Unique three-dimensional structure of a fish pharyngeal jaw subjected to unusually high mechanical loads.

We examine the structure of the bone of the pharyngeal jaws of a large fish, the black drum (Pogonias cromis), that uses its tooth-jaw complex to crush hard-shelled bivalve mollusks. During mastication huge compressive forces are concentrated in a tiny zone at the tooth-bone interface. We report on the structure of this bone, with emphasis on its contact with the teeth, at different hierarchical levels and in 3D. Micro-CT shows that the molariform teeth do not have roots and are supported by a circular narrow bony rim that surrounds the periphery of the tooth base. The lower pharyngeal jaw is highly porous, as seen by reflected light microscopy and secondary electron microscopy (SE-SEM). Porosity decreases close to the bone-tooth interface and back-scattered electron (BSE-SEM) microscopy shows a slight elevation in mineral density. Focused ion beam - scanning electron microscopy (FIB-SEM) in the serial surface view (SSV) mode reveals a most surprising organization at the nanoscale level: parallel arrays of mineralized collagen fibrils surrounding channels of ~100 nm diameter, both with their long axes oriented along the load direction. The channels are filled with organic matter. These fibril-channel arrays are surrounded by a highly disordered mineralized material. This unusual structure clearly functions efficiently under compression, but the precise way by which this unique arrangement achieves this function is unknown.

[The effectiveness of the application of mineral water from the Klyuchi health resort in the programs of combined spa-based rehabilitation of the patients presenting with acquired jaw defects].

The results of the present study give evidence of the well-apparent sanation effect of balneotherapy with the use of therapeutic mineral water (MW) extracted at the Klyuchi health resort. Irrigation of the oral cavity with this mineral water was shown to reduce the number of the strains of opportunistic microorganisms and their virulence potential. It is concluded that the proposed method may be recommended for the treatment and prevention of dysbiosis in the oral cavity of the patients presenting with acquired jaw defects.

Occlusion on implants - is there a problem?

Oral rehabilitation restores form and function and impacts on general health. Teeth provide a discriminating sense of touch and directional specificity for occlusal perception, management of food with mastication and swallowing, and awareness of its texture and hardness. Peripheral feedback for control of jaw muscles includes the enamel-dentine-pulp complex and mechanoreceptors in the periodontal tissues. The implications of feedback from periodontal and other intra-oral mechanoreceptors as well as changes in central representation are significant for function and adaptation to oral rehabilitation. With implants, in the absence of the periodontium and periodontal mechanoreceptor feedback, fine motor control of mastication is reduced, but patients are still able to function adequately. Further, there is no significant difference in function with full-arch fixed prostheses on teeth in comparison with implants. Predictable implant outcomes depend on bone support. Optimum restoration design appears to be significant for bone remodelling and bone strains around implants with occlusal loading. Finite element analysis data confirmed load concentrations at the coronal bone around the upper section of the implant where bone loss is commonly observed clinically. Load concentration increased with steeper cusp inclination and broader occlusal table and decreased with central fossa loading and narrower occlusal table size. It is recommended that occlusal design should follow a narrow occlusal table, with central fossa loading in intercuspal contact and low cusp inclination to minimise lateral loading in function and parafunction. Acknowledging these features should address potential problems associated with the occlusion in implant therapy.

Ultrastructural analysis of glutamate-immunopositive synapses onto the rat jaw-closing motoneurons during postnatal development.

The excitatory synapses on the jaw-closing (JC) motoneurons mediate the neuronal input that ensures smooth and rhythmic movements of the jaw. Recently, we have shown that the neurotransmitter phenotype of the inhibitory boutons onto JC motoneurons shifts from GABA to glycine, and new inhibitory synapses onto JC motoneurons are continuously formed during postnatal development (Paik et al. [2007] J. Comp. Neurol. 503:779­789). To test whether the developmental pattern of the excitatory synapses onto JC motoneurons differs from that of the inhibitory synapses, we studied the distribution of glutamate-immunopositive boutons onto the rat JC motoneurons during postnatal development by using a combination of retrograde labeling with horseradish peroxidase (HRP), postembedding immunogold staining, and quantitative ultrastructural analysis. The analysis of 175, 281, and 465 boutons contacting somata of JC motoneurons at postnatal days P2, P11, and P31, respectively, revealed that the number of glutamate-immunopositive (Glut(+)) boutons increased by 2.6 times from P2 to P11 and showed no significant change after that, whereas the length of apposition of these boutons increased continuously from P2 to P31, suggesting that the time course for the development of Glut(+) boutons differed from that for Glut(-) boutons, most of which were immunopositive for GABA and/or glycine. Our findings indicate that excitatory and inhibitory synapses onto JC motoneurons exhibit distinctly different developmental patterns that may be closely related to the maturation of the masticatory system.

Plate-like permanent dental laminae of upper jaw dentition in adult gobiid fish, Sicyopterus japonicus.

Sicyopterus japonicus (Teleostei, Gobiidae) possesses a unique upper jaw dentition different from that known for any other teleosts. In the adults, many (up to 30) replacement teeth, from initiation to attachment, are arranged orderly in a semicircular-like strand within a capsule of connective tissue on the labial side of each premaxillary bone. We have applied histological, ultrastructural, and three-dimensional imaging from serial sections to obtain insights into the distribution and morphological features of the dental lamina in the upper jaw dentition of adult S. japonicus. The adult fish has numerous permanent dental laminae, each of which is an infolding of the oral epithelium at the labial side of the functional tooth and forms a thin plate-like structure with a wavy contour. All replacement teeth of a semicircular-like strand are connected to the plate-like dental lamina by the outer dental epithelium and form a tooth family; neighboring tooth families are completely separated from each other. The new tooth germ directly buds off from the ventro-labial margin of the dental lamina, whereas no distinct free end of the dental lamina is present, even adjacent to this region. Cell proliferation concentrated at the ventro-labial margin of the dental lamina suggests that this region is the site for repeated tooth initiation. During tooth development, the replacement tooth migrates along a semicircular-like strand and eventually erupts through the dental lamina into the oral epithelium at the labial side of the functional tooth. This unique thin plate-like permanent dental lamina and the semicircular-like strand of replacement teeth in the upper jaw dentition of adult S. japonicus probably evolved as a dental adaptation related to the rapid replacement of teeth dictated by the specialized feeding habit of this algae-scraping fish.

Importance of microcracks in etiology of bisphosphonate-related osteonecrosis of the jaw: a possible pathogenetic model of symptomatic and non-symptomatic osteonecrosis of the jaw based on scanning electron microscopy findings.

The aim of this study was to evaluate a possible role of microcracks in the pathogenesis of bisphosphonate-related osteonecrosis of the jaw (ONJ) and to discuss an etiological model. Bone samples from 35 patients with ONJ were analyzed. Control samples were taken from five patients with osteomyelitis (OM), ten patients with osteoradionecrosis, seven patients with osteoporosis and bisphosphonate medication without signs of ONJ, and six osteoporotic elderly patients. Samples were examined using scanning electron microscopy. In 54% of the bone samples of patients with ONJ, microcracks were seen. Inflammatory and connective tissue reactions within the microcracks were evident in 82% of the cases, indicating that these cracks were not artificial. In contrast, only 29% of samples from patients with oral bisphosphonate medication without ONJ, no sample from patients with OM, none of the osteoradionecrosis group, and only 17% from patients with osteoporosis showed microcracks. Statistically significant differences could be found between the ONJ group and the group after irradiation and the group with OM, respectively. The evidence of microcracks could be a first step in the pathogenesis of bisphosphonate-related ONJ. The accumulation of these microcracks leads to a situation that could be named "non-symptomatic ONJ". Disruptions of the mucosal integrity may then allow bacterial invasion, leading to jawbone infection with exposed bone, fistulas, and pain. This state could be called "symptomatic ONJ". Furthermore, an assumed local immunosuppression as indicated by various studies could explain the severe courses of therapy-resistant ONJ as regularly observed.

Ontogeny of the tessellated skeleton: insight from the skeletal growth of the round stingray Urobatis halleri.

The majority of the skeleton of elasmobranch fishes (sharks, rays and relatives) is tessellated: uncalcified cartilage is overlain by a superficial rind of abutting, mineralized, hexagonal blocks called tesserae. We employed a diversity of imaging techniques on an ontogenetic series of jaw samples to investigate the development of the tessellated skeleton in a stingray (Urobatis halleri). We compared these data with the cellular changes that characterize cartilage calcification in bony skeletons. Skeletal growth is characterized by the appearance of tesserae as well as changes in chondrocyte shape, arrangement and density. Yolk sac embryos (35-56 mm disc width, DW) have untessellated lower jaw tissue wrapped in perichondrium and densely packed with chondrocytes. Chondrocyte density decreases dramatically after yolk sac absorption (histotroph stage: 57-80 mm DW) until the formation of tesserae, which are first visible using our techniques as thin (approximately 60 microm), sub-perichondral plaques. During the histotroph stage, flattened chondrocytes align parallel to the perichondrium at the tissue periphery, where we believe they are incorporated into developing tesserae to form the cell-rich laminae observed within tesserae; in older animals peripheral cells in the uncalcified phase are rounder and less uniformly oriented. By parturition (approximately 75 mm DW), cell density and the number of adjoining chondrocyte pairs (an indicator of cell division) have dropped to less than a third of their initial values; these remain low and tesserae continue to grow in size. The tessellated skeleton is a simple solution to the conundrum of growth in an endoskeleton with external mineralization and no remodeling. Although we see parallels with endochondral ossification (e.g. chondrocytes decreasing in density with age), the lack of chondrocyte hypertrophy and the fact that mineralization is sub-perichondral (not the case in mammalian cartilage) suggest that the similarities end there.

Ultrastructure and function of the mastax in Dicranophorus forcipatus (Rotifera: Monogononta).

Rotifers are characterized by a complex set of cuticularized jaw elements in the pharynx. The fine structure of the jaw elements has been the subject of SEM studies for some time, but only very limited information exists on the ultrastructure of the jaw elements and their function beyond taxonomic considerations. Drawing on SEM and TEM techniques, the present study presents a detailed analysis of the mastax in Dicranophorus forcipatus, a carnivorous monogonont rotifer species from freshwater habitats characterized by an extrusible, grasping jaw apparatus. Based on ultrathin serial sections, the jaw elements are reconstructed and, in total, nine paired and two unpaired muscles identified. Possibly homologous muscles in other rotifer species are discussed and functional considerations of the forcipate mastax are suggested.

Diet versus jaw bones: Lessons from experimental models and potential clinical implications.

The consumption of different types of diets influences not only body health but the bone remodeling process as well. Nutritional components can directly affect maxillary and mandibular alveolar bone microarchitecture. In this review, we focus on the current knowledge regarding the influence of diets and dietary supplementation on alveolar bone. Accumulating evidence from experimental models suggests that carbohydrate- and fat-rich diets are detrimental for alveolar bone, whereas protective effects are associated with consumption of calcium, ω-3, and bioactive compounds. Little is known about the effects of protein-free and protein-rich diets, boron, vitamin C, vitamin E, zinc, and caffeine on alveolar bone remodeling. Adipokines and direct effects of nutritional components on bone cells are proposed mechanisms linking diet and bone. Results from animal models substantiate the role of nutritional components on alveolar bone. It is a well-built starting point for clinical studies on nutritional monitoring and intervention for patients with alveolar bone disorders, especially those who are treatment refractory.

Ultrastructure of the maxillary organ of Scutigera coleoptrata (Chilopoda, Notostigmophora): description of a multifunctional head organ.

The maxillary organ of Scutigera coleoptrata was investigated using light microscopy, electron microscopy, and maceration techniques. Additionally, we compared the maxillary organ of S. coleoptrata with those of two other notostigmophoran centipedes, Parascutigera festiva and Allothereua maculata, using SEM. The maxillary organ is located inside the posterior coxal lobes of the first maxillae and extends posteriorly as sac-like pouches. The narrow epidermis of the maxillae is differentiated to form the epithelium of the maxillary organ. Two types of epithelia are distinguishable: a simple cuboidal epithelium of different height and differentiation (types I, II, IV) and a pseudostratified columnar epithelium (type III). These epithelia are covered by a highly specialized cuticle. The pseudostratified epithelium is the most prominent feature of the maxillary organ. It is covered with hundreds of setae, protruding deep into the maxillary organ. Two different types of setae can be distinguished, filiform and fusiform. The maxillary organ communicates with the oral cavity, the maxillary organ gland, the maxillary nephridium, and with a large number of epidermal glands that secrete into the maxillary organ. Epithelium III allows the extension of the maxillary organ when its pouches are filled with secretion. The maxillary organ is a complex multifunctional organ. The organ probably stores excretion from the maxillary nephridia and secretory fluid from the maxillary organ gland and other epidermal glands. The fluid is primarily required as preening fluid. The ammonia of the excretory fluid is thought to evaporate via the setae and the wide opening of the maxillary organ. It is likely that parts of the fluid can be reabsorbed by the animal via the oral cavity.

Central distribution of nociceptive intradental afferent nerve fibers in the rat.

The central distribution of intradental afferent nerve fibers was investigated by combining electron microscopic observations with a selective method for inducing degeneration of the A delta- and C-type afferent fibers. Degenerating terminals were found on the proprioceptive mesencephalic trigeminal neurons and on dendrites in the neuropil of the trigeminal motor nucleus after application of capsaicin to the rat's lower incisor tooth pulp. The results give anatomical evidence of new sites of central projection of intradental A delta- and C-type fibers whereby the nociceptive information from the tooth pulp can affect jaw muscle activity.

New characters in the Gnathostomulid mouth parts revealed by scanning electron microscopy.

An analysis with SEM of the mouth parts of 16 species belonging to 10 genera of Gnathostomulida resulted in the following new characters and conclusions: 1) At least in the genus Haplognathia, jaw teeth that are visible by conventional light microscopy are composed of the same aggregated needle-like denticles that are found, often in large numbers, on the basal plates of many filospermoid species. 2) Other new ultrastructural tooth features include the tricuspid basal plate teeth in Problognathia minima, tripartite teeth in Austrognathia and Austrognatharia, and the clear separation, in the Gnathostomula basal plate, of a mediodorsal set of teeth from a more extensive rostroventral set. 3) Three rows of teeth, as typical for Gnathostomulidae and Austrognathiidae, are also present in the filospermoid Haplognathia filum. 4) The wide range of geographic variation in Haplognathia ruberrima is confirmed by significant differences in jaw teeth between specimens from Belize and Bermuda. 5) A compartmentalized involucrum is present in Labidgonathia longicollis. 6) A pair of lamellae addentales, until now only known from Valvognathia pogonostoma, was found in Tenuignathia rikerae, Problognathia minima, and probably also Labidognathia rikerae. 7) In all gnathostomulids, the lamella symphysis is composed of identical rods that are considered homologous with those in the mouth parts of Rotifera and Micrognathozoa.

Further structures in the jaw apparatus of Limnognathia maerski (Micrognathozoa), with notes on the phylogeny of the Gnathifera.

The jaws of Limnognathia maerski, Micrognathozoa, were investigated with light- and scanning electron microscopy. The study yielded several new structures and sclerites, including the ventral part of main jaw, the pharyngeal lamellae, the manus, the dorsal and ventral fibularium teeth, and a reinterpretation of the fibularium compartmentalization. Furthermore, it was shown that several jaw elements are composed of densely packed rods. Comparison with Rotifera and Gnathostomulida suggested that the micrognathozoan main jaw is homologous with the rotifer incus and the gnathostomulid articularium and that the pseudophalangids (the ventral jaws) and their associated sclerites correspond to the rotifer mallei. These results imply that Micrognathozoa is more closely related to Rotifera than to Gnathostomulida.

Anatomy of the pharyngeal jaw apparatus of Zenarchopterus (gill) (Teleostei: Beloniformes).

The structure of the pharyngeal jaw apparatus (PJA) of Zenarchopterus dispar and Z. buffonis, carnivorous estuarine and freshwater West-Pacific halfbeaks, was investigated using dissection, light, and scanning electron microscopy as part of a comparison with estuarine and marine herbivorous confamilials. The Zenarchopterus PJA differs from published descriptions of hemiramphid PJAs in that the otic capsules are less pronounced; the pharyngocranial articulation facet is trough-like; the third pharyngobranchials are ankylosed; the second pharyngobranchial anterior processes are relatively hypotrophied; all pharyngeal teeth except the posterior teeth in the fifth ceratobranchial face posteriorly; the muscularis craniopharyngobranchialis 2 posterior is short; the muscularis craniopharyngobranchialis 2 anterior is lacking, as is its insertion site, the inferior parasphenoid apophysis; the protractor pectoralis is well developed; the pharyngocleithralis internus originates dorsal to the level of the fifth ceratobranchial bony process; the fifth ceratobranchial bony processes are directed ventrolaterally; the opposing upper and lower tooth fields appear not to occlude erosively; and the muscular portion of the pharyngohyoideus is well developed anteriorly. The extent of these differences and their implications for the function of the PJA support recent molecular studies that suggest that the Hemiramphidae is polyphyletic.

Laser irradiation of bone. I. An in vitro study concerning the effects of the CO2 laser on oral mucosa and subjacent bone.

The purpose of this study was twofold: first, to evaluate the histologic effects of CO2 laser irradiation on biopsies of porcine oral mucosa and underlying bone under conditions that simulate the applications of the laser during gingival surgery; and second, to evaluate the histologic effects on cortical bone following irradiation with increasing energy densities. Specimens consisting of mucosa and underlying bone were subjected to multiple passes of the laser beam in the same line of incision at energy densities ranging from 240 to 1,032 J/cm2. A second group of specimens consisting only of cortical bone was irradiated by a single pass of the laser at energy densities ranging from 40 to 2,062 J/cm2. In both groups the mean depth of ablation, width of surface damage, and widths of the zones of thermal necrosis and thermal damage were determined. Results showed a direct correlation between increasing energy density and/or number of energy beam passes and increasing depths of ablation and widths of surface damage. Further, more than three passes at 1,032 J/cm2 penetrated the mucosal layer to involve underlying bone. The mean depth of ablation for bone specimens following a single pass of the energy beam ranged from 0.02 mm at 160 J/cm2 to a maximum of 0.75 mm at 2,062 J/cm2. Using those energy densities most common to oral soft tissue surgery, the mean depth of ablation in bone specimens ranged from 0.17 mm at 240 J/cm2 to 0.28 mm at 640 J/cm2 to 0.35 mm at 1,032 J/cm2. All specimens regardless of tissue composition, energy density, or number of energy beam passes exhibited a distinct layer of residual carbonized tissue, a zone of thermal necrosis characterized by tissue coagulation, and a zone of tissue exhibiting thermal damage.

Silver methenamine staining for scanning electron microscopy of bone sections containing biomaterials.

Sections of tissue containing orthopedic materials are currently used to study the compatibility of those materials and to perform electron probe microanalysis at the material-tissue interface. Identification of the cells in contact with the material by Scanning electron microscopy (SEM) is of interest. We have developed a method for staining cells and tissue structures embedded in polymethyl methacrylate with silver methenamine once the sections have been obtained. Sections were prepared by grinding, and the silver methenamine was applied after oxidation with periodic acid. The procedure was carried out in a microwave oven. Backscatter SEM showed staining of the cell nucleus membrane, chromatin, the nuclear organizers, and the chromosomes of dividing cells. The cytoplasm and the cytoplasmic membrane were also stained. Collagen fibers of the extracellular matrix and the mineralized matrix of bone were labeled. Material particles in the macrophages were easily recognizable and Energy-Dispersive Spectrometer were not impaired by the presence of silver in the preparation.

Dentition and dentigerous bones in juveniles and adults of Polypterus senegalus (Cladistia, Actinopterygii).

Tooth types, their arrangement and the mode of tooth replacement were studied in juvenile and adult specimens of Polypterus senegalus by means of scanning electron microscopy of cleared and stained specimens as well as sections. All the dermal bones of the oropharynx are almost completely covered with teeth except for the angulare. The same is true for the branchial apparatus where only the hyoid skeleton is toothless. The teeth are uniformly monocuspid and conical, but can be classified according to shape and size into three types. These types and the mode of tooth replacement are characteristic for each dermal bone. In some of the jaw bones each tooth possesses a lingually situated replacement tooth. This is true for the teeth of the premaxillary, the maxillary, and the dentary which are arranged in a single line, and those of the dermopalatine, the coronoids, and the vomer which are in several lines and graded in size. Replacement teeth of all the other dentigerous elements develop on top of existing pulpal openings, forming an anastomosing common pulpal complex only after resorption of the previous tooth. The tooth plates of the dermal bones of the branchial apparatus are connected by syndesmosis only to the perichondrally ossified and to the cartilaginous or connective tissue material of the elements of the gill-arches. The dentition and its association with the bones of the head in Polypterus senegalus bear resemblances to advanced actinopterygians on the one hand (e.g. differentiation of tooth-types, arrangement), but also some similarities to living Amphibia (anchoring material and mode of replacement) on the other. The accentuation of a single marginal line of large teeth in both, the outer and the inner dental arcade of the jaws is a peculiarity of Polypterus that in a way parallels the derived state of similar monolinear tooth arrangements in Actinopterygii and Tetrapoda.

[Morphological study of histiocytosis from Langerhans cells in jaw bones]. / Rezul'taty morfologicheskogo issledovaniia gistiotsitoza iz kletok Langergansa v cheliustnykh kostiakh.

Histiocytosis of the jaws in biopsies from 17 patients aged 19 to 51 years were studied at light and electron microscopy level. Birbek's bodies were found in Langerhans cells in all the biopsies confirming the diagnosis of Langerhans cell histiocytoma. The structure of Birbek's bodies changed depending on the treatment--radiation or chemotherapy.

Electrically conductive bacterial nanowires in bisphosphonate-related osteonecrosis of the jaw biofilms.

OBJECTIVE: Bacterial biofilms play a role in the pathogenesis of bisphosphonate-related osteonecrosis of the jaw (BRONJ). The purpose of this preliminary study was to test the hypothesis that the extracellular filaments observed in biofilms associated with BRONJ contain electrically conductive nanowires. STUDY DESIGN: Bone samples of patients affected by BRONJ were evaluated for conductive nanowires by scanning electron microscopy (SEM) and conductive probe atomic force microscopy (CP-AFM). We created nanofabricated electrodes to measure electrical transport along putative nanowires. RESULTS: SEM revealed large-scale multispecies biofilms containing numerous filamentous structures throughout necrotic bone. CP-AFM analysis revealed that these structures were electrically conductive nanowires with resistivities on the order of 20 Ω·cm. Nanofabricated electrodes spaced along the nanowires confirmed their ability to transfer electrons over micron-scale lengths. CONCLUSIONS: Electrically conductive bacterial nanowires to date have been described only in environmental isolates. This study shows for the first time that these nanowires can also be found in clinically relevant biofilm-mediated diseases, such as BRONJ, and may represent an important target for therapy.

Cartilage, bone, and intermandibular connective tissue in the Australian lungfish, Neoceratodus forsteri (Osteichthyes: Dipnoi).

The connective tissue that links the bones of the mandible in the Australian lungfish, Neoceratodus forsteri, has been described as an intermandibular cartilage, and as such has been considered important for phylogenetic analyses among lower vertebrates. However, light and electron microscopy of developing lungfish jaws demonstrates that the intermandibular tissue, like the connective tissue that links the bones of the upper jaw, contains fibroblasts and numerous bundles of collagen fibrils, extending from the trabeculae of the bones supporting the tooth plates. It differs significantly in structure and in staining reactions from the cartilage and the bone found in this species. In common with the cladistian Polypterus and with actinopterygians and some amphibians, lungfish have no intermandibular cartilage. The connective tissue linking the mandibular bones has no phylogenetic significance for systematic grouping of lungfish, as it is present in a range of different groups among lower vertebrates.