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Biomedical subjects

B H Clarkson

Publications and source records attributed to B H Clarkson.

At least 37 records · Page 2Linked to original sources

Phosphoprotein extraction from the dentine/cementum complex of human tooth roots.

Root shards were placed in dialysis tubing and demineralized to completion in either 10% disodium EDTA, pH 7.4, 0.6 M HCl, 0.1 M HCl, 0.5 M acetic or 75 mM-25 mM lactic-acetic acids. The demineralized shards were then re-extracted with 0.05 M tris-HCl, 1.0 M NaCl. DEAE chromatography revealed that the major peak of the 0.6 M CHl and EDTA extracts contained organic phosphorus, whereas much less organic phosphorus was found in the major peak of the 0.1 M HCl extract. Analysis of the re-extracts gave a pattern opposite to that obtained from the initial extractions. Measurements of protein and organic phosphorus released during extraction and re-extraction confirmed these results. Staining of SDS-PAGE gels for phosphoprotein with Stains-All resulted in a blue smear in fractions containing organic phosphorus. Thus the extraction of phosphoproteins from human tooth roots differed depending upon the demineralizing conditions. This ability to remove phosphoprotein differentially will allow further investigation of the role of phosphoprotein in mineralization and remineralization.

Acetates↗

Effect of topical fluoride treatments on fluoride distribution during in vitro caries-like lesion formation.

Tooth sections were treated in vitro for 4 min with APF, SnF2, or received no treatment. Each treatment group then received washes of KOH, (24 h), an inorganic solution (24 h), or DDH2O (2 min) and were placed into dialyzed 15% w/v gel (pH 4.3) containing 0.15 mM hydroxyapatite, but no fluoride (less than 0.02 ppm F). Adjacent nontreated sound enamel acted as the control. Lesion microdissection, after 2 weeks exposure to the acidified gel, revealed an inverse relationship between lesion body fluoride concentration and lesion depth. The lesion depth was smallest, and the lesion body fluoride concentration was greatest in both fluoride-treated groups after a 2-min DDH2O wash, a 24-hour inorganic wash, and a 24-hour KOH wash, respectively. These data support the theory that the progress of carious lesions is related to the fluoride concentration in the lesion and that the fluoride concentration in the lesion is related to the acquired fluoride concentration in sound enamel.

Acidulated Phosphate Fluoride↗

In vitro caries-like lesion production by Streptococcus mutans and Actinomyces viscosus using sucrose and starch.

We investigated the formation of caries-like lesions on root and enamel sections by S. mutans and A. viscosus when grown in four different carbohydrate substrates. The substrates were: sucrose, starch + alpha-amylase, and a combination of starch and sucrose with and without alpha-amylase. Twenty-four sections with exposed windows on both the root and enamel surfaces were exposed to the individual bacterial species in each of the four substrates for three weeks. At three weeks, the sections were removed and lesion depths measured. When grown in sucrose alone, S. mutans produced significantly deeper lesions in both root and enamel surfaces when compared with A. viscosus. However, S. mutans failed to produce lesions when grown in "limited" sucrose with starch added, whereas A. viscosus in this same substrate produced lesions in the enamel and root surfaces. A viscosus also produced either similar or significantly deeper lesions in both root and enamel surfaces, when compared with S. mutans grown in the two substrates with amylase. Thus, under these in vitro conditions, it was demonstrated that A. viscosus could utilize starch, whereas utilization of starch by S. mutans was alpha-amylase-dependent.

Actinomyces↗

Effect of proteolytic enzymes on caries lesion formation in vitro.

This study investigates enzyme degradation of collagen at various pH levels and is designed to obtain information pertaining to the effect of proteolytic enzymes on caries-like lesion formation and progression. One hundred and eight sections of human teeth with exposed areas on enamel and root face, were placed into an enzyme/buffer system maintained at pH levels of 4.0, 5.5 and 7.0. Periodically, throughout the experimental period, sections were removed and examined histologically for caries-like lesion formation. Results showed consistent surface erosion occurring on the root face only at a pH level of 5.5 in buffer containing enzymes. Sections placed in pH 5.5 buffer only, which served as controls, showed obvious lesion formation on the root face, but with histologically intact surfaces. This interaction occurring between enzymatic activity and acid demineralization suggests that enzymes may have a contributory effect on caries lesion formation in root surfaces.

Dental Caries↗

Fluoride distribution in enamel after in vitro caries-like lesion formation.

Using a microdissection technique to obtain samples from carious and adjacent sound enamel, we compared the fluoride concentrations in surface and subsurface enamel. Undialyzed and dialyzed gels were used to create lesions with and without negatively birefringent surface zones, respectively. The fluoride concentration in the surface enamel of the lesions was always greater than that in the corresponding sound enamel, as was the subsurface fluoride concentrations of caries lesions compared with adjacent sound enamel at the same depth.

Dental Caries↗

Natural root caries: a histologic and microradiographic evaluation.

The purpose of this study was to obtain information about the detailed histopathology of naturally occurring root caries. Fifty extracted human teeth exhibiting some degree of root caries were sectioned longitudinally and examined with transmitted light, polarized light and microradiography. The occurrence of the histological zones of dentinal caries was tabulated and revealed a lesion body in all cases. Light microscopy showed the same basic features as microradiography and was most useful when the sections were imbibed in quinoline. A structureless area below the main body of the lesion was observed in 77% of the cases when using quinoline. This fluid more clearly defined the extent of the lesion and may show a "phenolic reaction" when using polarized light microscopy. Partial radiopaque surface layers were observed in almost 80% of the sections. This study has described the detailed histopathology of root caries, and aids in the development of model systems to evaluate this emerging dental health problem.

Dental Caries↗

The effect of polyvalent metal ion mordanting on caries-like lesion progression in enamel.

The ability of metal ion mordanting followed by APF application to inhibit the progression of caries-like lesions was compared with APF treatment alone. The progression of the lesion was investigated in vitro using an acidified gel system, and the lesions were compared using polarized light microscopy. Both ferric and zirconyl pre-treatments, but not titanium, were shown to inhibit lesion progression to a greater extent than did APF. This inhibition was thought to be due to the presence of an amorphous-type coating similar to those produced on sound enamel by metal ion mordanting.

Acidulated Phosphate Fluoride↗

Microprobe and SEM analysis of surface coatings on caries-like lesions in enamel after metal ion mordanting and APF application.

The production of tenacious surface coatings on enamel after metal ion mordanting followed by APF has been reported to prevent caries-like lesion initiation in vitro. In this study, similar surface coatings, which may be responsible for the inhibition of lesion progression, were examined using SEM and microprobe analysis. Relatively thick, even surface coatings which were in intimate contact with the enamel surface of caries-like lesions were seen after zirconium and iron (ferric) pre-treatment, but a thinner and unevenly distributed coating was seen after titanium pre-treatment. This latter coating was similar to that seen after APF treatment alone. Fluoride concentrations on the surface and in the subsurface of caries-like lesions were similar for all treatment groups. The surface coatings after zirconium and iron mordanting may be responsible for the greater inhibition of lesion progression than that seen after both titanium pre-treatment and APF treatment alone.

Acidulated Phosphate Fluoride↗

A model for producing caries-like lesions in enamel and dentin using oral bacteria in vitro.

Subsurface enamel lesions and root surface caries-like lesions were consistently produced in vitro using Streptococcus mutans FA1 cultured in thioglycollate broth containing 3.5% w/v dextrose and 2% w/v gelatin. When viewed in polarized light and after imbibition in water, the enamel lesions had a negatively birefringent surface zone and positively birefringent body of the lesion. Those lesions produced after six weeks, after imbibition in quinoline, exhibited a dark zone. The root surface caries-like lesions exhibited a less-radiolucent surface zone above a heavily demineralized body of the lesion. However, no reactionary dentin was seen in the in vitro lesions.

Dental Caries↗