Abstract
Endodontic treatment of immature permanent teeth with necrotic pulp poses several clinical challenges and is one of the most demanding interventions in endodontics. Recently, with new discoveries in the field of tissue engineering, novel treatment protocols have been established. The most promising treatment modality is revascularization, whose integral part is the exposure of collagen matrix and embedded growth factors. However, optimization of the treatment protocol requires a development of analytical procedures able to analyze growth factors directly on the sample surface. In this work, method based on surface-enhanced Raman spectroscopy (SERS) was developed to investigate the influence of the time of the medical treatment using EDTA on exposure and accessibility of the growth factors, namely TGF-ß1, BMP-2, and bFGF on the dentine surface. The nanotags, which consist of magnetic Fe3O4@Ag nanocomposite covalently functionalized by tagged antibodies (anti-TGF-ß1-Cy3, anti-BMP-2-Cy5, and anti-bFGF-Cy7), were employed as a SERS substrate. Each antibody was coupled with a unique label allowing us to perform a parallel analysis of all three growth factors within one analytical run. Developed methodology presents an interesting alternative to a fluorescence microscopy and in contrary allows evaluating a chemical composition and thus minimizing possible false-positive results.
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References
Dhillon H, Kaushik M, Sharma R. Regenerative endodontics-creating new horizons: regenerative endodontics. J Biomed Mater Res B Appl Biomater. 2016;104:676–85. https://doi.org/10.1002/jbm.b.33587.
Wigler R, Kaufman AY, Lin S, Steinbock N, Hazan-Molina H, Torneck CD. Revascularization: a treatment for permanent teeth with necrotic pulp and incomplete root development. J Endod. 2013;39:319–26. https://doi.org/10.1016/j.joen.2012.11.014.
Nosrat A, Homayounfar N, Oloomi K. Drawbacks and unfavorable outcomes of regenerative endodontic treatments of necrotic immature teeth: a literature review and report of a case. J Endod. 2012;38:1428–34. https://doi.org/10.1016/j.joen.2012.06.025.
Žižka R, Šedý J. Paradigm shift from stem cells to cell-free regenerative endodontic procedures: a critical review. Stem Cells Dev. 2017;26:147–53. https://doi.org/10.1089/scd.2016.0264.
Smith AJ, Scheven BA, Takahashi Y, Ferracane JL, Shelton RM, Cooper PR. Dentine as a bioactive extracellular matrix. Arch Oral Biol. 2012;57:109–21. https://doi.org/10.1016/j.archoralbio.2011.07.008.
Galler KM, Buchalla W, Hiller K-A, Federlin M, Eidt A, Schiefersteiner M, et al. Influence of root canal disinfectants on growth factor release from dentin. J Endod. 2015;41:363–8. https://doi.org/10.1016/j.joen.2014.11.021.
Kakaboura A. Smear layer on prepared dentin. Odontostomatol Proodos. 1989;43:211–23.
Ferracane JL, Cooper PR, Smith AJ. Dentin matrix component solubilization by solutions at pH relevant to self-etching dental adhesives. J Adhes Dent. 2013;15:407–12. https://doi.org/10.3290/j.jad.a29536.
Galler KM, Widbiller M, Buchalla W, Eidt A, Hiller K-A, Hoffer PC, et al. EDTA conditioning of dentine promotes adhesion, migration and differentiation of dental pulp stem cells. Int Endod J. 2016;49:581–90. https://doi.org/10.1111/iej.12492.
Zeng Q, Nguyen S, Zhang H, Chebrolu HP, Alzebdeh D, Badi MA, et al. Release of growth factors into root canal by irrigations in regenerative endodontics. J Endod. 2016;42:1760–6. https://doi.org/10.1016/j.joen.2016.04.029.
Duncan HF, Smith AJ, Fleming GJP, Reid C, Smith G, Cooper PR. Release of bio-active dentine extracellular matrix components by histone deacetylase inhibitors (HDACi). Int Endod J. 2017;50:24–38. https://doi.org/10.1111/iej.12588.
Hisamoto M, Goto M, Muto M, Nio-Kobayashi J, Iwanaga T, Yokoyama A. A systematic analysis for localization of predominant growth factors and their receptors involved in murine tooth germ differentiation using in situ hybridization technique. Biomed Res. 2015;36:205–17. https://doi.org/10.2220/biomedres.36.205.
Li S, Pan Y. Differential expression of transforming growth factor-beta1, connective tissue growth factor, phosphorylated-SMAD2/3 and phosphorylated-ERK1/2 during mouse tooth development. J Mol Histol. 2017;48:347–55. https://doi.org/10.1007/s10735-017-9733-4.
Niwa T, Yamakoshi Y, Yamazaki H, Karakida T, Chiba R, Hu JC-C, et al. The dynamics of TGF-β in dental pulp, odontoblasts and dentin. Sci Rep. 2018;8:4450. https://doi.org/10.1038/s41598-018-22823-7.
Zhao S, Sloan AJ, Murray PE, Lumley PJ, Smith AJ. Ultrastructural localisation of TGF-β exposure in dentine by chemical treatment. Histochem J. 2000;32:489–94.
Ionita I. Diagnosis of tooth decay using polarized micro-Raman confocal spectroscopy. Rom Rep Phys. 2009;61:567–74.
Zavala-Alonso V, Loyola-Rodríguez JP, Terrones H, Patiño-Marín N, Martínez-Castañón GA, Anusavice K. Analysis of the molecular structure of human enamel with fluorosis using micro-Raman spectroscopy. J Oral Sci. 2012;54:93–8.
Wang Y, Yao X. Morphological/chemical imaging of demineralized dentin layer in its natural, wet state. Dent Mater. 2010;26:433–42. https://doi.org/10.1016/j.dental.2010.01.002.
Beier BD, Quivey RG, Berger AJ. Raman microspectroscopy for species identification and mapping within bacterial biofilms. AMB Express. 2012;2:35.
Cepeda-Pérez E, Moreno-Hernández C, López-Luke T, Monzón-Hernández D, de la Rosa E. Evaluation of bacterial presence in the root canal by Raman spectroscopy: a preliminary study. Biomed Phys Eng Express. 2016;2:065006. https://doi.org/10.1088/2057-1976/2/6/065006.
Fleischmann M, Hendra PJ, McQuillan AJ. Raman spectra of pyridine adsorbed at a silver electrode. Chem Phys Lett. 1974;26:4.
Balzerova A, Fargasova A, Markova Z, Ranc V, Zboril R. Magnetically-assisted surface enhanced Raman spectroscopy (MA-SERS) for label-free determination of human immunoglobulin G (IgG) in blood using Fe3O4@Ag nanocomposite. Anal Chem. 2014;86:11107–14. https://doi.org/10.1021/ac503347h.
Ranc V, Markova Z, Hajduch M, Prucek R, Kvitek L, Kaslik J, et al. Magnetically assisted surface-enhanced Raman scattering selective determination of dopamine in an artificial cerebrospinal fluid and a mouse striatum using Fe3O4/Ag nanocomposite. Anal Chem. 2014;86:2939–46. https://doi.org/10.1021/ac500394g.
Chaloupková Z, Balzerová A, Bařinková J, Medříková Z, Šácha P, Beneš P, et al. Label-free determination of prostate specific membrane antigen in human whole blood at nanomolar levels by magnetically assisted surface enhanced Raman spectroscopy. Anal Chim Acta. 2017; https://doi.org/10.1016/j.aca.2017.10.008.
Panacek A, Balzerova A, Prucek R, Ranc V, Vecerova R, Husickova V, et al. Preparation, characterization and antimicrobial efficiency of Ag/PDDA-diatomite nanocomposite. Colloids Surf B-Biointerfaces. 2013;110:191–8. https://doi.org/10.1016/j.colsurfb.2013.04.031.
Marková Z, Šišková K, Filip J, Šafářová K, Prucek R, Panáček A, et al. Chitosan-based synthesis of magnetically-driven nanocomposites with biogenic magnetite core, controlled silver size, and high antimicrobial activity. Green Chem. 2012;14:2550. https://doi.org/10.1039/c2gc35545k.
Prucek R, Panacek A, Fargasova A, Ranc V, Masek V, Kvitek L, et al. Re-crystallization of silver nanoparticles in a highly concentrated NaCl environment-a new substrate for surface enhanced IR-visible Raman spectroscopy. CrystEngComm. 2011;13:2242–8. https://doi.org/10.1039/c0ce00776e.
Wang Y, Schlücker S. Rational design and synthesis of SERS labels. Analyst. 2013;138:2224–38. https://doi.org/10.1039/C3AN36866A.
Funding
The authors received financial support from project NPU LO1305 of the Ministry of Education, Youth and Sports of the Czech Republic; project IGA_PrF_2018_021, CZ.1.07/2.3.00/30.0004, and Research Infrastructure NanoEnviCz, supported by the Ministry of Education, Youth and Sports of the Czech Republic under Project No. LM2015073; and project 16-02938S of the Grant Agency of the Czech Republic.
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The approval of the ethic committee of the Faculty Hospital Olomouc and Medical Faculty of Palacký University for the study was obtained (NV-18-08-00328).
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Ranc, V., Žižka, R., Chaloupková, Z. et al. Imaging of growth factors on a human tooth root canal by surface-enhanced Raman spectroscopy. Anal Bioanal Chem 410, 7113–7120 (2018). https://doi.org/10.1007/s00216-018-1311-4
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DOI: https://doi.org/10.1007/s00216-018-1311-4