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01.05.2024 | Research

Acquired enamel pellicle and biofilm engineering with a combination of acid-resistant proteins (CaneCPI-5, StN15, and Hemoglobin) for enhanced protection against dental caries - in vivo and in vitro investigations

verfasst von: Tamara Teodoro Araujo, Aline Dionizio, Thamyris Souza Carvalho, Chelsea Maria Vilas Boas Feitosa, Mariele Vertuan, João Victor Frazão Câmara, Flavio Henrique-Silva, Reinaldo Marchetto, Marcos Roberto Chiaratti, Angélica Camargo Santos, Lindomar Oliveira Alves, Milene Ferro, Marília Afonso Rabelo Buzalaf

Erschienen in: Clinical Oral Investigations | Ausgabe 5/2024

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Abstract

Objective

This study was designed in two-legs. In the in vivo, we explored the potential of a rinse solution containing a combination (Comb) of 0.1 mg/mL CaneCPI-5 (sugarcane-derive cystatin), 1.88 × 10^− 5M StN15 (statherin-derived peptide) and 1.0 mg/mL hemoglobin (Hb) to change the protein profile of the acquired enamel pellicle(AEP) and the microbiome of the enamel biofilm. The in vitro, was designed to reveal the effects of Comb on the viability and bacterial composition of the microcosm biofilm, as well as on enamel demineralization.

Materials and methods

In vivo study, 10 participants rinsed (10mL,1 min) with either deionized water (H₂O-control) or Comb. AEP and biofilm were collected after 2 and 3 h, respectively, after rinsing. AEP samples underwent proteomics analysis, while biofilm microbiome was assessed via 16 S-rRNA Next Generation Sequencing(NGS). In vitro study, a microcosm biofilm protocol was employed. Ninety-six enamel specimens were treated with: 1)Phosphate-Buffered Solution-PBS(negative-control), 2)0.12%Chlorhexidine, 3)500ppmNaF and 4)Comb. Resazurin, colony-forming-units(CFU) and Transversal Microradiography(TMR) were performed.

Results

The proteomic results revealed higher quantity of proteins in the Comb compared to control associated with immune system response and oral microbial adhesion. Microbiome showed a significant increase in bacteria linked to a healthy microbiota, in the Comb group. In the in vitro study, Comb group was only efficient in reducing mineral-loss and lesion-depth compared to the PBS.

Conclusions

The AEP modification altered the subsequent layers, affecting the initial process of bacterial adhesion of pathogenic and commensal bacteria, as well as enamel demineralization.

Clinical relevance

Comb group shows promise in shaping oral health by potentially introducing innovative approaches to prevent enamel demineralization and deter tooth decay.

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Peres MA et al (2020) Oral diseases: a global public health challenge - authors’ reply. Lancet 395(10219):186–187PubMedCrossRef

Flemming J, Hannig C, Hannig M (2022) Caries Management-the role of surface interactions in De- and remineralization-processes. J Clin Med, 11(23)

Hay DI (1973) The interaction of human parotid salivary proteins with hydroxyapatite. Arch Oral Biol 18(12):1517–1529PubMedCrossRef

Lendenmann U, Grogan J, Oppenheim FG (2000) Saliva and dental pellicle–a review. Adv Dent Res 14:22–28PubMedCrossRef

Trautmann S et al (2019) Proteomic analysis of the initial oral pellicle in Caries-active and caries-Free individuals. Proteom Clin Appl 13(4):e1800143CrossRef

da Silva Ventura TM et al (2017) The proteomic profile of the acquired enamel pellicle according to its location in the dental arches. Arch Oral Biol 79:20–29CrossRef

Huang R, Li M, Gregory RL (2011) Bacterial interactions in dental biofilm. Virulence 2(5):435–444PubMedPubMedCentralCrossRef

Hannig M, Joiner A (2006) The structure, function and properties of the acquired pellicle. Monogr Oral Sci 19:29–64PubMed

Zimmerman JN et al (2013) Proteome and peptidome of human acquired enamel pellicle on deciduous teeth. Int J Mol Sci 14(1):920–934PubMedPubMedCentralCrossRef

10.

Siqueira WL, Custodio W, McDonald EE (2012) New insights into the composition and functions of the acquired enamel pellicle. J Dent Res 91(12):1110–1118PubMedCrossRef

11.

Siqueira WL et al (2007) Identification of protein components in in vivo human acquired enamel pellicle using LC-ESI-MS/MS. J Proteome Res 6(6):2152–2160PubMedCrossRef

12.

Siqueira WL et al (2007) Acquired enamel pellicle and its potential role in oral diagnostics. Ann N Y Acad Sci 1098:504–509PubMedCrossRef

13.

Delecrode TR et al (2015) Identification of acid-resistant proteins in acquired enamel pellicle. J Dent 43(12):1470–1475PubMedCrossRef

14.

Siqueira WL, Oppenheim FG (2009) Small molecular weight proteins/peptides present in the in vivo formed human acquired enamel pellicle. Arch Oral Biol 54(5):437–444PubMedPubMedCentralCrossRef

15.

Delecrode TR et al (2015) Exposure to acids changes the proteomic of acquired dentine pellicle. J Dent 43(5):583–588PubMedCrossRef

16.

Taira EA et al (2018) Changes in the Proteomic Profile of Acquired Enamel Pellicles as a function of their time of formation and hydrochloric acid exposure. Caries Res 52(5):367–377PubMedCrossRef

17.

Martini T et al (2019) Proteomics of acquired pellicle in gastroesophageal reflux disease patients with or without erosive tooth wear. J Dent 81:64–69PubMedCrossRef

18.

Santiago AC et al (2017) A New Sugarcane Cystatin strongly binds to Dental Enamel and reduces Erosion. J Dent Res 96(9):1051–1057PubMedCrossRef

19.

Pela VT et al (2021) Safety and in situ antierosive effect of CaneCPI-5 on Dental Enamel. J Dent Res 100(12):1344–1350PubMedCrossRef

20.

Araujo TT et al (2022) Protein-based engineering of the initial acquired enamel pellicle in vivo: proteomic evaluation. J Dent 116:103874PubMedCrossRef

21.

Martini T et al (2020) Salivary hemoglobin protects against erosive tooth wear in gastric reflux patients. Caries Res 54(5–6):466–474PubMedCrossRef

22.

Pela VT et al (2023) Use of Reflectometer Optipen to assess the preventive effect of a sugarcane cystatin on initial dental erosion in vivo. J Mech Behav Biomed Mater 141:105782PubMedCrossRef

23.

Araújo TT et al (2021) A sugarcane cystatin (CaneCPI-5) alters microcosm biofilm formation and reduces dental caries. Biofouling 37:109–116PubMedCrossRef

24.

Makrodimitris K et al (2007) Structure prediction of protein-solid surface interactions reveals a molecular recognition motif of statherin for hydroxyapatite. J Am Chem Soc 129(44):13713–13722PubMedCrossRef

25.

Raj PA et al (1992) Salivary statherin. Dependence on sequence, charge, hydrogen bonding potency, and helical conformation for adsorption to hydroxyapatite and inhibition of mineralization. J Biol Chem 267(9):5968–5976PubMedCrossRef

26.

Shah S et al (2011) An in vitro scanning microradiography study of the reduction in hydroxyapatite demineralization rate by statherin-like peptides as a function of increasing N-terminal length. Eur J Oral Sci 119(Suppl 1):13–18PubMedCrossRef

27.

Choi M et al (2014) MSstats: an R package for statistical analysis of quantitative mass spectrometry-based proteomic experiments. Bioinformatics 30(17):2524–2526PubMedCrossRef

28.

Dawes C (1972) Circadian rhythms in human salivary flow rate and composition. J Physiol 220(3):529–545PubMedPubMedCentralCrossRef

29.

Perez-Riverol Y, Bandla BJ, Hewapathirana C, García-Seisdedos S, Kamatchinathan D, Kundu S, Prakash D, Frericks-Zipper A, Eisenacher A, Walzer M, Wang M, Brazma S, Vizcaíno A (2022) The PRIDE database resources in 2022: a hub for mass spectrometry-based proteomics evidences. Nucleic Acids Res 50(D1):D543–D552PubMedCrossRef

30.

Araujo TT et al (2024) Acquired pellicle and Biofilm engineering by rinsing with hemoglobin solution. Caries Res

31.

Klindworth A et al (2013) Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic Acids Res 41(1):e1PubMedCrossRef

32.

Fernando JR et al (2019) The prebiotic effect of CPP-ACP sugar-free chewing gum. J Dent 91:103225PubMedCrossRef

33.

Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30(15):2114–2120PubMedPubMedCentralCrossRef

34.

Marinho VC et al (2016) Fluoride mouthrinses for preventing dental caries in children and adolescents. Cochrane Database Syst Rev 7(7):CD002284PubMed

35.

McBain AJ (2009) Chap. 4: in vitro biofilm models: an overview. Adv Appl Microbiol 69:99–132

36.

Maddi A, Scannapieco FA (2013) Oral biofilms, oral and periodontal infections, and systemic disease. Am J Dent 26(5):249–254PubMed

37.

Alonso B et al (2017) Comparison of the XTT and resazurin assays for quantification of the metabolic activity of Staphylococcus aureus biofilm. J Microbiol Methods 139:135–137PubMedCrossRef

38.

Braga AS, Pires JG, Magalhães AC (2018) Effect of a mouthrinse containing Malva sylvestris on the viability and activity of microcosm biofilm and on enamel demineralization compared to known antimicrobials mouthrinses. Biofouling 34(3):252–261PubMedCrossRef

39.

Cheng L et al (2012) Dental plaque microcosm biofilm behavior on calcium phosphate nanocomposite with quaternary ammonium. Dent Mater 28(8):853–862PubMedPubMedCentralCrossRef

40.

Dos Santos DMS et al (2019) Protective effect of 4% titanium tetrafluoride varnish on dentin demineralization using a microcosm biofilm model. Caries Res 53(5):576–583PubMedCrossRef

41.

Angmar B, Carlström D, Glas J-E (1963) Studies on the ultrastructure of dental enamel: IV. The mineralization of normal human enamel. J Ultrastruct Res 8(1–2):12–23PubMedCrossRef

42.

Pela VT et al (2022) Preventive effect of chitosan gel containing CaneCPI-5 against enamel erosive wear in situ. Clin Oral Investig 26(11):6511–6519PubMedCrossRef

43.

Carvalho TS et al (2024) Hemoglobin protects enamel against intrinsic enamel erosive demineralization. Caries Res

44.

Carvalho TS et al (2020) Acquired pellicle protein-based engineering protects against erosive demineralization. J Dent 102:103478PubMedCrossRef

45.

Taira EA et al (2021) Rinsing with statherin-derived peptide alters the Proteome of the Acquired Enamel Pellicle. Caries Res 55(4):333–340PubMedCrossRef

46.

Martini T et al (2023) Proteomic analysis of stimulated saliva in gastroesophageal reflux disease patients with and without erosive tooth wear: observational study. J Dent 139:104724PubMedCrossRef

47.

Reis FN et al (2023) Solutions containing a statherin-derived peptide reduce Enamel Erosion in vitro. Caries Res 57(1):52–58PubMedCrossRef

48.

Reis FN et al (2023) Gels containing statherin-derived peptide protect against enamel and dentin erosive tooth wear in vitro. J Mech Behav Biomed Mater 137:105549PubMedCrossRef

49.

Hope CK, Wilson M (2003) Measuring the thickness of an outer layer of viable bacteria in an oral biofilm by viability mapping. J Microbiol Methods 54(3):403–410PubMedCrossRef

50.

Valente MT et al (2018) Acquired Enamel Pellicle Engineered peptides: effects on Hydroxyapatite Crystal Growth. Sci Rep 8(1):3766PubMedPubMedCentralCrossRef

51.

Vukosavljevic D et al (2014) Nanoscale adhesion forces between enamel pellicle proteins and hydroxyapatite. J Dent Res 93(5):514–519PubMedPubMedCentralCrossRef

52.

Kawasaki T, Takahashi S, Ikeda K (1985) Hydroxyapatite high-performance liquid chromatography: column performance for proteins. Eur J Biochem 152(2):361–371PubMedCrossRef

53.

Gironda CC et al (2022) New insights into the anti-erosive property of a sugarcane-derived cystatin: different vehicle of application and potential mechanism of action. J Appl Oral Sci 30:e20210698PubMedPubMedCentralCrossRef

54.

van der Mei HC et al (2008) Bond strengthening in oral bacterial adhesion to salivary conditioning films. Appl Environ Microbiol 74(17):5511–5515PubMedPubMedCentralCrossRef

55.

Gell DA (2018) Structure and function of haemoglobins. Blood Cells Mol Dis 70:13–42PubMedCrossRef

56.

Huttlin EL et al (2017) Architecture of the human interactome defines protein communities and disease networks. Nature 545(7655):505–509PubMedPubMedCentralCrossRef

57.

Marsh PD (2004) Dental plaque as a microbial biofilm. Caries Res 38(3):204–211PubMedCrossRef

58.

Rupf S et al (2018) Comparison of initial oral microbiomes of young adults with and without cavitated dentin caries lesions using an in situ biofilm model. Sci Rep 8(1):14010PubMedPubMedCentralCrossRef

59.

Li T et al (2001) Different type 1 fimbrial genes and tropisms of commensal and potentially pathogenic Actinomyces spp. with different salivary acidic proline-rich protein and statherin ligand specificities. Infect Immun 69(12):7224–7233PubMedPubMedCentralCrossRef

60.

Aas JA et al (2008) Bacteria of dental caries in primary and permanent teeth in children and young adults. J Clin Microbiol 46(4):1407–1417PubMedPubMedCentralCrossRef

61.

Hajishengallis G (2011) Immune evasion strategies of Porphyromonas gingivalis. J Oral Biosci 53(3):233–240PubMedPubMedCentralCrossRef

62.

Bostanci N, Belibasakis GN (2012) Porphyromonas gingivalis: an invasive and evasive opportunistic oral pathogen. FEMS Microbiol Lett 333(1):1–9PubMedCrossRef

63.

Smalley JW, Olczak T (2017) Heme acquisition mechanisms of Porphyromonas gingivalis - strategies used in a polymicrobial community in a heme-limited host environment. Mol Oral Microbiol 32(1):1–23PubMedCrossRef

64.

Haeri-Araghi H et al (2018) Evaluating the relationship between dental caries number and salivary level of IgA in adults. J Clin Exp Dent 10(1):e66–e69PubMedPubMedCentral

65.

Wu Z et al (2020) Association between salivary s-IgA concentration and dental caries: a systematic review and meta-analysis. Biosci Rep, 40(12)

66.

Pudakalkatti PS, Baheti AS (2015) Correlation of salivary immunoglobulin A against lipopolysaccharide of Porphyromonas gingivalis with clinical periodontal parameters. Contemp Clin Dent 6(3):305–308PubMedPubMedCentralCrossRef

67.

Lynge Pedersen AM, Belstrom D (2019) The role of natural salivary defences in maintaining a healthy oral microbiota. J Dent 80(Suppl 1):S3–S12PubMedCrossRef

68.

Levine M (2011) Susceptibility to dental caries and the salivary proline-rich proteins. Int J Dent 2011:953412PubMedPubMedCentralCrossRef

69.

Laputkova G et al (2018) Salivary Protein Roles in oral health and as predictors of Caries Risk. Open Life Sci 13:174–200PubMedPubMedCentralCrossRef

70.

Azen E, Prakobphol A, Fisher SJ (1993) PRB3 null mutations result in absence of the proline-rich glycoprotein gl and abolish Fusobacterium nucleatum interactions with saliva in vitro. Infect Immun 61(10):4434–4439PubMedPubMedCentralCrossRef

71.

Lorenzon EN et al (2012) Effects of dimerization on the structure and biological activity of antimicrobial peptide Ctx-Ha. Antimicrob Agents Chemother 56(6):3004–3010PubMedPubMedCentralCrossRef

72.

Marinho VC et al (2016) Fluoride mouthrinses for preventing dental caries in children and adolescents. Cochrane Database Syst Rev 7:CD002284PubMed

Titel: Acquired enamel pellicle and biofilm engineering with a combination of acid-resistant proteins (CaneCPI-5, StN15, and Hemoglobin) for enhanced protection against dental caries - in vivo and in vitro investigations
verfasst von: Tamara Teodoro Araujo
Aline Dionizio
Thamyris Souza Carvalho
Chelsea Maria Vilas Boas Feitosa
Mariele Vertuan
João Victor Frazão Câmara
Flavio Henrique-Silva
Reinaldo Marchetto
Marcos Roberto Chiaratti
Angélica Camargo Santos
Lindomar Oliveira Alves
Milene Ferro
Marília Afonso Rabelo Buzalaf
Publikationsdatum: 01.05.2024
Verlag: Springer Berlin Heidelberg
Erschienen in: Clinical Oral Investigations / Ausgabe 5/2024
Print ISSN: 1432-6981
Elektronische ISSN: 1436-3771
DOI: https://doi.org/10.1007/s00784-024-05651-0

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Acquired enamel pellicle and biofilm engineering with a combination of acid-resistant proteins (CaneCPI-5, StN15, and Hemoglobin) for enhanced protection against dental caries - in vivo and in vitro investigations