Serotonergic modulation of slow inward rectification in mesencephalic trigeminal neurons.

Inward rectification in response to membrane hyperpolarization is a prominent feature of mesencephalic trigeminal (Mes V) neurons and the hyperpolarization-activated inward current (Ih), as the basis of this property, regulates the spike discharge characteristics and input frequency preference (resonance) in these neurons, suggesting that Ih modulation is an important regulator of oral motor activity. To examine a possible contribution of serotonin (5-HT) to the modulation of Ih activation characteristics, in the present study, we investigated the modulatory effects of 5-HT receptor activation on Ih in postnatal day (P) 2-12 rat Mes V neurons by whole-cell patch-clamp recording. Bath application of 5-HT suppressed the Ih-dependent voltage sag and Ih conductance, but induced only a modest shift in the voltage dependence of Ih activation. This 5-HT-induced suppression of Ih was greater in P10-12 than P2-4 neurons, and involved the cAMP/protein kinase A (PKA) signaling pathway but not the PKC pathway. Pharmacological activation of the 5-HT1A receptor mimicked the effect of 5-HT, while modulation of other receptor subtypes, including 5-HT1B,1D, 5-HT2, and 5-HT3, had little or no effect on Ih. Low-frequency (<10 Hz) resonance at membrane potentials below the resting potential were reduced by 5-HT, suggesting that serotonergic Ih modulation can substantially alter the frequency preference to synaptic inputs. These results suggest that changes in resonance properties through serotonergic modulation of Ih may tune the firing of Mes V neurons to different afferent input frequencies and alter motor outputs to the jaw, thereby regulating oral motor activity.

Nationwide survey for bisphosphonate-related osteonecrosis of the jaws in Japan.

PURPOSE: A nationwide retrospective cohort study was conducted by the Japanese Society of Oral and Maxillofacial Surgeons to assess the occurrence of bisphosphonate (BP)-related osteonecrosis of the jaws (BRONJ) during 2006 to 2008 and to elucidate the outcome and factors associated with remission of BRONJ. MATERIALS AND METHODS: A written questionnaire, including the clinical characteristics, management, and outcome of patients with BRONJ, was sent to 248 institutions certified as training facilities by the Japanese Society of Oral and Maxillofacial Surgeons in 2008. RESULTS: A total of 568 patients with BRONJ, including suspicious cases, were registered. Of these 568 patients, 263, including the maxilla in 81, the mandible in 160, and both in 22, met the working definition of BRONJ proposed by the American Association of Oral and Maxillofacial Surgeons. The patients included 219 women (83.3%) and 44 men (16.7%). Of these patients, 152 (57.8%) had received intravenous BPs, 104 (39.5%) had received oral BPs, and 7 (2.7%) had received both. The mean duration of administration until onset of BRONJ was 23.6 months for intravenous BPs and 33.2 months for oral BPs. BRONJ was stage 1 in 42 patients (16.0%), stage 2 in 187 (71.1%), stage 3 in 32 (12.2%), and unknown in 2. Of these patients, 34.2% had remission of BRONJ, 46.0% had persistent or progressive disease, and 19.7% died of malignancy or were lost to follow-up. Statistical analysis revealed that surgical treatment, including tooth extraction, sequestrectomy, and segmental mandibulectomy, contributed to the remission of BRONJ. In contrast, conservative treatment, concurrent anticancer drugs, poor oral hygiene, and the use of intravenous BPs did not. CONCLUSIONS: The relative ratio of BRONJ related to the use of oral BPs was greater in Japan than in the United States and European Union. Surgical treatment contributed to remission of BRONJ, and conservative treatment, concurrent anticancer drugs, poor oral hygiene, and intravenous BPs did not.

Aerial dispersal of blood-contaminated aerosols during dental procedures.

OBJECTIVES: Dental procedures with high-speed instruments produce large amounts of aerosols. The present study aimed to clarify whether blood-contaminated aerosols were existent and floating in air during dental procedures and to evaluate the effect of an extraoral evacuator system. METHOD AND MATERIALS: An extraoral evacuator system with a test filter was used for sample collection at distances from 50 cm (n = 102) and 100 cm (n =124) behind the patient. A leucomalachite green presumptive test was performed on the test filter. One or more positive reactions on the test filter was considered a positive result, and the positive ratio in each procedure was compared. To assess the effect of the extraoral evacuator, an additional extraoral evacuator was introduced, and the positive ratio during third molar extractions at 100 cm behind the patient was compared (n = 55). RESULTS: At 50 cm from the mouth of the patient during third molar surgery, full-crown preparation, inlay cavity (Black Class II) preparation, and scaling with an ultrasonic scaler, positive results were obtained in 92% (12/13), 70% (21/30), 35% (9/26), and 33% (11/33) of cases, respectively. At a distance of 100 cm, positive ratios sustained 90% (35/39), 48% (15/31), 29% (6/21), and 12% (4/33), respectively. When the second extraoral evacuator was utilized, the positive ratio, at a distance of 100 cm, decreased significantly from 90% (35/39) to 60% (33/55) (P = .0015, chi-square test). CONCLUSION: This study showed that blood-contaminated aerosols can be suspended in air, even in general dental settings, and that extraoral evacuators are useful for reducing contaminated aerosols.

Inferior alveolar nerve canal and branches detected with dental cone beam computed tomography in lower third molar region.

PURPOSE: To evaluate the course of the inferior alveolar nerve and its branches, the detectable branches were investigated with dental cone beam computed tomography (CBCT). MATERIALS AND METHODS: Patients in whom the lower third molar (M3) and inferior alveolar nerve canal showed overlapping in the initial panoramic image were included. One hundred twelve impacted lower M3s were extracted after examination with dental CBCT. The detection ratio, the course of the branches, and their relation with the M3 were retrospectively investigated. RESULTS: One hundred fifty-five branches were observed in 106 cases (94.6%, 106/112) around the M3. Most branches coursed under the M3 (55.5%, 86/155), and 85 branches (54.8%, 85/155) were in contact with the M3. The inferior alveolar nerve canal and branch(es) were mostly in contact with the M3 (57.5%, 61/106). CONCLUSIONS: Dental CBCT can detect most tubular structures representing branches in the impacted lower M3 region.

Bone formation without lamina dura in the middle-aged and elderly: possible dependence on enamel.

Bone formation below the crown of mandibular horizontal incompletely impacted third molar is frequently seen in the middle-aged and elderly. The phenomenon shows lamina dura loss without radiolucency and we hypothesized the participation of mature enamel without any influence on the environmental oral status. In order to investigate the characteristics of the phenomenon based on the presence/absence of the lamina dura and radiolucency below the crown, we studied the relationship between 58 men and 43 women with a lamina dura without radiolucency, 12 men and 8 women without a lamina dura with radiolucency, 34 men and 16 women without a lamina dura without radiolucency, and the status of teeth in the ipsilateral mandible. Subjects without a lamina dura without radiolucency were significantly older than those with a lamina dura without radiolucency in both men (P < 0.0001) and women (P <0.01), indicating different chronological causes. Men without lamina dura with radiolucency showed significantly more tooth loss than those with a lamina dura without radiolucency (P < 0.00001) and those without a lamina dura without radiolucency (P < 0.0001), indicating the influence of poor oral health. Thus, the phenomenon without a lamina dura without radiolucency may show the clinical importance of bone formation in the elderly.

A comparison of the postnatal development of muscle-spindle and periodontal-ligament neurons in the mesencephalic trigeminal nucleus of the rat.

The trigeminal mesencephalic nucleus (Vmes) is known to include primary afferent neurons of jaw muscle spindles (MS neurons) and periodontal ligament receptors (PL neurons). The aim of this study was to clarify the postnatal development of Vmes neurons by comparing MS neurons with PL neurons using horseradish peroxidase labeling. We measured somal diameter and somal shape of MS and PL neurons in rats from postnatal day (P)7 to P70. No significant changes were seen between postnatal day P7 and P70 in somal diameter or somal shape of MS neurons. Conversely, PL neurons showed a larger somal diameter at P7 than at P14, and in terms of somal profile, multipolar neurons comprised 0% at P7, but 4.8% at P14 and 16.9% at P70. These findings suggest that PL neurons develop with the eruption of teeth, taking into account the fact that tooth eruption occurs from around P14 in rats. Conversely, the lack of postnatal changes in MS neurons is due to the fact that these neurons have been active since the embryonic period, as swallowing starts in utero.

Bone formation with disruption of the lamina dura in the mandibular third molar.

The lamina dura is a healthy structural component in teeth. This study highlights the association of bone formation with disruption of lamina dura below the crown of the mandibular horizontal incompletely impacted third molar.

Occupational safety among dental health-care workers.

Compared to other health-care workers, dental health-care workers come in close contact with patients and use a variety of sharp and high-speed rotating instruments. It is important to understand the characteristics of the occupational accidents that occur. We reviewed incident reports from April 1, 2005, to March 31, 2010, at Matsumoto Dental University Hospital. In addition, questionnaires dealing with identification of occupational safety issues, especially splash exposures, were conducted for dentists, dental hygienists, and nurses. Thirty-two occupational injuries were reported during the study period, including 23 sharp instrument injuries (71.9%), 6 splash exposures (18.8%), and 3 others. Of the six splash exposures, only two cases involved potential contamination with blood or other potentially infectious patient material. Of the 66 workers who experienced sharps injuries, 20 workers (30.3%, 20/66) reported them to the hospital work safety team. The questionnaire revealed high incident of splash exposures and conjunctiva exposures: 87.9% (51/58) and 60.3% (35/58) in dentists and 88.6% (39/44) and 61.4% (27/44) in dental hygienists. The compliance rate for routine use of protective eyewear was 60.3% (35/58) for dentists and 34.1% (15/44) for hygienists. Of the presented informational items included in the questionnaire, those that strongly persuaded respondents to use protective eyewear were 'splatters from the patient's mouth contain blood' (90%, 99/110) and 'dental operations at our clinic are performed based only on a questionnaire without serious examinations for HBV, HCV, and HIV' (71.8%, 79/110). The reason of low compliance of protective eyewear among dentists might relate to fine dental procedures. Appropriate information is important for the motive of wearing personal protective equipment, and an early educational program may have a potential to increase compliance with the use of that equipment.

Age-related disruption of the lamina dura: evidence in the mandibular horizontal incompletely impacted third molar.

Changes in the lamina dura are associated with dental diseases around the root of the tooth and with systemic diseases; however, the lamina dura below the crown of horizontal, incompletely impacted third molars has not been studied. Using orthopantomography, we studied the age of subjects with and without the lamina dura in 419 subjects. The participants were between the ages of 21 and 89 years. Mean age in men with the lamina dura was 30.29 +/- 9.92 and without the lamina dura was 47.64 +/- 16.32 (P < 0.0001), and in women with a lamina dura it was 29.65 +/- 8.19 and without a lamina dura 41.97 +/- 11.07 (P < 0.0001). To study the effect of aging, the relationship between the lamina dura and dental status was assessed in subjects over the age of 31 years. Alveolar bone resorption in the canine and the first molar of the ipsilateral mandible in subjects without the lamina dura was not significantly higher than in those with the lamina dura. There were no significant differences in the number of teeth lost, except in men, the number of treated teeth and the number of decayed teeth differed between groups. Disruption of the lamina dura was related to age, but with no alveolar bone resorption in the mandible.

Acute inflammation in horizontal incompletely impacted third molar with radiolucency in the elderly.

Although radiolucency has been shown as a risk of infection, the poorly understood effects of aging on radiolucency correlate with acute pericoronitis, which has a high risk of infection extending any complications. We reviewed the records of 346 consecutive patients aged more than 41 years to evaluate whether pericoronal radiolucency below the crown in mandibular horizontal incompletely impacted third molars is related to acute inflammation. The frequency of acute inflammation in teeth with pericoronal radiolucency below the crown was similar to that in teeth without; however, the odds ratio of acute inflammation exhibited in women aged more than 61 years compared to women aged 41-50 years was 9.77 (95% confidence interval [CI]: 1.67-57.29; P < <0.05), and in women aged more than 61 years compared to women aged 51-60 years was 26.25 (95% CI: 2.94-234.38; P < 0.01). The odds ratio of severe acute inflammation exhibited in men aged more than 61 years compared to men aged 41-50 years was 16.67 (95% CI: 1.76-158.27; P < 0.01). These odds ratios indicate an association of acute pericoronitis, including the severe forms of acute inflammation that result from pericoronitis, with pericoronal radiolucency below the crown in the elderly.

Docetaxel inhibits bone resorption through suppression of osteoclast formation and function in different manners.

Osteoclasts are formed from the monocyte-macrophage lineage in response to receptor activator of nuclear factor kappaB ligand (RANKL) expressed by osteoblasts. Bone is the most common site of breast cancer metastasis, and osteoclasts play roles in the metastasis. The taxane-derived compounds paclitaxel and docetaxel are used for the treatment of malignant diseases, including breast cancer. Here we explored the effects of docetaxel on osteoclastic bone resorption in mouse culture systems. Osteoclasts were formed within 6 days in cocultures of osteoblasts and bone marrow cells treated with 1,25-dihydroxyvitamin D(3) plus prostaglandin E(2). Docetaxel at 10(-8) M inhibited osteoclast formation in the coculture when added for the entire culture period or for the first 3 days. Docetaxel, even at 10(-6) M added for the final 3 days, failed to inhibit osteoclast formation. Osteoprotegerin, a decoy receptor of RANKL, completely inhibited osteoclast formation when added for the final 3 days. Docetaxel at 10(-8) M inhibited the proliferation of osteoblasts and bone marrow cells. RANKL mRNA expression induced by 1,25-dihydroxyvitamin D(3) plus prostaglandin E(2) in osteoblasts was not affected by docetaxel even at 10(-6) M. Docetaxel at 10(-6) M, but not at 10(-8) M, inhibited pit-forming activity of osteoclasts cultured on dentine. Actin ring formation and L: -glutamate secretion by osteoclasts were also inhibited by docetaxel at 10(-6) M. Thus, docetaxel inhibits bone resorption in two different manners: inhibition of osteoclast formation at 10(-8) M and of osteoclast function at 10(-6) M. These results suggest that taxanes have beneficial effects in the treatment of bone metastatic cancers.

Bone resorption in incompletely impacted mandibular third molars and acute pericoronitis.

Acute pericoronitis (AP) arises frequently in incompletely impacted mandibular third molars, but it remains unknown whether bone resorption in aging is associated with acute inflammation of the third molar. We conducted an experiment to compare the ratio of bone resorption to root length in the distal surface of the second molar (A), the proximal surface (B), and distal surface (C) in mesio-angular, incompletely impacted third molars in 27 young and 58 older adults with AP and 77 young and 79 older adults without a history of AP. Bone resorption in A, B, and C in older adults with AP demonstrated a significantly higher ratio when compared to those without AP, whereas there was no difference between those with and without AP in young adults except for B in women. However, there were no differences between bone resorption in B with AP in young and older women, and between bone resorption in C with AP in young and older adults. These indicate that AP and bone resorption are associated with incompletely impacted mandibular third molars in older adults.

Radiolucency below the crown of mandibular horizontal incompletely impacted third molars and acute inflammation in men with diabetes.

Although mandibular third molar has a high risk of infection extending any complications, the influence of diabetes on radiolucency and acute inflammation in pericoronitis remains unclear. The present study was to evaluate whether radiolucency below the crown is related to acute inflammation in mandibular horizontal incompletely impacted third molars and to review the records of 140 men more than 45 years with and without diabetes. The odds ratio of exhibiting acute inflammation was 3.38 (95% CI: 1.13-10.16, p < 0.05) and that of exhibiting severe acute inflammation was 15.38 (95% CI: 3.56-66.49, p < 0.0001), indicating an association of acute pericoronitis in diabetes. The frequency of radiolucency below the crown and below the root in diabetics was similar to that in nondiabetics. However, the odds ratio of exhibiting both radiolucency below the crown and acute inflammation under the diabetic condition was 4.85 (95% CI: 1.60-14.73, p < 0.01), whereas that of diabetics showing both radiolucency below the root and acute inflammation was 0.46 (95% CI: 0.06-3.74, p = 0.74). Radiolucency below the crown and acute inflammation were associated with diabetes, but that below root and acute inflammation were not associated with diabetes, indicating that the region below the crown carries susceptibility to acute pericoronitis, whereas the periodontium shows a protective effect against acute pericoronitis.

Acute inflammation at a mandibular solitary horizontal incompletely impacted molar.

Acute inflammation is frequently seen in the elderly around incompletely impacted molars located apart from molars or premolars. To identify the factors causing acute inflammation in the solitary molars without second molars or without second and first molars, ages of patients and rates of acute inflammation in 75 horizontal incompletely impacted mandibular molars in contact or not in contact with molars in subjects 41 years old or older were studied using orthopantomographs. Acute inflammation was seen in nine third molars out of 48 third molars in contact with second molars (18.8%), whereas acute inflammation was seen in 11 molars out of 19 solitary molars without second molars or without first and second molars (57.9%) (p < 0.01). The mean age of 48 subjects with third molars in contact with the second molar was 50.42 +/- 7.62 years, and the mean age of 19 subjects with isolated molars was 65.16 +/- 10.41 years (p < 0.0001). These indicate that a solitary horizontal incompletely impacted molar leads more frequently to acute inflammation along with aging due to possible bone resorption resulting from teeth loss.

Effect of inorganic polyphosphate in periodontitis in the elderly.

AIM: Inorganic polyphosphate exists as chains of phosphate molecules and is distributed in osteoblasts, and regulates osteoblastic cell differentiation and bone matrix calcification. The purpose of this study was to clarify the effects of inorganic polyphosphate on periodontitis. MATERIAL AND METHODS: Subgingival local irrigation with inorganic polyphosphate was studied in a randomised double-blind study of 33 patients with periodontitis. Scaling and root planing were performed 1 week after the initial examination. RESULTS: No significant differences between the inorganic polyphosphate group and control were detected in each item except IL-1beta. Patients in whom both the bleeding on probing and gingival index at 1 week had improved were significantly older in the inorganic polyphosphate group than in the control group (p < 0.05). Bone regeneration was seen in one case of the inorganic polyphosphate group. CONCLUSIONS: Inorganic polyphosphate was useful in the treatment of periodontitis in the elderly, indicating a probable effect of anti-ageing, with similar bone regenerations occurring in both groups.

Postnatal changes in 5HT and NK1 receptors in rat trigeminal motor nucleus and surroundings.

Our previous study showed that developmental changes to serotonin and substance P coexist in the trigeminal motor nucleus (Vmo), dorsolateral subnucleus (Vmo.dl), ventromedial subnucleus (Vmo.vm) and the area within 300 microm surrounding Vmo (SVmo). This occurred in rats from embryonic day (E)19 to postnatal day (P)70, with density of these terminals peaking at P7 in these three areas. The present study examined postnatal development of serotonergic 1A receptor (5HT1A receptor) and substance P receptor (NK1 receptor) expression in Vmo and SVmo in rats from E19 to P70. No significant changes in percentages of 5HT1A and NK1 receptor-expressing cells were seen between E19 and P70 by immunohistochemical study or in situ hybridization. In a real-time PCR study, quantities of 5HT1A and NK1 receptor expression peaked at P7, indicating that expression of these receptors was maximized in each neuronal cell body in the Vmo and SVmo at P7. This result corresponds with postnatal changes in serotonin/substance P-coexisting terminals found in our previous study. Furthermore, 5HT1A and NK1 receptors displayed very similar patterns of expression, which may support the hypothesis that potentiation of serotonin and substance P are involved in excitability regulating trigeminal motor functions, including mastication and breathing.

Multidrug resistance-associated protein 7 expression is involved in cross-resistance to docetaxel in salivary gland adenocarcinoma cell lines.

The aim of the present study was to clarify whether ATP binding cassette transporters are refractory factors in head and neck cancers. For in vitro and in vivo chemotherapeutic studies, we used the following head and neck cancer cell lines: a mouse oral squamous cell carcinoma (SCC) cell line, Sq-1979; a human SCC cell line, SCCHA; a mouse salivary gland adenocarcinoma (SGA) cell line, NR-PG; and a human SGA cell line, HSY. We used a vinca alkaloid anticancer drug, vincristine (VCR), as a chemotherapeutic anticancer drug. To determine the cause of multidrug resistance, Western blot analysis, reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry of xenografted tumors in nude mice, drug efflux analysis, and drug efflux inhibitory assays were performed. VCR-treated cell lines, Sq-1979/VCR, SCCHA/VCR, NR-PG/VCR, and HSY/VCR, intensively expressed multidrug resistance (MDR) gene 1 mRNA and multidrug resistance associated protein (MRP) 1 mRNA. MRP7 mRNA and protein were expressed in NR-PG/VCR and HSY/VCR cells, but not in Sq-1979/VCR and SCCHA/VCR cells. In each cell clone of NR-PG/VCR and HSY/VCR, MRP7 mRNA was induced by VCR treatment, suggesting an acquired resistance to VCR in the context of MRP7 expression. In the in vivo chemotherapeutic nude mice model, VCR-treated xenografted SCCHA and HSY cells expressed MDR1 and MRP1. Moreover, MRP7 expression was immunohistochemically found in xenografted HSY cells of VCR-injected tumor-bearing mice, but not in SCCHA cells. Furthermore, doxorubicin accumulation was increased and drug cross-resistance to docetaxel decreased in HSY/VCR in the presence of a competitive MRP7 inhibitor, 17-beta-estradiol-(17-beta-D-glucuronide). These results indicate that MDR1 expression, MRP1 expression, and MRP7 expression are refractory factors in head and neck cancer chemotherapy and suggest that induction of MRP7 expression is involved in drug resistance to natural products, especially to docetaxel in SGA.

NK1 receptor activation by geniohyoid primary afferents modulates parasympathetic postganglionic neuronal excitability in the rat.

Previous studies have indicated that the geniohyoid (GH) muscle receives innervation via both the hypoglossal nerve (CNXII) and the ansa cervicalis. Our recent studies revealed that the efferent root that contributes to the ansa cervicalis is a parasympathetic pathway and contains postganglionic cell bodies. Afferent axons from the GH muscle also travel via the ansa cervicalis, and afferent cell bodies are located in spinal ganglia. The present study attempts to locate the central terminations of these afferents. From the peripheral cut end of the ansa cervicalis, we recorded afferent discharges that coincided with inspiration and these were elicited by stretch of the GH muscle. After cutting CNXII proximal to its union with the ansa cervicalis, we applied horseradish peroxidase to the branch of CNXII that innervates the GH muscle. This procedure labeled cells ipsilaterally in the C2 spinal ganglia but not in the brainstem or upper spinal cord. Substance P-reactive terminals in the peripheral CNXII trunk were in apparent contact with vasoactive intestinal peptide-reactive cell bodies. Addition of the NK1 receptor agonist SP(NK1) excited parasympathetic postganglionic neurons and the specific NK1 receptor antagonist GR82334 blocked these effects in vitro. These results suggest that GH primary afferents synapse on parasympathetic postganglionic neurons in the CNXII trunk and that activation of SP(NK1) receptors modulates activity in these neurons.

Colocalization of serotonin and substance P in the postnatal rat trigeminal motor nucleus and its surroundings.

Trigeminal motoneurons are involved in a variety of oral motor activities, including mastication and breathing, which must be adapted to postnatal environmental change. Serotonin has both an excitatory and an inhibitory effect on trigeminal motor function, whereas substance P has mainly an excitatory effect. In the present study, we measured the density of serotonin- and substance P-immunoreactive nerve terminals in the trigeminal motor nucleus and the area 300 microm surrounding it in rats from embryonic day 19 to postnatal day 70. The density of these terminals gradually increased from embryonic day 19 to postnatal day 7 and decreased thereafter. The density was greatest in the ventromedial subnucleus of the trigeminal motor nucleus at embryonic day 19 and postnatal day 0 and in the area 300 microm surrounding trigeminal motor nucleus at postnatal day 4 and older. Two-color fluorescence immunohistochemistry was used to identify nerve processes immunoreactive for both substance P and serotonin. Approximately, 90% of serotonergic terminals also contained substance P at all ages examined, which suggests that the physiological function of terminals in which these neurotransmitters are colocalized is similar throughout development.

Muramyl dipeptide enhances osteoclast formation induced by lipopolysaccharide, IL-1 alpha, and TNF-alpha through nucleotide-binding oligomerization domain 2-mediated signaling in osteoblasts.

Muramyl dipeptide (MDP) is the minimal essential structural unit responsible for the immunoadjuvant activity of peptidoglycan. As well as bone-resorbing factors such as 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3) and PGE2, LPS and IL-1alpha stimulate osteoclast formation in mouse cocultures of primary osteoblasts and hemopoietic cells. MDP alone could not induce osteoclast formation in the coculture, but enhanced osteoclast formation induced by LPS, IL-1alpha, or TNF-alpha but not 1alpha,25(OH)2D3 or PGE2. MDP failed to enhance osteoclast formation from osteoclast progenitors induced by receptor activator of NF-kappaB ligand (RANKL) or TNF-alpha. MDP up-regulated RANKL expression in osteoblasts treated with LPS or TNF-alpha but not 1alpha,25(OH)2D3. Osteoblasts expressed mRNA of nucleotide-binding oligomerization domain 2 (Nod2), an intracellular sensor of MDP, in response to LPS, IL-1alpha, or TNF-alpha but not 1alpha,25(OH)2D3. Induction of Nod2 mRNA expression by LPS but not by TNF-alpha in osteoblasts was dependent on TLR4 and MyD88. MDP also enhanced TNF-alpha-induced osteoclast formation in cocultures prepared from Toll/IL-1R domain-containing adapter protein (TIRAP)-deficient mice through the up-regulation of RANKL mRNA expression in osteoblasts, suggesting that TLR2 is not involved in the MDP-induced osteoclast formation. The depletion of intracellular Nod2 by small interfering RNA blocked MDP-induced up-regulation of RANKL mRNA in osteoblasts. LPS and RANKL stimulated the survival of osteoclasts, and this effect was not enhanced by MDP. These results suggest that MDP synergistically enhances osteoclast formation induced by LPS, IL-1alpha, and TNF-alpha through RANKL expression in osteoblasts, and that Nod2-mediated signals are involved in the MDP-induced RANKL expression in osteoblasts.