Kiyofumi Furusawa

Muramyl Dipeptide Enhances Osteoclast Formation Induced by Lipopolysaccharide, IL-1α, and TNF-α 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 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3) and PGE2, LPS and IL-1α 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-1α, or TNF-α but not 1α,25(OH)2D3 or PGE2. MDP failed to enhance osteoclast formation from osteoclast progenitors induced by receptor activator of NF-κB ligand (RANKL) or TNF-α. MDP up-regulated RANKL expression in osteoblasts treated with LPS or TNF-α but not 1α,25(OH)2D3. Osteoblasts expressed mRNA of nucleotide-binding oligomerization domain 2 (Nod2), an intracellular sensor of MDP, in response to LPS, IL-1α, or TNF-α but not 1α,25(OH)2D3. Induction of Nod2 mRNA expression by LPS but not by TNF-α in osteoblasts was dependent on TLR4 and MyD88. MDP also enhanced TNF-α-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-1α, and TNF-α through RANKL expression in osteoblasts, and that Nod2-mediated signals are involved in the MDP-induced RANKL expression in osteoblasts.

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.

Inorganic Polyphosphate: a Possible Stimulant of Bone Formation

Inorganic polyphosphates [Poly(P)] are often distributed in osteoblasts. We undertook the present study to verify the hypothesis that Poly(P) stimulates osteoblasts and facilitates bone formation. The osteoblast-like cell line MC 3T3-E1 was cultured with Poly(P), and gene expression and potential mineralization were evaluated by reverse-transcription polymerase chain-reaction. Alkaline phosphatase activity, von Kossa staining, and resorption pit formation analyses were also determined. The potential role of Poly(P) in bone formation was assessed in a rat alveolar bone regeneration model. Poly(P) induced osteopontin, osteocalcin, collagen 1α, and osteoprotegerin expression and increased alkaline phosphatase activity in MC 3T3-E1 cells. Dentin slice pit formation decreased with mouse osteoblast and bone marrow macrophage co-cultivation in the presence of Poly(P). Promotion of alveolar bone regeneration was observed locally in Poly(P)-treated rats. These findings suggest that Poly(P) plays a role in osteoblastic differentiation, activation, and bone mineralization. Thus, local poly(P) delivery may have a therapeutic benefit in periodontal disease.

Orthodontic Treatment of an Impacted Dilacerated Maxillary Central Incisor Combined with Surgical Exposure and Apicoectomy

The impacted incisor was moved into its proper position with surgical exposure and orthodontic traction. Although apicoectomy was performed during the orthodontic treatment, the incisor showed good stability after the long retention period.

Central distribution and peripheral functional properties of afferent and efferent components of the superior laryngeal nerve: morphological and electrophysiological studies in the rat.

The central distribution of the afferent and efferent components of the superior laryngeal nerve (SLN), which in the rat is ramified into the three branches of the rostral branch (R.Br), middle branch (M.Br), and caudal branch (C.Br), was examined after application of horseradish peroxidase conjugated with wheat germ agglutinin (HRP‐WGA) to the proximal cut end of each branch. In addition, the afferent and efferent neural activities of each branch were recorded to investigate the functional properties. The present study provided several new findings as to the distribution of each branch and the functional properties of the SLN. The following conclusions were drawn: 1) the R.Br, containing only afferent fibers projecting to the ipsilateral lateral region of the nucleus of the solitary tract (NST), extends between slightly below the obex and the region approximately 0.6 mm rostral from the obex, and it corresponds to the interstitial subnucleus of the NST; 2) the M.Br, innervating the cricothyroid muscle, contains only efferent fibers originating ipsilaterally from the motoneurons localized within the ambiguus nucleus (Amb) and in the area ventrolateral to the Amb; and 3) the C.Br, which innervates the inferior pharyngeal constrictor muscle, contains both efferent and afferent fibers. HRP‐WGA‐labeled cells are distributed within both the Amb and the dorsal motor nucleus of the vagus nerve, ipsilateral to the injection site. Afferent proprioceptive fibers project to the ipsilateral interstitial subnucleus of the NST. The present results provide evidence that each branch of the SLN has distinctive functional properties and contributes to the laryngeal functions.

Inorganic Polyphosphate Induces Osteoblastic Differentiation

Inorganic polyphosphate [Poly(P)] is especially prevalent in osteoblasts. We tested the hypothesis that Poly(P) stimulates osteoblastic differentiation and polyphosphate metabolism for bone formation. The osteoblast-like cell line, MC 3T3-E1, was cultured with Poly(P), and gene expression was evaluated by real-time reverse-transcription polymerase chain-reaction. Phosphatase activity and extracellular matrix mineralization were also determined. The role of Poly(P) was assessed in a beagle dog alveolar bone regeneration model. Poly(P) increased osteocalcin, osterix, bone sialoprotein, and tissue non-specific alkaline phosphatase gene expression, with a high level of end-polyphosphatase activity, resulting in low-chain-length Poly(P), inorganic pyrophosphate, and inorganic phosphate production. MC3T3-E1 cells differentiated into mature osteoblasts and showed expression of ectonucleotide pyrophosphatase phosphodiesterase 1, while mouse progressive ankylosis gene expression remained unchanged. Promotion of alveolar bone regeneration was observed in Poly(P)-treated beagle dogs. These findings suggest that Poly(P) induces osteoblastic differentiation and bone mineralization, and acts as a resource for mineralization.

The assessment of fracture of the mandibular condyle by use of computerized tomography. Incidence of sagittal split fracture

A survey was carried out to clarify the incidence of sagittal splitting fracture of the mandibular condyle using computerized tomography. There were 33 patients, between 11 and 67 years of age, with displaced or dislocated mandibular condylar process fractures (41 cases), seen at our clinic between 1986 and 1992. The incidence of no displacement was 4.9%; deviation and displacement, 34.1%; dislocation, 46.3%; and complete avulsion, 4.9%. A sagittal splitting fracture of condyle occurred with an incidence of 9.8%. Conservative treatment was effective in the treatment of sagittal splitting fracture. Therefore, classification of fracture of mandibular condyle should include the sagittal split fracture, and investigations should include computerized tomography.

Early evaluation of necrotizing fasciitis with use of CT

Gas bubbles are one of the important signs in the clinical diagnosis of necrotizing fasciitis, and radiographs are mandatory. An instance where gas bubbles not found on the plain radiographs were clearly shown by CT, which is a useful method for close monitoring and decompression of gas bubbles situated deeply in the spaces of the maxillo-facial and neck regions.

The innervation of the levator veil palatini muscle by the glossopharyngeal nerve

We developed a novel method which enables bloodless exposure of the levator veli palatini muscle in rat in order to investigate the physiological properties of this muscle. The levator veli palatini muscle which is innervated by a branch of the glossopharyngeal nerve showed rhythmic spontaneous movement in rats. Cutting the branch supplying LVP of the glossopharyngeal nerve caused cessation of the spontaneous movement of the levator veli palatini muscle. The spontaneous discharges of the glossopharyngeal nerve were synchronized with those of the phrenic nerve. A mixture of 95% oxygen and 5% room air influenced the efferent discharges from the branch of the glossopharyngeal nerve supplying the levator veli palatini muscle. These findings indicate that the motor nerve supply to the levator veli palatini muscle is the glossopharyngeal nerve, and the levator veli palatini muscle is related to the respiratory system, in particular with inspiration in rats.

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.