Skip to main content
Erschienen in: Clinical Oral Investigations 2/2014

01.03.2014 | Original Article

Behaviour of co-cultured human osteoclastic and osteoblastic cells exposed to endodontic sealers’ extracts

verfasst von: Cláudia Rodrigues, João Costa-Rodrigues, José A. Capelas, Maria H. Fernandes

Erschienen in: Clinical Oral Investigations | Ausgabe 2/2014

Einloggen, um Zugang zu erhalten

Abstract

Objectives

Bone tissue is constantly being moulded and shaped by the coordinated action of bone-resorbing osteoclasts and bone-synthesizing osteoblasts. This study addresses the long-term effects of endodontic sealers’ extracts (AH Plus™, GuttaFlow™, Tubliseal™, Sealapex™ and RealSeal™) on co-cultures of human osteoclastic and osteoblastic cells.

Methods

The sealers were mixed according to the manufacturer’s instructions, freshly extracted with culture medium (1.3 cm2/ml; 24 h; 37 °C, 5 % CO2/air) and diluted (1:20–1:2,500). Co-cultures of osteoclastic and osteoblastic cells, established from precursors present in human peripheral blood mononuclear cells and bone marrow cells, respectively, were exposed to the extracts for 21 days. Co-cultures were characterized for the osteoclastic and osteoblastic response.

Results

The sealers caused a dose-dependent decrease on TRAP and ALP activities, respectively, an osteoclastic and an osteoblastic marker. The resorbing ability of the osteoclastic cells and the expression of osteoclastic and osteoblastic genes were also decreased; in addition, the extracts affected several intracellular signalling pathways. Inhibition was higher during the two first weeks, followed by adaptive cell responses. Osteoblastic response was more sensitive to the extracts’ toxicity and showed lower adaptive ability.

Significance

A correlation to the clinical situation cannot be predicted; however, the results suggest that the sealers’ eluents might disrupt the highly regulated interaction between osteoblastic and osteoclastic cells, compromising the local bone metabolism. Also, the higher susceptibility of the osteoblastic response might be particularly relevant in the initial stages of the healing of periapical lesions, due to the significant role of the bone formation events.
Literatur
1.
Zurück zum Zitat Hauman CH, Love RM (2003) Biocompatibility of dental materials used in contemporary endodontic therapy: a review. Part 2. Root-canal-filling materials. Int Endod J 36:147–160PubMedCrossRef Hauman CH, Love RM (2003) Biocompatibility of dental materials used in contemporary endodontic therapy: a review. Part 2. Root-canal-filling materials. Int Endod J 36:147–160PubMedCrossRef
2.
Zurück zum Zitat Huang FM, Chou MY, Chang YC (2003) Induction of cyclooxygenase-2 mRNA and protein expression by epoxy resin and zinc oxide-eugenol based root canal sealers in human osteoblastic cells. Biomaterials 24:1869–1875PubMedCrossRef Huang FM, Chou MY, Chang YC (2003) Induction of cyclooxygenase-2 mRNA and protein expression by epoxy resin and zinc oxide-eugenol based root canal sealers in human osteoblastic cells. Biomaterials 24:1869–1875PubMedCrossRef
3.
Zurück zum Zitat Huang FM, Tsai CH, Yang SF, Chang YC (2005) Induction of interleukin-6 and interleukin-8 gene expression by root canal sealers in human osteoblastic cells. J Endod 31:679–683PubMedCrossRef Huang FM, Tsai CH, Yang SF, Chang YC (2005) Induction of interleukin-6 and interleukin-8 gene expression by root canal sealers in human osteoblastic cells. J Endod 31:679–683PubMedCrossRef
4.
Zurück zum Zitat Huang FM, Yang SF, Chang YC (2008) Up-regulation of gelatinases and tissue type plasminogen activator by root canal sealers in human osteoblastic cells. J Endod 34:291–294PubMedCrossRef Huang FM, Yang SF, Chang YC (2008) Up-regulation of gelatinases and tissue type plasminogen activator by root canal sealers in human osteoblastic cells. J Endod 34:291–294PubMedCrossRef
5.
Zurück zum Zitat Huang FM, Lee SS, Yang SF, Chang YC (2009) Up-regulation of receptor activator nuclear factor-kappaB ligand expression by root canal sealers in human osteoblastic cells. J Endod 35:363–366PubMedCrossRef Huang FM, Lee SS, Yang SF, Chang YC (2009) Up-regulation of receptor activator nuclear factor-kappaB ligand expression by root canal sealers in human osteoblastic cells. J Endod 35:363–366PubMedCrossRef
6.
Zurück zum Zitat Huang FM, Yang SF, Chang YC (2010) Effects of root canal sealers on alkaline phosphatase in human osteoblastic cells. J Endod 36:1230–1233PubMedCrossRef Huang FM, Yang SF, Chang YC (2010) Effects of root canal sealers on alkaline phosphatase in human osteoblastic cells. J Endod 36:1230–1233PubMedCrossRef
7.
Zurück zum Zitat Xu P, Liang J, Dong G, Zheng L, Ye L (2010) Cytotoxicity of RealSeal on human osteoblast-like MG63 cells. J Endod 36:40–44PubMedCrossRef Xu P, Liang J, Dong G, Zheng L, Ye L (2010) Cytotoxicity of RealSeal on human osteoblast-like MG63 cells. J Endod 36:40–44PubMedCrossRef
8.
Zurück zum Zitat Lee DH, Lim BS, Lee YK, Yang HC (2007) Mechanisms of root canal sealers cytotoxicity on osteoblastic cell line MC3T3-E1. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 104:717–721PubMedCrossRef Lee DH, Lim BS, Lee YK, Yang HC (2007) Mechanisms of root canal sealers cytotoxicity on osteoblastic cell line MC3T3-E1. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 104:717–721PubMedCrossRef
9.
Zurück zum Zitat Ames JM, Loushine RJ, Babb BR, Bryan TE, Lockwood PE, Sui M, Roberts S, Weller RN, Pashley DH, Tay FR (2009) Contemporary methacrylate resin-based root canal sealers exhibit different degrees of ex vivo cytotoxicity when cured in their self-cured mode. J Endod 35:225–228PubMedCrossRef Ames JM, Loushine RJ, Babb BR, Bryan TE, Lockwood PE, Sui M, Roberts S, Weller RN, Pashley DH, Tay FR (2009) Contemporary methacrylate resin-based root canal sealers exhibit different degrees of ex vivo cytotoxicity when cured in their self-cured mode. J Endod 35:225–228PubMedCrossRef
10.
Zurück zum Zitat Brackett MG, Messer RL, Lockwood PE, Bryan TE, Lewis JB, Bouillaguet S, Wataha JC (2010) Cytotoxic response of three cell lines exposed in vitro to dental endodontic sealers. J Biomed Mater Res B Appl Biomater 95:380–386PubMedCrossRef Brackett MG, Messer RL, Lockwood PE, Bryan TE, Lewis JB, Bouillaguet S, Wataha JC (2010) Cytotoxic response of three cell lines exposed in vitro to dental endodontic sealers. J Biomed Mater Res B Appl Biomater 95:380–386PubMedCrossRef
11.
Zurück zum Zitat Bryan TE, Khechen K, Brackett MG, Messer RL, El-Awady A, Primus CM, Gutmann JL, Tay FR (2010) In vitro osteogenic potential of an experimental calcium silicate-based root canal sealer. J Endod 36:1163–1169PubMedCrossRef Bryan TE, Khechen K, Brackett MG, Messer RL, El-Awady A, Primus CM, Gutmann JL, Tay FR (2010) In vitro osteogenic potential of an experimental calcium silicate-based root canal sealer. J Endod 36:1163–1169PubMedCrossRef
12.
Zurück zum Zitat Al-Awadhi S, Spears R, Gutmann JL, Opperman LA (2004) Cultured primary osteoblast viability and apoptosis in the presence of root canal sealers. J Endod 30:527–533PubMedCrossRef Al-Awadhi S, Spears R, Gutmann JL, Opperman LA (2004) Cultured primary osteoblast viability and apoptosis in the presence of root canal sealers. J Endod 30:527–533PubMedCrossRef
13.
Zurück zum Zitat Datta HK, Ng WF, Walker JA, Tuck SP, Varanasi SS (2008) The cell biology of bone metabolism. J Clin Pathol 61:577–587PubMedCrossRef Datta HK, Ng WF, Walker JA, Tuck SP, Varanasi SS (2008) The cell biology of bone metabolism. J Clin Pathol 61:577–587PubMedCrossRef
14.
Zurück zum Zitat Matsuo K, Irie N (2008) Osteoclast-osteoblast communication. Arch Biochem Biophys 473:201–209PubMedCrossRef Matsuo K, Irie N (2008) Osteoclast-osteoblast communication. Arch Biochem Biophys 473:201–209PubMedCrossRef
15.
Zurück zum Zitat Heinemann C, Heinemann S, Worch H, Hanke T (2011) Development of an osteoblast/osteoclast co-culture derived by human bone marrow stromal cells and human monocytes for biomaterials testing. Eur Cell Mater 21:80–93PubMed Heinemann C, Heinemann S, Worch H, Hanke T (2011) Development of an osteoblast/osteoclast co-culture derived by human bone marrow stromal cells and human monocytes for biomaterials testing. Eur Cell Mater 21:80–93PubMed
16.
Zurück zum Zitat Costa-Rodrigues J, Fernandes A, Fernandes MH (2011) Reciprocal osteoblastic and osteoclastic modulation in co-cultured MG63 osteosarcoma cells and human osteoclast precursors. J Cell Biochem 112:3704–3713PubMedCrossRef Costa-Rodrigues J, Fernandes A, Fernandes MH (2011) Reciprocal osteoblastic and osteoclastic modulation in co-cultured MG63 osteosarcoma cells and human osteoclast precursors. J Cell Biochem 112:3704–3713PubMedCrossRef
17.
Zurück zum Zitat International Organization for Standardization (2009) Biological evaluation of medical devices—part 5: tests for in vitro cytotoxicity. ISO 10993–5, Geneva, Switzerland International Organization for Standardization (2009) Biological evaluation of medical devices—part 5: tests for in vitro cytotoxicity. ISO 10993–5, Geneva, Switzerland
18.
Zurück zum Zitat Coelho MJ, Fernandes MH (2000) Human bone cell cultures in biocompatibility testing. Part II: effect of ascorbic acid, beta-glycerophosphate and dexamethasone on osteoblastic differentiation. Biomaterials 21:1095–1102PubMedCrossRef Coelho MJ, Fernandes MH (2000) Human bone cell cultures in biocompatibility testing. Part II: effect of ascorbic acid, beta-glycerophosphate and dexamethasone on osteoblastic differentiation. Biomaterials 21:1095–1102PubMedCrossRef
19.
Zurück zum Zitat Vaananen HK, Laitala-Leinonen T (2008) Osteoclast lineage and function. Arch Biochem Biophys 473:132–138PubMedCrossRef Vaananen HK, Laitala-Leinonen T (2008) Osteoclast lineage and function. Arch Biochem Biophys 473:132–138PubMedCrossRef
20.
Zurück zum Zitat Zhao Q, Shao J, Chen W, Li YP (2007) Osteoclast differentiation and gene regulation. Front Biosci 12:2519–2529PubMedCrossRef Zhao Q, Shao J, Chen W, Li YP (2007) Osteoclast differentiation and gene regulation. Front Biosci 12:2519–2529PubMedCrossRef
21.
Zurück zum Zitat Lian JB, Stein GS, Javed A, van Wijnen AJ, Stein JL, Montecino M, Hassan MQ, Gaur T, Lengner CJ, Young DW (2006) Networks and hubs for the transcriptional control of osteoblastogenesis. Rev Endocr Metab Disord 7:1–16PubMedCrossRef Lian JB, Stein GS, Javed A, van Wijnen AJ, Stein JL, Montecino M, Hassan MQ, Gaur T, Lengner CJ, Young DW (2006) Networks and hubs for the transcriptional control of osteoblastogenesis. Rev Endocr Metab Disord 7:1–16PubMedCrossRef
22.
Zurück zum Zitat Costa-Rodrigues J, Fernandes A, Fernandes MH (2011) Spontaneous and induced osteoclastogenic behaviour of human peripheral blood mononuclear cells and their CD14(+) and CD14(−) cell fractions. Cell Prolif 44:410–419PubMedCrossRef Costa-Rodrigues J, Fernandes A, Fernandes MH (2011) Spontaneous and induced osteoclastogenic behaviour of human peripheral blood mononuclear cells and their CD14(+) and CD14(−) cell fractions. Cell Prolif 44:410–419PubMedCrossRef
Metadaten
Titel
Behaviour of co-cultured human osteoclastic and osteoblastic cells exposed to endodontic sealers’ extracts
verfasst von
Cláudia Rodrigues
João Costa-Rodrigues
José A. Capelas
Maria H. Fernandes
Publikationsdatum
01.03.2014
Verlag
Springer Berlin Heidelberg
Erschienen in
Clinical Oral Investigations / Ausgabe 2/2014
Print ISSN: 1432-6981
Elektronische ISSN: 1436-3771
DOI
https://doi.org/10.1007/s00784-013-0983-6

Weitere Artikel der Ausgabe 2/2014

Clinical Oral Investigations 2/2014 Zur Ausgabe

Newsletter

Bestellen Sie unseren kostenlosen Newsletter Update Zahnmedizin und bleiben Sie gut informiert – ganz bequem per eMail.