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In vivo model for microbial invasion of tooth root dentinal tubules.

Brittan JL, Sprague SV, Macdonald EL, Love RM, Jenkinson HF, West NX - J Appl Oral Sci (2016)

Bottom Line: A spectrum of Gram-positive and Gram-negative cell morphotypes were visualized, and molecular typing identified species of Granulicatella, Streptococcus, Klebsiella, Enterobacter, Acinetobacter, and Pseudomonas as dentinal tubule residents.A range of bacteria were able to initially invade dentinal tubules within exposed dentine.The model will be useful for testing the effectiveness of antiseptics, irrigants, and potential tubule occluding agents in preventing bacterial invasion of dentine.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral and Dental Sciences, University of Bristol, Bristol, United Kingdom.

ABSTRACT
Objective Bacterial penetration of dentinal tubules via exposed dentine can lead to root caries and promote infections of the pulp and root canal system. The aim of this work was to develop a new experimental model for studying bacterial invasion of dentinal tubules within the human oral cavity. Material and Methods Sections of human root dentine were mounted into lower oral appliances that were worn by four human subjects for 15 d. Roots were then fixed, sectioned, stained and examined microscopically for evidence of bacterial invasion. Levels of invasion were expressed as Tubule Invasion Factor (TIF). DNA was extracted from root samples, subjected to polymerase chain reaction amplification of 16S rRNA genes, and invading bacteria were identified by comparison of sequences with GenBank database. Results All root dentine samples with patent tubules showed evidence of bacterial cell invasion (TIF value range from 5.7 to 9.0) to depths of 200 mm or more. A spectrum of Gram-positive and Gram-negative cell morphotypes were visualized, and molecular typing identified species of Granulicatella, Streptococcus, Klebsiella, Enterobacter, Acinetobacter, and Pseudomonas as dentinal tubule residents. Conclusion A novel in vivo model is described, which provides for human root dentine to be efficiently infected by oral microorganisms. A range of bacteria were able to initially invade dentinal tubules within exposed dentine. The model will be useful for testing the effectiveness of antiseptics, irrigants, and potential tubule occluding agents in preventing bacterial invasion of dentine.

No MeSH data available.


Related in: MedlinePlus

Transverse sections of human roots after 15 days incubation in situ in subject 2. Sections were prepared as described in Material and Methods, and stained by Brown & Brenn method. Panels: A, sample A, Gram-positive cocci invading to a depth of >150 mm and showing a 10 mm depth dense biofilm at the surface (arrowed); B, sample B, disintegration of internal dentine structure meant that sections from this sample could not be analyzed; C, sample C, Gram-positive bacteria at the surface and deep penetration by smaller Gram-negative bacteria ≥200 mm (arrowed); sample D, Gram-positive and Gram-negative bacteria penetration with accumulation of Gram-positive cocci biofilm at the surface of the sectioned sample (arrowed). TIF scores for specimens are shown in Figure 3
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f05: Transverse sections of human roots after 15 days incubation in situ in subject 2. Sections were prepared as described in Material and Methods, and stained by Brown & Brenn method. Panels: A, sample A, Gram-positive cocci invading to a depth of >150 mm and showing a 10 mm depth dense biofilm at the surface (arrowed); B, sample B, disintegration of internal dentine structure meant that sections from this sample could not be analyzed; C, sample C, Gram-positive bacteria at the surface and deep penetration by smaller Gram-negative bacteria ≥200 mm (arrowed); sample D, Gram-positive and Gram-negative bacteria penetration with accumulation of Gram-positive cocci biofilm at the surface of the sectioned sample (arrowed). TIF scores for specimens are shown in Figure 3

Mentions: In subject 2, sample A carried a dense invasive biofilm of Gram-positive stained material at the surface (Figure 5A) and there was invasion of tubules >150 mm by Gram-positive cocci. A similar pattern was seen for root sample D (Figure 5D), while sample C from subject 2 showed Gram-positive bacteria at the surface and deep penetration ≥200 mm by small Gram-negative organisms. Block B from this subject was one of the samples that could not be properly analyzed, as the internal dentine structure was disintegrated (Figure 5B).


In vivo model for microbial invasion of tooth root dentinal tubules.

Brittan JL, Sprague SV, Macdonald EL, Love RM, Jenkinson HF, West NX - J Appl Oral Sci (2016)

Transverse sections of human roots after 15 days incubation in situ in subject 2. Sections were prepared as described in Material and Methods, and stained by Brown & Brenn method. Panels: A, sample A, Gram-positive cocci invading to a depth of >150 mm and showing a 10 mm depth dense biofilm at the surface (arrowed); B, sample B, disintegration of internal dentine structure meant that sections from this sample could not be analyzed; C, sample C, Gram-positive bacteria at the surface and deep penetration by smaller Gram-negative bacteria ≥200 mm (arrowed); sample D, Gram-positive and Gram-negative bacteria penetration with accumulation of Gram-positive cocci biofilm at the surface of the sectioned sample (arrowed). TIF scores for specimens are shown in Figure 3
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836919&req=5

f05: Transverse sections of human roots after 15 days incubation in situ in subject 2. Sections were prepared as described in Material and Methods, and stained by Brown & Brenn method. Panels: A, sample A, Gram-positive cocci invading to a depth of >150 mm and showing a 10 mm depth dense biofilm at the surface (arrowed); B, sample B, disintegration of internal dentine structure meant that sections from this sample could not be analyzed; C, sample C, Gram-positive bacteria at the surface and deep penetration by smaller Gram-negative bacteria ≥200 mm (arrowed); sample D, Gram-positive and Gram-negative bacteria penetration with accumulation of Gram-positive cocci biofilm at the surface of the sectioned sample (arrowed). TIF scores for specimens are shown in Figure 3
Mentions: In subject 2, sample A carried a dense invasive biofilm of Gram-positive stained material at the surface (Figure 5A) and there was invasion of tubules >150 mm by Gram-positive cocci. A similar pattern was seen for root sample D (Figure 5D), while sample C from subject 2 showed Gram-positive bacteria at the surface and deep penetration ≥200 mm by small Gram-negative organisms. Block B from this subject was one of the samples that could not be properly analyzed, as the internal dentine structure was disintegrated (Figure 5B).

Bottom Line: A spectrum of Gram-positive and Gram-negative cell morphotypes were visualized, and molecular typing identified species of Granulicatella, Streptococcus, Klebsiella, Enterobacter, Acinetobacter, and Pseudomonas as dentinal tubule residents.A range of bacteria were able to initially invade dentinal tubules within exposed dentine.The model will be useful for testing the effectiveness of antiseptics, irrigants, and potential tubule occluding agents in preventing bacterial invasion of dentine.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral and Dental Sciences, University of Bristol, Bristol, United Kingdom.

ABSTRACT
Objective Bacterial penetration of dentinal tubules via exposed dentine can lead to root caries and promote infections of the pulp and root canal system. The aim of this work was to develop a new experimental model for studying bacterial invasion of dentinal tubules within the human oral cavity. Material and Methods Sections of human root dentine were mounted into lower oral appliances that were worn by four human subjects for 15 d. Roots were then fixed, sectioned, stained and examined microscopically for evidence of bacterial invasion. Levels of invasion were expressed as Tubule Invasion Factor (TIF). DNA was extracted from root samples, subjected to polymerase chain reaction amplification of 16S rRNA genes, and invading bacteria were identified by comparison of sequences with GenBank database. Results All root dentine samples with patent tubules showed evidence of bacterial cell invasion (TIF value range from 5.7 to 9.0) to depths of 200 mm or more. A spectrum of Gram-positive and Gram-negative cell morphotypes were visualized, and molecular typing identified species of Granulicatella, Streptococcus, Klebsiella, Enterobacter, Acinetobacter, and Pseudomonas as dentinal tubule residents. Conclusion A novel in vivo model is described, which provides for human root dentine to be efficiently infected by oral microorganisms. A range of bacteria were able to initially invade dentinal tubules within exposed dentine. The model will be useful for testing the effectiveness of antiseptics, irrigants, and potential tubule occluding agents in preventing bacterial invasion of dentine.

No MeSH data available.


Related in: MedlinePlus