Fanny Hontoir

Systematic Review of Efficacy of Nutraceuticals to Alleviate Clinical Signs of Osteoarthritis

BACKGROUND Various treatments of osteoarthritis (OA) have been described, including use of nutraceuticals. OBJECTIVES To review systematically the literature about the effects of nutraceuticals on clinical signs of pain or abnormal locomotion in horses, dogs, and cats, and to discuss methodological aspects of trials and systematic reviews. METHODS A systematic search of controlled trials evaluating the impact of nutraceuticals on OA in horses, dogs, and cats was performed, using Medline, CAB Abstracts, and Google Scholar. Scientific evidence was evaluated by means of criteria proposed by the Food and Drug Administration (FDA), and a scoring system adapted from both the CONsolidated Standards of Reporting Trials (CONSORT) statement and recommendations for assessing trials by the Center of Evidence Based Medicine of Oxford. RESULTS Twenty-two papers were selected and reviewed, with 5 studies performed in horses, 16 in dogs, and 1 in cats. The strength of evidence was low for all nutraceuticals except for omega-3 fatty acid in dogs. There were limited numbers of rigorous randomized controlled trials and of participants in clinical trials. CONCLUSIONS AND CLINICAL IMPORTANCE The evidence of efficacy of nutraceuticals is poor, with the exception of diets supplemented with omega-3 fatty acids in dogs. Greater access to systematic reviews must be part of the objectives of the veterinary science in the future. Their reporting would be improved by internationally agreed-upon criteria for standards and guidelines.

A comparison of 3-T magnetic resonance imaging and computed tomography arthrography to identify structural cartilage defects of the fetlock joint in the horse

Articular cartilage defects are prevalent in metacarpo/metatarsophalangeal (MCP/MTP) joints of horses. The aim of this study was to determine and compare the sensitivity and specificity of 3-Tesla magnetic resonance imaging (3-T MRI) and computed tomography arthrography (CTA) to identify structural cartilage defects in the equine MCP/MTP joint. Forty distal cadaver limbs were imaged by CTA (after injection of contrast medium) and by 3-T MRI using specific sequences, namely, dual-echo in the steady-state (DESS), and sampling perfection with application-optimised contrast using different flip-angle evolutions (SPACE). Gross anatomy was used as the gold standard to evaluate sensitivity and specificity of both imaging techniques. CTA sensitivity and specificity were 0.82 and 0.96, respectively, and were significantly higher than those of MRI (0.41 and 0.93, respectively) in detecting overall cartilage defects (no defect vs. defect). The intra and inter-rater agreements were 0.96 and 0.92, respectively, and 0.82 and 0.88, respectively, for CT and MRI. The positive predictive value for MRI was low (0.57). CTA was considered a valuable tool for assessing cartilage defects in the MCP/MTP joint due to its short acquisition time, its specificity and sensitivity, and it was also more accurate than MRI. However, MRI permits assessment of soft tissues and subchondral bone and is a useful technique for joint evaluation, although clinicians should be aware of the limitations of this diagnostic technique, including reduced accuracy.

Hyaluronan does not regulate human epidermal keratinocyte proliferation and differentiation

Hyaluronan (HA) is synthesized by three HA synthases (HAS1, HAS2, and HAS3) and secreted in the extracellular matrix. In human skin, large amounts of HA are found in the dermis. HA is also synthesized by keratinocytes in the epidermis, although its epidermal functions are not clearly identified yet. To investigate HA functions, we studied the effects of HA depletion on human keratinocyte physiology within in vitro reconstructed human epidermis. Inhibition of HA synthesis with 4-methylumbelliferone (4MU) did not modify the expression profile of the epidermal differentiation markers involucrin, keratin 10, and filaggrin during tissue reconstruction. In contrast, when keratinocytes were incubated with 4MU, cell proliferation was decreased. In an attempt to rescue the proliferation function, HA samples of various mean molecular masses were added to keratinocyte cultures treated with 4MU. These samples were unable to rescue the initial proliferation rate. Furthermore, treatments with HA-specific hyaluronidase, although removing almost all HA from keratinocyte cultures, did not alter the differentiation or proliferation processes. The differences between 4MU and hyaluronidase effects did not result from differences in intracellular HA, sulfated glycosaminoglycan concentration, apoptosis, or levels of HA receptors, all of which remained unchanged. Similarly, knockdown of UDP-glucose 6-dehydrogenase (UGDH) using lentiviral shRNA effectively decreased HA production but did not affect proliferation rate. Overall, these data suggest that HA levels in the human epidermis are not directly correlated with keratinocyte proliferation and differentiation and that incubation of cells with 4MU cannot equate with HA removal.

Magnetic resonance compositional imaging of articular cartilage: What can we expect in veterinary medicine?

Since cartilage has limited ability to repair itself, it is useful to determine its biochemical composition early in clinical cases. It is also important to assess cartilage content in research animals in longitudinal studies in vivo. In recent years, compositional imaging techniques using magnetic resonance imaging (MRI) have been developed to assess the biochemical composition of cartilage. This article describes MR compositional imaging techniques, and discusses their use and interpretation. Technical concerns still limit the use of some techniques for research and clinical use, especially in veterinary medicine. Glycosaminoglycan chemical-exchange saturation transfer and sodium imaging are better used with high field magnets, which have limited availability. Long acquisition times are sometimes required, for instance in T1rho (ρ) and diffusion-weighted imaging, and necessitate general anaesthesia. Even in human medicine, some techniques such as ultra-short echo T2 are not fully validated, and nearly all techniques require validation for veterinary research and clinical practice. Delayed gadolinium-enhanced MRI of cartilage and T2 mapping appear to be the most applicable methods for compositional imaging of animal cartilage. Combining T2 mapping and T1ρ allows for the assessment of proteoglycans and the collagen network, respectively.

Computed Tomography and Magnetic Resonance Anatomy of the Normal Orbit and Eye of the Horse

Traumatic and infectious diseases of the eye and orbit can occur in horses. For diagnosis and monitoring of such diseases, medical imaging is useful including computed tomography (CT) and magnetic resonance imaging (MRI). The aim of the current study was to describe CT and MRI anatomy of the equine orbit and ocular globe. The heads from four adult horses were scanned with a 6‐slice Emotion 6 CT (Siemens, Erlangen), and a 3.0 Tesla Siemens Verio 6 MRI using T1 and T2‐weighted sequences. To validate CT and MR reference images, these were compared with anatomical models and gross anatomical sections. The bony limits of the orbital cavity, the relationship of the orbit with sinuses and foramina of the skull were well identified by CT. MRI was useful to observe soft tissues and was able to identify adnexae of the ocular globe (eyelids, periorbital fat, extraocular muscles, lacrymal and tarsal glands). Although MRI was able to identify all components of the eye (including the posterior chamber), it could not differentiate sclera from choroid and retina. The only nerve identified was the optic nerve. Vessels were not seen in this series of cadaver heads. This study showed that CT and MRI are useful techniques to image the equine orbit and eye that can have clinical applications.

Accuracy of computed tomographic arthrography for assessment of articular cartilage defects in the ovine stifle.

Articular cartilage defects are one of the features of osteoarthritis in animals and humans. Early detection of cartilage defects is a challenge in clinical veterinary practice and also in translational research studies. An accurate, diagnostic imaging method would be desirable for detecting and following up lesions in specific anatomical regions of the articular surface. The current prospective experimental study aimed to describe the accuracy of computed tomographic arthrography (CTA) for detecting cartilage defects in a common animal model used for osteoarthritis research, the ovine stifle (knee, femoropatellar/femorotibial) joint. Joints in cadaver limbs (n = 42) and in living animals under anesthesia (n = 13) were injected with a contrast medium and imaged using a standardized CT protocol. Gross anatomy and histological assessment of specific anatomic regions were used as a gold standard for the evaluation of sensitivity, specificity, negative predictive value, and positive predictive value for CTA identification of articular cartilage defects in those regions. Pooled estimated sensitivity and specificity were 90.32% and 97.30%, respectively, in cadaver limbs, and 81.82% and 95.24%, respectively, in living animals. Pooled estimated positive predictive value and negative predictive values were 98.25% and 85.71%, respectively, in cadaver limbs, and 81.82% and 95.24%, respectively, in living animals. The delineation of cartilage surface was good for anatomical regions most frequently affected by cartilage defects in the ovine stifle: medial femoral condyle, medial tibial condyle, and patella. This study supported the use of CTA as an imaging technique for detecting and monitoring articular cartilage defects in the ovine stifle joint.

Magnetic resonance imaging (MRI) anatomy of the ovine stifle.

OBJECTIVE To describe magnetic resonance imaging (MRI) anatomy of the ovine stifle and investigate meniscotibial and cruciate ligaments anatomy. STUDY DESIGN Descriptive ex vivo study. ANIMALS Pelvic limbs (n = 44) from 22 adult Texel ewes. METHODS Forty limbs (n = 40) were scanned using 3 Tesla MRI before gross anatomic dissection. Two other limb pairs were frozen and transected to obtain sections that were compared with MRI images for identification of anatomic structures. RESULTS In all stifles, the craniomedial bundle of the cranial cruciate ligament inserted caudally to the cranial attachment of the medial meniscus. No transverse intermeniscal ligament was identified in 80% of stifles, whereas a few small ligamentous fibers were seen crossing from 1 cranial horn to the other in 20% of stifles. There was good differentiation of menisci, ligaments, and synovial cavities on MRI images. Two bundles were identified in all cranial cruciate ligaments on MRI. Sensitivity and specificity of 3T MRI for detection of transverse intermeniscal ligament were 42% and 84%, respectively. CONCLUSION 3T MRI provided well defined reference images for menisci, synovial cavities, and most ligaments.

Angiographic Anatomy of External Iliac Arteries in the Sheep.

External iliac artery atherosclerotic disease and aneurism occur in man. For treatment, imaging is required to facilitate minimally invasive introduction and advancement of stents within the intended vessels. Sheep are commonly used to test and improve stents. However, little information is published regarding the angiographic anatomy of the iliac arteries in the ovine species. The objective of this study was to describe the angiographic anatomy of the iliac arteries in the sheep. Computed tomography (CT) angiography and gross anatomical dissection were performed in, respectively, 10 and 43 adult ewes. Diameters and lengths of the arteries were measured. In comparison with man, salient anatomical differences were identified in the sheep: (1) the absence of common iliac arteries, (2) the common trunk at the origin of internal iliac arteries and (3) the location of the bifurcation of the external iliac arteries into femoral arteries in the pelvis (not in the limb). External iliac arteries in this series of sheep were 86 mm long in average and had a mean diameter of 7.5 mm. Lengths of arteries are only slightly different between man and sheep, while diameters are rather similar. Therefore, the sheep model appears to be sufficiently similar to man to test stent properties. This study provides useful reference images and measures of lengths and diameters of relevant arteries that could be applied to research with ovine models.

Magnetic resonance imaging (MRI) anatomy of the ovine lumbar spine

Although the ovine spine is a useful research model for intervertebral disc pathology and vertebral surgery, there is little peer‐reviewed information regarding the MRI anatomy of the ovine spine. To describe the lumbar spine MRI anatomy, 10 lumbar segments of cadaver ewes were imaged by 1.5‐Tesla MR. Sagittal and transverse sequences were performed in T1 and T2 weighting (T1W, T2W), and the images were compared to gross anatomic sagittal and transverse sections performed through frozen spines. MRI was able to define most anatomic structures of the ovine spine in a similar way as can be imaged in humans. In both T1W and T2W, the signals of ovine IVDs were similar to those observed in humans. Salient anatomic features were identified: (1) a 2‐ to 3‐mm linear zone of hypersignal was noticed on both extremities of the vertebral body parallel to the vertebral plates in sagittal planes; (2) the tendon of the crura of the diaphragm appeared as a hypointense circular structure between hypaxial muscles and the aorta and caudal vena cava; (3) dorsal and ventral longitudinal ligaments and ligamentum flavum were poorly imaged; (4) no ilio‐lumbar ligament was present; (5) the spinal cord ended between S1–S2 level, and the peripheral white matter and central grey matter were easily distinguished on T1W and T2W images. This study provides useful reference images to researchers working with ovine models.

Retrograde Neuroanatomical Tracing of Phrenic Motor Neurons in Mice

Phrenic motor neurons are cervical motor neurons originating from C3 to C6 levels in most mammalian species. Axonal projections converge into phrenic nerves innervating the respiratory diaphragm. In spinal cord slices, phrenic motor neurons cannot be identified from other motor neurons on morphological or biochemical criteria. We provide the description of procedures for visualizing phrenic motor neuron cell bodies in mice, following intrapleural injections of cholera toxin subunit beta (CTB) conjugated to a fluorophore. This fluorescent neuroanatomical tracer has the ability to be caught up at the diaphragm neuromuscular junction, be carried retrogradely along the phrenic axons and reach the phrenic cell bodies. Two methodological approaches of intrapleural CTB delivery are compared: transdiaphragmatic versus transthoracic injections. Both approaches are successful and result in similar number of CTB-labeled phrenic motor neurons. In conclusion, these techniques can be applied to visualize or quantify the phrenic motor neurons in various experimental studies such as those focused on the diaphragm-phrenic circuitry.