Original Article

Effect of Proanthocyanidin, Fluoride and Casein Phosphopeptide Amorphous Calcium Phosphate Remineralizing Agents on Microhardness of Demineralized Dentin


Objectives: The aim of this study was to evaluate the effect of dentin remineralization using proanthocyanidin (PA), fluoride varnish and casein phosphopeptide amorphous calcium phosphate (CPP-ACP) paste and their various combinations on microhardness of demineralized root dentin.
Materials and Methods: One-hundred and twenty freshly extracted sound human premolars were selected and randomly divided into eight groups for dentin treatment as follows. C: Deionized water (control); PA: 6.5% PA solution; F: fluoride varnish (5% NaF, 22600 ppm fluoride); CP: CCP-ACP; PAF: 6.5% PA + fluoride varnish; PACP: 6.5% PA + CCP-ACP; FCP: fluoride varnish + CCP-ACP and PAFCP: 6.5% PA + fluoride varnish + CCP-ACP. All specimens were subjected to Vickers microhardness test (500 g, 10 seconds, 3 points). Data were analyzed using one-way ANOVA and Tukey’s post hoc test. The significance level was set at 0.05.
Results: The mean and standard deviation (SD) values of Vickers hardness number (VHN) in groups C, PA, F, CP, PAF, PACP, FCP and PAFCP were 37.39±4.97, 38.68±4.62, 48.28±2.68 ,41.91±3.32, 48.59±2.55, 53.34±2.57, 48.413±4.00 and 55.20±1.82, respectively. Pairwise comparisons of the groups revealed that there was no significant difference between groups C and PA, PA and CP, F and PAF, F and FCP, PAF and FCP, and PACP and FPACP (P>0.05); but significant differences were observed between other groups (P<0.05).
Conclusions: The results of this study showed that the tested dentin treatments increased the microhardness of demineralized root dentin except for PA application.




- Hayes M, Da Mata C, Cole M, McKenna G, Burke F, Allen PF. Risk indicators associated with root caries in independently living older adults. J Dent. 2016 Aug;51:8-14.

- Kinney JH, Habelitz S, Marshall SJ, Marshall GW. The importance of intrafibrillar mineralization of collagen on the mechanical properties of dentin. J Dent Res. 2003 Dec;82(12):957-61.

- Cao CY, Mei ML, Li QL, Lo ECM, Chu CH. Methods for Biomimetic Remineralization of Human Dentine: A Systematic Review. Int J Mol Sci. 2015 Mar;16(3):4615-27.

- Jose P, Sanjeev K, Sekar M. Effect of green and white tea pretreatment on remineralization of demineralized dentin by CPP-ACFP- An in vitro microhardness analysis. J Clin Diagn Res. 2016 Apr; 10(4):ZC85-9.

- Savas S, Kavrìk F, Kucukyìlmaz E. Evaluation of the remineralization capacity of quantitative methods. J Appl Oral Sci. 2016 May-Jun;24(3):198-203.

- Chokshi K, Chokshi A, Konde S, Shetty SR, Chandra KN, Jana S, et al. An in vitro comparative evaluation of three remineralizing agents using confocal microscopy. J Clin Diagn Res. 2016 Jun;10(6): ZC39-ZC42.

- Xin W, Leung KC, Lo EC, Mok MY, Leung MH. A randomized, double-blind, placebo controlled clinical trial of fluoride varnish in preventing dental caries of Sjögren’s syndrome patients. BMC Oral Health. 2016 Sep;16(1):102.

- Elsayad I, Sakr A, Badr Y. Combining casein phosphopeptide-amorphous calcium phosphate with fluoride: synergistic remineralization potential of artificially demineralized enamel or not? J Biomed Opt. 2009 Jul-Aug;14(4):044039.

- Cochrane NJ, Saranathan S, Cai F, Cross KJ, Reynolds EC. Enamel subsurface lesion remineralisation with casein phosphopeptide stabilised solutions of calcium, phosphate and fluoride. Caries Res. 2008;42(2):88-97.

- Reynolds EC. Calcium phosphate-based remineralization systems: scientific evidence? Aust Dent J. 2008 Sep;53(3):268-73.

- Petti S, Scully C. Polyphenols, oral health and disease: A review. J Dent. 2009 Jun;37(6):413-23.

- Xie Q, Bedran-Russo AK, Wu CD. In vitro remineralization effects of grape seed extract on artificial root caries. J Dent. 2008 Nov;36(11):900-6.

- Aron PM, Kennedy JA. Flavan-3-ols: nature, occurrence and biological activity. Mol Nutr Food Res. 2008 Jan;52(1):79-104.

- Bedran-Russo AK, Pashley DH, Agee K, Drummond JL, Miescke KJ. Changes in stiffness of demineralized dentin following application of collagen crosslinkers. J Biomed Mater Res B Appl Biomater. 2008 Aug;86(2):330-4.

- Castellan CS, Pereira PN, Grande RH, Bedran-Russo AK. Mechanical characterization of proanthocyanidin-dentin matrix interaction. Dent Mater. 2010 Oct;26(10):968-73.

- Walter R, Miguez PA, Arnold RR, Pereira PN, Duarte WR, Yamauchi M. Effects of natural crosslinkers on the stability of dentin collagen and the inhibition of root caries in vitro. Caries Res. 2008;42(4):263-8.

- Somasundaram P, Vimala N, Mandke LG. Protective potential of casein phosphopeptide amorphous calcium phosphate containing paste on enamel surfaces. J Conserv Dent. 2013 Mar;16(2): 152-6.

- Pavan S, Xie Q, Hara AT, Bedran-Russo AK. Biomimetic approach for root caries prevention using a proanthocyanidin-rich agent. Caries Res. 2011; 45(5):443-7.

- Broyles AC, Pavan S, Bedran-Russo AK. Effect of dentin surface modification on the microtensile bond strength of self-adhesive resin cements. J Prosthodont. 2013 Jan;22(1):59-62.

- Liu Y, Chen M, Yao X, Xu C, Zhang Y, Wang Y. Enhancement in dentin collagen's biological stability after proanthocyanidins treatment in clinically relevant time periods. Dent Mater. 2013 Apr;29 (4):485-92.

- Salehzadeh Esfahani K, Mazaheri R, Pishevar L. Effects of treatment with various remineralizing agents on the microhardness of demineralized enamel surface. J Dent Res Dent Clin Dent Prospects. 2015 Fall;9(4):239-45.

- Takahashi N, Nyvad B. Ecological hypothesis of dentin and root caries. Caries Res. 2016;50(4):422-31.

- Cao CY, Mei ML, Li QL, Lo EC, Chu CH. Methods for biomimetic remineralization of human dentine: a systematic review. Int J Mol Sci. 2015 Mar;16(3):4615-27.

- Chen Z, Cao S, Wang H, Li Y, Kishen A, Deng X, et al. Biomimetic remineralization of demineralized dentine using scaffold of CMC/ACP nanocomplexes in an in vitro tooth model of deep caries. PLoS One. 2015 Jan;10(1):e0116553.

- Chaussain-Miller C, Fioretti F, Goldberg M, Menashi S. The role of matrix metalloproteinases (MMPs)in human caries. J Dent Res. 2006 Jan;85 (1):22-32.

- Fine AM. Oligomeric proanthocyanidin complexes: history, structure, and phytopharmaceutical applications. Altern Med Rev. 2000 Apr;5(2):144-51.

- Panich M, Poolthong S. The effect of casein phosphopeptide-amorphous calcium phosphate and a cola soft drink on in vitro enamel hardness. J Am Dent Assoc. 2009 Apr;140(4):455-60.

- Silva AP, Gonçalves RS, Borges AF, Bedran-Russo AK, Shinohara MS. Effectiveness of plant-derived proanthocyanidins on demineralization on enamel and dentin under artificial cariogenic challenge. J Appl Oral Sci. 2015 May-Jun;23(3):302-9.

- Wu CD. Grape products and oral health. J Nutr. 2009 Sep;139(9):1818S-23S.

- Hattori M, Kusumoto IT, NambaT, Ishigami T, Yukihiko HA. Effect of tea polyphenols on glucan synthesis by glucosyltransferase from Streptococcus mutans. Chem Pharm Bull (Tokyo). 1990 Mar;38(3):717-20.

- Karlinsey RL, Mackey AC. Solid-state preparation and dental application of an organically modified calcium phosphate. J Mater Sci. 2009 Jan;44(1):346-9.

- Karlinsey RL, Mackey AC, Walker ER, Frederick KE. Preparation, characterization and in vitro efficacy of an acid-modified β-TCP material for dental hard-tissue remineralization. Acta Biomater. 2010 Mar;6(3):969-78.

- Shirahatti RV, Ankola AV, Nagesh L, Hallikerimath S. The effects of three different pastes on enamel caries formation and lesion depth progression- an in vitro study. J Oral Health Comm Dent. 2007;1(1):1-6.

- Ahmadi Zenouz G, Ezoji F, Enderami SA, Khafri S. Effect of fluoride, casein phosphopeptide-amorphous calcium phosphate and casein phosphopeptide-amorphous calcium phosphate fluoride on enamel surface microhardness after microabrasion: An in vitro study. J Dent (Tehran). 2015 Oct;12(10):705-11.

- Lata S, Varghese NO, Varughese JM. Remineralization potential of fluoride and amorphous calcium phosphate-casein phosphopeptide on enamel lesions: An in vitro comparative evaluation. J Conserv Dent. 2010 Jan-Mar;13(1):42-6.

- Theodore M, Harald O, Edward J. Sturdevant’s art and science of operative dentistry. Missouri, Mosby, 2006:807-40.

- Pulido MT, Wefel JS, Hernandez MM, Denehy GE, Guzman-Armstrong S, Chalmers JM, et al. The inhibitory effect of MI paste, fluoride and a combination of both on the progression of artificial caries-like lesions in enamel. Oper Dent. 2008 Sep-Oct;33(5):550-5.

- Ten Cate JM. Remineralization of deep enamel dentine caries lesions. Aust Dent J. 2008 Sep;53(3): 281-5.

- Shen P, Manton DJ, Cochrane NJ, Walker GD, Yuan Y, Reynolds C, et al. Effect of added calcium phosphate on enamel remineralization by fluoride in a randomized controlled in situ trial. J Dent. 2011 Jul;39(7):518-25.

- Shen P, Manton DJ, Cochrane NJ, Walker GD, Yuan Y, Reynolds C, et al. Effect of added calcium phosphate on enamel remineralization by fluoride in a randomized controlled in situ trial. J Dent. 2011 Jul;39(7):518-25.

- Bedran-Russo AK, Pereira PN, Duarte WR, Drummond JL, Yamauchi M. Application of crosslinkers to dentin collagen enhances the ultimate tensile strength. J Biomed Mater Res B Appl Biomater. 2007 Jan;80(1):268-72.

- Shi JH, Li H, Wang YN. [In vitro remineralization effect of grape seed extract on artificial dentin caries]. Shanghai Kou Qiang Yi Xue.2015 Feb;24(1):18-22.

- Benjamin S, Sharma R, Thomas SS, Nainan MT. Grape seed extract as a potential remineralizing agent: a comparative in vitro study. J Contemp Dent Pract. 2012 Jul;13(4):425-30.

- Loomis WD. Overcoming problems of phenolics and quinones in the isolation of plant enzymes and organelles. Methods Enzymol. 1974 Dec;31:528-44.

- Arumugam MT, Nesamani R, Kittappa K, Sanjeev K, Sekar M. Effect of various antioxidants on the shear bond strength of composite resin to bleached enamel: An in vitro study. J Conserv Dent. 2014 Jan;17(1):22-6.

- Bertassoni LE, Habelitz S, Marshall SJ, Marshall GW. Mechanical recovery of dentin following remineralization in vitro—an indentation study. J Biomech. 2011 Jan;44(1):176-81.

- Epasinghe DJ, Yiu C, Burrow MF. Synergistic effect of proanthocyanidin and CPP-ACFP on remineralization of artificial root caries. Aust Dent J. 2015 Dec;60(4):463-70.

- Besinis A, van Noort R, Martin N. Infiltration of demineralized dentin with silica and hydroxyapatite nanoparticles. Dent Mater. 2012 Sep;28(9):1012-23.

IssueVol 14, No 2 (2017) QRcode
SectionOriginal Article
Dentin Fluoride Proanthocyanidin Remineralization

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How to Cite
Khamverdi Z, Kordestani M, Soltanian A. Effect of Proanthocyanidin, Fluoride and Casein Phosphopeptide Amorphous Calcium Phosphate Remineralizing Agents on Microhardness of Demineralized Dentin. Front Dent. 2017;14(2):76-83.