Characterisation of a sucrose-independent in vitro biofilm model of supragingival plaque.

OBJECTIVES: Sugar consumption has been decreasing in Japan, suggesting higher rates of sucrose-independent supragingival plaque formation. For developing an in vitro biofilm model of sucrose-independent supragingival plaque, this study aimed to investigate the compositions and functions on contributing to cariogenicity in comparison with sucrose-dependent biofilm. MATERIALS AND METHODS: An in vitro multispecies biofilm containing Actinomyces naeslundii, Streptococcus gordonii, S. mutans, Veillonella parvula and Fusobacterium nucleatum was formed on 24-well plates in the absence or presence of 1% sucrose. Compositions were assessed by plate culture, scanning electron microscopy and confocal laser scanning microscopy after fluorescent in situ hybridisation or labelling of extracellular polymeric substances (EPS). Functions were assessed by acidogenicity, adherence strength and sensitivities to anticaries agents. RESULTS: Although both biofilms exhibited a Streptococcus predominant bacterial composition, there were differences in bacterial and EPS compositions; in particular, little glucan EPS was observed in sucrose-independent biofilm. Compared with sucrose-dependent biofilm, acidogenicity, adherence strength and antimicrobial resistance of sucrose-independent biofilm were only slightly lower. However, dextranase degradation was substantially lower in sucrose-independent biofilm. CONCLUSION: Our findings suggest that sucrose-independent biofilm may have cariogenicity as with sucrose-dependent biofilm. These in vitro models can help further elucidate plaque-induced caries aetiology and develop new anticaries agents.

Manometric evaluation of oral function with a hand-held balloon probe.

Tongue pressure measured with a disposable hand-held balloon probe has been used for assessing tongue function; however, no diagnostic standard for assessing other oral functions exists currently. In this study, to develop a method for multifactorial manometric evaluation of oral functions, 20 men and 20 women (21-32 years of age) were instructed to apply 7-s maximal voluntary muscular effort on a hand-held balloon probe placed against the anterior and lateral parts of the palate, buccal surface of the molars on the habitual chewing side and labial surface of the anterior teeth for measuring anterior and posterior tongue, cheek and lip pressures (LPs), respectively. Intra-session reproducibility was determined by three repeated measurements, and associations between the obtained and the conventional data on oral diadochokinesis were tested. The men exhibited higher values of all the pressure types. Further, both genders showed a positive correlation between anterior and posterior tongue pressures (PTPs) (P < 0·05), but only the women exhibited positive correlations between anterior tongue and cheek pressures, cheek and PTPs, anterior tongue and LPs, and cheek and LPs (P < 0·05). No statistically significant correlation was found between the pressures and the number of syllabic articulations, except between LP and the number of /pa/ articulations in the women (r = 0·524, P < 0·05). In conclusion, the balloon probe method enables objective manometric evaluation of oral functions and could be an effective tool for clinical epidemiological studies and evidence-based decision-making in nursing care.

Resting T cells negatively regulate osteoclast generation from peripheral blood monocytes.

There is accumulating evidence that T cells may be involved in osteoclastogenesis in a variety of murine systems. However, the precise role of human T cells in the regulation of osteoclast generation is still unclear. To address this issue, we investigated the effect of resting peripheral T cells on receptor activator of NF-kappaB ligand (RANKL)-induced osteoclast generation from human peripheral monocytes. Although osteoclasts were not generated in the culture of human peripheral blood mononuclear cells (PBMC) in the presence of RANKL and macrophage colony-stimulating factor (M-CSF), the addition of cyclosporine A (CsA), a potent inhibitor of T-cell function, resulted in the formation of an increasing number of lacunae resorption on dentine, suggesting T cells may inhibit osteoclast formation. In a coculture of T cells and monocytes, which were isolated from PBMC, T cells inhibited the osteoclast generation from monocytes, as determined by tartrate-resistant acid phosphatase (TRAP) staining and a pit assay using dentine. This inhibition of osteoclast generation by T cells was also observed in a culture of the parathyroid hormone-stimulated SaOS4/3 osteoblast cell line and monocytes. The culture in Transwell plates revealed that the cell-to-cell interaction was not required for the inhibition, suggesting that T-cell cytokines may be responsible for the inhibition. Among inhibitory T-cell cytokines on osteoclastogenesis, granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (IFN-gamma) were actively produced by CD4 T cells but not CD8 T cells in the coculture of T cells with monocytes, and the neutralizing antibodies to these cytokines partially rescued the T-cell-induced inhibition of osteoclast formation. Although CsA did not affect RANKL-induced osteoclast generation in the culture of monocytes alone, it completely rescued the T-cell-induced inhibition of osteoclast formation and strongly inhibited the production of GM-CSF and IFN-gamma. Thus, we demonstrate that resting T cells negatively regulate the osteoclast generation via production of GM-CSF and IFN-gamma by CD4 T cells and that CsA stimulates the osteoclast generation through the inhibition of the production of these cytokines. These findings provide new insight into therapeutic strategies for immunosuppression-induced bone loss in transplant and other diseases.

The detection of a unique antigen associated with papillary thyroid carcinoma.

We produced antibodies against a thyroid papillary carcinoma homogenate (PCAb) and analyzed antigens recognized by this antibody using western blotting. Fifty-four thyroid tissue specimens and 6 control tissue specimens obtained from non-thyroid carcinoma (gastric tissue, colon and liver) were analyzed. Consequently, an antigen of 40 kDa in size was found in 16 of 16 (100%) of papillary thyroid carcinoma from primary lesions and in 2 of 2 (100%) papillary thyroid carcinoma from metastatic foci, whereas it was not detected in thyroid tissue samples from follicular carcinoma, anaplastic carcinoma, follicular adenoma, adenomatous goiter, Graves' disease and normal thyroid tissues. The reactivity of thyroglobulin antiserum (TgAb) to this 40 kDa antigen was tested by western blotting and showed that TgAb did not appear to recognize the 40 kDa antigen. Moreover PCAb, after treatment with Tg, still reacted with this 40 kDa antigen. Therefore, this 40 kDa antigen might be different from Tg. Furthermore, to inspect the structure of this antigen, the effect of some chemicals and enzymes such as 2-mercaptoethanol, sodium dodecyl sulfate, ethanol and protease on the reactivity of PCAb to the 40 kDa antigen were analyzed. The results of these experiments suggested that this 40 kDa antigen may have a peptide structure. To our knowledge, the finding reported here represents the first demonstration of the protein specifically present in papillary thyroid carcinoma. Further investigations should elucidate the characteristics of this antigen and may contribute to definitive diagnosis of thyroid carcinoma as well as improving the understanding of the mechanisms involved in developing the thyroid carcinoma.

Hydroxyapatite clay used to fill the gap between implant and bone.

A clay containing hydroxyapatite (HA) was developed for use as a filling material between an uncemented implant and bone. It consists of 55% HA granules greater than 0.1 mm in size with a homogeneous pore distribution and a porosity of 35% to 48% in a saline solution of sodium alginate (6%). Ti-6A1-4V alloy rods with smooth surfaces were implanted into the distal medullary canal of one osteotomised tibia of 32 Japanese white rabbits. Sixteen rods were inserted with HA clay and 16 without the clay to act as a control group. Six of each group were killed at one week and ten at 12 weeks postoperatively. The pull-out strength of the implants with HA clay was significantly greater at 12 weeks (p < 0.05), as was the percentage of the area of the new bone (p < 0.05). The study suggests that HA clay has an osteoconductive property, allowing adequate bone fixation across a gap at an early stage. The use of HA clay to enhance the early stability of uncemented components may help to improve the functional outcome of total joint arthroplasty.

[Oro-pharyngeal burn during electrodissection of the adenoid and tonsil].

We present a case of oro-pharyngeal burn which occurred during electrodissection of the adenoid and tonsil in a 5-year-old boy. We intubated the patient with an uncuffed spiral tube of appropriate size and noticed a slight gas leak during positive-pressure ventilation. Anesthesia was maintained with a mixture of 60% nitrous oxide, 40% oxygen and 2.5% sevoflurane. During manipulations of the right tonsil, orange-colored flame blew out about 5 cm from the mouth. Fortunately, the patient underwent the operative procedures without any further troubles, recovered fully from the grade 1 burn in the oral mucosa, and was discharged 23 days after surgery. The surgeons speculated that sevoflurane had been ignited. Although it is well known that sevoflurane is nonflammable in the concentration of clinical use, several reports show that sevoflurane is flammable in concentration of 10% under pure oxygen or nitrous oxide. We concluded that this accident was caused by electrocautery-induced ignition of the gauze packed into the larynx under a high concentration of oxygen which leaked through an uncuffed endotracheal tube. We have to bear in mind that any flammable substance may ignite when using electrocautery in a small space such as the mouth under oxygen-rich environment.

[Chronic necrotizing pulmonary aspergillosis resistant to antimycotic drugs and treated by partial pulmonary resection].

A 46-year-old man with a history of left upper lobectomy for pulmonary tuberculosis was admitted to our hospital because of dilated cardiomyopathy. During hospitalization, fever and weight loss developed. The cause was suspected to be a round mass inside a cavity and a neighboring infiltrative shadow in the left upper lung field as seen on chest radiography. A percutaneous needle biopsy was done, and examination of the specimen showed an aggregate of Aspergillus fumigatus hyphae. Fluconazole (FCZ) was injected through an intracavitary catheter every day, and was then given by mouth. Treatment with FCZ was effective temporarily. However, he was again admitted to our hospital because of lower extension of the cavity and deteriorated inflammatory findings. From the clinical course, chronic necrotizing pulmonary aspergillosis was diagnosed. Treatment with all available antifungal agents did not improve his condition. Although he had decreased cardiac function due to dilated cardiomyopathy, partial pulmonary resection was done. The cavity with the fungus ball was resected completely. As of the time of this writing, he remains free of aspergillosis.

Hydroxyapatite-clay bone fixation for loaded implants.

A clay containing hydroxyapatite (HA clay), which was made by mixing HA granules (range of sizes: 0.1-0.3 mm) and a saline solution of sodium alginate, was inserted into the medullary canal of an osteotomized rabbit's tibia with a Ti-6Al-4V titanium alloy implant. Each implant had a conical portion for bearing load. The shear strength of the bone-implant interface for the implant with HA clay was significantly greater than that for the implant without HA clay 3 months postoperatively (P < .02), while there was no significant difference between the two strengths 1 week postoperatively. Under microscopic observation, the percentage of area of newly developed bone was also significantly greater for the implant with HA clay than for the implant without HA clay 3 months postoperatively (P < .04). This study suggests that HA clay encouraged adequate bone fixation of the loaded implant in 3 months, while the clay was not effective for immediate fixation.

In vivo properties of an intramedullary hydroxyapatite plug to improve femoral stem fixation.

An intramedullary hydroxyapatite (HA) plug was inserted in each tibia of five rabbits. Histological observation showed the presence of newly formed bone tissue bridging between the inner cortex and the HA plug at 3 months postoperatively. This new bone was in direct contact with the HA plug, and there was no fibrous tissue intervention. Based on these results, the HA plug was used in a hip hemiarthroplasty with bone cement. The radiograph taken 4 years postoperatively showed that the plug-bone interface was indistinct, and some of the plug's edges became rounded by replacement with newly developed bone. The femoral component was stable without subsidence. The HA plug stabilized by bone ingrowth in the femoral canal would certainly be useful to prevent the femoral component from subsiding, in addition to increasing the cement-bone interface pressure at the time of surgery.

In vitro properties of a chitosan-bonded self-hardening paste with hydroxyapatite granules.

A new self-hardening paste was made by using a combination of chitosan, hydroxyapatite (HA) granules, ZnO, and CaO. The sol was made by dissolving 0.1 g of chitosan in a solution of 0.1 g malic acid and 2.0 mL physiological saline solution. Mixed with 0.03 g of CaO and 0.04 g of ZnO powders was 2.77 g (55 wt %) of HA granules which had a homogeneous pore distribution and a porosity of 35-48%. The size of the granules was set for 0.1-0.3 mm. Kneading and setting of the paste generated a little amount of heat (32.8 degrees C) as compared with the heat produced by polymethyl-methacrylate (PMMA) bone cement (114.5 degrees C). The pH value of chitosan-HA-hardened composite after setting was nearly equal to that of human plasma (pH 7.4), while that of PMMA bone cement maintained an acid pH of 4.7. Hydroxyapatite granules less than 0.1 mm, 0.1-0.3 mm, or 0.3-0.6 mm were set using chitosan sol. The size of the granules did not influence the compressive strength of the set chitosan-HA-hardened composite. The greatest compressive strength of chitosan-HA-hardened composite was obtained by using 55 wt % of HA granules. The strength of the chitosan-HA-hardened composite was comparable to that of the cancellous bone derived from tibial eminentia, but was considerably lower than that of the PMMA bone cement.

Current progress and perspectives in immunoisolated islet transplantation.

This review summarizes the recent results of immunoioslated islet allo- and xenotransplants, especially in large animal species, and presents certain issues that would be important for the advancement of this technology toward clinical application. To date, the best results have been obtained with alginate microcapsules in both allo- and xenogeneic (porcine) islet transplantation in the spontaneously diabetic dogs and monkeys. Reversal of diabetes was also achieved in mice with cryopreserved, microencapsulated rat islets. However, results with these microcapsules have been highly variable and inconsistent from one laboratory to another. The causes of these discrepancies are multiple, but not totally understood. Immunoisolation devices have been investigated as an alternate approach to house a large number of islets in a space constructed by membranes of a defined pore size. Vascularized bioartificial devices were among the few cases that obtained long-term allo- and xenogeneic islet survival in totally pancreatectomized dogs. Thrombosis and associated problems were the major cause of failure with these devices. Diffusion chambers that have no vascular connection have been tested successfully in rodents, but no report has been available in large animal species. We have learned many important lessons and made considerable progress in islet immunoisolation. However, immunoisolated islet transplants have been only minimally successful in large animal models, and such technologies must be improved to achieve consistent success in large animal models prior to clinical trials. This will requires strict quality control of the islets, the membrane material, and the device construction. The membrane pore size that allows the permeation of molecules necessary for islet survival and function also permits the entry of cytokines, oxygen-radicals, and other small-size inflammatory products. Thus, the key to success appears to be to minimize inflammatory reactions generated by immuneisolation devices/capsules.

Fracture of the tibial tray following PCA knee replacement. A report of two cases.

We report two cases in which fracture of the tibial metal tray was thought to be due to improper design of the tray and heavy polyethylene wear. The bone beneath the fractured portion of the tray was deficient and had been replaced by granulomatous fibrous tissue including numerous polyethylene wear particles and foreign-body giant cells. Osteolysis occurred as a tissue reaction to these particles.

[Relationship of TMJ sounds and mandibular movements].

TMJ sounds are very important for diagnosing the state of TMJ and for evaluation of the effect of treatment. We made a report on a recording system of TMJ sounds and mandibular movement which objectively can evaluate the relationship between TMJ sounds and mandibular movement. This time we recorded and analyzed the TMJ sounds and mandibular movement under following condition. 1. Opening and closing the mandibular on some regulated velocity. 2. Opening and closing the mandibular after clenching. Conclusion 1. In case of changing habitual opening velocity, there were significantly different position of clicking in the most of TMJ disorders patients. Especially with the increasing of the velocity, the position of closing clicking tended to come near the closed condyle position. 2. The amplitude of clicking sound increased very much at the time of opening stage after clenching on the non clicking side.

Effect of low-friction ion-treated femoral heads on polyethylene wear rates.

Polyethylene wear is a major contributor to osteolysis and subsequent aseptic loosening of prosthetic components in total hip arthroplasty. Use of ion implantation as a surface modification to the metallic bearing component of orthopaedic implants may be an effective means of reducing wear debris at the bearing interface. In July 1991, low friction ion treated femoral heads were introduced. This study evaluates the effect of the low friction ion treated femoral head on polyethylene wear. Fifty-five total hip arthroplasties (53 patients) with low friction ion treated femoral heads followed up a minimum of 3 years were matched with 55 total hip arthroplasties (47 patients) without low friction ion treated femoral heads for the same postoperative period. Socket wear was evaluated radiographically. Case matching and strict inclusion criteria were used to control for known factors influencing polyethylene wear. These criteria included: (1) cases matched for gender and age within 2 years; (2) diagnosis limited to osteoarthritis or avascular necrosis of the femoral head only; (3) femoral head diameter limited to 26 or 28 mm only; (4) hydroxyapatite coated femoral stem of the same design and a metal backed socket of the one of two designs with the same polyethylene insert; and (5) minimum followup of 3 years. The linear wear rate of polyethylene was 0.161 +/- 0.095 mm per year in the group without the low friction ion treated heads and 0.116 +/- 0.101 mm per year in the low friction ion treated group. The volumetric wear rates were 74.5 +/- 44.3 mm3 per year for the group without the low friction ion treated heads and 57.8 +/- 51.1 mm3 per year for the low friction ion treated group. Assuming the sensitivity of these measurements can detect these small differences in wear accurately, these results suggest low friction ion treated prosthetic heads are useful in reducing polyethylene wear at 3-year minimum followup.

Preparation and characteristics of antibacterial sepiolite containing povidone-iodine as a useful pharmaceutical product for patient care.

Povidone-iodine (PVP-I), an antibacterial medicine, was infiltrated in sepiolite (SPL). The available iodine content in this new pharmaceutical product, a sepiolite preparation containing povidone-iodine (PVP-I-SPL), was retained at 98.9 and 98.3% during storage at 40 degrees C for 3 and 6 months, respectively. The effective removal of various gasses, including ammonia, hydrogen sulfide, ethylmercaptan and acetaldehyde, was achieved by use of PVP-I-SPL. Especially, the concentration of ammonia gas was reduced more than half after 30 min of exposure, suggesting that PVP-I-SPL has excellent ability to adsorb ammonia gas. The satisfactory antibacterial effect of PVP-I-SPL was also obtained by testing its minimum bactericidal concentration (MBC). No irritation reactions to the rabbit auricle or ophthalmic mucosa or to human skin were observed by the skin irritation test. The PVP-I-SPL preparation has bactericidal activity and gas-adsorbing ability; therefore, this pharmaceutical product should be useful for the prevention of infections and deodorization in hospital rooms and houses, as well as in nursing homes for elderly people.

[Two cases of mediastinitis as a complication of odontogenic infection and tonsillitis].

We experienced two cases of descending necrotizing mediastinitis with different etiology. Case 1: A 59-year-old woman presented with chief complaints of dyspnea and swallowing disturbance. She had been diagnosed as having tonsillitis one week before. She was very pyrexic, and laboratory examination indicated acute inflammation. Chest X-ray and CT-scan showed enlargement of the mediastinum and pleural effusion. We diagnosed the mediastinitis to be a complication of tonsillitis. Case 2: A 54-year-old man had a tooth extracted 3 weeks prior to admission. His chief complaints were craniomandibular disturbance and neck swelling. Laboratory examination disclosed multiple organ failure and DIC. Chest X-ray and CT-scan showed enlargement of the mediastinum and pleural effusion. We diagnosed the mediastinitis in this case to be a consequence of an odontogenic infection following tooth extraction. Both patients received continuous drainage and irrigation of the abscesses and recovered in about 2 months. Case 1 showed an impaired glucose tolerance after recovery from mediastinitis. Although the main causes of mediastinitis are cardiac surgery and esophageal perforation, our cases demonstrate that mediastinitis may occur as a complication of deep neck infection.