Abstract
Root demineralization is used in Periodontics as an adjuvant for mechanical treatment. The aim of this study was to evaluate the effects of root surface modification with mechanic, chemical, and photodynamic treatments on adhesion and proliferation of human gingival fibroblasts and osteoblasts. Root fragments were treated by scaling and root planing (C—control group), EDTA (pH 7), citric acid plus tetracycline (CA—pH 1), and antimicrobial photodynamic therapy (aPDT) with toluidine blue O and red laser (pH 4). Cells were seeded (104 cells/well, 6th passage) on root fragments of each experimental group and cultured for 24, 48, and 72 h. Cells were counted in scanning electron microscopy images by a calibrated examiner. For fibroblasts, the highest number of cells were present at 72-h period (p < 0.05). EDTA group showed a very low number of cells in relation to CA group (p < 0.05). CA and aPDT group presented higher number of cells in all periods, but without differences between other treatment groups (p > 0.05). For osteoblasts, there was a significant increase in cell numbers for aPDT group at 72 h (p < 0.05). In conclusion, aPDT treatment provided a positive stimulus to osteoblast growth, while for fibroblasts, aPDT and CA had a tendency for higher cell growth.
Similar content being viewed by others
References
Hatfield CG, Baumhammers A (1971) Cytotoxic effects of periodontally involved surfaces of human teeth. Arch Oral Biol 16:465–468
Adriaens PA, Edwards CA, De Boever JA, Loesche WJ (1988) Ultrastructural observations on bacterial invasion in cementum and radicular dentin of periodontally diseased human teeth. J Periodontol 59:493–503
Alyahya A, Alqareer A (2017) Does the removal of cementum facilitate bacterial penetration into dentinal tubules in vitro? J Endod 43:1111–1115
Sudhakar R, Pratebha B (2015) Fibrous architecture of cementodentinal junction in disease: a scanning electron microscopic study. J Oral Maxillofac Pathol 19:325–329
Polson AM, Caton J (1982) Factors influencing periodontal repair and regeneration. J Periodontol 53:617–625
Sant'Ana AC, Marques MM, Barroso TE, Passanezi E, de Rezende ML (2007) Effects of TGF-beta1, PDGF-BB, and IGF-1 on the rate of proliferation and adhesion of a periodontal ligament cell lineage in vitro. J Periodontol 78:2007–2017
Karam PS, Sant'Ana AC, de Rezende ML, Greghi SL, Damante CA, Zangrando MS (2016) Root surface modifiers and subepithelial connective tissue graft for treatment of gingival recessions: a systematic review. J Periodontal Res 51:175–185
Blomlöf J, Blomlöf L, Lindskog S (1997) Effect of different concentrations of EDTA on smear removal and collagen exposure in periodontitis-affected root surfaces. J Clin Periodontol 24:534–537
Feist IS, Gde M, Carneiro SRS, Eduardo CP, Myiagi SPH, Marques MM (2003) Adhesion and growth of cultured human gingival fibroblast and periodontally involved root surfaces treated by Er:YAG laser. J Periodontol 74:1368–1375
Passanezi E, Damante CA, de Rezende ML, Greghi SL (2015) Lasers in periodontal therapy. Periodontol 67:268–291
Damante CA, Ducati P, Ferreira R, Salmeron S, Zangrando MS, de Rezende ML, Sant’Ana AC, Greghi SL, Magalhães AC (2016) In vitro evaluation of adhesion/proliferation of human gingival fibroblasts on demineralized root surfaces by toluidine blue O in antimicrobial photodynamic therapy. Photodiagn Photodyn Ther 13:303–307
Karam PSBH, Ferreira R, Oliveira RC, Greghi SLA, de Rezende MLR, Sant’Ana ACP, Zangrando MSR, Damante CA (2017) Stimulation of human gingival fibroblasts viability and growth by roots treated with high intensity lasers, photodynamic therapy and citric acid. Arch Oral Biol 81:1–6
Register AA (1973) Bone and cementum induction by dentin, demineralized in situ. J Periodontol 44:49–54
Register AA, Burdick FA (1975) Accelerated reattachment with cementogenesis to dentin, demineralized in situ. I. Optimum range. J Periodontol 46:646–655
Chaves E, Cox CF, Morrison E, Caffesse R (1993) The effect of citric acid application on periodontally involved root surfaces. II. An in vitro scanning electron microscopic study. Int J Periodontics Restorative Dent 13:188–196
Wen CR, Caffesse RG, Morrison EC, Nasjleti CE, Parikh UK (1992) In vitro effects of citric acid application techniques on dentin surfaces. J Periodontol 63:883–889
Garret JS, Crigger M, Egelberg J (1978) The effects of citric acid on diseased root surfaces. J Periodontal Res 13:155–163
Daly CG (1982) Anti-bacterial effect of citric acid treatment of periodontally diseased root surfaces in vitro. J Clin Periodontol 9:386–392
Polson AM, Hanes PJ (1989) Cell and fiber responses to cementum from periodontitis- affected root surfaces after citric acid treatment. J Clin Periodontol 16:489–497
Hanes PJ, Polson AM, Ladenheim S (1985) Cell and fiber attachment to demineralized dentin from normal root surfaces. J Periodontol 56:752–765
Larjava H, Salonen J, Häkkinen L, Närhi T (1988) Effect of citric acid treatment on the migration of epithelium on root surfaces in vitro. J Periodontol 59:95–99
Blomlof J, Lindskog S (1995) Periodontal tissue vitality after different etching modalities. J Clin Periodontol 22:464–468
Mayfield L, Söderholm G, Norderyd O, Attström R (1998) Root conditioning using EDTA gel as an adjunct to surgical therapy for the treatment of intraosseous periodontal defects. J Clin Periodontol 25:707–714
Scelza MF, Pierro P, Scelza P, Pereira M (2004) Effect of three different time periods of irrigation with EDTA-T, EDTA and citric acid on smear layer removal. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 98:499–503
Golub LM, Ramamurthy N, McNamara TF, Gomes B, Wolff M, Casino A, Kapoor A, Zambon J, Ciancio S, Schneir M et al (1984) Tetracyclines inhibit tissue collagenase activity. A new mechanism in the treatment of periodontal disease. J Periodont Res 19:651–555
Lafferty TA, Gher MA, Gray IL (1993) Comparative SEM study on the effect of acid etching with tetracycline HCl or citric acido on instrumented periodontally involved human root surfaces. J Periodontol 64:689–693
Isik AG, Tarem B, Hafez AA, Yalcin FS, Onan U, Cox CF (2000) A comparative scanning electron microscopic study on the characteristics of demineralized dentin root surface using different tetracycline HCl concentrations and application times. J Periodontol 71:219–225
Amaral NG, Rezende ML, Hirata F, Rodrigues MG, Sant'ana AC, Greghi SL, Passanezi E (2011) Comparison among four commonly used demineralizing agents for root conditioning: a scanning electron microscopy. J Appl Oral Sci 19:469–475
Theodoro LH, Zezell DM, Garcia VG, Haypek P, Nagata MJH, Almeida JM, Eduardo CP (2010) Comparative analysis of root surface smear layer removal by different etching modalities or erbium:yttrium-aluminum-garnet laser irradiation. A scanning electron microscopy study. Laser Med Sci 25:485–491
Sgolastra F, Petrucci A, Severino M, Graziani F, Gatto R, Monaco A (2013) Adjunctive photodynamic therapy to non-surgical treatment of chronic periodontitis: a systematic review and meta-analysis. J Clin Periodontol 40:514–526
Xue D, Zhao Y (2017) Clinical effectiveness of adjunctive antimicrobial photodynamic therapy for residual pockets during supportive periodontal therapy: a systematic review and meta-analysis. Photodiagn Photodyn Ther 17:127–133
Passanezi E, Janson WA, Nahas D, Campos Júnior A (1989) Newly forming bone autografts to treat periodontal infrabony pockets: clinical and histological events. Int J Periodontics Restorative Dent 9:140–153
Sant'ana AC, Ferraz BF, Rezende ML, Greghi SL, Damante CA, Passanezi E (2012) Newly forming bone graft: a novel surgical approach to the treatment of denuded roots. J Appl Oral Sci 20:392–398
Salmeron S, Rezende ML, Consolaro A, Sant'ana AC, Damante CA, Greghi SL, Passanezi E (2013) Laser therapy as an effective method for implant surface decontamination: a histomorphometric study in rats. J Periodontol 84:641–649
Goulart RC, Thedei-Jr G, Souza SL, Tedesco AC, Ciancaglini P (2010) Study of methylene blue and erythrosine dyes employed in photodynamic therapy for inactivation of planktonic and biofilm-cultivated Aggregatibacter actinomycetemcomitans. Photomed Laser Surg 28:85–90
Giannelli M, Formigli L, Lorenzini L, Bani D (2012) Combined photoablative and photodynamic diode laser therapy as an adjunct to non-surgical periodontal treatment: a randomized split-mouth clinical trial. J Clin Periodontol 39:962–970
Petelin M, Perkič K, Seme K, Gašpirc B (2015) Effect of repeated adjunctive antimicrobial photodynamic therapy on subgingival periodontal pathogens in the treatment of chronic periodontitis. Lasers Med Sci 30:1647–1656
Polson AM, Proye MP (1983) Fibrin linkage: a precursor for new attachment. J Periodontol 54:141–147
Leite FR, Sampaio JE, Zandim DL, Dantas AA, Leite ER, Leite AA (2010) Influence of root-surface conditioning with acid and chelating agents on clot stabilization. Quintessence Int 41:341–349
George RP, Kumar S, Ramakrishna T, Emmadi P, Ambalavanan N (2013) Effects of tetracycline-containing gel and a mixture of tetracycline and citric acid-containing gel on non-surgical periodontal therapy. Indian J Dent Res 24:52–59
Pessoa L, Galvão V, Damante C, Sant’Ana ACP (2015) Removal of black stains from teeth by photodynamic therapy: clinical and microbiological analysis. BMJ Case Reports. https://doi.org/10.1136/bcr-2015-212276
Decrock E, Hoorelbeke D, Ramadan R, Delvaeye T, De Bock M, Wang N, Krysko DV, Baatout S, Bultynck G, Aerts A, Vinken M, Leybaert L (2017) Calcium, oxidative stress and connexin channels, a harmonious orchestra directing the response to radiotherapy treatment? Biochim Biophys Acta 1864:1099–1120
Zayzafoon M (2006) Calcium/calmodulin signaling controls osteoblast growth and differentiation. J Cell Biochem 97:56–70
Funding
This study was funded by grant #2010/15667-2 and #2012/06738-9 São Paulo Research Foundation (FAPESP).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This paper was approved by Bauru School of Dentistry Ethical Committee in Human Research (Protocols 721.759 and 1.710.789).
Informed consent
Informed consent was obtained from all individual participants that donated teeth for the study.
Rights and permissions
About this article
Cite this article
Ferreira, R., de Toledo Barros, R.T., Karam, P.S.B.H. et al. Comparison of the effect of root surface modification with citric acid, EDTA, and aPDT on adhesion and proliferation of human gingival fibroblasts and osteoblasts: an in vitro study. Lasers Med Sci 33, 533–538 (2018). https://doi.org/10.1007/s10103-017-2395-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10103-017-2395-3