EXTRAVASATION OF CONTRAST MEDIA AFTER SUBCARAPACIAL VESSEL INJECTION IN THREE CHELONIAN SPECIES.

The subcarapacial vessel is a popular site for venipuncture and intravenous medication administration in chelonians. Reports of adverse effects when using this site have increased, prompting evaluation of its safety. This study aimed to evaluate the anatomy of the subcarapacial vessel in 25 individual chelonians (2 box turtles, 3 red-eared sliders, and 20 red-footed tortoises) using computed tomography (CT). Individuals were sedated and administered contrast in the subcarapacial vessel. The vessel was visualized in 50% of the box turtles and red-footed tortoises, and 100% of the red-eared sliders. All species had contrast extravasation in the subarachnoid space, with red-footed tortoises having the largest percentage (70% compared to 50% and 33% of box turtles and red-eared sliders, respectively). Extravasation of contrast in the trachea or bronchi (70%) and lungs (80%) was seen in the red-footed tortoises only. Higher prevalence of contrast extravasation in the red-footed tortoises is likely because of anatomical differences, including a more cranially extending lung field and domed-shaped carapace compared to the other species. These findings highlight the risk associated with using the subcarapacial vessel for intravenous medication administration in certain species of chelonian.

The equine navicular apparatus as a premier enthesis organ: Functional implications.

Navicular syndrome has been traditionally characterized by progressive lameness with chronic degeneration of the navicular bone. Advances in imaging techniques have revealed that its associated soft tissue structures are also affected. This distribution of lesions is explained by conceptualizing the equine navicular apparatus as an enthesis organ that facilitates the dissemination of mechanical stress throughout the tissues of the foot. The navicular apparatus has the same structural adaptations to mechanical stress as the human Achilles tendon complex. These adaptations efficiently dissipate mechanical force away from the tendon's bony attachment site, thereby protecting it from failure. The comparison of these two anatomically distinct structural systems demonstrates their similar adaptations to mechanical forces, and illustrates that important functional insights can be gained from studying anatomic convergences and cross-species comparisons of function. Such a functional conceptualization of the equine navicular apparatus resolves confusion about the diagnosis of navicular syndrome and offers insights for the development of mechanically based therapies. Through comparison with the human Achilles complex, this review (1) re-conceptualizes the equine navicular apparatus as an enthesis organ in which mechanical forces are distributed throughout the structures of the organ; (2) describes the relationship between failure of the navicular enthesis organ and lesions of navicular syndrome; (3) considers the therapeutic implications of navicular enthesis organ degeneration as a form of chronic osteoarthritis; and based upon these implications (4) proposes a focus on whole body posture/motion for the development of prehabilitative and rehabilitative therapies similar to those that have already proven effective in humans.

Chronic Ethanol Feeding in Mice Decreases Expression of Genes for Major Structural Bone Proteins in a Nox4-Independent Manner.

Bone loss in response to alcohol intake has previously been hypothesized to be mediated by excessive production of reactive oxygen species via NADPH oxidase (Nox) enzymes. Nox4 is one of several Nox enzymes expressed in bone. We investigated the role of Nox4 in the chondro-osteoblastic lineage of the long bones in mice during normal chow feeding and during chronic ethanol feeding for 90 days. We generated mice with a genotype (PrxCre +/- Nox4 fl/fl) allowing conditional knockout of Nox4 in the limb bud mesenchyme. Adult mice had 95% knockdown of Nox4 expression in the femoral shafts. For mice on regular chow, only whole-body Nox4 knockout mice had clearly increased cortical thickness and bone mineral density in the tibiae. When chronically fed a liquid diet with and without ethanol, conditional Nox4 knockout mice had slightly reduced dimensions of the cortical and trabecular regions of the tibiae (P < 0.1). The ethanol diet caused a significant reduction in cortical bone area and cortical thickness relative to a control diet without ethanol (P < 0.05). The ethanol diet further reduced gene expression of Frizzled related protein (Frzb), myosin heavy chain 3, and several genes encoding collagen and other major structural bone proteins (P < 0.05), whereas the Nox4 genotype had no effects on these genes. In conclusion, Nox4 expression from both mesenchymal and nonmesenchymal cell lineages appears to exert subtle effects on bone. However, chronic ethanol feeding reduces cortical bone mass and cortical gene expression of major structural bone proteins in a Nox4-independent manner. SIGNIFICANCE STATEMENT: Excessive alcohol intake contributes to osteopenia and osteoporosis, with oxidative stress caused by the activity of NADPH oxidases hypothesized to be a mediator. We tested the role of NADPH oxidase (Nox) 4 in osteoblast precursors in the long bones of mice with a conditional Nox4 knockout model. We found that Nox4 exerted effects independent of alcohol intake, and ethanol effects on bone were Nox4-independent.

Mouthwash as a non-invasive method of indocyanine green delivery for near-infrared fluorescence dental imaging.

SIGNIFICANCE: X-ray imaging serves as the mainstream imaging in dentistry, but it involves risk of ionizing radiation. AIM: This study presents the feasibility of indocyanine green-assisted near-infrared fluorescence (ICG-NIRF) dental imaging with 785-nm NIR laser in the first (ICG-NIRF-I: 700 to 1000 nm) and second (ICG-NIRF-II: 1000 to 1700 nm) NIR wavelengths. APPROACH: Sprague Dawley rats with different postnatal days were used as animal models. ICG, as a fluorescence agent, was delivered to dental structures by subcutaneous injection (SC) and oral administration (OA). RESULTS: For SC method, erupted and unerupted molars could be observed from ICG-NIRF images at a short imaging time (<1 min). ICG-NIRF-II could achieve a better image contrast in unerupted molars at 24 h after ICG injection. The OA could serve as a non-invasive method for ICG delivery; it could also cause the glow-in-dark effect in unerupted molars. For erupted molars, OA can be considered as mouthwash and exhibits outstanding performance for delivery of ICG dye; erupted molar structures could be observed at a short imaging time (<1 min) and low ICG dose (0.05 mg / kg). CONCLUSIONS: Overall, ICG-NIRF with mouthwash could perform in-vivo dental imaging in two NIR wavelengths at a short time and low ICG dose.

Architecture of the bronchial tree in Cuvier's dwarf caiman (Paleosuchus palpebrosus).

We imaged the lungs of five Cuvier's dwarf caiman (Paleosuchus palpebrosus) via computed tomography (CT) and micro-computed tomography (µCT) and compared these data to the lungs of the American alligator (Alligator mississippiensis). These data demonstrate anatomical commonalities between the lungs of P. palpebrosus and A. mississippiensis, and a few notable differences. The structural similarities are (a) a proximally narrow, distally widened, hook-shaped primary bronchus; (b) a cervical ventral bronchus that branches of the primary bronchus and immediately makes a hairpin turn toward the apex of the lung; (c) a sequential series of dorsobronchi arising from the primary bronchus caudal to the cervical ventral bronchus; (d) intraspecifically highly variable medial sequence of secondary airways; (e) sac-like laterobronchi; and (f) grossly dead-ended caudal group bronchi in the caudal and ventral aspects of the lung. The primary differences between the two taxa are in the overall number of large bronchi (fewer in P. palpebrosus), and the number of branches that contribute to the cardiac regions. Imaging data of both a live and deceased specimen under varying states (postprandial, fasting, total lung capacity, open to atmosphere) indicate that the caudal margin and position of the lungs shift craniocaudally relative to the vertebral column. These imaging data suggest that the smooth thoracic ceiling may be correlated to visceral movement during ventilation, but this hypothesis warrants validation. These results provide the scaffolding for future comparisons between crocodilians, for generating preliminary reconstructions of the ancestral crocodilian bronchial tree, and establishing new hypotheses of bronchial homology across Archosauria.

Binge Ethanol Exposure in Mice Represses Expression of Genes Involved in Osteoblast Function and Induces Expression of Genes Involved in Osteoclast Differentiation Independently of Endogenous Catalase.

Excessive ethanol consumption is a risk factor for osteopenia. Since a previous study showed that transgenic female mice with overexpression of catalase are partially protected from ethanol-mediated trabecular bone loss, we investigated the role of endogenous catalase in skeletal ethanol toxicity comparing catalase knockout to wild-type mice. We hypothesized that catalase depletion would exacerbate ethanol effects. The mice were tested in a newly designed binge ethanol model, in which 12-week-old mice were exposed to 4 consecutive days of gavage with ethanol at 3, 3, 4, and 4.5 g ethanol/kg body weight. Binge ethanol decreased the concentration of serum osteocalcin, a marker of bone formation. The catalase genotype did not affect the osteocalcin levels. RNA sequencing of femoral shaft RNA from males was conducted. Ethanol exposure led to significant downregulation of genes expressed in cells of the osteoblastic lineage with a role in osteoblastic function and collagen synthesis, including the genes encoding major structural bone proteins. Binge ethanol further induced a smaller set of genes with a role in osteoclastic differentiation. Catalase depletion affected genes with expression in erythroblasts and erythrocytes. There was no clear interaction between binge ethanol and the catalase genotype. In an independent experiment, we confirmed that the binge ethanol effects on gene expression were reproducible and occurred throughout the skeleton in males. In conclusion, the binge ethanol exposure, independently of endogenous catalase, reduces expression of genes involved in osteoblastic function and induces expression of genes involved in osteoclast differentiation throughout the skeleton in males.

Construction of a Realistic, Whole-Body, Three-Dimensional Equine Skeletal Model using Computed Tomography Data.

Therapies based upon whole-body biomechanical assessments are successful for injury prevention and rehabilitation in human athletes. Similar approaches have rarely been used to study equine athletic injury. Degenerative osteoarthritis caused by mechanical stress can originate from chronic postural dysfunction, which, because the primary dysfunction is often distant from the site of tissue injury, is best identified through modeling whole-body biomechanics. To characterize whole-body equine kinematics, a realistic skeletal model of a horse was created from equine computed tomography (CT) data that can be used for functional anatomical and biomechanical modeling. Equine CT data were reconstructed into individual three-dimensional (3D) data sets (i.e., bones) using 3D visualization software and assembled into a complete 3D skeletal model. The model was then rigged and animated using 3D animation and modeling software. The resulting 3D skeletal model can be used to characterize equine postures associated with degenerative tissue changes as well as to identify postures that reduce mechanical stress at the sites of tissue injury. In addition, when animated into 4D, the model can be used to demonstrate unhealthy and healthy skeletal movements and can be used to develop preventative and rehabilitative individualized therapies for horses with degenerative lamenesses. Although the model will soon be available for download, it is currently in a format that requires access to the 3D animation and modeling software, which has quite a learning curve for new users. This protocol will guide users in (1) developing such a model for any organism of interest and (2) using this specific equine model for their own research questions.

Optimal imaging windows of indocyanine green-assisted near-infrared dental imaging with rat model and its comparison to X-ray imaging.

In this study, we used rat animal model to compare the efficiency of indocyanine green (ICG)-assisted dental near-infrared fluorescence imaging with X-ray imaging, and we optimized the imaging window for both unerupted and erupted molars. The results show that the morphology of the dental structures was observed clearly from ICG-assisted dental images (especially through the endoscope). A better image contrast was easily acquired at the short imaging windows (<10 minutes) for unerupted and erupted molars. For unerupted molars, there is another optimized imaging window (48-96 hours) with a prominent glow-in-the-dark effect: only the molars remain bright. This study also revealed that the laser ablation of dental follicles can disrupt the molar development, and our method is able to efficiently detect laser-treated molars and acquire the precise morphology. Thus, ICG-assisted dental imaging has the potential to be a safer and more efficient imaging modality for the real-time diagnosis of dental diseases.

Detection and analysis of enamel cracks by ICG-NIR fluorescence dental imaging.

Cracked teeth are the third most common cause of tooth loss, but there is no reliable imaging tool for the diagnosis of cracks. Here, we demonstrate the feasibility of indocyanine green near-infrared fluorescence (ICG-NIRF) dental imaging for the detection of enamel cracks and enamel-dentin cracks in vitro in the first (ICG-NIRF-I, 700-950 nm) and second (ICG-NIRF-II, 950-1700 nm) imaging windows with transmission excitation light, and compared ICG-NIRF with conventional NIR illumination-II (NIRi-II) and X-ray imaging. Dentin cracks were detected by CT scan, while most enamel cracks, undetectable under X-ray imaging, were clearly visible in NIR images. We found that ICG-NIRF-II detected cracks more effectively than NIRi-II, and that light orientation is an important factor for crack detection: an angled exposure obtained better image contrast of cracks than parallel exposure, as it created a shadow under the crack. Crack depth could be evaluated from the crack shadow in ICG-NIRF and NIRi-II images; from this shadow we could determine crack depth and discriminate enamel-dentin cracks from craze lines. Cracks could be observed clearly from ICG-NIRF images with 1-min ICG tooth immersion, although longer ICG immersion produced images with greater contrast. Overall, our data show that ICG-NIRF dental imaging is a useful tool for diagnosing cracked teeth at an early stage.

The structure of the cornified claw sheath in the domesticated cat (Felis catus): implications for the claw-shedding mechanism and the evolution of cornified digital end organs.

The morphology of cornified structures is notoriously difficult to analyse because of the extreme range of hardness of their component tissues. Hence, a correlative approach using light microscopy, scanning electron microscopy, three-dimensional reconstructions based on x-ray computed tomography data, and graphic modeling was applied to study the morphology of the cornified claw sheath of the domesticated cat as a model for cornified digital end organs. The highly complex architecture of the cornified claw sheath is generated by the living epidermis that is supported by the dermis and distal phalanx. The latter is characterized by an ossified unguicular hood, which overhangs the bony articular base and unguicular process of the distal phalanx and creates an unguicular recess. The dermis covers the complex surface of the bony distal phalanx but also creates special structures, such as a dorsal dermal papilla that points distally and a curved ledge on the medial and lateral sides of the unguicular process. The hard-cornified external coronary horn and proximal cone horn form the root of the cornified claw sheath within the unguicular recess, which is deeper on the dorsal side than on the medial and lateral sides. As a consequence, their rate of horn production is greater dorsally, which contributes to the overall palmo-apical curvature of the cornified claw sheath. The external coronary and proximal cone horn is worn down through normal use as it is pushed apically. The hard-cornified apical cone horn is generated by the living epidermis enveloping the base and free part of the dorsal dermal papilla. It forms nested horn cones that eventually form the core of the hardened tip of the cornified claw. The sides of the cornified claw sheath are formed by the newly described hard-cornified blade horn, which originates from the living epidermis located on the slanted face of the curved ledge. As the blade horn is moved apically, it entrains and integrates the hard-cornified parietal horn on its internal side. It is covered by the external coronary and proximal cone horn on its external side. The soft-cornified terminal horn extends distally from the parietal horn and covers the dermal claw bed at the tip of the uniguicular process, thereby filling the space created by the converging apical cone and blade horn. The soft-cornified sole horn fills the space between the cutting edges of blade horn on the palmar side of the cornified claw sheath. The superficial soft-cornified perioplic horn is produced on the internal side of the unguicular pleat, which surrounds the root of the cornified claw sheath. The shedding of apical horn caps is made possible by the appearance of microcracks in the superficial layers of the external coronary and proximal cone horn in the course of deformations of the cornified claw sheath, which is subjected to tensile forces during climbing or prey catching. These microcracks propagate tangentially through the coronary horn and do not injure the underlying living epidermal and dermal tissues. This built-in shedding mechanism maintains sharp claw tips and ensures the freeing of the claws from the substrate.

Indocyanine green-assisted dental imaging in the first and second near-infrared windows as compared with X-ray imaging.

Indocyanine green (ICG) has been widely used in medical imaging, such as in retinal angiography. Here, we describe a pilot ex vivo study of ICG-assisted near-infrared fluorescence (NIRF) dental imaging in the first (700-950 nm for ICG-NIRF-I) and second (1000-1700 nm for ICG-NIRF-II) NIR windows using human extracted teeth; our study is compared with the traditional prevalent X-ray imaging and NIR II illumination (NIRi-II, 1310 nm) without ICG enhancement. The results show that ICG fluorescence has much better imaging contrast in both windows compared with NIRi-II (by quantitatively comparing NIR intensity of the critical neighboring structures, such as enamel and dentin). Cracked teeth, notoriously hard to diagnose by dental X-ray and computed tomography, were clearly profiled in NIRF dental imaging. An insidious occlusal caries, missing in X-ray imaging, became a bright dot that was readily observed in ICG-NIRF-I images. For dental decay, NIRF imaging with ICG enhancement could clearly delineate the decay boundary. NIRF in both windows distinguished interproximal and occlusal superficial caries. Overall, ICG-assisted NIRF dental imaging has unique advantages in identifying cracked teeth and insidious caries. The two NIR imaging windows used in our study might one day serve as noninvasive and nonionizing-radiation methods for the diagnosis of critical dental diseases in situ.

Improvement of Tribological and Biocompatibility Properties of Orthopedic Materials Using Piezoelectric Direct Discharge Plasma Surface Modification.

Various types of alloys and polymers are utilized in orthopedic implants. However, there are still several issues accompanied by the use of prosthetic materials, such as low wear performance and catastrophic failure. Surface enhancement of biomaterials is a promising method that can improve the success rate of prosthetic operations without negatively affecting their bulk properties while improving the biocompatibility of implants and reducing infections. Nonthermal plasma treatment has become a ubiquitous surface modification method in sterilization and healthcare applications. However, the clinical applications of such an approach have been limited due to the lack of detailed studies delineating the wear behavior and biocompatibility of implants after plasma treatment. In this study, we have employed a handheld piezoelectric direct discharge (PDD) plasma generator to modify the surface of two common metallic (Ti6Al4V) and nonmetallic (GUR1020 polymer) biomaterials used typically in joint and disc replacements. We have observed an approximately 60-fold reduction in tribological wear rate along with a 2- to 3-fold increase in the biocompatibility properties of plasma coated samples compared to noncoated (untreated) surfaces, respectively. Our study introduces a novel application of nonthermal PDD plasma technology that is capable of increasing the quality and success rate of joint and disc replacements.

The Human Shoulder Suspension Apparatus: A Causal Explanation for Bilateral Asymmetry and a Fresh Look at the Evolution of Human Bipedality.

The combination of large mastoid processes and clavicles is unique to humans, but the biomechanical and evolutionary significance of their special configuration is poorly understood. As part of the newly conceptualized shoulder suspension apparatus, the mastoid processes and clavicles are shaped by forces exerted by the musculo-fascial components of the cleidomastoid and clavotrapezius muscles as they suspend the shoulders from the head. Because both skeletal elements develop during infancy in tandem with the attainment of an upright posture, increased manual dexterity, and the capacity for walking, we hypothesized that the same forces would have shaped them as the shoulder suspension apparatus evolved in ancestral humans in tandem with an upright posture, increased manual dexterity, and bipedality with swinging arms. Because the shoulder suspension apparatus is subjected to asymmetrical forces from handedness, we predicted that its skeletal features would grow asymmetrically. We used this prediction to test our hypothesis in a natural experiment to correlate the size of the skeletal features with the forces exerted on them. We (1) measured biomechanically relevant bony features within the shoulder suspension apparatus in 101 male human specimens (62 of known handedness); and (2) modeled and analyzed the forces within the shoulder suspension apparatus from X-ray CT data. We identified eight right-handed characters and demonstrated the causal relationship between these right-handed characters and the magnitude and direction of forces acting on them. Our data suggest that the presence of the shoulder suspension apparatus in humans was a necessary precondition for human bipedality.