Japanese Dental Science Review (2011) 47, 161—166

a v a i l a b l e a t w w w. s c i e n c e d i r e c t . c o m

journal homepage: www.elsevier.com/locate/jdsr

Mini Review

Roles of dental pulp fibroblasts in the recognition

of bacterium-related factors and subsequent
development of pulpitis
Tadashi Nakanishi *, Daisuke Takegawa, Kouji Hirao, Kanako Takahashi,
Hiromichi Yumoto, Takashi Matsuo

Department of Conservative Dentistry, Institute of Health Biosciences, The University of Tokushima Graduate School,
3-18-15 Kuramoto-cho, Tokushima 770-8504, Japan

Received 13 December 2010; received in revised form 14 February 2011; accepted 18 February 2011

KEY WORDS Summary As caries-related bacteria invade deeply into dentin and come into close proximity
Toll-like receptor (TLR); to the pulp, inflammatory cells (such as lymphocytes, macrophages and neutrophils) infiltrate
Nucleotide-binding into the bacterium-invaded area and consequently pulpitis develops. Many types of cytokines and
oligomerization domain adhesion molecules are responsible for the initiation and progression of pulpitis. Dental pulp
(NOD); fibroblasts, a major cell type in the dental pulp, also have capacity to produce pro-inflammatory
Dental pulp fibroblast; cytokines and express adhesion molecules in response to pathogen-associated molecular patterns
Cytokine; (PAMPs), including lipopolysaccharide. The innate immune system senses microbial infection
Pulpitis using pattern recognition receptors, such as Toll-like receptors (TLRs) and nucleotide-binding
oligomerization domain (NOD), for PAMPs. In this review, we summarize the roles of dental pulp
fibroblasts in the recognition of invaded bacterium-related factors via TLR and NOD pathways,
and the subsequent pulpal immune responses, leading to progressive pulpitis.
# 2011 Japanese Association for Dental Science. Published by Elsevier Ltd. All rights reserved.

Introduction [1]. Pulpal DCs expressing class II major histocompatibility

complex (MHC) molecules localize in the para-odontoblastic
Pulpitis is characterized as the immune response that is mainly and perivascular regions, where these cells capture foreign
triggered by the invasion of caries-related microorganisms into antigens [2—4]. An increased accumulation of pulpal DCs in the
dentinal tubules and pulp (Fig. 1). In the innate immune para-odontoblastic area corresponding to the carious dentinal
response of dental pulp to shallow caries, pulpal dendritic tubules is observed, even in the early stage of dentinal caries
cells (DCs) are considered important in immunosurveillance [5]. In addition to pulpal DCs, odontoblasts also play a pivotal
role in the pulpal innate immune response against caries
invasion. Normal odontoblasts express beta-defensin, which
induces antimicrobial activity [6], and interleukin-8, which is a
* Corresponding author. Tel.: +81 88 633 7340; fax: +81 88 631 4215. pro-inflammatory cytokine [7,8]. Transforming growth factor
E-mail address: [email protected] (T. Nakanishi). (TGF)-beta, which is important in anti-inflammatory activity

1882-7616/$ — see front matter # 2011 Japanese Association for Dental Science. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.jdsr.2011.02.001
162 T. Nakanishi et al.

divided into two subpopulations with regard to their cellular

localization. TLR1, TLR2, TLR4, TLR5, TLR6 and TLR11 are
expressed on the cell surface and recognize microbial mem-
brane components, whereas TLR3, TLR7, TLR8 and TLR9 are
expressed in intracellular vesicles such as the endosome and
the endoplasmic reticulum and predominantly recognize
microbial nucleic acid species. Of the cell-surface TLRs,
TLR4 is essential for responses to lipopolysaccharide (LPS),
a major constituent of the outer membrane of Gram-negative
bacteria, which is a potent immunostimulatory molecule
[38]. TLR2 recognizes a wide range of PAMPs derived from
various pathogens; for example, triacyl lipopeptides from
bacteria and mycobacteria, peptidoglycan and lipoteichoic
acid (LTA) from Gram-positive bacteria and zymosan from
fungi [39,40]. TLR5 recognizes flagellin, a protein component
of bacterial flagella [41]. On the other hand, of the intra-
cellular TLRs, TLR3 is implicated in triggering anti-viral
immune response, upon recognition of RNA species, such
Fig. 1 A schematic illustration of dental pulp responses to as double-stranded RNA (dsRNA) of viruses and a synthetic
dental caries. analogue of dsRNA:polyinosinic-polycytidylic acid (poly I:C)
[42,43]. TLR9 recognizes unmethylated CpG DNA motifs from
bacteria and homozoin from Plasmodium [44,45].
as well as dentinogenesis and repair, is also secreted by
In addition to TLRs, other cytosolic PRRs such as NOD-like
odontoblasts [9,10]. These two cell types cooperatively con-
receptors (NLRs) [46] and retinoic acid-inducible gene-I (RIG-
tribute to pulpal responses against carious irritation [11].
I)-like receptors (RLRs) for intracellular PAMPs exist [47].
As carious infection progresses to the pulp-dentin inter-
NOD1 and NOD2 are well-characterized members of the NLR
face, a decrease in the proportion of Gram-positive aerobic
family, which recognize the monomeric structure of pepti-
bacteria and an increase of Gram-negative anaerobic bac-
doglycan [48]. NOD1 recognizes g-D-glutamyl-meso-diamino-
teria occur [12], and marked infiltration of inflammatory cells
pimelic acid (iE-DAP), which is a motif found in peptidoglycan
is observed in the dental pulp [13—15]. In particular, sig-
from Gram-negative bacteria. In contrast, NOD2 recognizes
nificantly higher numbers of B cells and plasma cells are
muramyl dipeptides (MDP), which are minimal motifs present
found in severe pulpitis together with an increased CD4/CD8
in all peptidoglycans.
ratio of T cells [13,16]. Various pro-inflammatory mediators
such as cytokines and prostaglandins (PGs) are also expressed
in the inflamed pulp [7,14,17—25]. With the development of Odontoblasts
exposure to bacterial components, partial destruction of the
Immunohistochemical analysis demonstrated that TLR2 and
odontoblast layer along with severe damage or death of
TLR4 are mainly expressed on the odontoblast layer of normal
odontoblasts can be observed, and the underlying dental
pulp [49,50]. One of these reports shows that LPS-mediated
pulp cells including fibroblasts and undifferentiated
TLR4 activation increased pro-inflammatory cytokines, IL-1b
mesenchymal or stem cells in the cell-rich zone are activated
and TNF-a, in the odontoblasts using organotypic tooth crown
to participate in the host response and initiate reparative
odontoblast cultures, but TLR2 stimulation with TLR2 ligand
dentin formation [26—28]. Thus, the dental pulp cells, a
(Pam3CSK4, a synthetic lipopeptide) decreased these pro-
major cell type in the dental pulp, play a crucial role in
inflammatory markers, which suggest that pro-inflammatory
maintaining the structural integrity of connective tissues,
cytokines and innate immune responses in decayed teeth may
and they also have capacity to produce pro-inflammatory
result from TLR4 signaling [50]. Moreover, cultured human
cytokines and express adhesion molecules in response to
odontoblast-like cells are highly responsive to Gram-negative
pathogen-associated molecular patterns (PAMPs), which
bacteria, such as Prevotella intermedia and Fusobacterium
are structures expressed by microorganisms [29—34]. Gen-
nucleatum, compared with Gram-positive bacteria, such as
erally, the initial sensing of microbial pathogens is mediated
Streptococcus mutans and Lactobacillus casei, despite hetero-
by pattern recognition receptors (PRRs) for PAMPs. The PRRs,
geneity of TLR2 and TLR4 cell-surface expression [51]. On the
such as Toll-like receptor (TLR) and nucleotide-binding oli-
other hand, experimentally inflamed pulp in a murine model
gomerization domain (NOD), have been shown to recognize a
showed that the TLR2 mRNA level was 30-fold higher than the
number of PAMPs [35]. In this review, we describe the roles of
TLR4 mRNA level at 9 h after infection, and the TLR2-positive
odontoblasts and dental pulp cells in the recognition of
cells were observed in and around the odontoblast layer and
invaded bacterium-related factors via TLR and NOD path-
the area infiltrated by inflammatory cells [52]. This report
ways, and the subsequent host responses of dental pulp,
suggested that TLR2 may be mainly regulated during the early
leading to progressive pulpitis.
stage of pulp inflammation triggered by bacterial infection.
Other in vitro studies with odontoblast-like cells in culture
TLRs and NODs in dental pulp have also demonstrated that odontoblasts stimulated with
LTA, a Gram-positive bacterium-derived component recog-
In mammals, the TLR family comprises more than 12 mem- nized at the cell surface through TLR2, initiate an immune
bers [36,37]. The TLR family members can be conveniently response by triggering up-regulation of TLR2 and production of
Roles of dental pulp fibroblasts in pulpitis 163

Fig. 2 A possible role of NOD2 in TLR2- and TLR4-mediated signal pathways. NOD2 against MDP acts synergistically with TLR2, not
TLR4, agonist to stimulate the production of pro-inflammatory mediators in human dental pulp fibroblasts.

chemokines such as CCL2 and CXCL10 [53,54]. Conversely, LTA- In our study, flow cytometric analysis showed that the
dependent TLR2 activation in odontoblast-like cells did not expression level of TLR2 was higher than that of TLR4 in
lead to significant IL-1b and TNF-a production [55], similar to human dental pulp fibroblasts [59]. In accordance with this
another report with engagement of TLR2 by Pam3CSK4 [50]. analysis, the levels of pro-inflammatory mediators, such as
These findings suggest a possible role for TLR2-mediated CXCL10, IL-8 and PGE2, induced by LPS stimulation were
innate immunotolerance to prevent an uncontrolled inflam- much lower than those induced by Pam3CSK4. On the other
matory reaction as well as immunostimulation to produce hand, another group has shown that CXCL10 expression in
chemokines in odontoblasts. dental pulp fibroblasts was up-regulated by LPS but not LTA, a
Besides TLR2 and TLR4, in vitro-differentiated odonto- TLR2-agonist [54]. These results suggest differences of reac-
blasts constitutively express TLR1, 3, 5, 6 and 9 genes [53]. A tivity between Pam3CSK4 and LTA to elicit chemokine pro-
recent study has reported that TLR9 is expressed in the duction. In fact, our previous report indicates that the dental
mouse odontoblast-like cell line MDPC-23, a spontaneously pulp fibroblasts can produce CXCL10 in response to peptido-
immortalized cell line derived from fetal mouse molar dental glycan, but not LTA [24].
papillae, and that CpG DNA induces potent pro-inflammatory MAPKs have been implicated in many physiologic pro-
cytokine expression via the activation of TLR9 [56]. Addi- cesses, including cell proliferation, differentiation and death
tionally, an immunohistochemical report revealed that the [62—64]. Three major types of MAPKs in mammalian cells are
NOD2 protein expression was localized in odontoblasts and extracellular signal regulated kinase (ERK) 1/2, p38 MAPK
some vascular endothelial cells in normal human dental pulp and c-Jun NH2-terminal kinases (SAP/JNK). NF-kB is an oxi-
57]. However, more detailed studies on the functional role of dation-sensitive transcription factor that plays a critical role
these PRRs in the odontoblasts to recognize intradentinal in the regulation of various genes that are important in
irritation of caries-related bacteria are required in future. cellular responses, including inflammation, innate immunity,
growth and cell death [65]. In TLR2 ligand-stimulated human
Dental pulp cells dental pulp fibroblasts, the phosphorylations of ERK 1/2, p38
MAPK, SAP/JNK and NF-kB were increased, and specific
Dental pulp cells, especially dental pulp fibroblasts, are inhibitors of MAPKs or NF-kB markedly reduced the level
known to produce various inflammatory mediators, such as of pro-inflammatory mediators [66].
IL-8, IL-6 and vascular endothelial growth factor (VEGF), in We also hypothesized in the previous study that the
response to the components of caries-related bacteria, prior differences in capacity between peptidoglycan and LTA might
to the discovery and establishment of the innate immune be due to NOD proteins, which are intracellular PAMPs, as
system feature that PRRs including TLRs recognize various well as TLR2, and then examined NOD1 and NOD2 expression
PAMPs [29,31,32,58]. TLR2, TLR3, TLR4 and TLR5 expressions in human dental pulp tissues and cultured dental pulp fibro-
have been determined in dental pulp fibroblasts and their blasts. We first examined whether the human cultured dental
specific agonists can induce TLR-mediated inflammatory sig- pulp fibroblasts expressed NOD1 and NOD2, and consequently
nals [54,55,59,60], although immunohistological detection of clear expressions of NOD1 and NOD2 in human dental pulp
TLRs was not clear in the fibroblasts of dental pulp tissues fibroblasts were found by RT-PCR and flow cytometry [59].
[49,52]. A recent study has shown that the dental pulp stem Moreover, both of them constitutively expressed in the dental
cells as well as the dental pulp fibroblasts express TLR4, and pulp fibroblasts actually functioned to produce IL-8, IL-6 and
that LPS-induced VEGF is dependent upon mitogen-activated monocyte chemoattractant protein-1 (MCP-1). Next, we
protein kinase (MAPK) activation [61]. investigated whether healthy human dental pulp tissues
164 T. Nakanishi et al.

expressed NOD1 and NOD2 at the mRNA level. As a conse- Conflict of interest statement
quence, NOD2 expression was clearly detected, but NOD1
expression was non-detectable or hardly detected at the The authors declare no conflict of interest.
mRNA level in the dental pulp tissues; thereafter, using
immunohistochemistry, we confirmed whether health dental
pulp tissues expressed NOD2 at the protein level. In healthy Acknowledgement
dental pulp, NOD2 expression was observed in the area just
under the odontoblast layer, unlike in the other study This work was supported in part by the Japan Society for the
described above [57]. Further investigation to resolve this Promotion of Science [Grants-in-Aid for Scientific Research]
discrepancy remains to be carried out. (16591915, 20592228 and 20592229).
Several reports indicated that NOD2 is specifically respon-
sible for the cooperative effect with agonists for selective References
TLRs, including TLR2 and TLR4 [67—69]. Interestingly, we
demonstrated that both TLR2- and NOD2-mediated signals [1] Jontell M, Gunraj MN, Bergenholtz G. Immunocompetent cells
synergize at the production level of pro-inflammatory med- in the normal dental pulp. J Dent Res 1987;66:1149—53.
iators such as CXCL10, IL-8 and PGE2 [59]. In contrast, we [2] Jontell M, Bergenholtz G, Scheynius A, Ambrose W. Dendritic
observed that the additive effect was mediated by interac- cells and macrophages expressing class II antigens in the normal
tions between TLR4 and NOD2 intracellular pathways (Fig. 2). rat incisor pulp. J Dent Res 1988;67:1263—6.
This difference between the cooperative effects of TLR2 and [3] Okiji T, Kawashima N, Kosaka T, Matsumoto A, Kobayashi C, Suda
TLR4 with NOD2 might be due to their different expression H. An immunohistochemical study of the distribution of immu-
nocompetent cells, especially macrophages and Ia antigen-
levels in the dental pulp fibroblasts. Since pulpitis is char-
expressing cells of heterogeneous populations, in normal rat
acterized as the immune response triggered mainly by the molar pulp. J Dent Res 1992;71:1196—202.
invasion of caries-related bacteria into dentinal tubules and [4] Okiji T, Jontell M, Belichenko P, Bergenholtz G, Dahlstrom A.
pulp, pathogen recognition by multiple PRR engagement, Perivascular dendritic cells of the human dental pulp. Acta
including TLR-NOD as shown here, might constitute a key Physiol Scand 1997;159:163—9.
event for the onset of resulting exacerbated pulpal inflam- [5] Bergenholtz G, Nagaoka S, Jontell M. Class II antigen expressing
matory response. In particular, peptidoglycan from both cells in experimentally induced pulpitis. Int Endod J 1991;24:
Gram-positive and Gram-negative bacteria is recognized by 8—14.
both TLR2 and NOD2. Our findings may lead to the identifica- [6] Dommisch H, Winter J, Acil Y, Dunsche A, Tiemann M, Jepsen S.
tion of cooperative mechanisms by these multiple PRRs and Human beta-defensin (hBD-1, -2) expression in dental pulp. Oral
Microbiol Immunol 2005;20:163—6.
the selective blocking or inhibition of pulpal inflammation
[7] Huang GT, Potente AP, Kim JW, Chugal N, Zhang X. Increased
caused by caries-related bacteria. interleukin-8 expression in inflamed human dental pulps. Oral
Surg Oral Med Oral Pathol Oral Radiol Endod 1999;88:214—20.
Conclusion [8] Levin LG, Rudd A, Bletsa A, Reisner H. Expression of IL-8 by cells
of the odontoblast layer in vitro. Eur J Oral Sci 1999;107:131—7.
[9] Sloan AJ, Matthews JB, Smith AJ. TGF-beta receptor expression
This review article describes recent findings about the immune
in human odontoblasts and pulpal cells. Histochem J 1999;
system of dental pulp for the recognition of bacterium-related 31:565—9.
factors. TLRs and cytosolic sensing systems such as NLRs play a [10] Piattelli A, Rubini C, Fioroni M, Tripodi D, Strocchi R. Transform-
crucial role in defending against pathogenic microbial infec- ing growth factor-beta 1 (TGF-beta 1) expression in normal
tion through the induction of inflammatory cytokines. Further- healthy pulps and in those with irreversible pulpitis. Int Endod J
more, the responses of the innate immune system are 2004;37:114—9.
important not only to eliminate pathogens but also to develop [11] Ohshima H, Nakakura-Ohshima K, Takeuchi K, Hoshino M,
pathogen-specific adaptive immunity. Generally, immune cells Takano Y, Maeda T. Pulpal regeneration after cavity prepara-
such as dendritic cells, macrophages and polymorphonuclear tion, with special reference to close spatio-relationships be-
tween odontoblasts and immunocompetent cells. Microsc Res
leukocytes are regarded as key players in the innate immune
Tech 2003;60:483—90.
responses. In the dental pulp, odontoblasts and dental pulp
[12] Hoshino E. Predominant obligate anaerobes in human carious
fibroblasts are also shown to express several TLRs and NODs. dentin. J Dent Res 1985;64:1195—8.
The role of these cells in the progression of pulpitis has not [13] Izumi T, Kobayashi I, Okamura K, Sakai H. Immunohistochemical
been clarified in detail, but they might act as modulators of study on the immunocompetent cells of the pulp in human non-
both innate and adaptive immune responses. A recent report carious and carious teeth. Arch Oral Biol 1995;40:609—14.
reveals that TGF-b1 inhibited TLR2 and TLR4 expression and [14] Hahn CL, Best AM, Tew JG. Cytokine induction by Streptococcus
attenuated odontoblast responses, and thus suggests the mutans and pulpal pathogenesis. Infect Immun 2000;68:
potential use of TGF-b1 in the clinical treatment of pulpal 6785—9.
inflammation [51]. Our recent research demonstrates the anti- [15] Martin FE, Nadkarni MA, Jacques NA, Hunter N. Quantitative
microbiological study of human carious dentine by culture and
inflammatory effect of catechin (bioactive polyphenols in
real-time PCR: association of anaerobes with histopathological
green tea) on human dental pulp cells affected by bacter-
changes in chronic pulpitis. J Clin Microbiol 2002;40:1698—704.
ium-derived factors, especially TLR2 ligand [66,70]. There- [16] Hahn CL, Falkler Jr WA, Siegel MA. A study of T and B cells in
fore, understanding of the mechanism underlying PAMP- pulpal pathosis. J Endodont 1989;15:20—6.
induced pro-inflammatory reactions in the dental pulp is [17] D’Souza R, Brown LR, Newland JR, Levy BM, Lachman LB.
important for the development of future therapeutic strate- Detection and characterization of interleukin-1 in human dental
gies and treatments for pulpitis. pulps. Arch Oral Biol 1989;34:307—13.
Roles of dental pulp fibroblasts in pulpitis 165

[18] Tani-Ishii N, Wang CY, Stashenko P. Immunolocalization of bone- [40] Schwandner R, Dziarski R, Wesche H, Rothe M, Kirschning CJ.
resorptive cytokines in rat pulp and periapical lesions following Peptidoglycan- and lipoteichoic acid-induced cell activation is
surgical pulp exposure. Oral Microbiol Immunol 1995;10:213—9. mediated by toll-like receptor 2. J Biol Chem 1999;274:
[19] Rauschenberger CR, Bailey JC, Cootauco CJ. Detection of hu- 17406—9.
man IL-2 in normal and inflamed dental pulps. J Endodont [41] Hayashi F, Smith KD, Ozinsky A, Hawn TR, Yi EC, Goodlett DR,
1997;23:366—70. et al. The innate immune response to bacterial flagellin is
[20] Barkhordar RA, Hayashi C, Hussain MZ. Detection of interleukin- mediated by Toll-like receptor 5. Nature 2001;410:1099—103.
6 in human dental pulp and periapical lesions. Endod Dent [42] Alexopoulou L, Holt AC, Medzhitov R, Flavell RA. Recognition of
Traumatol 1999;15:26—7. double-stranded RNA and activation of NF-kappaB by Toll-like
[21] Nakanishi T, Matsuo T, Ebisu S. Quantitative analysis of immu- receptor 3. Nature 2001;413:732—8.
noglobulins and inflammatory factors in human pulpal blood [43] Wang T, Town T, Alexopoulou L, Anderson JF, Fikrig E, Flavell RA.
from exposed pulps. J Endodont 1995;21:131—6. Toll-like receptor 3 mediates West Nile virus entry into the brain
[22] Nakanishi T, Shimizu H, Hosokawa Y, Matsuo T. An immunohis- causing lethal encephalitis. Nat Med 2004;10:1366—73.
tological study on cyclooxygenase-2 in human dental pulp. J [44] Hemmi H, Takeuchi O, Kawai T, Kaisho T, Sato S, Sanjo H, et al. A
Endodont 2001;27:385—8. Toll-like receptor recognizes bacterial DNA. Nature 2000;408:
[23] Nakanishi T, Takahashi K, Hosokawa Y, Adachi T, Nakae H, Matsuo 740—5.
T. Expression of macrophage inflammatory protein 3alpha in [45] Coban C, Ishii KJ, Kawai T, Hemmi H, Sato S, Uematsu S, et al.
human inflamed dental pulp tissue. J Endodont 2005;31:84—7. Toll-like receptor 9 mediates innate immune activation by the
[24] Adachi T, Nakanishi T, Yumoto H, Hirao K, Takahashi K, Mukai K, malaria pigment hemozoin. J Exp Med 2005;201:19—25.
et al. Caries-related bacteria and cytokines induce CXCL10 in [46] Ting JP, Lovering RC, Alnemri ES, Bertin J, Boss JM, Davis BK,
dental pulp. J Dent Res 2007;86:1217—22. et al. The NLR gene family: a standard nomenclature. Immunity
[25] Takahashi K, Nakanishi T, Yumoto H, Adachi T, Matsuo T. CCL20 2008;28:285—7.
production is induced in human dental pulp upon stimulation by [47] Yoneyama M, Fujita T. Structural mechanism of RNA recognition
Streptococcus mutans and proinflammatory cytokines. Oral by the RIG-I-like receptors. Immunity 2008;29:178—81.
Microbiol Immunol 2008;23:320—7. [48] Girardin SE, Travassos LH, Herve M, Blanot D, Boneca IG,
[26] Warfvinge J. Morphometric analysis of teeth with inflamed pulp. Philpott DJ, et al. Peptidoglycan molecular requirements allow-
J Dent Res 1987;66:78—83. ing detection by Nod1 and Nod2. J Biol Chem 2003;278:
[27] Fitzgerald M, Chiego Jr DJ, Heys DR. Autoradiographic analysis 41702—8.
of odontoblast replacement following pulp exposure in primate [49] Jiang HW, Zhang W, Ren BP, Zeng JF, Ling JQ. Expression of toll
teeth. Arch Oral Biol 1990;35:707—15. like receptor 4 in normal human odontoblasts and dental pulp
[28] Goldberg M, Smith AJ. Cells and extracellular matrices of dentin tissue. J Endodont 2006;32:747—51.
and pulp: a biological basis for repair and tissue engineering. [50] Veerayutthwilai O, Byers MR, Pham TT, Darveau RP, Dale BA.
Crit Rev Oral Biol Med 2004;15:13—27. Differential regulation of immune responses by odontoblasts.
[29] Nagaoka S, Tokuda M, Sakuta T, Taketoshi Y, Tamura M, Takada Oral Microbiol Immunol 2007;22:5—13.
H, et al. Interleukin-8 gene expression by human dental pulp [51] Horst OV, Tompkins KA, Coats SR, Braham PH, Darveau RP, Dale
fibroblast in cultures stimulated with Prevotella intermedia BA. TGF-beta1 inhibits TLR-mediated odontoblast responses to
lipopolysaccharide. J Endodont 1996;22:9—12. oral bacteria. J Dent Res 2009;88:333—8.
[30] Hosoya S, Matsushima K. Stimulation of interleukin-1 beta [52] Mutoh N, Tani-Ishii N, Tsukinoki K, Chieda K, Watanabe K.
production of human dental pulp cells by Porphyromonas endo- Expression of Toll-like receptor 2 and 4 in dental pulp. J
dontalis lipopolysaccharide. J Endodont 1997;23:39—42. Endodont 2007;33:1183—6.
[31] Matsushima K, Ohbayashi E, Takeuchi H, Hosoya S, Abiko Y, [53] Durand SH, Flacher V, Romeas A, Carrouel F, Colomb E, Vincent
Yamazaki M. Stimulation of interleukin-6 production in human C, et al. Lipoteichoic acid increases TLR and functional chemo-
dental pulp cells by peptidoglycans from Lactobacillus casei. J kine expression while reducing dentin formation in in vitro
Endodont 1998;24:252—5. differentiated human odontoblasts. J Immunol 2006;176:
[32] Tokuda M, Sakuta T, Fushuku A, Torii M, Nagaoka S. Regulation of 2880—7.
interleukin-6 expression in human dental pulp cell cultures [54] Staquet MJ, Durand SH, Colomb E, Romeas A, Vincent C,
stimulated with Prevotella intermedia lipopolysaccharide. J Bleicher F, et al. Different roles of odontoblasts and fibroblasts
Endodont 2001;27:273—7. in immunity. J Dent Res 2008;87:256—61.
[33] Barkhordar RA, Ghani QP, Russell TR, Hussain MZ. Interleukin- [55] Keller JF, Carrouel F, Colomb E, Durand SH, Baudouin C, Msika P,
1beta activity and collagen synthesis in human dental pulp et al. Toll-like receptor 2 activation by lipoteichoic acid induces
fibroblasts. J Endodont 2002;28:157—9. differential production of pro-inflammatory cytokines in human
[34] Yang LC, Tsai CH, Huang FM, Liu CM, Lai CC, Chang YC. Induction odontoblasts, dental pulp fibroblasts and immature dendritic
of interleukin-6 gene expression by pro-inflammatory cytokines cells. Immunobiology 2010;215:53—9.
and black-pigmented bacteroides in human pulp cell cultures. [56] He W, Yu Q, Zhou Z, Wang P. CpG oligonucleotides induce an
Int Endod J 2003;36:352—7. immune response of odontoblasts through the TLR9, MyD88 and
[35] Takeda K, Akira S. Toll-like receptors in innate immunity. Int NF-kappaB pathways. Biochem Biophys Res Commun 2010;399:
Immunol 2005;17:1—14. 274—8.
[36] Akira S, Uematsu S, Takeuchi O. Pathogen recognition and [57] Lin ZM, Song Z, Qin W, Li J, Li WJ, Zhu HY, et al. Expression of
innate immunity. Cell 2006;124:783—801. nucleotide-binding oligomerization domain 2 in normal human
[37] Beutler BA. TLRs and innate immunity. Blood 2009;113: dental pulp cells and dental pulp tissues. J Endodont 2009;35:
1399—407. 838—42.
[38] Tapping RI, Akashi S, Miyake K, Godowski PJ, Tobias PS. Toll-like [58] Matsushita K, Motani R, Sakuta T, Nagaoka S, Matsuyama T,
receptor 4, but not toll-like receptor 2, is a signaling receptor Abeyama K, et al. Lipopolysaccharide enhances the production
for Escherichia and Salmonella lipopolysaccharides. J Immunol of vascular endothelial growth factor by human pulp cells in
2000;165:5780—7. culture. Infect Immun 1999;67:1633—9.
[39] Aliprantis AO, Yang RB, Mark MR, Suggett S, Devaux B, Radolf JD, [59] Hirao K, Yumoto H, Takahashi K, Mukai K, Nakanishi T, Matsuo T.
et al. Cell activation and apoptosis by bacterial lipoproteins Roles of TLR2, TLR4, NOD2, and NOD1 in pulp fibroblasts. J Dent
through toll-like receptor-2. Science 1999;285:736—9. Res 2009;88:762—7.
166 T. Nakanishi et al.

[60] Park C, Lee SY, Kim HJ, Park K, Kim JS, Lee SJ. Synergy of TLR2 and mitogen-activated protein kinase pathways in toll-like receptor
H1R on Cox-2 activation in pulpal cells. J Dent Res 2010;89:180—5. 2 ligand-stimulated dental pulp cells. Life Sci 2010;86:654—60.
[61] Botero TM, Son JS, Vodopyanov D, Hasegawa M, Shelburne CE, [67] Fritz JH, Girardin SE, Fitting C, Werts C, Mengin-Lecreulx D,
Nor JE. MAPK signaling is required for LPS-induced VEGF in pulp Caroff M, et al. Synergistic stimulation of human monocytes and
stem cells. J Dent Res 2010;89:264—9. dendritic cells by Toll-like receptor 4 and NOD1- and NOD2-
[62] Han J, Lee JD, Bibbs L, Ulevitch RJ. A MAP kinase targeted by activating agonists. Eur J Immunol 2005;35:2459—70.
endotoxin and hyperosmolarity in mammalian cells. Science [68] Netea MG, Ferwerda G, de Jong DJ, Jansen T, Jacobs L, Kramer
1994;265:808—11. M, et al. Nucleotide-binding oligomerization domain-2 modu-
[63] Kyriakis JM, Banerjee P, Nikolakaki E, Dai T, Rubie EA, Ahmad MF, lates specific TLR pathways for the induction of cytokine re-
et al. The stress-activated protein kinase subfamily of c-Jun lease. J Immunol 2005;174:6518—23.
kinases. Nature 1994;369:156—60. [69] Tada H, Aiba S, Shibata K, Ohteki T, Takada H. Synergistic effect
[64] Lee JC, Laydon JT, McDonnell PC, Gallagher TF, Kumar S, Green of Nod1 and Nod2 agonists with toll-like receptor agonists on
D, et al. A protein kinase involved in the regulation of inflam- human dendritic cells to generate interleukin-12 and T helper
matory cytokine biosynthesis. Nature 1994;372:739—46. type 1 cells. Infect Immun 2005;73:7967—76.
[65] Karin M, Lin A. NF-kappaB at the crossroads of life and death. [70] Nakanishi T, Mukai K, Yumoto H, Hirao K, Hosokawa Y, Matsuo T.
Nat Immunol 2002;3:221—7. Anti-inflammatory effect of catechin on cultured human dental
[66] Hirao K, Yumoto H, Nakanishi T, Mukai K, Takahashi K, Takegawa pulp cells affected by bacteria-derived factors. Eur J Oral Sci
D, et al. Tea catechins reduce inflammatory reactions via 2010;118:145—50.