Skip to main content
Top
Published in: Clinical Oral Investigations 1/2012

01-02-2012 | Original Article

Human saliva exposure modulates bone cell performance in vitro

Authors: Susanne Proksch, Thorsten Steinberg, Constantin Keller, Martin Wolkewitz, Margit Wiedmann-Al-Ahmad, Guenter Finkenzeller, Christian Hannig, Elmar Hellwig, Ali Al-Ahmad

Published in: Clinical Oral Investigations | Issue 1/2012

Login to get access

Abstract

Various situations encountered by a clinician during the daily routine including surgical periodontitis therapy, dental implant insertion, or tooth extraction involve the contact of saliva with the jaw bone. However, there are only sparse data concerning the influence of saliva on bone cells. Saliva specimens were incorporated within culture medium and administered to murine MC3T3 osteoblasts, of which the morphology (REM), proliferation (EZ4U), and differentiation (qRT-PCR, alkaline phosphatase activity, extracellular matrix calcification) were assessed. Simultaneously, the composition of saliva media was analyzed with respect to the content of lactoferrin, activities of classical salivary enzymes, and the ability to provoke inflammatory cytokine production (enzyme-linked immunosorbent assay) in MC3T3 osteoblasts. The morphology, proliferation, and expression of differentiation-associated genes were seriously handicapped by saliva contact. Saliva-touched cells exhibited less alkaline phosphatase but normal levels of extracellular matrix mineralization. Saliva-containing culture media featured physiological activities of salivary enzymes and considerable amounts of lactoferrin but almost completely lacked salivary alkaline phosphatase and unspecific proteases. Upon saliva incubation, MC3T3 osteoblasts did not release noteworthy levels of interleukin-1beta or tumor necrosis factor alpha. Although saliva is generally considered to vitalize oral tissues, this study reveals that it harms osteoblast-like cells more due to the presence of salivary enzymes than by triggering of inflammation. This issue is clinically relevant because it broadens the understanding of the bone cell fate within the rather complex cosmos of the oral cavity thereby providing a basis for clinical decision making and treatment guidelines. It seems to be reasonable to restrict the contact period between saliva and bone.
Appendix
Available only for authorised users
Literature
1.
2.
4.
go back to reference Hakkinen L, Uitto VJ, Larjava H (2000) Cell biology of gingival wound healing. Periodontol 2000 24:127–152PubMedCrossRef Hakkinen L, Uitto VJ, Larjava H (2000) Cell biology of gingival wound healing. Periodontol 2000 24:127–152PubMedCrossRef
5.
go back to reference Tenovuo J (1998) Antimicrobial function of human saliva—how important is it for oral health? Acta Odontol Scand 56(5):250–256PubMedCrossRef Tenovuo J (1998) Antimicrobial function of human saliva—how important is it for oral health? Acta Odontol Scand 56(5):250–256PubMedCrossRef
6.
go back to reference Dawes C (2008) Salivary flow patterns and the health of hard and soft oral tissues. J Am Dent Assoc 139(Suppl):18S–24SPubMed Dawes C (2008) Salivary flow patterns and the health of hard and soft oral tissues. J Am Dent Assoc 139(Suppl):18S–24SPubMed
7.
go back to reference Oudhoff MJ, Bolscher JG, Nazmi K et al (2008) Histatins are the major wound-closure stimulating factors in human saliva as identified in a cell culture assay. FASEB J 22(11):3805–3812PubMedCrossRef Oudhoff MJ, Bolscher JG, Nazmi K et al (2008) Histatins are the major wound-closure stimulating factors in human saliva as identified in a cell culture assay. FASEB J 22(11):3805–3812PubMedCrossRef
8.
go back to reference Hannig C, Spitzmuller B, Hannig M (2009) Characterisation of lysozyme activity in the in situ pellicle using a fluorimetric assay. Clin Oral Investig 13(1):15–21PubMedCrossRef Hannig C, Spitzmuller B, Hannig M (2009) Characterisation of lysozyme activity in the in situ pellicle using a fluorimetric assay. Clin Oral Investig 13(1):15–21PubMedCrossRef
9.
go back to reference Hannig C, Spitzmuller B, Miller M et al (2008) Intrinsic enzymatic crosslinking and maturation of the in situ pellicle. Arch Oral Biol 53(5):416–422PubMedCrossRef Hannig C, Spitzmuller B, Miller M et al (2008) Intrinsic enzymatic crosslinking and maturation of the in situ pellicle. Arch Oral Biol 53(5):416–422PubMedCrossRef
10.
go back to reference Morishita Y, Iinuma Y, Nakashima N et al (2000) Total and pancreatic amylase measured with 2-chloro-4-nitrophenyl-4-O-beta-D-galactopyranosylmaltoside. Clin Chem 46(7):928–933PubMed Morishita Y, Iinuma Y, Nakashima N et al (2000) Total and pancreatic amylase measured with 2-chloro-4-nitrophenyl-4-O-beta-D-galactopyranosylmaltoside. Clin Chem 46(7):928–933PubMed
11.
go back to reference Proctor GB, Chan KM (1994) A fluorometric assay of peroxidase activity utilizing 2′, 7′-dichlorofluorescein with thiocyanate: application to the study of salivary secretion. J Biochem Biophys Methods 28(4):329–336PubMedCrossRef Proctor GB, Chan KM (1994) A fluorometric assay of peroxidase activity utilizing 2′, 7′-dichlorofluorescein with thiocyanate: application to the study of salivary secretion. J Biochem Biophys Methods 28(4):329–336PubMedCrossRef
12.
go back to reference Brown H, Prescott R (1999) Applied mixed models in medicine. Wiley, Hoboken Brown H, Prescott R (1999) Applied mixed models in medicine. Wiley, Hoboken
13.
go back to reference Oudhoff MJ, van den Keijbus PA, Kroeze KL et al (2009) Histatins enhance wound closure with oral and non-oral cells. J Dent Res 88(9):846–850PubMedCrossRef Oudhoff MJ, van den Keijbus PA, Kroeze KL et al (2009) Histatins enhance wound closure with oral and non-oral cells. J Dent Res 88(9):846–850PubMedCrossRef
14.
go back to reference Boskey AL (1995) Osteopontin and related phosphorylated sialoproteins: effects on mineralization. Ann NY Acad Sci 760:249–256PubMedCrossRef Boskey AL (1995) Osteopontin and related phosphorylated sialoproteins: effects on mineralization. Ann NY Acad Sci 760:249–256PubMedCrossRef
15.
go back to reference Chabas D (2005) L’ostéopontine, une molecule aux multiples facettes. Méd Sci 21(10):832–838 Chabas D (2005) L’ostéopontine, une molecule aux multiples facettes. Méd Sci 21(10):832–838
16.
go back to reference Bar-Shavit Z (2008) Taking a toll on the bones: regulation of bone metabolism by innate immune regulators. Autoimmunity 41(3):195–203PubMedCrossRef Bar-Shavit Z (2008) Taking a toll on the bones: regulation of bone metabolism by innate immune regulators. Autoimmunity 41(3):195–203PubMedCrossRef
17.
go back to reference Ikeda E, Kusaka M, Hakeda Y et al (1988) Effect of interleukin 1 beta on osteoblastic clone MC3T3-E1 cells. Calcif Tissue Int 43(3):162–166PubMedCrossRef Ikeda E, Kusaka M, Hakeda Y et al (1988) Effect of interleukin 1 beta on osteoblastic clone MC3T3-E1 cells. Calcif Tissue Int 43(3):162–166PubMedCrossRef
18.
go back to reference Jin CH, Miyaura C, Ishimi Y et al (1990) Interleukin 1 regulates the expression of osteopontin mRNA by osteoblasts. Mol Cell Endocrinol 74(3):221–228PubMedCrossRef Jin CH, Miyaura C, Ishimi Y et al (1990) Interleukin 1 regulates the expression of osteopontin mRNA by osteoblasts. Mol Cell Endocrinol 74(3):221–228PubMedCrossRef
19.
go back to reference Lacey DC, Simmons PJ, Graves SE et al (2009) Proinflammatory cytokines inhibit osteogenic differentiation from stem cells: implications for bone repair during inflammation. Osteoarthritis Cartilage 17(6):735–742PubMedCrossRef Lacey DC, Simmons PJ, Graves SE et al (2009) Proinflammatory cytokines inhibit osteogenic differentiation from stem cells: implications for bone repair during inflammation. Osteoarthritis Cartilage 17(6):735–742PubMedCrossRef
20.
go back to reference Naot D, Grey A, Reid IR et al (2005) Lactoferrin—a novel bone growth factor. Clin Med Res 3(2):93–101PubMedCrossRef Naot D, Grey A, Reid IR et al (2005) Lactoferrin—a novel bone growth factor. Clin Med Res 3(2):93–101PubMedCrossRef
21.
go back to reference Syed F, Khosla S (2005) Mechanisms of sex steroid effects on bone. Biochem Biophys Res Commun 328(3):688–696PubMedCrossRef Syed F, Khosla S (2005) Mechanisms of sex steroid effects on bone. Biochem Biophys Res Commun 328(3):688–696PubMedCrossRef
22.
go back to reference Abiko Y, Saitoh M (2007) Salivary defensins and their importance in oral health and disease. Curr Pharm Des 13(30):3065–3072PubMedCrossRef Abiko Y, Saitoh M (2007) Salivary defensins and their importance in oral health and disease. Curr Pharm Des 13(30):3065–3072PubMedCrossRef
23.
24.
go back to reference Iijima K, Ando K, Kishi M et al (1983) Collagenase activity in human saliva. J Dent Res 62(6):709–712PubMedCrossRef Iijima K, Ando K, Kishi M et al (1983) Collagenase activity in human saliva. J Dent Res 62(6):709–712PubMedCrossRef
25.
go back to reference Hannig C, Hannig M, Attin T (2005) Enzymes in the acquired enamel pellicle. Eur J Oral Sci 113(1):2–13PubMedCrossRef Hannig C, Hannig M, Attin T (2005) Enzymes in the acquired enamel pellicle. Eur J Oral Sci 113(1):2–13PubMedCrossRef
26.
go back to reference Wang D, Christensen K, Chawla K et al (1999) Isolation and characterization of MC3T3-E1 preosteoblast subclones with distinct in vitro and in vivo differentiation/mineralization potential. J Bone Miner Res 14(6):893–903PubMedCrossRef Wang D, Christensen K, Chawla K et al (1999) Isolation and characterization of MC3T3-E1 preosteoblast subclones with distinct in vitro and in vivo differentiation/mineralization potential. J Bone Miner Res 14(6):893–903PubMedCrossRef
27.
go back to reference Quarles LD, Yohay DA, Lever LW et al (1992) Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: an in vitro model of osteoblast development. J Bone Miner Res 7(6):683–692PubMedCrossRef Quarles LD, Yohay DA, Lever LW et al (1992) Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: an in vitro model of osteoblast development. J Bone Miner Res 7(6):683–692PubMedCrossRef
28.
go back to reference Choi JY, Lee BH, Song KB et al (1996) Expression patterns of bone-related proteins during osteoblastic differentiation in MC3T3-E1 cells. J Cell Biochem 61(4):609–618PubMedCrossRef Choi JY, Lee BH, Song KB et al (1996) Expression patterns of bone-related proteins during osteoblastic differentiation in MC3T3-E1 cells. J Cell Biochem 61(4):609–618PubMedCrossRef
29.
go back to reference Tenovuo J (2002) Antimicrobial agents in saliva—protection for the whole body. J Dent Res 81(12):807–809PubMedCrossRef Tenovuo J (2002) Antimicrobial agents in saliva—protection for the whole body. J Dent Res 81(12):807–809PubMedCrossRef
Metadata
Title
Human saliva exposure modulates bone cell performance in vitro
Authors
Susanne Proksch
Thorsten Steinberg
Constantin Keller
Martin Wolkewitz
Margit Wiedmann-Al-Ahmad
Guenter Finkenzeller
Christian Hannig
Elmar Hellwig
Ali Al-Ahmad
Publication date
01-02-2012
Publisher
Springer-Verlag
Published in
Clinical Oral Investigations / Issue 1/2012
Print ISSN: 1432-6981
Electronic ISSN: 1436-3771
DOI
https://doi.org/10.1007/s00784-010-0506-7

Other articles of this Issue 1/2012

Clinical Oral Investigations 1/2012 Go to the issue