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Published in: Clinical Oral Investigations 9/2014

Open Access 01-12-2014 | Original Article

Development and validation of an in vitro model for measurements of cervical root dentine permeability

Authors: Holger Jungbluth, Thomas Attin, Wolfgang Buchalla

Published in: Clinical Oral Investigations | Issue 9/2014

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Abstract

Objectives

The aim of this series of studies was the development and validation of a new model for evaluation of dentinal hypersensitivity (DH) therapies.

Materials and methods

Roots from extracted human teeth were sealed with a flowable composite. In the cervical area, a 3-mm-wide circular window was ground through the seal 1 mm deep into dentine. The pulp lumen was connected to a reservoir of artificial dentinal fluid (ADF) containing protein, mineral salts and methylene blue. At increased pulpal pressure, the ADF released through the said window was collected in containers each with 20 ml of physiologic saline for a consecutive series of 30-min intervals and ADF concentration (absorption) was determined photometrically. The model was verified by three experiments. In experiment 1, the lower limit of quantification (LLoQ, coefficient of variation = 20 % and difference of 5 standard deviations (SD) from blank) of ADF in physiologic saline was determined by measuring the absorption of 15 dilutions of ADF in physiologic saline (containing 0.625 ng to 12.5 μg methylene blue/ml) photometrically for ten times. In experiment 2, long-term linearity of ADF perfusion/outflow was investigated using 11 specimens. The ADF released through the window was collected in the said containers separately for each consecutive interval of 30 min for up to 240 min. Absorption was determined and analysed by linear regression over time. In experiment 3, perfusion before (2×) and after single treatment according to the following three groups was measured: BisGMA-based sealant (Seal&Protect®), an acidic fluoride solution (elmex fluid®) and control (no treatment).

Results

In experiment 1, the LLoQ was 0.005 μg methylene blue/ml. In experiment 2, permeability was different within the specimens and decreased highly linearly with time, allowing the prediction of future values. In experiment 3, Seal&Protect® completely occluded dentinal tubules. elmex fluid® increased tubular permeability by about 30 % compared to control.

Conclusions

A model comprising the use of artificial dentinal fluid was developed and validated allowing screening of therapeutic agents for the treatment of DH through reliable measurement of permeability of cervical root dentine.

Clinical relevance

The described in vitro model allows evaluation of potential agents for the treatment of DH at the clinically relevant cervical region of human teeth.
Literature
1.
go back to reference Chabanski MB, Gillam DG (1997) Aetiology, prevalence and clinical features of cervical dentine sensitivity. J Oral Rehabil 24:15–19PubMedCrossRef Chabanski MB, Gillam DG (1997) Aetiology, prevalence and clinical features of cervical dentine sensitivity. J Oral Rehabil 24:15–19PubMedCrossRef
2.
go back to reference (2003) Consensus-based recommendations for the diagnosis and management of dentin hypersensitivity. J Can Dent Assoc 69:221–226 (2003) Consensus-based recommendations for the diagnosis and management of dentin hypersensitivity. J Can Dent Assoc 69:221–226
3.
go back to reference Addy M (1992) Clinical aspects of dentine hypersensitivity. Proc Finn Dent Soc 88(Suppl 1):23–30PubMed Addy M (1992) Clinical aspects of dentine hypersensitivity. Proc Finn Dent Soc 88(Suppl 1):23–30PubMed
4.
go back to reference Holland GR, Narhi MN, Addy M, Gangarosa L, Orchardson R (1997) Guidelines for the design and conduct of clinical trials on dentine hypersensitivity. J Clin Periodontol 24:808–813PubMedCrossRef Holland GR, Narhi MN, Addy M, Gangarosa L, Orchardson R (1997) Guidelines for the design and conduct of clinical trials on dentine hypersensitivity. J Clin Periodontol 24:808–813PubMedCrossRef
5.
6.
go back to reference Gysi A (1900) An attempt to explain the sensitiveness of dentine. Br J Dent Sci 785:865–868 Gysi A (1900) An attempt to explain the sensitiveness of dentine. Br J Dent Sci 785:865–868
8.
go back to reference Absi EG, Addy M, Adams D (1987) Dentine hypersensitivity. A study of the patency of dentinal tubules in sensitive and non-sensitive cervical dentine. J Clin Periodontol 14:280–284PubMedCrossRef Absi EG, Addy M, Adams D (1987) Dentine hypersensitivity. A study of the patency of dentinal tubules in sensitive and non-sensitive cervical dentine. J Clin Periodontol 14:280–284PubMedCrossRef
9.
go back to reference Yoshiyama M, Masada J, Uchida A, Ishida H (1989) Scanning electron microscopic characterization of sensitive vs. insensitive human radicular dentin. J Dent Res 68:1498–1502PubMedCrossRef Yoshiyama M, Masada J, Uchida A, Ishida H (1989) Scanning electron microscopic characterization of sensitive vs. insensitive human radicular dentin. J Dent Res 68:1498–1502PubMedCrossRef
10.
go back to reference Amarasena N, Spencer J, Ou Y, Brennan D (2011) Dentine hypersensitivity in a private practice patient population in Australia. J Oral Rehabil 38:52–60PubMedCrossRef Amarasena N, Spencer J, Ou Y, Brennan D (2011) Dentine hypersensitivity in a private practice patient population in Australia. J Oral Rehabil 38:52–60PubMedCrossRef
11.
go back to reference von Troil B, Needleman I, Sanz M (2002) A systematic review of the prevalence of root sensitivity following periodontal therapy. J Clin Periodontol 29(Suppl 3):173–7, discussion 195-6CrossRef von Troil B, Needleman I, Sanz M (2002) A systematic review of the prevalence of root sensitivity following periodontal therapy. J Clin Periodontol 29(Suppl 3):173–7, discussion 195-6CrossRef
12.
go back to reference Attin T, Siegel S, Buchalla W, Lennon AM, Hannig C, Becker K (2004) Brushing abrasion of softened and remineralised dentin: an in situ study. Caries Res 38:62–66PubMedCrossRef Attin T, Siegel S, Buchalla W, Lennon AM, Hannig C, Becker K (2004) Brushing abrasion of softened and remineralised dentin: an in situ study. Caries Res 38:62–66PubMedCrossRef
13.
go back to reference Attin T, Weiss K, Becker K, Buchalla W, Wiegand A (2005) Impact of modified acidic soft drinks on enamel erosion. Oral Dis 11:7–12PubMedCrossRef Attin T, Weiss K, Becker K, Buchalla W, Wiegand A (2005) Impact of modified acidic soft drinks on enamel erosion. Oral Dis 11:7–12PubMedCrossRef
14.
go back to reference Davis WB, Winter PJ (1980) The effect of abrasion on enamel and dentine and exposure to dietary acid. Br Dent J 148:253–256PubMedCrossRef Davis WB, Winter PJ (1980) The effect of abrasion on enamel and dentine and exposure to dietary acid. Br Dent J 148:253–256PubMedCrossRef
15.
go back to reference Nagata T, Ishida H, Shinohara H et al (1994) Clinical evaluation of a potassium nitrate dentifrice for the treatment of dentinal hypersensitivity. J Clin Periodontol 21:217–221PubMedCrossRef Nagata T, Ishida H, Shinohara H et al (1994) Clinical evaluation of a potassium nitrate dentifrice for the treatment of dentinal hypersensitivity. J Clin Periodontol 21:217–221PubMedCrossRef
16.
go back to reference Schiff T, Dos Santos M, Laffi S et al (1998) Efficacy of a dentifrice containing 5 % potassium nitrate and 1500 PPM sodium monofluorophosphate in a precipitated calcium carbonate base on dentinal hypersensitivity. J Clin Dent 9:22–25PubMed Schiff T, Dos Santos M, Laffi S et al (1998) Efficacy of a dentifrice containing 5 % potassium nitrate and 1500 PPM sodium monofluorophosphate in a precipitated calcium carbonate base on dentinal hypersensitivity. J Clin Dent 9:22–25PubMed
17.
go back to reference Seong J, Macdonald E, Newcombe RG et al (2013) In situ randomised trial to investigate the occluding properties of two desensitising toothpastes on dentine after subsequent acid challenge. Clin Oral Investig 17:195–203 Seong J, Macdonald E, Newcombe RG et al (2013) In situ randomised trial to investigate the occluding properties of two desensitising toothpastes on dentine after subsequent acid challenge. Clin Oral Investig 17:195–203
18.
go back to reference Addy M, Dowell P (1983) Dentine hypersensitivity—a review. Clinical and in vitro evaluation of treatment agents. J Clin Periodontol 10:351–363PubMedCrossRef Addy M, Dowell P (1983) Dentine hypersensitivity—a review. Clinical and in vitro evaluation of treatment agents. J Clin Periodontol 10:351–363PubMedCrossRef
19.
go back to reference Pashley DH, Matthews WG, Zhang Y, Johnson M (1996) Fluid shifts across human dentine in vitro in response to hydrodynamic stimuli. Arch Oral Biol 41:1065–1072PubMedCrossRef Pashley DH, Matthews WG, Zhang Y, Johnson M (1996) Fluid shifts across human dentine in vitro in response to hydrodynamic stimuli. Arch Oral Biol 41:1065–1072PubMedCrossRef
20.
go back to reference Mjör IA, Nordahl I (1996) The density and branching of dentinal tubules in human teeth. Arch Oral Biol 41:401–412PubMedCrossRef Mjör IA, Nordahl I (1996) The density and branching of dentinal tubules in human teeth. Arch Oral Biol 41:401–412PubMedCrossRef
21.
go back to reference Schellenberg U, Krey G, Bosshardt D, Nair PN (1992) Numerical density of dentinal tubules at the pulpal wall of human permanent premolars and third molars. J Endod 18:104–109PubMedCrossRef Schellenberg U, Krey G, Bosshardt D, Nair PN (1992) Numerical density of dentinal tubules at the pulpal wall of human permanent premolars and third molars. J Endod 18:104–109PubMedCrossRef
22.
go back to reference Ozok AR, Wu MK, Wesselink PR (2002) Comparison of the in vitro permeability of human dentine according to the dentinal region and the composition of the simulated dentinal fluid. J Dent 30:107–111PubMedCrossRef Ozok AR, Wu MK, Wesselink PR (2002) Comparison of the in vitro permeability of human dentine according to the dentinal region and the composition of the simulated dentinal fluid. J Dent 30:107–111PubMedCrossRef
23.
go back to reference Pashley DH, Nelson R, Kepler EE (1982) The effects of plasma and salivary constituents on dentin permeability. J Dent Res 61:978–981PubMedCrossRef Pashley DH, Nelson R, Kepler EE (1982) The effects of plasma and salivary constituents on dentin permeability. J Dent Res 61:978–981PubMedCrossRef
24.
go back to reference Suge T, Ishikawa K, Kawasaki A et al (2002) Calcium phosphate precipitation method for the treatment of dentin hypersensitivity. Am J Dent 15:220–226PubMed Suge T, Ishikawa K, Kawasaki A et al (2002) Calcium phosphate precipitation method for the treatment of dentin hypersensitivity. Am J Dent 15:220–226PubMed
25.
go back to reference Shah VP, Midha KK, Findlay JW et al (2000) Bioanalytical method validation—a revisit with a decade of progress. Pharm Res 17:1551–1557PubMedCrossRef Shah VP, Midha KK, Findlay JW et al (2000) Bioanalytical method validation—a revisit with a decade of progress. Pharm Res 17:1551–1557PubMedCrossRef
26.
go back to reference Dohme GmbH, München: MSD-Manual der Diagnostik und Therapie. 5. 1993 Dohme GmbH, München: MSD-Manual der Diagnostik und Therapie. 5. 1993
27.
go back to reference Coffey CT, Ingram MJ, Bjorndal AM (1970) Analysis of human dentinal fluid. Oral Surg Oral Med Oral Pathol 30:835–837PubMedCrossRef Coffey CT, Ingram MJ, Bjorndal AM (1970) Analysis of human dentinal fluid. Oral Surg Oral Med Oral Pathol 30:835–837PubMedCrossRef
28.
go back to reference Knutsson G, Jontell M, Bergenholtz G (1994) Determination of plasma proteins in dentinal fluid from cavities prepared in healthy young human teeth. Arch Oral Biol 39:185–190PubMedCrossRef Knutsson G, Jontell M, Bergenholtz G (1994) Determination of plasma proteins in dentinal fluid from cavities prepared in healthy young human teeth. Arch Oral Biol 39:185–190PubMedCrossRef
29.
go back to reference Spreter von Kreudenstein T, Stüben J (1956) Dentinstoffwechselstudien VI. Mitteilung: Papierelektrophoretische Untersuchungen über die Zusammensetzung der Dentinliquorproteine. Dtsch Zahnärztl Z 6:500–507 Spreter von Kreudenstein T, Stüben J (1956) Dentinstoffwechselstudien VI. Mitteilung: Papierelektrophoretische Untersuchungen über die Zusammensetzung der Dentinliquorproteine. Dtsch Zahnärztl Z 6:500–507
30.
go back to reference Spreter von Kreudenstein T, Stüben J (1956) Dentinstoffwechselstudien IV. Mitteilung: Über die Gewinnung des Dentinliquors durch Elution zum Zweck der Bestimmung seiner Bestandteile sowie Untersuchungen über den Wassergehalt des Dentins und über das Dentinliquorspatium. Dtsch Zahnärztl Z 11:1214–1220 Spreter von Kreudenstein T, Stüben J (1956) Dentinstoffwechselstudien IV. Mitteilung: Über die Gewinnung des Dentinliquors durch Elution zum Zweck der Bestimmung seiner Bestandteile sowie Untersuchungen über den Wassergehalt des Dentins und über das Dentinliquorspatium. Dtsch Zahnärztl Z 11:1214–1220
Metadata
Title
Development and validation of an in vitro model for measurements of cervical root dentine permeability
Authors
Holger Jungbluth
Thomas Attin
Wolfgang Buchalla
Publication date
01-12-2014
Publisher
Springer Berlin Heidelberg
Published in
Clinical Oral Investigations / Issue 9/2014
Print ISSN: 1432-6981
Electronic ISSN: 1436-3771
DOI
https://doi.org/10.1007/s00784-014-1194-5

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