Nobuyoshi Mizukawa

Levels of human defensin-1, an antimicrobial peptide, in saliva of patients with oral inflammation

OBJECTIVE The purpose of this study was to investigate the presence of an antimicrobial peptide, human defensin-1, in the saliva of patients with oral inflammation. STUDY DESIGN Whole saliva samples were collected from patients with oral inflammation and from healthy volunteers. Human defensin-1 in saliva was isolated and purified by reversed-phase high-performance liquid chromatography. The amino acid sequence and molecular weight of defensin-1 were determined. The concentration of defensin-1 in saliva was quantified by high-performance liquid chromatography. Serum C-reactive protein concentration was measured by particle-enhanced turbidimetric immunoassay. RESULTS The salivary defensin-1 concentration was significantly higher in patients with oral inflammation than in healthy volunteers; furthermore, in patients with oral inflammation, the concentration was significantly higher before treatment than after treatment. In the patients with oral inflammation, there was a strong positive correlation between salivary defensin-1 concentration and serum C-reactive protein concentration. CONCLUSIONS The findings suggest that defensin-1 in saliva may be a convenient marker of inflammation associated with oral disease.

Regulation of bone metabolism in immunosuppressant (FK506)-treated rats

After organ transplantation, severe osteoporosis is occasionally seen, and the use of immunosuppressants is thought to be one of the causes of such osteoporosis. In the present study, we investigated the effects of FK506 monotherapy on bones and determined the mechanism of onset of osteoporosis, both by assessing chronological changes in bone metabolism and by identifying factors that facilitate bone resorption. In 8-week-old male Sprague-Dawley rats, FK506 (1 mg/kg) was injected intraperitoneally every day for 5 weeks (FK506-treated group), and for comparison, physiological saline was administered in the same manner in a control group of rats. Serum and urine samples were collected at weeks 0, 1, 3, and 5 of administration. The femur and tibia were collected within 24 h of the final administration. When compared to the control group, findings on three-dimensional micro-computed tomography of the femur for the FK506-treated group showed a significant decrease in trabecular bone volume. The level of serum osteocalcin in the FK506-treated group at week 1 of administration was significantly higher than the control. Throughout the administration period, the sum of urinary pyridinoline (PYD) and deoxypyridinoline (Dpd) was significantly higher in the FK506-treated group. Of the various bone resorption factors tested, the level of serum parathyroid hormone (PTH) in the FK506-treated group was significantly higher than the control at week 3 of administration. The results of the present study confirmed that FK506 monotherapy in rats induced high-turnover osteoporosis. Soon after the start of FK506 administration, bone formation and resorption were elevated, and PTH appeared to have been involved in the maintenance of the elevated bone resorption.

Immobilized recombinant human bone morphogenetic protein-2 enhances the phosphorylation of receptor-activated Smads.

Bone morphogenetic protein (BMP)-2 plays an important role in bone growth and regeneration; however, BMP-2 is easily lost by diffusion through body fluid and has some inhibitory pathways. To address this problem, we previously immobilized recombinant human BMP-2 (rhBMP-2) on succinylated type I atelocollagen. Here, we examined the effect of immobilized rhBMP-2 in vitro and vivo. In ST2, MC3T3-E1, and C2C12 cells, alkaline phosphatase activity, which is a marker of osteoblast differentiation, was enhanced more by immobilized than nonimmobilized rhBMP-2. In addition, the phosphorylation of receptor-activated Smads, part of the signaling pathway activated by BMP-2, was prolonged by immobilized rhBMP-2 in these cells. Furthermore, implantation of immobilized rhBMP-2 into the backs of rats promoted the formation of mature bone-like structure. These results demonstrate that immobilized rhBMP-2 has higher bioactivity than nonimmobilized rhBMP-2, and, therefore, immobilization of rhBMP-2 can prolong BMP signaling.

Basic fibroblast growth factor supports expansion of mouse compact bone-derived mesenchymal stem cells (MSCs) and regeneration of bone from MSC in vivo

Some progress has been made in development of methods to regenerate bone from cultured cells, however no method is put to practical use. Here, we developed methods to isolate, purify, and expand mesenchymal stem cells (MSCs) from mouse compact bone that may be used to regenerate bone in vivo. These cells were maintained in long-term culture and were capable of differentiating along multiple lineages, including chondrocyte, osteocyte, and adipocyte trajectories. We used standard cell isolation and culture methods to establish cell cultures from mouse compact bone and bone marrow. Cultures were grown in four distinct media to determine the optimal composition of culture medium for bone-derived MSCs. Putative MSCs were subjected to flow cytometry, alkaline phosphatase assays, immunohistochemical staining, and several differentiation assays to assess cell identity, protein expression, and developmental potential. Finally, we used an in vivo bone formation assay to determine whether putative MSCs were capable of regenerating bone. We found that compact bone of mice was a better source of MCSs than the bone marrow, that growth in plastic flasks served to purify MSCs from hematopoietic cells, and that MSCs grown in basic fibroblast growth factor (bFGF)-conditioned medium were, based on multiple criteria, superior to those grown in leukemia inhibitory factor-conditioned medium. Moreover, we found that the MSCs isolated from compact bone and grown in bFGF-conditioned medium were capable of supporting bone formation in vivo. The methods and results described here have implications for understanding MSC biology and for clinical purpose.

Quantitative analysis of free flap volume changes in head and neck reconstruction.

The purpose of this study was to determine whether free flap volume decreases or increases in the long‐term postoperative period.

Regeneration of the mandibular head from grafted periosteum.

Grafted periosteum has a rich potential to induce heterotopic bone formation. In the current study the authors investigate whether autogenous periosteal grafts can regenerate the mandibular head in a rabbit model. They removed the mandibular head of Japanese white rabbits and grafted tibial periosteum to the cut surface of the mandible. Grafted periosteum was observed histologically and radiographically at day 7, 14, 21, and 45 after surgery. At day 7 after grafting, grafted tissue showed remarkable cell proliferation. By 14 days these cells had differentiated into chondrocytes to form cartilage, and endochondral ossification took place after 21 days. At 45 days after surgery, soft X-ray findings showed a newly formed mandibular head, which was similar histologically to that of a normal mandibular head. The cut mandible without periosteal graft showed no regeneration. These findings indicate that grafted periosteum can regenerate the mandibular head without special procedures such as bone fixation in a rabbit model, and suggest that this technique may be useful clinically.

Treatment of a plunging ranula with fenestration and continuous pressure

We present a new method of fenestration and continuous pressure as a simple, effective and uninvasive procedure for the treatment of plunging ranulas. We have recently used in four female patients, aged 10-29 years old. After treatment, the patients remained symptom-free and assessment by magnetic resonance imaging (MRI) showed regression of the ranula in all cases. The procedure resulted in satisfactory healing and we advocate it as a simple and effective treatment that is better for patients than conventional treatment.

Evaluation of osteogenic/chondrogenic cellular proliferation and differentiation in the xenogeneic periosteal graft.

To determine whether grafted young periosteum can induce new bone formation in elderly patients, this preliminary study evaluated cell proliferation and differentiation in xenogeneic periosteal grafts in old rats radiographically, histologically, and immunohistochemically. Periosteum harvested from the tibia of young Japanese white rabbits were grafted into old Sprague–Dawley rats with or without administration of 1.0 mg per kilogram per day immunosuppressant FK506. Autogenous old periosteal tissue grafts were also evaluated as a control. Grafted tissue was extirpated after 7, 14, 21, and 45 days. In the xenogeneic group, proliferative cell nuclear antigen-positive cells were observed 7 days after surgery, which differentiated into chondroblasts with bone morphogenetic protein-2 expression and finally formed cartilage by 14 days. Endochondral ossification was observed at 21 days, and bone replacement was completed by 45 days. No osteogenic cell activity was observed in the two other groups. Xenogeneic young periosteum thus maintained its osteogenic/chondrogenic potentiality in older rats.

Prevention points for plate exposure in the mandibular reconstruction.

INTRODUCTION The rate of complications for mandibular reconstruction after segmental mandibulectomy is higher with reconstruction plates than with vascularised bone grafts. We have experience of over 100 patients using reconstructive plates for reconstruction immediately after segmental mandibulectomy and have considered factors contributing to plate exposure. PATIENTS AND METHODS Seventeen cases utilised our prevention methods in which reconstructive plates were used for mandibular reconstruction were reviewed. The flaps used with reconstruction plates were rectus abdominis myocutanenous flaps in 10 cases, anterolateral thigh flaps combined vastus lateralis muscle in four cases, and the omentum in one case; no flap was transferred in two cases. RESULTS In only one of 17 cases was a plate exposed at 3 months postoperatively. No plate exposure occurred during the follow-up period in the other 16 cases. Because no flap had been transferred in the patient with plate exposure, a possible contributing factor was the persistence of dead space beneath the plate. CONCLUSION This series suggests that factors other than flap selection contribute to the exposure of reconstructive plates. Use of a reconstruction plate is a useful reconstructive method, especially for patients who cannot tolerate transfer of a vascularised bone graft.

Bone quality and quantity of the anterior maxillary trabecular bone in dental implant sites

OBJECTIVES The aim of this study was to investigate the characteristics of implant sites on the edentulous alveolar ridge in the anterior maxilla. We studied the bone quantity and quality of implant sites at the anterior maxilla using CT images for the 33 implant sites on patients who underwent dental implant therapy in our Department since 2006. MATERIALS AND METHODS Computed tomography (CT) images of 33 patients (20 women: 13 men) encompassing 33 implant sites were chosen and examined. The recipient sites for implant placement were determined based on CT data using an implant planning software (Simplant 11.0). The mean bone density values in Hounsfield unit (HU) were recorded using Simplant for both the simulated implant areas and the trabecular bone width. We classified the edentulous alveolar ridge and bone quality according to a classification based on Lekholm and Zarb (1985). RESULTS Incisors had higher bone densities than canines. Women had lower bone densities than men. Canines displayed greater trabecular bone density and alveolar bone widths than incisors. No maxillary sites were judged to have a bone quality of 1 in this group. Quality 3 accounted for 69.7% of the total samples. CONCLUSIONS An assessment of bone quality in the anterior alveolar ridge may well reflect age-related systemic pathological conditions and should be used in dental implant treatment planning to avoid associated risk factors.