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Erschienen in: Clinical Oral Investigations 3/2017

25.04.2016 | Original Article

Ultrasonic activation of irrigants increases growth factor release from human dentine

verfasst von: M. Widbiller, A. Eidt, K.-A. Hiller, W. Buchalla, G. Schmalz, K. M. Galler

Erschienen in: Clinical Oral Investigations | Ausgabe 3/2017

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Abstract

Objectives

Bioactive proteins are sequestered in human dentine and play a decisive role in dental pulp regeneration and repair. They can be released and exposed on the dentine surface by acids, but also chelators, such as ethylenediaminetetraacetic acid (EDTA). The objectives of this study were (i) to evaluate whether ultrasonic activation of irrigants in the root canal will promote growth factor release from dentine and (ii) to collect bioactive proteins in a physiological solution.

Materials and methods

Human dentine disks underwent irrigation with and without ultrasonic activation. The protocols included treatment by either a single or two consecutive steps with 10 % EDTA and phosphate-buffered saline (PBS), where each sample was treated three times. To mimic clinical conditions, selected irrigation regimens were applied to root canals of extracted human teeth after preparation. Amounts of transforming growth factor β1 (TGF-β1) in solution were quantified using enzyme-linked immunosorbent assays. Nonparametric statistical analysis was performed to compare different groups as well as repetitions within a group (Mann-Whitney U test, α = 0.05). Additionally, morphological changes of dentine surfaces were visualized by scanning electron microscopy (SEM).

Results

TGF-β1 was not detectable after irrigation of dentine with PBS, neither with nor without ultrasonic activation. Irrigation with EDTA released TGF-β1, and ultrasonic activation of EDTA enhanced this effect. However, preceding EDTA conditioning enabled the release of bioactive proteins into PBS solution. Similar results were observed in dentine disks and root canals. Visualization of dentine surfaces after different treatment revealed superficial erosion after ultrasonic activation irrespective of the irrigant solution, but different degrees of exposure of organic substance.

Conclusions

Ultrasonic activation enhances growth factor release from human dentine. Bioactive proteins can be isolated in physiological solvents and may act as autologous supplements for regenerative endodontic treatment or pulp tissue engineering.

Clinical relevance

Autologous growth factors from human dentine can advance treatment strategies in dental pulp tissue engineering.
Literatur
1.
Zurück zum Zitat Smith AJ, Scheven BA, Takahashi Y, et al. (2012) Dentine as a bioactive extracellular matrix. Arch Oral Biol 57:109–121CrossRefPubMed Smith AJ, Scheven BA, Takahashi Y, et al. (2012) Dentine as a bioactive extracellular matrix. Arch Oral Biol 57:109–121CrossRefPubMed
2.
Zurück zum Zitat Goldberg M, Smith AJ (2004) Cells and extracellular matrices of dentin and pulp: a biological basis for repair and tissue engineering. Crit Rev Oral Biol Med 15:13–27CrossRefPubMed Goldberg M, Smith AJ (2004) Cells and extracellular matrices of dentin and pulp: a biological basis for repair and tissue engineering. Crit Rev Oral Biol Med 15:13–27CrossRefPubMed
3.
Zurück zum Zitat Dung SZ, Gregory RL, Li Y, Stookey GK (1995) Effect of lactic acid and proteolytic enzymes on the release of organic matrix components from human root dentin. Caries Res 29:483–489CrossRefPubMed Dung SZ, Gregory RL, Li Y, Stookey GK (1995) Effect of lactic acid and proteolytic enzymes on the release of organic matrix components from human root dentin. Caries Res 29:483–489CrossRefPubMed
4.
Zurück zum Zitat Tomson PL, Grover LM, Lumley PJ, et al. (2007) Dissolution of bio-active dentine matrix components by mineral trioxide aggregate. J Dent 35:636–642CrossRefPubMed Tomson PL, Grover LM, Lumley PJ, et al. (2007) Dissolution of bio-active dentine matrix components by mineral trioxide aggregate. J Dent 35:636–642CrossRefPubMed
5.
Zurück zum Zitat Graham LW, Cooper PR, Cassidy N, et al. (2006) The effect of calcium hydroxide on solubilisation of bio-active dentine matrix components. Biomaterials 27:2865–2873CrossRefPubMed Graham LW, Cooper PR, Cassidy N, et al. (2006) The effect of calcium hydroxide on solubilisation of bio-active dentine matrix components. Biomaterials 27:2865–2873CrossRefPubMed
6.
Zurück zum Zitat Ferracane JL, Cooper PR, Smith AJ (2013) Dentin matrix component solubilization by solutions of pH relevant to self-etching dental adhesives. J Adhes Dent 15:407–412PubMed Ferracane JL, Cooper PR, Smith AJ (2013) Dentin matrix component solubilization by solutions of pH relevant to self-etching dental adhesives. J Adhes Dent 15:407–412PubMed
7.
Zurück zum Zitat Galler KM, Widbiller M, Buchalla W, et al. (2015) EDTA conditioning of dentine promotes adhesion, migration and differentiation of dental pulp stem cells. Int Endod J. doi:10.1111/iej.12492 Galler KM, Widbiller M, Buchalla W, et al. (2015) EDTA conditioning of dentine promotes adhesion, migration and differentiation of dental pulp stem cells. Int Endod J. doi:10.​1111/​iej.​12492
9.
Zurück zum Zitat Chun SY, Lee HJ, Choi YA, et al. (2011) Analysis of the soluble human tooth proteome and its ability to induce dentin/tooth regeneration. Tissue Eng Part A 17:181–191CrossRefPubMed Chun SY, Lee HJ, Choi YA, et al. (2011) Analysis of the soluble human tooth proteome and its ability to induce dentin/tooth regeneration. Tissue Eng Part A 17:181–191CrossRefPubMed
10.
Zurück zum Zitat Baker SM, Sugars RV, Wendel M, et al. (2009) TGF-beta/extracellular matrix interactions in dentin matrix: a role in regulating sequestration and protection of bioactivity. Calcif Tissue Int 85:66–74CrossRefPubMed Baker SM, Sugars RV, Wendel M, et al. (2009) TGF-beta/extracellular matrix interactions in dentin matrix: a role in regulating sequestration and protection of bioactivity. Calcif Tissue Int 85:66–74CrossRefPubMed
11.
Zurück zum Zitat Finkelman RD, Mohan S, Jennings JC, et al. (1990) Quantitation of growth factors IGF-I, SGF/IGF-II, and TGF-beta in human dentin. J Bone Miner Res 5:717–723CrossRefPubMed Finkelman RD, Mohan S, Jennings JC, et al. (1990) Quantitation of growth factors IGF-I, SGF/IGF-II, and TGF-beta in human dentin. J Bone Miner Res 5:717–723CrossRefPubMed
12.
Zurück zum Zitat Roberts-Clark DJ, Smith AJ (2000) Angiogenic growth factors in human dentine matrix. Arch Oral Biol 45:1013–1016CrossRefPubMed Roberts-Clark DJ, Smith AJ (2000) Angiogenic growth factors in human dentine matrix. Arch Oral Biol 45:1013–1016CrossRefPubMed
13.
Zurück zum Zitat Zhang R, Smith AJ, Cooper PR, et al. (2011) Angiogenic activity of dentin matrix components. J Endod 37:26–30CrossRefPubMed Zhang R, Smith AJ, Cooper PR, et al. (2011) Angiogenic activity of dentin matrix components. J Endod 37:26–30CrossRefPubMed
14.
Zurück zum Zitat Suzuki T, Lee CH, Chen M, et al. (2011) Induced migration of dental pulp stem cells for in vivo pulp regeneration. J Dent Res 90:1013–1018CrossRefPubMed Suzuki T, Lee CH, Chen M, et al. (2011) Induced migration of dental pulp stem cells for in vivo pulp regeneration. J Dent Res 90:1013–1018CrossRefPubMed
15.
Zurück zum Zitat Howard C, Murray PE, Namerow KN (2010) Dental pulp stem cell migration. J Endod 36:1963–1966CrossRefPubMed Howard C, Murray PE, Namerow KN (2010) Dental pulp stem cell migration. J Endod 36:1963–1966CrossRefPubMed
16.
Zurück zum Zitat Tziafas D, Alvanou A, Panagiotakopoulos N, et al. (1995) Induction of odontoblast-like cell differentiation in dog dental pulps after in vivo implantation of dentine matrix components. Arch Oral Biol 40:883–893CrossRefPubMed Tziafas D, Alvanou A, Panagiotakopoulos N, et al. (1995) Induction of odontoblast-like cell differentiation in dog dental pulps after in vivo implantation of dentine matrix components. Arch Oral Biol 40:883–893CrossRefPubMed
17.
Zurück zum Zitat He H, Yu J, Liu Y, et al. (2008) Effects of FGF2 and TGFbeta1 on the differentiation of human dental pulp stem cells in vitro. Cell Biol Int 32:827–834CrossRefPubMed He H, Yu J, Liu Y, et al. (2008) Effects of FGF2 and TGFbeta1 on the differentiation of human dental pulp stem cells in vitro. Cell Biol Int 32:827–834CrossRefPubMed
18.
Zurück zum Zitat Melin M, Joffre-Romeas A, Farges JC, et al. (2000) Effects of TGFbeta1 on dental pulp cells in cultured human tooth slices. J Dent Res 79:1689–1696CrossRefPubMed Melin M, Joffre-Romeas A, Farges JC, et al. (2000) Effects of TGFbeta1 on dental pulp cells in cultured human tooth slices. J Dent Res 79:1689–1696CrossRefPubMed
19.
Zurück zum Zitat Kalyva M, Papadimitriou S, Tziafas D (2010) Transdentinal stimulation of tertiary dentine formation and intratubular mineralization by growth factors. Int Endod J 43:382–392CrossRefPubMed Kalyva M, Papadimitriou S, Tziafas D (2010) Transdentinal stimulation of tertiary dentine formation and intratubular mineralization by growth factors. Int Endod J 43:382–392CrossRefPubMed
20.
Zurück zum Zitat Mullane EM, Dong Z, Sedgley CM, et al. (2008) Effects of VEGF and FGF2 on the revascularization of severed human dental pulps. J Dent Res 87:1144–1148CrossRefPubMed Mullane EM, Dong Z, Sedgley CM, et al. (2008) Effects of VEGF and FGF2 on the revascularization of severed human dental pulps. J Dent Res 87:1144–1148CrossRefPubMed
21.
Zurück zum Zitat Galler KM, D'Souza RN, Federlin M, et al. (2011) Dentin conditioning codetermines cell fate in regenerative endodontics. J Endod 37:1536–1541CrossRefPubMed Galler KM, D'Souza RN, Federlin M, et al. (2011) Dentin conditioning codetermines cell fate in regenerative endodontics. J Endod 37:1536–1541CrossRefPubMed
22.
Zurück zum Zitat Lovelace TW, Henry MA, Hargreaves KM, Diogenes AR (2011) Evaluation of the delivery of mesenchymal stem cells into the root canal space of necrotic immature teeth after clinical regenerative endodontic procedure. J Endod 37:133–138CrossRefPubMed Lovelace TW, Henry MA, Hargreaves KM, Diogenes AR (2011) Evaluation of the delivery of mesenchymal stem cells into the root canal space of necrotic immature teeth after clinical regenerative endodontic procedure. J Endod 37:133–138CrossRefPubMed
23.
Zurück zum Zitat Chrepa V, Henry MA, Daniel BJ, Diogenes AR (2015) Delivery of apical mesenchymal stem cells into root canals of mature teeth. J Dent Res 94:1653–1659CrossRefPubMed Chrepa V, Henry MA, Daniel BJ, Diogenes AR (2015) Delivery of apical mesenchymal stem cells into root canals of mature teeth. J Dent Res 94:1653–1659CrossRefPubMed
24.
Zurück zum Zitat Casagrande L, Demarco FF, Zhang Z, et al. (2010) Dentin-derived BMP-2 and odontoblast differentiation. J Dent Res 89:603–608CrossRefPubMed Casagrande L, Demarco FF, Zhang Z, et al. (2010) Dentin-derived BMP-2 and odontoblast differentiation. J Dent Res 89:603–608CrossRefPubMed
25.
Zurück zum Zitat Pang NS, Lee SJ, Kim E, et al. (2014) Effect of EDTA on attachment and differentiation of dental pulp stem cells. J Endod 40:811–817CrossRefPubMed Pang NS, Lee SJ, Kim E, et al. (2014) Effect of EDTA on attachment and differentiation of dental pulp stem cells. J Endod 40:811–817CrossRefPubMed
26.
Zurück zum Zitat Carreira AC, Lojudice FH, Halcsik E, et al. (2014) Bone morphogenetic proteins: facts, challenges, and future perspectives. J Dent Res 93:335–345CrossRefPubMed Carreira AC, Lojudice FH, Halcsik E, et al. (2014) Bone morphogenetic proteins: facts, challenges, and future perspectives. J Dent Res 93:335–345CrossRefPubMed
27.
Zurück zum Zitat Lin LM, Ricucci D, Huang GTJ (2014) Regeneration of the dentine-pulp complex with revitalization/revascularization therapy: challenges and hopes. Int Endod J 47:713–724CrossRefPubMed Lin LM, Ricucci D, Huang GTJ (2014) Regeneration of the dentine-pulp complex with revitalization/revascularization therapy: challenges and hopes. Int Endod J 47:713–724CrossRefPubMed
28.
Zurück zum Zitat Diogenes AR, Henry MA, Teixeira FB, Hargreaves KM (2013) An update on clinical regenerative endodontics. Endod Top 28:2–23CrossRef Diogenes AR, Henry MA, Teixeira FB, Hargreaves KM (2013) An update on clinical regenerative endodontics. Endod Top 28:2–23CrossRef
29.
Zurück zum Zitat Becerra P, Ricucci D, Loghin S, et al. (2014) Histologic study of a human immature permanent premolar with chronic apical abscess after revascularization/revitalization. J Endod 40:133–139CrossRefPubMed Becerra P, Ricucci D, Loghin S, et al. (2014) Histologic study of a human immature permanent premolar with chronic apical abscess after revascularization/revitalization. J Endod 40:133–139CrossRefPubMed
30.
Zurück zum Zitat Ballal NV, Kundabala M, Bhat S, et al. (2009) A comparative in vitro evaluation of cytotoxic effects of EDTA and maleic acid: root canal irrigants. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 108:633–638CrossRefPubMed Ballal NV, Kundabala M, Bhat S, et al. (2009) A comparative in vitro evaluation of cytotoxic effects of EDTA and maleic acid: root canal irrigants. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 108:633–638CrossRefPubMed
31.
Zurück zum Zitat Malheiros CF, Marques MM, Gavini G (2005) In vitro evaluation of the cytotoxic effects of acid solutions used as canal irrigants. J Endod 31:746–748CrossRefPubMed Malheiros CF, Marques MM, Gavini G (2005) In vitro evaluation of the cytotoxic effects of acid solutions used as canal irrigants. J Endod 31:746–748CrossRefPubMed
32.
Zurück zum Zitat Sceiza MF, Daniel RL, Santos EM, Jaeger MM (2001) Cytotoxic effects of 10 % citric acid and EDTA-T used as root canal irrigants: an in vitro analysis. J Endod 27:741–743CrossRefPubMed Sceiza MF, Daniel RL, Santos EM, Jaeger MM (2001) Cytotoxic effects of 10 % citric acid and EDTA-T used as root canal irrigants: an in vitro analysis. J Endod 27:741–743CrossRefPubMed
33.
Zurück zum Zitat Schmidt TF, Teixeira CS, Felippe MCS, et al. (2015) Effect of ultrasonic activation of irrigants on smear layer removal. J Endod 41:1359–1363CrossRefPubMed Schmidt TF, Teixeira CS, Felippe MCS, et al. (2015) Effect of ultrasonic activation of irrigants on smear layer removal. J Endod 41:1359–1363CrossRefPubMed
34.
Zurück zum Zitat Mozo S, Llena C, Forner L (2012) Review of ultrasonic irrigation in endodontics: increasing action of irrigating solutions. Med Oral Patol Oral Cir Bucal 17:e512–e516CrossRefPubMed Mozo S, Llena C, Forner L (2012) Review of ultrasonic irrigation in endodontics: increasing action of irrigating solutions. Med Oral Patol Oral Cir Bucal 17:e512–e516CrossRefPubMed
35.
Zurück zum Zitat Guerisoli DMZ, Marchesan MA, Walmsley AD, et al. (2002) Evaluation of smear layer removal by EDTAC and sodium hypochlorite with ultrasonic agitation. Int Endod J 35:418–421CrossRefPubMed Guerisoli DMZ, Marchesan MA, Walmsley AD, et al. (2002) Evaluation of smear layer removal by EDTAC and sodium hypochlorite with ultrasonic agitation. Int Endod J 35:418–421CrossRefPubMed
36.
Zurück zum Zitat Lui JN, Kuah HG, Chen NN (2007) Effect of EDTA with and without surfactants or ultrasonics on removal of smear layer. J Endod 33:472–475CrossRefPubMed Lui JN, Kuah HG, Chen NN (2007) Effect of EDTA with and without surfactants or ultrasonics on removal of smear layer. J Endod 33:472–475CrossRefPubMed
37.
Zurück zum Zitat Castagna F, Rizzon P, da Rosa RA, et al. (2013) Effect of passive ultrassonic instrumentation as a final irrigation protocol on debris and smear layer removal—a SEM analysis. Microsc Res Tech 76:496–502CrossRefPubMed Castagna F, Rizzon P, da Rosa RA, et al. (2013) Effect of passive ultrassonic instrumentation as a final irrigation protocol on debris and smear layer removal—a SEM analysis. Microsc Res Tech 76:496–502CrossRefPubMed
38.
Zurück zum Zitat Ruggeri A, Prati C, Mazzoni A, et al. (2007) Effects of citric acid and EDTA conditioning on exposed root dentin: an immunohistochemical analysis of collagen and proteoglycans. Arch Oral Biol 52:1–8CrossRefPubMed Ruggeri A, Prati C, Mazzoni A, et al. (2007) Effects of citric acid and EDTA conditioning on exposed root dentin: an immunohistochemical analysis of collagen and proteoglycans. Arch Oral Biol 52:1–8CrossRefPubMed
39.
Zurück zum Zitat Higashi T, Okamoto H (1995) The effect of ultrasonic irrigation before and after citric acid treatment on collagen fibril exposure: an in vitro SEM study. J Periodontol 66:887–891CrossRefPubMed Higashi T, Okamoto H (1995) The effect of ultrasonic irrigation before and after citric acid treatment on collagen fibril exposure: an in vitro SEM study. J Periodontol 66:887–891CrossRefPubMed
40.
Zurück zum Zitat Smith AJ, Duncan HF, Diogenes AR, et al. (2016) Exploiting the bioactive properties of the dentin-pulp complex in regenerative endodontics. J Endod 42:47–56CrossRefPubMed Smith AJ, Duncan HF, Diogenes AR, et al. (2016) Exploiting the bioactive properties of the dentin-pulp complex in regenerative endodontics. J Endod 42:47–56CrossRefPubMed
41.
Zurück zum Zitat Galler KM, Buchalla W, Hiller KA, et al. (2015) Influence of root canal disinfectants on growth factor release from dentin. J Endod 41:363–368CrossRefPubMed Galler KM, Buchalla W, Hiller KA, et al. (2015) Influence of root canal disinfectants on growth factor release from dentin. J Endod 41:363–368CrossRefPubMed
42.
Zurück zum Zitat Galler KM, Eidt A, Schmalz G (2014) Cell-free approaches for dental pulp tissue engineering. J Endod 40:S41–S45CrossRefPubMed Galler KM, Eidt A, Schmalz G (2014) Cell-free approaches for dental pulp tissue engineering. J Endod 40:S41–S45CrossRefPubMed
43.
Zurück zum Zitat Diogenes AR, Ruparel NB, Teixeira FB, Hargreaves KM (2014) Translational science in disinfection for regenerative endodontics. J Endod 40:S52–S57CrossRefPubMed Diogenes AR, Ruparel NB, Teixeira FB, Hargreaves KM (2014) Translational science in disinfection for regenerative endodontics. J Endod 40:S52–S57CrossRefPubMed
44.
Zurück zum Zitat Althumairy RI, Teixeira FB, Diogenes AR (2014) Effect of dentin conditioning with intracanal medicaments on survival of stem cells of apical papilla. J Endod 40:521–525CrossRefPubMed Althumairy RI, Teixeira FB, Diogenes AR (2014) Effect of dentin conditioning with intracanal medicaments on survival of stem cells of apical papilla. J Endod 40:521–525CrossRefPubMed
45.
Zurück zum Zitat Carrigan PJ, Morse DR, Furst ML, Sinai IH (1984) A scanning electron microscopic evaluation of human dentinal tubules according to age and location. J Endod 10:359–363CrossRefPubMed Carrigan PJ, Morse DR, Furst ML, Sinai IH (1984) A scanning electron microscopic evaluation of human dentinal tubules according to age and location. J Endod 10:359–363CrossRefPubMed
46.
Zurück zum Zitat Radlanski RJ (2011) Curriculum Orale Struktur- und Entwicklungsbiologie. Quintessenz, Berlin Radlanski RJ (2011) Curriculum Orale Struktur- und Entwicklungsbiologie. Quintessenz, Berlin
47.
Zurück zum Zitat de Gregorio C, Arias A, Navarrete N, et al. (2013) Effect of apical size and taper on volume of irrigant delivered at working length with apical negative pressure at different root curvatures. J Endod 39:119–124CrossRefPubMed de Gregorio C, Arias A, Navarrete N, et al. (2013) Effect of apical size and taper on volume of irrigant delivered at working length with apical negative pressure at different root curvatures. J Endod 39:119–124CrossRefPubMed
48.
Zurück zum Zitat Tay FR, Gu LS, Schoeffel GJ, et al. (2010) Effect of vapor lock on root canal debridement by using a side-vented needle for positive-pressure irrigant delivery. J Endod 36:745–750CrossRefPubMedPubMedCentral Tay FR, Gu LS, Schoeffel GJ, et al. (2010) Effect of vapor lock on root canal debridement by using a side-vented needle for positive-pressure irrigant delivery. J Endod 36:745–750CrossRefPubMedPubMedCentral
49.
Zurück zum Zitat Lumley PJ, Walmsley AD, Laird WR (1991) Streaming patterns produced around endosonic files. Int Endod J 24:290–297CrossRefPubMed Lumley PJ, Walmsley AD, Laird WR (1991) Streaming patterns produced around endosonic files. Int Endod J 24:290–297CrossRefPubMed
50.
Zurück zum Zitat Ahmad M, Pitt Ford TJ, Crum LA (1987) Ultrasonic debridement of root canals: acoustic streaming and its possible role. J Endod 13:490–499CrossRefPubMed Ahmad M, Pitt Ford TJ, Crum LA (1987) Ultrasonic debridement of root canals: acoustic streaming and its possible role. J Endod 13:490–499CrossRefPubMed
51.
Zurück zum Zitat Niu W, Yoshioka T, Kobayashi C, Suda H (2002) A scanning electron microscopic study of dentinal erosion by final irrigation with EDTA and NaOCl solutions. Int Endod J 35:934–939CrossRefPubMed Niu W, Yoshioka T, Kobayashi C, Suda H (2002) A scanning electron microscopic study of dentinal erosion by final irrigation with EDTA and NaOCl solutions. Int Endod J 35:934–939CrossRefPubMed
Metadaten
Titel
Ultrasonic activation of irrigants increases growth factor release from human dentine
verfasst von
M. Widbiller
A. Eidt
K.-A. Hiller
W. Buchalla
G. Schmalz
K. M. Galler
Publikationsdatum
25.04.2016
Verlag
Springer Berlin Heidelberg
Erschienen in
Clinical Oral Investigations / Ausgabe 3/2017
Print ISSN: 1432-6981
Elektronische ISSN: 1436-3771
DOI
https://doi.org/10.1007/s00784-016-1824-1

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