Measurement of Femoral Trochlear Morphology in Dogs Using Ultrasonography.

OBJECTIVE: The aim of this study was to examine a method for measuring femoral trochlear morphology in dogs using ultrasonography, and survey femoral trochlear morphology in skeletally normal dogs. STUDY DESIGN: To examine the validity of ultrasonography, three raters measured the sulcus angle (SA; the angle between the medial and lateral facets) five times in five skeletally normal dogs. Intra- and interrater reliabilities were then calculated. We also calculated the correlation coefficient between SAs measured using ultrasonography and computed tomography (CT) or magnetic resonance imaging (MRI). Then, we surveyed the femoral trochlear morphology in skeletally normal dogs using ultrasonography. RESULTS: The intra- and interrater reliability for SAs measured using ultrasonography was good, and the correlation coefficient between SAs measured using ultrasonography and CT, or MRI, was strong. In skeletally normal dogs, the SAs for osseous contours decreased until 6 months of age. However, the SAs for cartilaginous contours did not change during the growth period. CONCLUSION: The ultrasonographic measurement method is reliable for the evaluation of femoral trochlear morphology in dogs. The contour of the articular cartilage surface of the femoral trochlea is already determined early in life, and ossification of the articular cartilage of the femoral trochlea proceeds until 6 months of age. The results of this study will provide crucial information for the evaluation of femoral trochlear morphology in dogs.

Computed Tomographic Measurements of the Sulcus Angle of the Femoral Trochlea in Small-Breed Dogs with and without Medial Patellar Luxation.

Trochleoplasty is often performed in dogs with medial patellar luxation (MPL); however, the current guidelines on when to perform a trochleoplasty in dogs are vague. The sulcus angle (SA) is used to assess the femoral trochlear morphology in humans. The aim of this study is to describe a method to measure the SA and other parameters of trochlea morphology in dogs using computed tomography. First, we searched for a suitable measuring location for the SA. Transverse images of the femurs were obtained as perpendicular planes to the tangent of the femoral trochlea which was 0 to 60 degrees (every 5 degrees) to the anatomical axis of the femur. The deepest point of the femoral trochlea was found in the transverse images perpendicular to the tangent of the femoral trochlea which was at 15 degrees to the anatomical axis of the femur. The SA and the other parameters of femoral trochlea morphology were measured at the deepest point of the femoral trochlea. The SA of the stifle joints with grade 3 and 4 MPL was significantly higher than the SA of stifle joints not affected by MPL. There was no significant difference in the SA between dogs affected by grade 1 and 2 MPL and dogs not affected by MPL. Further studies are needed to establish whether the SA can be used as selection criteria for trochleoplasty.

Leptin Modulates the mRNA Expression of Follicle Development Markers in Post-hatch Chicks in an Age-Dependent Manner.

Leptin is involved in regulating reproductive function in chickens, and the development of the leptin system is initiated during the early embryonic stage; however, whether leptin has a specific role in regulating the ovarian development in early post-hatch days is still not fully understood. This study investigated the expression of ovarian functional markers in growing juvenile chickens, along with the effects of leptin on gene expression in the hypothalamus-pituitary-gonadal (HPG) axis on specific ovarian-remodeling days. Leptin receptor (LEPR), follicle-stimulating hormone receptor (FSHR), and the mRNA expression of aromatase (CYP19A1) tended to increase with age in the ovaries of growing chicks. In the ovaries of 7-day-old chicks, intraperitoneally injected leptin significantly increased the mRNA expressions of LEPR, FSHR, and CYP19A1, and this resulted in the increased serum estradiol levels. However, leptin had no effect on hypothalamic LEPR, gonadotropin-releasing hormone 1 (GnRH1), or gonadotropin-inhibitory hormone (GnIH) mRNAs; however, in the pituitary gland, leptin significantly increased the mRNA expression of luteinizing hormone beta subunit (LHB) but had no effect on the mRNA expression of follicle-stimulating hormone beta subunit (FSHB). In 28-day-old chicks, hypothalamic and pituitary mRNAs were unaffected by leptin administration, except hypothalamic LEPR mRNA that was upregulated by a high dose of leptin. In the ovary, leptin dose-dependently decreased the mRNA expression of LEPR; low doses of leptin significantly increased the mRNA expression of FSHR, whereas high doses significantly decreased this expression; leptin did not affect the mRNA expression of CYP19A1; and high leptin doses significantly reduced the serum estradiol levels. Collectively, the results of this study show that leptin modulates ovarian development and folliculogenesis marker genes by primarily acting on ovaries on the specific ovarian-remodeling days in post-hatch chicks, which may alter folliculogenesis and ovarian development toward puberty in chicken.

Traumatic medial luxation of the triceps brachii tendon with medial subluxation of the elbow joint in a dog.

OBJECTIVE: To describe the surgical reduction of luxation of the triceps brachii tendon in a dog. ANIMAL: One 2.5-year-old 2.58 kg castrated male toy poodle. STUDY DESIGN: Clinical case report. METHODS: The dog displayed intermittent, non-weight bearing lameness of the right forelimb for approximately 18 months before presenting at the veterinary medical center. Medial subluxation of the right elbow joint was detected by palpation. The Campbell test was consistent with an increased range of motion during supination. At ultrasonographic examination, medial luxation of the triceps brachii tendon was noted, whereas collateral ligaments appeared normal. No skeletal deformities were found on radiographs of the right forelimb. The luxation of the triceps brachii tendon was surgically corrected with antirotational suture, a stopper pin, medial retinaculum release, and imbrication of the lateral retinaculum. RESULTS: The right triceps brachii tendon and elbow joint were successfully reduced. Gait returned to normal by 55 days postoperatively. No implant failure or recurrence were observed 3.5 years after surgery. CONCLUSION: Surgical reduction of a luxation of the triceps brachii tendon in a dog resolved lameness and restored the range of motion of the affected elbow, leading to good long-term outcome.

A Biomechanical Comparison of Three Miniature Locking Plate Systems in a Rabbit Radial and Ulnar Fracture Model.

OBJECTIVE: The aim of this study was to evaluate the biomechanical properties of three different miniature locking plate systems used to fixate radial and ulnar fractures in toy breed dogs. Implant size, shape, material and locking systems differ, and their influence on the fracture healing process is unknown. In the present study, we aimed to investigate this matter in vivo using rabbit radial and ulnar fracture models. STUDY DESIGN: Eighteen rabbits were randomly divided into three groups, and the left radius and ulna were osteotomized to create fracture models. The osteotomies were then fixated using either the TITAN LOCK 1.5, Fixin micro or LCP 1.5 system. Radiographs were obtained 2, 3 and 4 weeks after surgery. Four weeks after surgery, the radiuses were collected and used for biomechanical testing or histological examinations. RESULTS: During the 4 weeks of observation, no adverse effects due to the implants occurred. The radiographic scores in each group did not differ significantly at any time point. The maximum load in the LCP group was significantly higher than that in the TITAN and Fixin groups. There was no significant difference in bending stiffness or work to failure among the groups. Initial fracture healing via woven bone was evident at histological evaluation. CONCLUSIONS: All three miniature locking plate systems provided adequate fracture stabilization 4 weeks after surgery, despite their differences, in rabbit models.

The significance of lipid accumulation in breast carcinoma cells through perilipin 2 and its clinicopathological significance.

Both systemic and intratumoral lipid metabolism have been recently reported to play pivotal roles in both tumor development and progression in various human malignancies including breast cancer. However, its details have remained largely unknown in breast cancer patients. Therefore, in this study, we focused on perilipin 2, which is involved in constituting the intracellular lipid composition. Perilipin 2 was first immunolocalized in 105 cases of breast cancer. The status of perilipin 2 immunoreactivity was significantly positively associated with histological grade, Ki-67 labeling index and HER2 status and negatively with estrogen receptor status of these patients. Subsequent in vitro study also revealed that its mRNA expression in triple negative breast carcinoma cells was higher than cells of other subtypes. We then examined the correlation between perilipin 2 immunoreactivity and intracellular lipid droplet evaluated by Oil-red O stating in 13 cases of breast carcinoma tissues. A significantly positive correlation was detected between the status of perilipin 2 and Oil-red O staining. These findings above did indicate that perilipin 2 could represent the status of intracellular lipid droplets in surgical pathology specimens of breast cancer and perilipin 2 was also associated with its more aggressive biological phenotypes.

Mesenchymal stem cells for cartilage regeneration in dogs.

Articular cartilage damage and osteoarthritis (OA) are common orthopedic diseases in both humans and dogs. Once damaged, the articular cartilage seldom undergoes spontaneous repair because of its avascular, aneural, and alymphatic state, and the damage progresses to a chronic and painful situation. Dogs have distinctive characteristics compared to other laboratory animal species in that they share an OA pathology with humans. Dogs can also require treatment for naturally developed OA; therefore, effective treatment methods for OA are desired in veterinary medicine as well as in human medicine. Recently, interest has grown in regenerative medicine that includes the use of mesenchymal stem cells (MSCs). In cartilage repair, MSCs are a promising therapeutic tool due to their self-renewal capacity, ability to differentiate into cartilage, potential for trophic factor production, and capacity for immunomodulation. The MSCs from dogs (canine MSCs; cMSCs) share various characteristics with MSCs from other animal species, but they show some deviations, particularly in their differentiation ability and surface epitope expression. In vivo studies of cMSCs have demonstrated that intraarticular cMSC injection into cartilage lesions results in excellent hyaline cartilage regeneration. In clinical situations, cMSCs have shown great therapeutic effects, including amelioration of pain and lameness in dogs suffering from OA. However, some issues remain, such as a lack of regulations or guidelines and a need for unified methods for the use of cMSCs. This review summarizes what is known about cMSCs, including their in vitro characteristics, their therapeutic effects in cartilage lesion treatment in preclinical in vivo studies, their clinical efficacy for treatment of naturally developed OA in dogs, and the current limitations of cMSC studies.

Bone regeneration by calcium phosphate-loaded carboxymethyl cellulose nonwoven sheets in canine femoral condyle defects.

The bone regeneration capacities of calcium phosphate (CaP)-loaded carboxymethyl cellulose (CMC) nonwoven sheet (CMC/CaP) were evaluated using a dog lateral femoral condyle defect model. In addition, the effect of bFGF on bone regeneration when added to CMC/CaP sheet was investigated. The CMC and CMC/CaP sheets have high operability. The new bone formation rate in the CMC/CaP group was significantly higher than that in the control and CMC groups based on micro-computed tomography and histological evaluation. In contrast, there was no significant difference between the CMC/CaP group and the CMC/CaP/f group. In conclusion, the CMC/CaP sheet has the ability to promote new bone formation and seems to be useful as a sheet-shaped bone graft substitute. The effect of the auditioning signaling molecules to the CMC/CaP sheet, such as bFGF, requires further investigation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1516-1521, 2019.

Canine mesenchymal stem cells from synovium have a higher chondrogenic potential than those from infrapatellar fat pad, adipose tissue, and bone marrow.

Osteoarthritis (OA), a common chronic joint disorder in both humans and canines, is characterized by a progressive loss of articular cartilage. Canines can serve as an animal model of OA for human medicine, and this research can simultaneously establish effective veterinary treatments for canine OA. One attractive treatment that can lead to cartilage regeneration is the use of mesenchymal stem cells (MSCs). However, for canine OA, little information is available regarding the best source of MSCs. The purpose of this study was to identify a promising MSC source for canine cartilage regeneration. We collected synovial, infrapatellar fat pad, inguinal adipose, and bone marrow tissues from six canines and then conducted a donor-matched comparison of the properties of MSCs derived from these four tissues. We examined the surface epitope expression, proliferation capacity, and trilineage differentiation potential of all four populations. Adherent cells derived from all four tissue sources exhibited positivity for CD90 and CD44 and negativity for CD45 and CD11b. The positive rate for CD90 was higher for synovium-derived than for adipose-derived and bone marrow-derived MSCs. Synovium-derived and infrapatellar fat pad-derived MSCs displayed substantial proliferation ability, and all four populations underwent trilineage differentiation. During chondrogenesis, the wet weight was heavier for cartilage pellets derived from synovium MSCs than from the other three sources. The synovium is therefore a promising source for MSCs for canine cartilage regeneration. Our findings provide useful information about canine MSCs that may be applicable to regenerative medicine for treatment of OA.

A zebrafish chemical suppressor screening identifies small molecule inhibitors of the Wnt/ß-catenin pathway.

Genetic screening for suppressor mutants has been successfully used to identify important signaling regulators. Using an analogy to genetic suppressor screening, we developed a chemical suppressor screening method to identify inhibitors of the Wnt/ß-catenin signaling pathway. We used zebrafish embryos in which chemically induced ß-catenin accumulation led to an "eyeless" phenotype and conducted a pilot screening for compounds that restored eye development. This approach allowed us to identify geranylgeranyltransferase inhibitor 286 (GGTI-286), a geranylgeranyltransferase (GGTase) inhibitor. Our follow-up studies showed that GGTI-286 reduces nuclear localization of ß-catenin and transcription dependent on ß-catenin/T cell factor in mammalian cells. In addition to pharmacological inhibition, GGTase gene knockdown also attenuates the nuclear function of ß-catenin. Overall, we validate our chemical suppressor screening as a method for identifying Wnt/ß-catenin pathway inhibitors and implicate GGTase as a potential therapeutic target for Wnt-activated cancers.