Evaluación de la mineralización en mandíbula y diente de trucha arcoíris (Oncorhynchus mykiss) mediante microscopía electrónica de barrido de presión variable acoplada a detector de espectroscopía de rayos X de energía dispersiva / Evaluation of mineralization in jaw and teeth of rainbow trout (Oncorhynchus mykiss) using variable pressure scanning electron microscopy coupled to energy-dispersive X-ray spectroscopy detector

RESUMEN: En salmonicultura se utilizan imágenes de rayos X, para evaluar la columna vertebral y las aletas, pero estas no permiten cuantificar los minerales que constituyen el hueso, para esto se utilizan otras técnicas que son destructivas. La Microscopía Electrónica de Barrido de presión variable (VP SEM) acoplada al detector espectroscopia de rayos X de energía dispersiva (EDX) acoplado, nos permite analizar la microestructura y a la vez determinar elementos químicos, porcentaje y distribución presentes en puntos específicos en una muestra. Se utilizaron 5 truchas control y 5 con deformidad mandibular, de agua dulce en etapa de smolt, se analizó la estructura y mineralización del hueso, se eligieron en promedio 6 puntos de cuantificación por región de interés y se analizaron mediante Microscopía Electrónica de Barrido (VP SEM-EDX). Los datos obtenidos muestran porcentaje en masa de Ca promedio en hueso articular de truchas fueron de 8.07 % y de 14.48 % en truchas con deformidad mandibular y control respectivamente y el porcentaje en masa promedio de P es de 4.07 % y 7.60 %, en truchas con deformidad mandibular y control respectivamente. Se identificó además otros elementos presentes en la muestra como Mg, Na, C, O, N, S, F, Zn, Al y Fe, con especial interés en el aumento de carbono en las muestras analizadas con deformidad mandibular y la presencia de aluminio en todas las muestras. La Técnica de VP SEM-EDX, permite evaluar de forma directa, sin destrucción de la muestra y con una preparación mínima de la muestra. En el hueso, la aplicación más frecuente de SEM-EDX es la medición del contenido de Ca y P y la relación que existe entre estos elementos Ca/P, en la muestra. Paralelamente la técnica nos permite la detección de otros microelementos provenientes del agua o de la alimentación y que eventualmente pueden provocar alteraciones en los peces, confirmando la hipótesis que el microanálisis elemental tiene utilidad para la salmonicultura.

SUMMARY: In salmon farming systems, X-ray images are used to evaluate the spine and fins, but these do not allow quantifying the minerals that make up the bone, for different techniques that are destructive are used. Variable Pressure Scanning Electron Microscopy (VP SEM) coupled to an Energy Dispersive X-ray spectroscopy detector (EDX), allows us to analyze the microstructure and at the same time determine chemical elements, percentages, and distribution present at specific points in a sample. Five control and five jaw deformity trout, from freshwater and in the smolt stage were used. The structure and mineralization of the bone were analyzed, an average six quantification points were chosen per region of interest (ROI) and then they were analyzed by Scanning Electron Microscopy (VP SEM-EDX). The data obtained have shown the average mass percentage of calcium in trout joint bone was 8.07 % and 14.48 % in jaw deformity and control trout, respectively; and the average mass percentage of phosphorus is 4.07 % and 7.60 %, in jaw deformity and control trout, respectively. Other elements present in the sample were also identified, such as magnesium, sodium, carbon, oxygen, nitrogen, sulphur, fluorine, zinc, aluminium, and iron, with special interest the increase of carbon in the analyzed samples with mandibular deformity and the presence of aluminum in all samples. The VP SEM-EDX Technique allows direct evaluation, without destruction of the sample and with minimal sample preparation. In bone, the most frequent application of SEM-EDX is the measurement of the content of calcium (Ca) and phosphorus (P) and the relationship that exists between these elements, calcium/phosphorus (Ca/P), in the sample. At the same time, the technique allows us to detect other microelements from water or food that can eventually cause alterations in fish, confirming the hypothesis that elemental microanalysis is useful for salmon farming.

Is the Jaw Bone Micro-Structure Altered in Response to Osteoporosis and Bisphosphonate Treatment? A Micro-CT Analysis.

The aim of the study was to quantify the micro-architectural changes of the jaw bone in response to ovariectomy, exposed or not to bisphosphonate treatment. A total of 47 Wistar rats were ovariectomized (OVX) or sham-operated (shOVX) and exposed to osteoporosis preventive treatment for eight weeks either with bisphosphonates (alendronate, ALN; group OVX-ALN) three days/week at a dose of 2 mg/kg or with saline solution (untreated control condition; group OVX). The bone morphometric parameters of the trabecular jaw bone were assessed using ex vivo micro-computed tomography. The regions of interest investigated in the maxilla were the inter-radicular septum of the second molar and the tuber. The regions quantified in the mandible included the three molar regions and the condyle. A one-way analysis of variance followed by pairwise comparison using Tukey's HSD and the Games-Howell test was conducted to explore significant differences between the groups. In the maxilla, OVX decreased the bone volume in the inter-radicular septum of the second molar. Bisphosphonate treatment was able to prevent this deterioration of the jaw bone. The other investigated maxillary regions were not affected by (un)treated ovariectomy. In the mandible, OVX had a significant negative impact on the jaw bone in the buccal region of the first molar and the inter-radicular region of the third molar. Treatment with ALN was able to prevent this jaw bone loss. At the condyle site, OVX significantly deteriorated the trabecular connectivity and shape, whereas preventive bisphosphonate treatment showed a positive effect on this trabecular bone region. No significant results between the groups were observed for the remaining regions of interest. In summary, our results showed that the effects of ovariectomy-induced osteoporosis are manifested at selected jaw bone regions and that bisphosphonate treatment is capable to prevent these oral bone changes.

Development of a Rat Model of Mandibular Irradiation Sequelae for Preclinical Studies of Bone Repair.

Repairing mandibular bone defects after radiotherapy of the upper aerodigestive tract is clinically challenging. Although bone tissue engineering has recently generated a number of innovative treatment approaches for osteoradionecrosis (ORN), these modalities must be evaluated preclinically in a relevant, reproducible, animal model. The objective of this study was to evaluate a novel rat model of mandibular irradiation sequelae, with a focus on the adverse effects of radiotherapy on bone structure, intraosseous vascularization, and bone regeneration. Rats were irradiated with a single 80 Gy dose to the jaws. Three weeks after irradiation, mandibular bone defects of different sizes (0, 1, 3, or 5 mm) were produced in each hemimandible. Five weeks after the surgical procedure, the animals were euthanized. Explanted mandibular samples were qualitatively and quantitatively assessed for bone formation, bone structure, and intraosseous vascular volume by using micro-computed tomography, scanning electron microscopy, and histology. Twenty irradiated hemimandibles and 20 nonirradiated hemimandibles were included in the study. The bone and vessel volumes were significantly lower in the irradiated group. The extent of bone remodeling was inversely related to the defect size. In the irradiated group, scanning electron microscopy revealed a large number of polycyclic gaps consistent with periosteocytic lysis (described as being pathognomonic for ORN). This feature was correlated with elevated osteoclastic activity in a histological assessment. In the irradiated areas, the critical-sized defect was 3 mm. Hence, our rat model of mandibular irradiation sequelae showed hypovascularization and osteopenia. Impact statement Repairing mandibular bone defects after radiotherapy of the upper aerodigestive tract is clinically challenging. Novel tissue engineering approaches for healing irradiated bone must first be assessed in animal models. The current rat model of mandibular irradiation sequelae is based on tooth extraction after radiotherapy. However, the mucosal sequelae of radiotherapy often prevent the retention of tissue-engineered biomaterials within the bone defect. We used a submandibular approach to create a new rat model of mandibular irradiation sequelae, which enables the stable retention of biomaterials within the bone defect and should thus facilitate the assessment of bone regeneration.

A structural biorhythm related to human sexual dimorphism.

Life on earth is regulated by biological rhythms, some of which oscillate with a circadian, monthly or lunar cycle. Recent research suggests that there is a near weekly biorhythm that may exert an influence on human skeletal growth. Evidence for the timing of this biorhythm is retained in tooth enamel as the periodicity of Retzius lines. Studies report that Retzius periodicity (RP) relates to adult human stature and enamel thickness. Adult human stature is sexually dimorphic, and so is enamel thickness of maxillary third molars (M3) but not mandibular M3. Yet, previous studies report sex differences in RP are apparent in some populations but not others, and it is unknown if dimorphism in enamel thickness relates to RP. To further our understanding of this biorhythm we analysed sex-related variation in RP and its relationship with enamel thickness in a sample of M3's (n = 94) from adults in Northern Britain. Results reveal RP was significantly higher in our sample of female molars compared to those of males, which is consistent with the previously reported correlation between the biorhythm and adult stature. The RP of maxillary M3 related to sex differences in enamel thickness, but this relationship was not present in mandibular M3. Our results support previous findings suggesting that this biorhythm is sexually dimorphic and provide the first evidence that RP may be one factor influencing sex differences in enamel thickness. Our study also shows that correlations between RP and enamel thickness appear to be most readily detected for tooth types with sufficiently wide ranges of enamel thickness variation, as is the case for maxillary but not mandibular M3. Achieving a sufficient sample size was critical for detecting a sex difference in periodicity.

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.

A case with multiple accessory foramina of mandible in a fetus.

It is aimed to better recognize the mandibular variations by understanding the diversity and positions of accessory foramina better. Accessory formations on a full-term fetal mandible dissected for a mandibular study were examined under a microscope. To observe these formations more clearly, they were photographed with the help of a camera and microscope. In one of the mandibles dissected for a fetal mandibular study, a lateral accessory foramen (LAF1) was detected in the right half just near the mental foramen, and also a medial accessory foramen (MAF1) was detected over the mandibular foramen. In the left half, on the lateral surface relative to the mental foramen, one in the medial (LAF3) and one just above it (LAF2), and last one is near to the ramus of mandible (LAF4), three lateral accessory foramina, were detected. Again, a medial accessory foramen just above the left mandibular foramen (MAF3), and another foramen is near to mandibular symphysis (MAF2), also two medial accessory foramina on the lateral surface were detected. Detection and recognition of such variations are quite important for clinicians in the diagnostic methods and prevention of possible surgical complications.

Histochemical and Ultrastructural Study of Developing Gonial Bone With Reference to Initial Ossification of the Malleus and Reduction of Meckel's Cartilage in Mice.

Development of mouse gonial bone and initial ossification process of malleus were investigated. Before the formation of the gonial bone, the osteogenic area expressing alkaline phosphatase and Runx2 mRNA was widely recognized inferior to Meckel's cartilage. The gonial bone was first formed within the perichondrium at E16.0 via intramembranous ossification, surrounded the lower part of Meckel's cartilage, and then continued to extend anteriorly and medially until postnatal day (P) 3.0. At P0, multinucleated chondroclasts started to resorb the mineralized cartilage matrix with ruffled borders at the initial ossification site of the malleus (most posterior part of Meckel's cartilage). Almost all CD31-positive capillaries did not run through the gonial bone but entered the cartilage through the site where the gonial bone was not attached, indicating the forms of the initial ossification site of the malleus are similar to those at the secondary ossification center rather than the primary ossification center in the long bone. Then, the reducing process of the posterior part of Meckel's cartilage with extending gonial bone was investigated. Numerous tartrate-resistant acid phosphatase-positive mononuclear cells invaded the reducing Meckel's cartilage, and the continuity between the malleus and Meckel's cartilage was completely lost by P3.5. Both the cartilage matrix and the perichondrium were degraded, and they seemed to be incorporated into the periosteum of the gonial bone. The tensor tympani and tensor veli palatini muscles were attached to the ligament extending from the gonial bone. These findings indicated that the gonial bone has multiple functions and plays important roles in cranial formation. Anat Rec, 302:1916-1933, 2019. © 2019 American Association for Anatomy.

Using ImageJ (Fiji) to Analyze and Present X-Ray CT Images of Enamel.

X-ray micro CT has become a popular methodology for the nondestructive analysis of dental tissues and has been used extensively in the amelogenesis field. The aim of this chapter is to introduce ImageJ/Fiji to researchers new to CT scanning and the analysis of CT image data. The program can be applied to analyzing X-ray CT images of enamel but can be extrapolated to other tissues as well.

Microcomputed Tomography Imaging in Odontogenesis Studies.

3D analysis of animal or human whole teeth and alveolar bone can be performed with high sensitivity in a nondestructive manner by microcomputed tomography. Here we describe the protocols to be followed for the most common applications in the developmental studies of dental and craniofacial tissues. Emphasis is placed on the basis of choosing settings for image acquisition, such as voxel resolution (Fig. 1), or beam energy (Fig. 2) and for processing, such as segmentation method (Fig. 3), parameters. The limitations to take into account for optimal efficiency and image quality are also explained.

Three-Dimensional Assessment of Crown Size and Eruption Space for Developing Third Molars: Data Collection Techniques Based on Cone-Beam Computed Tomography (CBCT).

Third molar development and eruption are two related areas of major interest in dental research into the etiology of "wisdom tooth" impaction. Third molars are not only an excellent model for studying dental development but also of fundamental clinical importance because they are very frequently impacted. Because the third molar is located in the distal-most region of the oral cavity, clinical access is relatively challenging. With the increasingly widespread use of cone-beam computed tomography (CBCT) in dentistry, studies and measurements of the third molar and its eruption area have become considerably easier to do. Here we present a novel CBCT-based measurement methodology we developed for our recent investigations that we hope will also be useful for the broader dental research community.

Functional morphology of the mouthparts of lady beetle Coccinella transversoguttata (Coccinellidae, Coleoptera), with reference to their feeding mechanism.

The lady beetle Coccinella transversoguttata is an important biocontrol agent of aphids. As the main the feeding apparatus, mouthparts play essential roles in feeding process, and the morphological variation of mouthparts is correlated with variation in food source and feeding behavior. To better understand the feeding behavior of C. transversoguttata, we studied the functional morphology of mouthparts, with special attention to the fine morphology of each part of mouthpart and various kinds of sensilla. The mouthpart of C. transversoguttata is typical mandibulate type that composed of labrum, mandible, maxillae, labium, and hypopharynx. Detailed descriptions were given about the fine morphology of each part, and various types of sensilla were identified and classified. In total, two types of sensilla chaetica, four types of sensilla basiconica, two types of sensilla styloconica, two types of sensilla placodea, and one type of cuticular pore were identified according to their length, morphology and distribution. Specially, the putative function of each part of mouthpart as well as each kind of sensilla and their coordinative mechanisms in feeding process were discussed. These results would lay a solid foundation for understanding the feeding mechanism of lady beetles.

An unusual disordered alveolar bone material in the upper furcation region of minipig mandibles: A 3D hierarchical structural study.

Teeth are subjected to compressive loads during mastication. Under small loads the soft tissue periodontal ligament (PDL) deforms most. However when the loads increase and the PDL is highly compressed, the tooth and the alveolar bone supporting the tooth, begin to deform. Here we report on the structure of this alveolar bone in the upper furcation region of the first molars of mature minipigs. Using light microscopy and scanning electron microscopy (SEM) of bone cross-sections, we show that this bone is hypermineralized, containing abundant small pores around 1-5 µm in diameter, lacunae around 10-20 µm as well as larger spaces. This bone does not possess the typical lamellar motif or other repeating structures normally found in cortical or trabecular mammalian bone. We also use high resolution focused ion beam scanning electron microscopy (FIB-SEM) in the serial surface mode to image the 3D organization of the demineralized bone matrix. We show that the upper furcation bone matrix has a disordered isotropic structure composed mainly of individual collagen fibrils with no preferred orientation, as well as highly staining material that is probably proteoglycans. Much larger aligned arrays of collagen fibers - presumably Sharpey's fibers - are embedded in this material. This unusual furcation bone material is similar to the disordered material found in human lamellar bone. In the upper furcation region this disordered bone comprises almost all the volume excluding Sharpey's fibers. We surmise that this most unusual bone type functions to resist the repeating compressive loads incurred by molars during mastication.

Ultrastructure features of the surface of the gills and the lower pharyngeal jaw of the Tilapia Zilli, redbelly tilapia (Coptodon Zillii, Gervais, 1848).

The present investigation was designed to describe the surface ultrastructure of the gill system of tilapia Zilli. The gill system is formed from four gill arches and each gill arch carries a row of gill filaments on its convex border and two rows of the gill rakers on its concave border. The quadrilateral interbranchial septum has elevated part at the level of the third gill arch. By SEM observations, the gill arch was divided into three regions: rostral, middle and caudal region. The caudal region contained two characteristic structures: oval leaf-like structure and rounded-shaped structure. Each oval leaf-like structure carried two lateral rows of the triangular pointed spines separated by a median groove. All surfaces of gill arches, rakers and filaments were covered with a mosaic of the polygonal pavement cells, in addition to the opening of chloride cells and mucous cells. The gill arch and gill raker had only one appearance of taste buds named type I. Meanwhile, the filaments contained two types of different appearance of the taste buds named: type I and type II. Type I was the main common and similar to that present in gill arch and raker and characterized by its blunt end, while type II had hair-like structures that projected from the volcano-shaped depression. The gill rakers were formed from central axis surrounded by two lateral lobulated regions which carry pointed spines, taste buds and the opening of chloride cells. The surface of triangular lower pharyngeal jaw carries numerous teeth-like papillae which originated from the socket-like depression.

Morphology of the mandibles and the first maxillae in the family Lithobiidae (Myriapoda, Chilopoda), with remarks on their phylogenetic significance.

The mandibles and the first maxillae of 37 species of the family Lithobiidae (Myriapoda, Chilopoda) were investigated and compared to provide a structural overview and evaluate their significance for the systematics of the family. The species sampling focused on the genus Lithobius, examining 33 species of four subgenera (Lithobius, Monotarsobius, Sigibius, Ezembius), as this genus represents about half of the known diversity of Lithobiidae, including more than 500 assigned species and subspecies. The microstructures on the mandibular gnathal edge and the first maxillary telopodites and coxal projections were studied using scanning electron microscopy. Although having a similar structural pattern, we demonstrate that the microstructures are variable within and between species of adult specimens and commonly show intergradation. To check for intraspecific variability of microstructures and character stability, specimen sampling was extended for the two common Austrian species Lithobius dentatus and Lithobius validus, for which seven specimens depicted no major differences in the mandibular gnathal edge and the first maxillae. Our data suggest the presence of three characters in the mandibular gnathal edge and the first maxillae useful for lithobiid phylogeny. These characters were tested in a phylogenetic analysis together with previously described and novel morphological characters. Subgenera of Lithobius are mostly non-monophyletic, and several other genera of Lithobiinae as well as other subfamilies group with particular species or clades of Lithobius. The results corroborate a close relationship between Disphaerobius loricatus and Lithobius (Ezembius) giganteus, strengthening the hypothesis that Pterygoterginae is nested within Lithobiinae and specifically within Lithobius, allied to L. (Ezembius) and Hessebius.

Molecular alterations of newly formed mandibular bone caused by zoledronate.

Bone quality is defined by structural and material characteristics. Most studies on the mandible have focused on the analysis of structural characteristics, with insufficient investigation of material characteristics. This study tested whether zoledronate affects the material characteristics of newly formed mandibular bone. Thirty-six female Wistar rats were assigned to three groups: sham-ovariectomized rats (SHAM, n=12), ovariectomized rats (OVX, n=12), and ovariectomized rats treated with zoledronate (ZOL, n=12). The left side of the mandibular ramus of all rats was drilled bicortically. Twenty-eight days after surgery, all surviving rats were euthanized and all mandibles were removed. Raman microspectroscopy was performed, and five spectra per specimen of newly formed mandibular bone were analysed. Compared with OVX rats, the mineral/matrix ratio in ZOL rats was significantly increased (5.43±1.88 vs. 7.86±2.05), while crystallinity (0.055±0.002 vs. 0.050±0.002), relative proteoglycan content (0.43±0.10 vs. 0.31±0.05), and collagen structural integrity (1.16±0.21 vs. 0.72±0.06) were significantly decreased. These changes in material characteristics may explain why rats that received zoledronate exhibited peculiar biological phenomena such as bisphosphonate-related osteonecrosis of the jaw.

Alveolar bone remodeling after tooth extraction in irradiated mandible: An experimental study with canine model.

OBJECTIVES: The aim of the present study is to investigate the morphological and cellular changes in dental extraction socket that has been irradiated after the tooth extraction and to describe morphological characteristics of the osteocytes and osteocyte-lacunar-canalicular network (LCN) by scanning electron microscopy (SEM). MATERIAL AND METHODS: Five beagle dogs aged 1-2 years were used in this study. One side of each mandible was irradiated in two sessions and the other side of mandible (non-irradiated) served as a control. The mandible bone blocks were processed by bulk staining en bloc in basic fuchsin and the specimens were embedded routinely in polymethyl methacrylate resin without preliminary decalcification. All blocks were subjected to micro-CT imaging, after that the specimens were prepared for light microscopy and SEM. RESULTS: Alterations in bone macrostructure are minimal in irradiated bone, but the changes in LCN are clear. In the area of the tooth extraction socket, the connections of osteocytes to the vessels and to neighboring osteocytes were not observed both in irradiated and nonirradiated bone. However, osteoclasts were located in the bone surface entering inside to the bone between osteons. In the lamellar bone of lateral sides, a decrease in canalicular connections between osteocytes and periosteum was found in irradiated bone as compared to the non-irradiated side. CONCLUSIONS: The novelty of the present study is that radiation disrupts osteocytes and their dendrites.

Calcification in rat developing mandibular bone demonstrated by whole mount staining, microcomputed tomography and scanning electron microscopy with energy dispersive X-ray spectroscopy.

The study was designed to investigate calcification in developing rat mandibular bone using whole mount staining, micro-computed tomography (micro-CT) and scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX). Wistar rats at embryonic days 16, 18, and 20 and postnatal weeks 1 and 6 were used. Rats were fixed with 4% paraformaldehyde and heads were resected, frozen and sectioned for histology, then analysed with SEM-EDX. Some of the specimens were observed with micro-CT. Other rats were fixed and stained with alcian blue and alizarin red for whole mount staining. Histology and whole mount staining showed that osteoid was deposited around Meckel's cartilage at day 15 and developed into bone at day 16. Accumulation of Ca and P was identified in the bone matrix with SEM-EDX. The area of bone expanded until week 6. The Ca/P ratio increased, whereas the C/Ca and C/P ratios decreased during development. Micro-CT demonstrated an increase in radio-opacity with bone development. The results suggest that rat mandibular bone formation is initiated around Meckel's cartilage at day 15. Deposition and maturation of the calcium phosphate mineral increase gradually with decrease in the organic component as the rat mandible develops.

A robust methodology for the quantitative assessment of the rat jawbone microstructure.

Micro-computed tomography can be applied for the assessment of the micro-architectural characteristics of the cortical and trabecular bones in either physiological or disease conditions. However, reports often lack a detailed description of the methodological steps used to analyse these images, such as the volumes of interest, the algorithms used for image filtration, the approach used for image segmentation, and the bone parameters quantified, thereby making it difficult to compare or reproduce the studies. This study addresses this critical need and aims to provide standardized assessment and consistent parameter reporting related to quantitative jawbone image analysis. Various regions of the rat jawbones were screened for their potential for standardized micro-computed tomography analysis. Furthermore, the volumes of interest that were anticipated to be most susceptible to bone structural changes in response to experimental interventions were defined. In the mandible, two volumes of interest were selected, namely, the condyle and the trabecular bone surrounding the three molars. In the maxilla, the maxillary tuberosity region and the inter-radicular septum of the second molar were considered as volumes of interest. The presented protocol provides a standardized and reproducible methodology for the analysis of relevant jawbone volumes of interest and is intended to ensure global, accurate, and consistent reporting of its morphometry. Furthermore, the proposed methodology has potential, as a variety of rodent animal models would benefit from its implementation.

Morphology and ultrastructure of the allomone and sex-pheromone producing mandibular gland of the parasitoid wasp Leptopilina heterotoma (Hymenoptera: Figitidae).

Chemical communication by the parasitoid wasp Leptopilina heterotoma is based largely on (-)-iridomyrmecin. The female wasps use (-)-iridomyrmecin as a defensive allomone, a chemical cue to avoid competition with con- and heterospecific females, and as a major component of their sex pheromone to attract males. Males of L. heterotoma produce (+)-isoiridomyrmecin, which is also used for chemical defense. In this study we show that females and males of L. heterotoma produce the iridomyrmecins in a pair of mandibular glands. Each gland consists of a secretory part composed of class 3 gland cells and their accompanying duct cells, as well as a reservoir bordered by a thin intima. The gland discharges between the mandible base and the clypeus. Males have considerably smaller glands than females, which corresponds to the lower amount of iridomyrmecins produced by males. Chemical analyses of the mandibular gland contents showed that the gland of females contained mainly (-)-iridomyrmecin, as well as low amounts of the other previously described iridoid pheromone compounds, while the glands of males contained only (+)-isoiridomyrmecin. The morphology and sizes of the mandibular glands of males and females of L. heterotoma have evolved to the multi-functional use of iridomyrmecin.

Effects on Bone Tissue After Osteotomy with Different High-Energy Lasers: An Ex Vivo Study.

OBJECTIVE: The purpose of this study was to evaluate the application of Er:YAG and Er,Cr:YSGG lasers during osteotomy in bovine bone blocks. BACKGROUND DATA: Er:YAG/Er,Cr:YSGG lasers for osteotomy procedures have been used to induce tissue ablation without thermal damage. However, even these lasers can induce thermal damages, which can cause a delay on the bone healing. MATERIALS AND METHODS: Sixty standardized bovine bone mandible samples were divided into three groups with 20 samples each: Group 1, Bur; Group 2, Er:YAG laser; and Group 3, Er,Cr:YSGG laser. Two irradiations/cuts were made in each sample. The first was performed until the sample was completely separated into two fragments, and the time required for this procedure was recorded in seconds. The second irradiation/cut was made the same way as the first one; however, it was made partially, without separating the sample into two fragments, but rather making sulci ∼1.5-2 mm deep. Ten samples in each group were used to evaluate the surface morphology of the osteotomy by scanning electron microscopy (SEM), while the other 10 samples were submitted to descriptive histological analysis. RESULTS: The bur was the instrument that performed the osteotomy in the shortest amount of time (p < 0.05). All the instruments caused thermal damage; however, the Er,Cr:YSGG laser was the only type that induced carbonization. CONCLUSIONS: The Er:YAG laser induced the lowest degree of thermal damage in bone tissue after osteotomy; however, a shorter clinical time was required to perform the osteotomy in the bur group.