (2SEM), 2-succinoxypropyl methacrylate (2PrM) and 2-succinoxy-3-phenoxypropyl methacrylate (SPPM)] were synthesized in order to develop a new direct-bonding adhesive which has sufficient bond strength to mild etched enamel and stainless steel brackets for occlusal and orthodontic force. Experimental direct-bonding adhesives consisted of a MMA solution containing synthesized monomers (5-10 mol%), PMMA powder and an oxidized tri-n-butyl borane as an initiator. A MMA solution without synthesized monomers was used as the control. Preparation of 10% and 40% phosphoric acid solutions and 1M maleic acid solution was carried out for use as etchants. The bond strength to unetched and etched enamel, polished stainless steel, simulated brackets and brackets was measured. The enamel surfaces etched with each etchant and the fractured resin and enamel surfaces after the bond test were observed with SEM and the ease of debonding of adhesive was investigated to totally evaluate the experimental direct-bonding adhesives. The results were as follows: 1) The amount of decalcification for 10% phosphoric acid and 1M meleic acid solutions was less than that for 40% phosphoric acid solution. The SEM micrograph showed most clearly the appearance of enamel prisms on the enamel surface treated with 40% phosphoric acid solution. 2) The experimental direct-bonding adhesives had higher bond strength to the polished enamel, the polished stainless steel and the simulated bracket than the control. 3) The bond strength to the etched enamel was comparable to that of the control. 4) The treatment of 1M maleic acid solution provided the same bond strength to enamel as that of 40% phosphoric acid solution and the higher bond strength than of 10% phosphoric acid solution. The bond strength to enamel treated with 1M maleic acid solution and 40% phosphoric acid solution decreased very slightly by immersion in water and thermal cycling. 5) The MMA solutions containing each 20 mol% synthesized monomer were adequate to direct-bonding adhesive monomers on the basis of the results of bond strength and manipulation. 6) The strongest bond strength between bracket and etched enamel was obtained with the direct-bonding adhesive containing 2SEM under all conditions. 7) The SEM micrographs showed that the tooth-resin interface failure for each adhesive occurred during bond testing. 8) There was little remaining of the resin on the tooth surface after debonding regardless of types of direct bonding adhesives.

Dental Bonding↗
PubMed2002

Interfacial micromorphological differences in hybrid layer formation between water- and solvent-based dentin bonding systems.

STATEMENT OF PROBLEM: Many dentin bonding systems of different compositions, and in particular containing different solvents, have been introduced to the market. Their effect on the quality of the interface requires clarification by means of comparative trials. PURPOSE: This study investigated micromorphological differences in hybrid layer formation with a variety of commercially available water- or solvent-based dentin bonding products and their recommended compomers. MATERIAL AND METHODS: Five bonding systems were used on groups of 10 teeth each as follows: group I, acetone-based system used with 36% phosphoric acid; group II, a different acetone-based system containing nano-sized particles for filler loading and used with a non-rinsing conditioner containing maleic acid; group III, the acetone-based system of group II used with 36% phosphoric acid (the only difference in the treatment for groups II and III was the acid etching system); group IV, a mixed-solvent-based system (water/ethanol) used with 37% phosphoric acid; and group V, a water-based system used with 37% phosphoric acid. Each bonding system was covered with the recommended compomer. Class I occlusal preparations were made in extracted teeth and restored with one of the above systems. Five specimens of each group were studied with optical microscopy after staining. Scanning electron microscopy was used to examine the interface of the bonding system/dentin of the other 5 teeth in each group. The optical microscopy measurements were made with a 10 x 10 reticle. A micron mark with scale was used for the scanning electron microscope. All measurements were made in microm. The following criteria were used to define a good interface: absence of voids between the different parts of the interface, uniformity of the hybrid layer, good opening of the tubuli orifices, and tag adherence to the tubuli walls. RESULTS: Morphological differences were found at the interface depending on dentin treatment and adhesive composition. The acetone-containing systems were associated with a continuous, gap-free hybrid layer that was linked intimately with the dentin. The tags adhered well to the tubuli walls and were often joined by side branches. In the water-based solvent systems, a lack of contact was visible between the resin tags and the tubuli walls, with some incompletely filled tubuli and some gaps in the hybrid layer. The 2 observational methods used, optical and scanning electron microscopy, proved to be complementary. CONCLUSION: Within the limitations of this study, use of the acetone-based systems after phosphoric acid etching resulted in a continuous, thick hybrid layer with reverse-cone-shaped tags in close contact with the tubuli walls. Use of the water-based systems resulted in a thinner hybrid layer with some incompletely sealed dentinal tubules.

Acetone↗
PubMed1992

Soft drink consumption and urinary stone recurrence: a randomized prevention trial.

The object of this study was to determine if a strong association between soft drink (soda) consumption and recurrence of urinary stone disease, found in an earlier case-control study of adult males, had a causal component. The study sample consisted of 1009 male subjects, who completed an episode of urinary stone disease, who were aged 18-75 at that time, and who reported consuming at least 160 ml per day of soft drinks. Half of the subjects were randomized to refrain from consuming soft drinks, while the remaining subjects served as controls. The intervention group had an observed 6.4% advantage in actuarial 3 yr freedom from recurrence (p = 0.023 one-sided) over the control group. One important secondary finding was that for those who reported at the time of the index stone that their most consumed drink was acidified by phosphoric acid but not citric acid, the experimental group had a 15% higher 3 yr recurrence-free rate than the controls, p = 0.002, while for those who reported at the time of the index stone that their most consumed drink was acidified by citric acid with or without phosphoric acid, the experimental group had a similar 3 yr recurrence-free rate to the controls, p = 0.55. This interaction was significant, p = 0.019.

Actuarial Analysis↗
PubMed2004

Four-year water degradation of a resin-modified glass-ionomer adhesive bonded to dentin.

Glass-ionomers are auto-adhesive to tooth tissue through combined micro-mechanical and chemical bonding. How much each of the two bonding components contributes to the actual bonding effectiveness is, however, not known and there is not much information available on long-term stability. The objective of this study was to assess the bonding effectiveness of a resin-modified glass-ionomer adhesive to dentin after 4 yr of water storage. Fuji Bond LC (GC) was applied without (i) and with pretreatment using (ii) a polyalkenoic acid conditioner and (iii) a 37.5% phosphoric acid etchant. The etchant was used to exclude any chemical interaction with hydroxyapatite. The micro-tensile bond strength ( microTBS) to dentin decreased significantly over the 4 yr period in all three experimental groups. After 24 h and 4 yr, the lowest micro TBS was recorded when dentin was not pretreated. The highest micro TBS was obtained following polyalkenoic acid pretreatment, although this was not significantly different from specimens that were pretreated using phosphoric acid. Pretreating dentin with phosphoric acid intensified micromechanical interlocking at the expense of chemical bonding potential to hydroxyapatite. Nevertheless, correlating the micro TBS data with failure analysis through scanning electron microscopy and transmission electron microscopy indicated that combined micro-mechanical and chemical bonding involving pretreatment with the polyalkenoic acid conditioner yielded the most durable bond.

Acid Etching, Dental↗
PubMed2001

Evaluation of the F2000 bonding procedure: a high resolution SEM study.

OBJECTIVES: The aim of this study was to define the morphological bonding of the F2000 Restorative System (3M Dental Product) to enamel and dentin. For this purpose, high resolution scanning electron microscope (SEM) in conjunction with a tensile bond strength analysis was used to correlate the morphological findings to the bonding potential. Scotchbond 1 (SB1) adhesive system was utilized as a control. MATERIALS AND METHODS: Teeth were prepared in vivo and prepared as: (1) F2000 PA for 15s, (2) F2000 PA for 30s, (3) 35% phosphoric acid for 15s and SB1 and (4) 35% phosphoric acid for 30s and SB1. All teeth were then restored with the F2000 compomer restorative material, extracted, sectioned and analyzed by means of a field emission in-lens SEM (FEISEM). The conditioning/etching patterns of the two adhesive systems were also evaluated in vitro on dentin disks in order to show high resolution details of every single step of the bonding procedure. RESULTS: FEISEM images showed a repetitive pattern of the enamel crystals with several microporosities after the 30s application of F2000 PA; moreover a good adaptation of the adhesive resin on the conditioned enamel was observable. FEISEM analysis of the dentin surface revealed no evidence of a smear layer after the application of F2000 PA for 30s. Residual smear plugs were partially obliterating the tubule orifices when F2000 PA was used, while they were completely absent after etching with 35% phosphoric acid. The total etching technique revealed open tubules and resin tags formation after the application of SB1. CONCLUSIONS: The one step etching/priming/bonding procedure of the F2000 PA showed favorable results at ultra-structural level when applied on both the enamel and the dentin surface for 30s (as per manufacturers' instructions), but the bond strength analysis showed higher values of bonding of the F2000 compomer when the SB1 adhesive agent (after etching with 35% phosphoric acid) was used (7.7+/-2.2 vs 19.0+/-4.4MPa).

Acid Etching, Dental↗
PubMed1998

Morphological effect of the type, concentration and etching time of acid solutions on enamel and dentin surfaces.

Thirty-six noncarious human molars were extracted and stored in saline. The buccal surfaces were ground wet on 400-grit silicon carbide paper to create a flat surface. The enamel and dentin surfaces were treated with 35% phosphoric acid gel, 10% phosphoric acid gel and 10% maleic acid gel for 15 and 60 seconds. Scanning electron microscope analysis showed that all the acid etchants modified the micromorphological appearance of enamel and dentin surfaces independent of the type of acid, the etching time and the concentration. On the enamel surface, the 35% and 10% phosphoric acid gels and the 10% maleic acid gel applied for 15 or 60 seconds preferentially removed the prism core material. In the specimens treated for 15 seconds with 10% maleic acid the prism core material was partially removed. On the dentin surface, the phosphoric acid gels (35% and 10%) and the 10% maleic acid gel applied for 15 and 60 seconds removed the smear layer and opened the dentinal tubule orifices; however, the dentinal surface etched for 15 or 60 seconds with 10% maleic acid gel showed residues of the smear layer.

Acid Etching, Dental↗
PubMedDate not supplied

Bond strengths of single-bottle dentin adhesives to caries-affected dentin.

There is concern that some acidic conditioners may not be strong enough to adequately etch sclerotic or caries-affected dentin. The hypothesis that was tested was that there were no significant differences in the bond strengths of single-bottle bonding systems to normal or caries-affected dentin, regardless of the strength of the phosphoric-acid conditioner. Extracted teeth with coronal caries extending into mid-dentin were prepared by grinding the occlusal surface flat. This left a central region of caries-affected dentin surrounded by normal dentin. The One-Step bonding system was used to bond dentin following etching with 10 or 32% phosphoric acid. The Single Bond system was used after etching dentin with 10 or 35% phosphoric acid. After 24 hours in water, serial vertical sections were made through the bonded teeth to create slabs 0.7 mm thick. Each tooth yielded four to five slabs, some of which included normal dentin, while others included caries-affected dentin. Each slab was trimmed into an hourglass configuration to limit the test area to normal or caries-affected dentin. The results obtained with One-Step following etching with 10% phosphoric acid showed lower (P < 0.05) tensile bond strengths to caries-affected dentin compared to normal dentin (36.9 +/- 8.0 MPa vs 47.7 +/- 6.5 MPa, respectively). This difference disappeared when using 32% phosphoric acid (49.7 +/- 6.1 MPa vs 45.0 +/- 7.2 MPa, respectively). Bonds made to caries-affected dentin with Single Bond were always lower than bonds to normal dentin regardless of the strength of the phosphoric acid. Scanning electron microscopy of polished cross sections sequentially challenged with acid and NaOCl revealed loss of the middle of the hybrid layers created by either bonding system in caries-affected dentin etched with 10% phosphoric acid. It is clear that 32-35% phosphoric acid is required to adequately etch caries-affected dentin in order to produce high bond strengths and well-infiltrated demineralized dentin.

Acid Etching, Dental↗
PubMed2006

Swine teeth as potential substitutes for in vitro studies in tooth adhesion: a SEM observation.

Animal teeth are routinely used in studies that test dental materials; however, no studies have yet proposed the use of swine teeth in laboratory tests. The purpose of this study was to compare the effects of acid etching on the dentine and enamel of human and swine teeth, aiming at the use of swine teeth as substitutes for human teeth in adhesion tests. Four human molars and four swine molars free of caries were used. For the study of enamel, two human and two swine teeth were longitudinally sectioned with a dental handpiece and a carborundum disk. For evaluation of dentine, the remaining teeth had the occlusal third of their crowns removed using a low speed diamond saw. All specimens underwent acid etching with 37% phosphoric acid for 30s. The etched surfaces were then washed with air/water spray for 10s. The specimens were then prepared for analysis in a Sterioscan 440 scanning electron microscope. It was observed a similar morphology in human and swine teeth, despite swine enamel was thinner than human enamel. Acid etching of swine enamel promoted a demineralization similar to that observed in human enamel in which the three types of Silverstone's patterns were observed. Acid etching of dentine exposed the opening of dentinal tubules in both swine and human teeth. Peritubular dentine was removed by phosphoric acid, while collagen fibrils appeared discernible in intertubular dentine. Thus, this study showed that enamel and dentine from swine and human teeth share many structural similarities after acid etching.

Acid Etching, Dental↗
PubMedDate not supplied

An in vitro shear bond strength study of enamel/dentin bonding systems on enamel.

The purpose of this study was to compare the enamel shear bond strengths achieved with four acid conditioners employed by current enamel/dentin bonding systems (maleic acid, citric acid, nitric acid, oxalic acid) with a 37% phosphoric acid etching technique. The study also compared enamel shear bond strengths between the manufacturers' enamel/dentin conditioner used with an unfilled enamel bonding resin. The facial surfaces of 135 bovine incisors were ground flat and divided into nine test groups of n=15. Conditioning and bonding procedures were carried out following manufacturers' instructions. The phosphoric acid groups were etched for 15 seconds, rinsed for 30 seconds, and dried for 20 seconds with compressed air. All bonding was accomplished at a constant temperature of 21 degrees C and a relative humidity of 60%. A single composite restorative resin (Z-100) was used with all specimens to eliminate variables between composite materials. All specimens were thermocycled 2500 times (5 to 45 degrees C) and had a total storage time of 21 days prior to shear testing on an Instron at a crosshead speed of 1.0 mm/minute. Shear strengths were calculated by dividing load at failure by specimen area. A light microscope was used to determine failure mode. The data were subjected to Bartlett's test for homogeneity of variance and were found not to be homogeneous. The Welch Test was applied and indicated that the treatments used had influence on bond strength at P<0.05. There was no significant difference in bond strength values between traditional phosphoric acid/enamel bonding resin and Mirage Bond Dentin and Enamel Adhesive, Clearfil Liner Bond System, and Scotchbond Multi-Purpose Dental Adhesive System.

Acid Etching, Dental↗
PubMed2002

Dentin proteoglycans: an immunocytochemical FEISEM study.

Dentin proteoglycans are fundamental constituents of the dentin matrix and are distributed ubiquitously both in dentin and cement. They have several important functional properties; in particular, they have a fundamental role in the maintenance and the correct stabilization of collagen fibers. The use of phosphoric acid on dentin, as proposed in most common dental adhesive systems to establish a reliable bond, may affect the molecular structure of proteoglycans. The aim of this study was to evaluate, after the application of EDTA or phosphoric acid on dentin, the dentin proteoglycans with an immunocytochemical approach with high resolution SEM. For this purpose, dentin disks obtained from recently extracted human molars were etched with a 35% water solution of phosphoric acid for 15 s, 30 s, and 60 s. Control specimens were conditioned with EDTA. Specimens were immunolabeled with a monoclonal antibody antichondroitin sulfate and visualized with a gold-conjugated secondary antibody. Conditioning dentin with EDTA resulted in a distinct labeling of the proteoglycans, as visualized on branching fibrillar structures in the order of 10-20 nm. The use of 35% phosphoric acid on dentin revealed a coagulation of proteoglycans after etching for 15 s while a very low labeling signal was detectable after 30 s. No labeling was obtained after etching dentin with phosphoric acid for 60 s. These results suggest that the use of 35% phosphoric acid on dentin is able to produce significant structural modifications of the dentin proteoglycans even after short application times. Additionally, when applied on the dentin surface for more than 30 s, phosphoric acid produces a dramatic decrease in proteoglycans' antigenicity, probably due to structural modifications of the three-dimensional conformation of these molecules.

Dentin↗