Evaluation of remineralisation potential of experimental nano hydroxyapatite pastes using scanning electron microscope with energy dispersive X-ray analysis: an in-vitro trial

  • V. Vijayasankari
  • S. AsokanEmail author
  • P. R. GeethaPriya
Original Scientific Article



To analyse the remineralisation potential of experimental nano hydroxyapatite (nHAP) paste on artificial caries lesions using scanning electron microscope (SEM) with energy dispersive X-ray analysis (EDX).


Sixty-five enamel specimens were randomly divided into 4 test groups (n = 15): 1% commercially available nHAP paste (CnHAP) group, 1% experimental nHAP paste (EnHAP) group, 10% EnHAP group, casein phosphopeptide–amorphous calcium phosphate (CPP–ACP) group and control group (n = 5). Artificial carious lesions were induced in all the specimens and treated with respective paste twice daily for 14 days. The specimens were subjected to EDX analysis at baseline, after demineralisation and after remineralisation. The surface characteristics of the remineralised enamel specimens were evaluated by SEM.


All the test groups showed significant change in calcium and phosphorus weight percentage (wt%) after remineralisation. The 10 % EnHAP showed higher mean value of calcium and phosphorus wt% followed by 1% CnHAP, 1% EnHAP and CPP–ACP. After remineralisation, both 10% EnHAP and CPP–ACP showed favourable enamel surface changes in SEM analysis.


Commercially available and experimental nHAP have the potential to remineralise artificially induced carious lesions.


Tooth remineralisation Nano hydroxyapatite paste Energy dispersive X-ray Scanning electron microscope 


Compliance with ethical standards

Conflict of interest

All three authors declare no conflict of interest.


  1. Bailey DL, Adams GG, Tsao CE, et al. Regression of post-orthodontic lesions by a remineralizing cream. J Dent Res. 2009;88(12):1148–53.CrossRefGoogle Scholar
  2. Beerens MW, Vanderveen MH, Vanbeek H, Tencate JM. Effects of casein phosphopeptide amorphous calcium fluoride phosphate paste on white spot lesions and dental plaque after orthodontic treatment: a 3-month follow-up. Eur J Oral Sci. 2010;118:610–17.CrossRefGoogle Scholar
  3. Cai F, Manton DJ, Shen P, et al. Effect of addition of citric acid and casein phosphopeptide–amorphous calcium phosphate to a sugar-free chewing gum on enamel remineralization in situ. Caries Res. 2007;41(5):377–83.CrossRefGoogle Scholar
  4. Cao Y, Mei ML, Xu J, et al. Biomimetic mineralisation of phosphorylated dentine by CPP–ACP. J Dent. 2013;41:818–25.CrossRefGoogle Scholar
  5. Chen F, Wang ZC, Lin CJ. Preparation and characterization of nano-sized hydroxyapatite particles and hydroxyapatite/chitosan nano-composite for use in biomedical materials. Mater Lett. 2002;57(4):858–61.CrossRefGoogle Scholar
  6. Chen H, Liu X, Dai J, et al. Effect of remineralizing agents on white spot lesions after orthodontic treatment: a systematic review. Am J Orthod Dentofac Orthop. 2013;143:376–82.CrossRefGoogle Scholar
  7. Cochrane NJ, Reynolds EC. Calcium phosphopeptides-mechanisms of action and evidence for clinical efficacy. Adv Dent Res. 2012;24(2):41–7.CrossRefGoogle Scholar
  8. Comar LP, Souza BM, Gracindo LF, et al. Impact of experimental nano-HAP pastes on bovine enamel and dentin submitted to a pH cycling model. Braz Dent J. 2013;24(3):273–8.CrossRefGoogle Scholar
  9. de Carvalho FG, Vieira BR, Santos RL, et al. In vitro effects of nano-hydroxyapatite paste on initial enamel carious lesions. Pediatr Dent. 2014;36(3):85–9.Google Scholar
  10. Fejerskov O, Kidd AME. Defining the disease: an introduction. In: Fejerskov O, Kidd EA, Nyvad B, Baelum V, editors. Dental caries: the disease and its clinical management. 2nd ed. Copenhagen: Blackwell Munksgaard publication; 2008.Google Scholar
  11. Haghgoo R, Rezvani MB, SalehiZeinabadi M. Comparison of nano-hydroxyapatite and sodium fluoride mouthrinse for remineralization of incipient carious lesions. J Dent. 2014;11(4):406–10.Google Scholar
  12. Hedge S, Roma M, Shetty D. Non fluoridated remineralizing agents in dentistry. J Pharm Sci Res. 2016;8(8):884–7.Google Scholar
  13. Huang SB, Gao SS, Yu HY. Effect of nano-hydroxyapatite concentration on remineralization of initial enamel lesion in vitro. Biomed Mater. 2009;4(3):034104.CrossRefGoogle Scholar
  14. Kamath P, Nayak R, Kamath SU, Pai D. A comparative evaluation of the remineralization potential of three commercially available remineralizing agents on white spot lesions in primary teeth: an in vitro study. J Indian Soc Pedod Prev Dent. 2017;35:229–37.CrossRefGoogle Scholar
  15. Keerthi V, Manish R. Remineralizing agents in dentistry: a review. IOSR J Dent Med Sci. 2014;13(4):57–60.CrossRefGoogle Scholar
  16. Kidd AME. Introduction: what is caries. In: Kidd EA, editors. Essentials of dental caries: the disease and its management. 3rd ed. Oxford: Oxford medical Publication; 2005.Google Scholar
  17. Kim MY, Kwon HK, Choi CH, Kim BI. Combined effects of nano-hydroxyapatite and NaF on remineralization of early caries lesion. Key Eng Mater. 2007;1347:330–2.Google Scholar
  18. Kumar VL, Itthagarun A, King NM. The effect of casein phosphopeptide-amorphous calcium phosphate on remineralization of artificial caries-like lesions: an in vitro study. Aust Dent J. 2008;53(1):34–40.CrossRefGoogle Scholar
  19. Li J, Xie X, Wang Y, et al. Long-term remineralizing effect of casein phosphopeptide-amorphous calcium phosphate (CPP–ACP) on early caries lesions in vivo: a systematic review. J Dent. 2014;42:769–77.CrossRefGoogle Scholar
  20. McKnight-Hanes C, Whitford GM. Fluoride release from three glass ionomer materials and the effects of varnishing with or without finishing. Caries Res. 1992;26:345–50.CrossRefGoogle Scholar
  21. Najibfard K, Ramalingam K, Chedjieu I, Amaechi BT. Remineralization of early caries by a nano-hydroxyapatite dentifrice. J Clin Dent. 2011;22(5):139–43.Google Scholar
  22. Naveena P, Nagarathana C, Sakunthala BK. Remineralizing agent—then and now—an update. Dentistry. 2014;4(9):1–5.Google Scholar
  23. Patil N, Choudhari S, Kulkarni S, Joshi SR. Comparative evaluation of remineralizing potential of three agents on artificially demineralized human enamel: an in vitro study. J Conserv Dent. 2013;16:116–20.CrossRefGoogle Scholar
  24. Rao SK, Bhat GS, Aradhya S, et al. Study of the efficacy of toothpaste containing casein phosphopeptide in the prevention of dental caries: a randomized controlled trial in 12- to 15-year-old high caries risk children in Bangalore, India. Caries Res. 2009;43:430–5.CrossRefGoogle Scholar
  25. Roveri N, Battistella E, Bianchi CL, et al. Surface enamel remineralization: biomimetic apatite nanocrystals and fluoride ions different effects. J Nanomater. 2009;746383:1–9.CrossRefGoogle Scholar
  26. Shetty PA, Shetty D, Shetty S. A comparison of clinical efficacy of dentifrices containing calcium sodium phosphosilicate, nanoparticle hydroxyapatite and a dentifrice containing casein phosphopeptide amorphous calcium phosphate on dentinal hypersensitivity—a comparative triple blind randomized study. Adv Hum Biol. 2014;4:57–64.Google Scholar
  27. Singh A, Shetty B, Mahesh CM, et al. Evaluation of efficiency of two nano hydroxyl apatite remineralizing agents with a hydroxyapatite and a conventional dentifrice: a comparative in vitro study. J Indian Orthod Soc. 2017;51:92–102.CrossRefGoogle Scholar
  28. Souza BM, Comar LP, Vertuan M, et al. Effect of an experimental paste with hydroxyapatite nanoparticles and fluoride on dental demineralisation and remineralisation in situ. Caries Res. 2015;49(5):499–507.CrossRefGoogle Scholar
  29. TenCate JM, Duijsters PP. Influence of fluoride in solution on tooth demineralization. Caries Res. 1983;17:193–9.CrossRefGoogle Scholar
  30. Verma P, Gupta U, Dodwad V, et al. Evaluation of the clinical efficacy of a new desensitizing toothpaste containing nano-crystalline hydroxyapatite in dentine hypersensitivity patients: a double blind randomized controlled clinical trial. J Dent Spec. 2013;1(2):47–54.Google Scholar
  31. Yengopal V, Mickenautsch S. Caries preventive effect of casein phosphopeptide–amorphous calcium phosphate (CPP–ACP): a meta-analysis. Acta Odonto Scand. 2009;67(6):321–32.CrossRefGoogle Scholar
  32. Zhou C, Zhang D, Bai Y, Li S. Casein phosphopeptide–amorphous calcium phosphate remineralization of primary teeth early enamel lesions. J Dent. 2014;42:21–9.CrossRefGoogle Scholar

Copyright information

© European Academy of Paediatric Dentistry 2018

Authors and Affiliations

  • V. Vijayasankari
    • 1
  • S. Asokan
    • 1
    Email author
  • P. R. GeethaPriya
    • 1
  1. 1.Department of Pedodontics and Preventive DentistryKSR Institute of Dental Science and ResearchTiruchengodeIndia

Personalised recommendations