Platelet-rich plasma improves the therapeutic efficacy of mesenchymal stem cells by enhancing their secretion of angiogenic factors in a combined radiation and wound injury model.

Delayed wound healing after radiation exposure can cause serious cutaneous damage, and its treatment is a major clinical challenge. Although mesenchymal stem cells (MSCs) have emerged as a promising therapeutic agent in regenerative medicine, they alone do not produce satisfactory effects in a combined radiation and wound injury (CRWI) model. Here, we investigated the therapeutic effect of combined umbilical cord blood-derived (UCB)-MSCs and platelet-rich plasma (PRP) treatment on wound healing in a CRWI mouse model. First, we assessed the release of cytokines from UCB-MSCs cultured with PRP and observed changes in the expression of angiogenic factors. The angiogenic paracrine factors from UCB-MSCs cultured with PRP were assessed in human umbilical vein endothelial cells (HUVECs). To assess therapeutic efficacy, UCB-MSCs and PRP were topically implanted into a CRWT mouse model. Vascular endothelial growth factor (VEGF), a pro-angiogenic growth factor, urokinase-type plasminogen activator and contributor to VEGF-induced signalling were more highly expressed in conditioned media of UCB-MSCs cultured with PRP than in that of UCB-MSCs alone. Furthermore, conditioned media of UCB-MSCs cultured with PRP increased the formation of tube-like structures in HUVECs. Co-treatment of UCB-MSCs and PRP in a CRWI mouse model increased the wound closure rate and angiogenesis compared with an untreated irradiated group. Moreover, increased expression of VEGF and CD31 were observed in the wound tissue of co-treated mice compared with untreated irradiated mice. PRP stimulates the release of angiogenic factors from UCB-MSCs, and combined therapy of UCB-MSCs and PRP improves regeneration efficacy by enhancing angiogenesis in a CRWI model.

Transhiatal esophagogastric anastomosis and postoperative monitoring of thoracic esophageal leiomyosarcoma in a dog.

A 12-year-old Maltese dog was referred to the Veterinary Teaching Hospital at Konkuk University because of severe regurgitation. Radiography, ultrasonography, and computed tomography showed a mass in the thoracic esophagus. Localization of the tumor, its extraluminal nature, the positioning and involvement of the stomach, and the lack of diffuse metastasis to the lung were factors considered when developing a surgical plan. A successful surgical procedure was performed. The final diagnosis was leiomyosarcoma. Following surgery, clinical signs were significantly reduced and postoperative complications were not observed. The dog died 25 days after surgery; we suspected that the death was due to postoperative stricture. Key clinical message: Surgical approaches that prioritize maintenance of low tension on the thoracic esophagus are important to prevent arrhythmia, bradycardia, and ventricular premature complex during esophagogastric anastomosis. In dogs with a small esophageal lumen anastomosis may lead to postoperative stricture.

Anastomose oesophago-gastrique trans-hiatal et suivi post-opératoire d'un léiomyosarcome oesophagien thoracique chez un chien. Un Bichon maltais âgé de 12 ans fut référé à l'hôpital vétérinaire d'enseignement de la Konkuk University à cause de régurgitations sévères. Les radiographies, l'échographie et la tomodensitométrie ont montré la présence d'une masse dans l'oesophage thoracique. La localisation de la tumeur, sa nature extra-luminaire, le positionnement et l'implication de l'estomac et l'absence de métastase diffuse au poumon étaient des facteurs considérés lors du développement d'un plan chirurgical. Une procédure chirurgicale réussie fut réalisée. Le diagnostic final était un léiomyosarcome. Suite à la chirurgie, les signes cliniques étaient significativement réduits et aucune complication post-opératoire ne fut observée. Le chien est décédé 25 jours après la chirurgie, nous soupçonnons que la mort était due à un rétrécissement post-opératoire.Message clinique clé:Les approches chirurgicales qui priorisent le maintien de faible tension sur l'oesophage thoracique sont importantes pour prévenir l'arythmie, la bradycardie et un complexe ventriculaire prématuré lors d'une une anastomose oesophagogastrique. Chez les chiens avec un petit lumen oesophagien l'anastomose peut entraîner un rétrécissement post-opératoire.(Traduit par Dr Serge Messier).

3D Bioprinted Artificial Trachea with Epithelial Cells and Chondrogenic-Differentiated Bone Marrow-Derived Mesenchymal Stem Cells.

Tracheal resection has limited applicability. Although various tracheal replacement strategies were performed using artificial prosthesis, synthetic stents and tissue transplantation, the best method in tracheal reconstruction remains to be identified. Recent advances in tissue engineering enabled 3D bioprinting using various biocompatible materials including living cells, thereby making the product clinically applicable. Moreover, clinical interest in mesenchymal stem cell has dramatically increased. Here, rabbit bone marrow-derived mesenchymal stem cells (bMSC) and rabbit respiratory epithelial cells were cultured. The chondrogenic differentiation level of bMSC cultured in regular media (MSC) and that in chondrogenic media (d-MSC) were compared. Dual cell-containing artificial trachea were manufactured using a 3D bioprinting method with epithelial cells and undifferentiated bMSC (MSC group, n = 6) or with epithelial cells and chondrogenic-differentiated bMSC (d-MSC group, n = 6). d-MSC showed a relatively higher level of glycosaminoglycan (GAG) accumulation and chondrogenic marker gene expression than MSC in vitro. Neo-epithelialization and neo-vascularization were observed in all groups in vivo but neo-cartilage formation was only noted in d-MSC. The epithelial cells in the 3D bioprinted artificial trachea were effective in respiratory epithelium regeneration. Chondrogenic-differentiated bMSC had more neo-cartilage formation potential in a short period. Nevertheless, the cartilage formation was observed only in a localized area.

Human umbilical cord blood-derived mesenchymal stromal cells and small intestinal submucosa hydrogel composite promotes combined radiation-wound healing of mice.

BACKGROUND AIMS: Mesenchymal stromal cells (MSCs) are a promising agent for treating impaired wound healing, and their therapeutic potential may be enhanced by employing extracellular matrix scaffolds as cell culture scaffolds or transplant cell carriers. Here, we evaluated the effect of human umbilical cord blood-derived (hUCB)-MSCs and a porcine small intestinal submucosa (SIS)-derived extracellular matrix scaffold in a combined radiation-wound mouse model of impaired wound healing. METHODS: hUCB-MSCs and SIS hydrogel composite was applied to the excisional wound of whole-body irradiated mice. Assessment of wound closing and histological evaluation were performed in vivo. We also cultured hUCB-MSCs on SIS gel and examined the angiogenic effect of conditioned medium on irradiated human umbilical vein endothelial cells (HUVECs) in vitro. RESULTS: hUCB-MSCs and SIS hydrogel composite treatment enhanced wound healing and angiogenesis in the wound site of mice. Conditioned medium from hUCB-MSCs cultured on SIS hydrogel promoted the chemotaxis of irradiated HUVECs more than their proliferation. The secretion of angiogenic growth factors hepatocyte growth factor, vascular endothelial growth factor-A and angiopoietin-1 from hUCB-MSCs was significantly increased by SIS hydrogel, with HGF being the predominant angiogenic factor of irradiated HUVECs. CONCLUSIONS: Our results suggest that the wound healing effect of hUCB-MSCs is enhanced by SIS hydrogel via a paracrine factor-mediated recruitment of vascular endothelial cells in a combined radiation-wound mouse model.

Abnormal changes in both mandibular salivary glands in a dog: Non-mineral radiopaque sialoliths.

A 10-year-old Maltese dog was presented with a firm mass on the left side of his neck. Physical examination confirmed a firm mass in the left and a submandibular swelling in the right cervical region. Sialolithiasis and associated sialocele in both mandibular salivary glands were suspected and bilateral sialoadenectomy was performed. The stones were identified as non-mineral sialoliths.

Changements anormaux dans les deux glandes salivaires mandibulaires chez un chien : sialolithes radiopaques non minéraux. Un chien Maltais âgé de 10 ans a été présenté avec une masse ferme du côté droit du cou. L'examen physique a confirmé une masse ferme dans l'enflure gauche et sous-mandibulaire dans la région cervicale droite. La sialolithiase et une sialocèle connexe dans les deux glandes salivaires mandibulaires ont été suspectées et une sialoadénectomie bilatérale a été réalisée. Les pierres ont été identifiées comme des sialolithes non minéraux.(Traduit par Isabelle Vallières).

The use of fully threaded headless cannulated screws for femoral neck fracture fixation in small-breed dogs show promise in cadaveric study.

OBJECTIVE: To biomechanically evaluate the stability of internal fixation methods for femoral neck fractures in small-breed dogs. Furthermore, the possibility of replacing the headed screw with fully threaded headless cannulated screws in the fixation method was assessed. METHODS: The study was conducted from December 12, 2023, to February 7, 2024. A total of 18 femurs from 9 canine cadavers were used in this study. After a simple neck fracture was created, in group A (n = 6), the fracture was stabilized with three 1.1-mm parallel Kirschner wires (K-wires). In group B (n = 6), a 3.0-mm partially threaded cannulated screw and an antirotation pin were used. In group C (n = 6), a 2.5-mm fully threaded headless cannulated screw and an antirotation pin were used. A mechanical test was conducted to apply a single axial compressive load to the femoral head. RESULTS: 9 adult small-breed dogs weighing 3.6 to 8.3 kg (mean ± SD; 5.9 ± 1.6). The mean maximum failure load was highest in group C (495 ± 81 N), followed by group B (454 ± 50.4 N), and then group A (222 ± 21.6 N). Significant differences in maximum failure load were observed between groups A and B as well as groups A and C but not between groups B and C. CONCLUSION: The use of fully threaded headless cannulated screws presents a promising method for internal fixation of canine femoral neck fractures. CLINICAL RELEVANCE: To demonstrate the potential stability and reliability of fully threaded headless cannulated screws.

Biomechanical evaluation of three fixation methods in a feline transverse patella fracture model.

OBJECTIVES: This study aimed to compare the biomechanical properties of three stabilisation techniques in feline patella transverse fractures and select the strongest method with potentially minimal complications. METHODS: Patella fracture was simulated in 27 feline cadaveric pelvic limbs (mean weight of the cadavers 3.78 kg), and the limbs were randomly grouped to be stabilised with one of the three stabilisation methods. The modified tension band wiring technique with a single Kirschner wire (0.9 mm) and figure-of-eight wiring (20 G) was applied to group 1 (n = 9). Group 2 (n = 9) was stabilised with a combination of circumferential and figure-of-eight wiring techniques with orthopaedic wire (20 G). Group 3 (n = 9) was stabilised with the same technique as group 2, but with #2 FiberWire. The knee joints were positioned and fixed in the neutral standing angle (135°) and tested by applying tensile force. The loads at gap formations of 1, 2 and 3 mm were recorded, and the maximum failure load was measured in each group. RESULTS: In all the loads at displacement (1, 2 and 3 mm), group 3 was significantly stronger than groups 1 and 2, respectively (P <0.017). Group 3 (261.0 ± 52.8 N) showed significantly stronger fixation in the maximum load compared with group 1 (172.9 ± 45.6 N) (P <0.017). No significant difference was observed between groups 1 and 2 (204.9 ± 68.4 N) or between groups 2 and 3. CONCLUSIONS AND RELEVANCE: This study shows that the combination of circumferential and figure-of-eight techniques with FiberWire is more resistant to displacement than metal wire in this ex vivo feline patella fracture model.

Evaluation of LigaTie device for total lung lobectomy in small breed dogs - An ex vivo study.

This study aimed to evaluate an effectiveness of airway-sealing techniques used in total lung lobectomies in small dog breeds. Total lung lobectomies were performed on five canine cadavers weighing 2.5-8 kg. Airway-sealing techniques were performed on two randomly selected lobes per cadaver. The airway-sealing techniques consisted of traditional suture ligation, Endoloop, and LigaTie. After applying all surgical methods, bronchial stump was collected from the tongue and prepared. The effectiveness of each airway-sealing technique was evaluated by submerging the bronchial stumps in saline, followed by intubation to gradually increase the airway pressure to 80 cmH2O to identify the presence of air leakage in each lobe. Seven of the ten lobes in the traditional suture ligation group had a leakage; the leakages were fatal in three lobes. A single fatal leak occurred in the Endoloop group, while no leakage was noted in the LigaTie group. Therefore, the LigaTie technique had a higher statistical efficacy than the traditional suture ligation (p = 0.007). In conclusion, the LigaTie application reduced the incidence of air leakage following total lung lobectomies in small dog breeds.

Anterior cruciate ligament reconstruction in a rabbit model using canine small intestinal submucosa and autologous platelet-rich plasma.

BACKGROUND: The bone-ligament interface is the main point of failure after anterior cruciate ligament reconstruction. Synthetic ligament materials have problems such as a greater failure rate of the bone-ligament insertion than autografts. Small intestinal submucosa (SIS) is a biologic scaffold that has been used to repair musculoskeletal tissue and has been shown to promote cell migration and enhance collagen fiber regeneration. Autologous platelet-rich plasma (PRP) has also been investigated as a potential promoter of tendon healing. We investigated SIS and PRP as biomaterials that might strengthen the bone-tunnel interface and improve tendon structure formation. METHODS: Anterior cruciate ligament grafts were formed of braid-twist canine SIS. These canine SIS ligament grafts were used for anterior cruciate ligament reconstruction in 20 New Zealand white rabbits. The rabbits were divided into 2 treatment groups. In 1 group (SIS group; n = 10), we only implanted the canine SIS grafts. In the second group (PRP group; n = 10), we applied autologous PRP to the surgical area after implantation of canine SIS grafts. We determined the cytokine level of the autologous PRP using a transforming growth factor-ß1 enzyme-linked immunosorbent assay kit. At 1 and 4 wk after surgery, magnetic resonance imaging was performed to evaluate the grafts. The femur-graft-tibia complex was assessed histologically and biomechanically at 8 wk after surgery. RESULTS: At 1 wk after surgery, the magnetic resonance imaging scans of the PRP group showed high signal-intensity lesions. In biomechanical tests, the SIS group had a significantly greater maximum load, maximum stress, and ultimate load and strain than the PRP group. The histologic findings of the PRP group revealed a greater cellular response, fibrotic tissue regeneration around the graft, broad chondrocyte cell infiltration, and collagen fibers that were loosely attached to the bone. CONCLUSIONS: The PRP group had significantly lower tension load values than the SIS group, and there was greater cellular response in a broad area around the grafts of the rabbits in the PRP group compared with those in the SIS group. The early inflammatory responses around the canine SIS grafts in the PRP group and the altered cytokine or growth factor concentration in the intra-articular capsule of the rabbits in PRP group might explain their relatively low tensile strength results.

Tracheal replacement with fresh and cryopreserved aortic allograft in adult dog.

BACKGROUND: Many reports have described tracheal replacement using aortic allografts, with varying results and minimal understanding of the mechanism of tracheal regeneration. The present study attempted tracheal regeneration in adult dogs using fresh aortic allografts (FAA) and cryopreserved aortic allografts (CAA). MATERIALS AND METHODS: Twelve adult beagles underwent tracheal resection and were transplanted with FAA (n = 5) or CAA (n = 7). Animals were followed-up with serial radiography and magnetic resonance imaging, and were euthanized at predetermined times up to 16 mo post-surgery. RESULTS: There were no procedural deaths, but two animals died due to stent migration. Stent migration occurred in seven of the 12 animals. Evidence of regeneration of tracheal epithelium was observed in the surviving animals, with the transformation of squamous metaplasia to mucociliary epithelium being time-dependent. Islet of cartilage were observed in animals after 6 mo, but ring-like cartilage structures were absent, even after 16 mo. During autopsy, axial graft contractions up to 68% were observed. Serial radiographs show that most of the contraction occurred within 1 mo. The results of the MRI showed that the graft area was strongly enhanced for up to 2 mo, but was clearly reduced after 3 mo. CONCLUSIONS: Tracheal replacement in adult dogs using FAA or CAA is feasible. However, immaturity of the neotracheal cartilage did not allow the tissue to function as native tracheal tissue. Prolonged stenting should be considered in adult if the procedure is to be clinically contemplated.

Efficacy of milrinone versus nitroglycerin in controlling pulmonary arterial hypertension induced by intravenous injections of absolute ethanol in anesthetized dogs.

PURPOSE: To compare the attenuating effects of milrinone versus nitroglycerin on pulmonary artery pressure (PAP) increase caused by repeat intravenous bolus injection of absolute ethanol in anesthetized dogs. MATERIALS AND METHODS: Twenty-four dogs were randomly distributed into three groups: a control group (group C), a nitroglycerin group (treated with a continuous infusion of nitroglycerin at 5.0 microg/kg/min; group N), and a milrinone group (treated with a continuous infusion of milrinone at 3.0 microg/kg/min after an initial bolus dose of 100 microg/kg; group M). Absolute ethanol (0.1 mL/kg) was injected at 10-minute intervals via a 5-F angiographic catheter advanced into the proximal portion of the inferior vena cava. Immediately before and after each intravenous injection of absolute ethanol, hemodynamic values were obtained through a pulmonary arterial catheter. RESULTS: Group M showed more stable hemodynamic values of systolic, mean, and diastolic PAP and pulmonary vascular resistance (PVR) compared with groups C and N. In addition, group M showed significantly higher values of cardiac output versus group C at 10 minutes after the 10th bolus of ethanol. After restoration of spontaneous breathing, group M also showed the least hemodynamic changes in systolic, mean, and diastolic PAPs and PVR among the groups. CONCLUSIONS: Although a continuous infusion of nitroglycerin or milrinone effectively prevented cardiovascular deterioration, milrinone was superior to nitroglycerin in managing the acute hemodynamic changes that resulted from repeated intravenous bolus injections of absolute ethanol (0.1 mL/kg) in healthy anesthetized dogs.

Differential neurochemical responses of the canine striatum with pentobarbital or ketamine anesthesia: a 3T proton MRS study.

Although anesthetic agents are known to affect cerebral metabolism, pentobarbital and ketamine have been widely used for animal imaging studies. The purpose of this study is to evaluate alterations in striatum metabolites in dogs between anesthetized with pentobarbital and with ketamine in proton magnetic resonance spectroscopy ((1)H-MRS). (1)H-MRS was performed to ten healthy adult beagle dogs (9-11 kg) at a field strength of 3 T in order to identify metabolic changes after pentobarbital or ketamine administration in the striatum in vivo. Ten dogs were divided into 2 groups as follows: 5 as the pentobarbital-administered group (P group) and 5 as the ketamine-administered group (K group). We found that levels of Glx of the P group was significantly lower than that of the K group (6.90 +/- 0.99 (SD) vs 9.77 +/- 1.14 in 5 dogs, p= 0.003). In addition, the P group also has lower levels of Cr (6.29 +/- 0.44 vs 7.89 +/- 0.91 in 5 dogs, p=0.009) and NAA (5.02 +/- 0.65 vs 6.45 +/- 1.13 in 5 dogs, p=0.041) compared to the K group. However, there were no significant difference between the P group and the K group in striatal levels of Cho and Ins (p>0.1). We demonstrated that MRS-measured metabolites in the specific regions of the brain can be influenced by anesthetic agents.

Up-regulation of MicroRNAs-21 and -223 in a Sprague-Dawley Rat Model of Traumatic Spinal Cord Injury.

In this experimental animal study, we examined alterations in the degree of transcription of two microRNAs (miRs)-miR-21 and -223-in a Sprague-Dawley (SD) rat model of traumatic spinal cord injury (TSCI). Depending on the volume of the balloon catheter (V), a total of 75 male SD rats were divided into the three experimental groups: the sham group (n = 25; V = 0 µL), the mild group (n = 25; V = 20 µL), and the severe group (n = 25; V = 50 µL). Successful induction of TSCI was confirmed on both locomotor rating scale at 4 h and 1, 3 and 7 days post-lesion and histopathologic examinations. Then, RNA isolation and quantitative polymerase chain reaction (PCR) were performed. No differences in the level of miR-21 expression were found at the first time point studied (4 h post-lesion) between the three experimental groups, whereas such differences were significant at all the other time points (P < 0.05). Moreover, there were significant alterations in the level of miR-223 expression at all time points studied through all the experimental groups (P < 0.05). Furthermore, locomotor rating scale scores had a linear relationship with the level of miR-21 expression (R2 = 0.4363, Y = 1.661X + 3.096) and that of miR-223 one (R2 = 0.9104, Y = 0.8385X + 2.328). Taken together, we conclude that up-regulation of miR-21 and -223 might be closely associated with progression and the early course of TSCI, respectively.

Intraarterially delivered human umbilical cord blood-derived mesenchymal stem cells in canine cerebral ischemia.

The present study examined the effects of human umbilical cord blood-derived mesenchymal stem cells (HUCB-derived MSCs) delivered through the basilar artery in a canine thromboembolic brain ischemia model. Cerebral ischemia was induced through occlusion of the middle cerebral artery by injecting thrombus emboli into 10 beagles. In the HUCBC group (n = 5), 1 x 10(6) HUCB-derived MSCs were transplanted through the basilar artery 1 day after ischemic induction using an endovascular interventional approach. In the control group (n = 5), phosphate-buffered saline (PBS) was injected in the same manner in as the HUCBC group. Upon neurobehavioral examination, earlier recovery was observed in the HUCBC group. The HUCBC group showed a decrease in the infarction volume at 1 week after cerebral ischemic induction, whereas the control group showed an increase in the infarction volume at 1 week, by magnetic resonance image analysis. Transplanted cells had differentiated into neurons and astrocytes and were observed in and around endothelial cells that were positive for von Willebrand factor (vWF). HUCB-derived MSCs expressed neuroprotective factors, such as brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF), at 4 weeks after the transplantation. The transplanted cells demonstrated their efficacy by reducing the infarction lesion volume and through earlier recovery from the neurological deficit. These results suggest that intraarterial transplantation of HUCB-derived MSCs could be useful in clinical treatment of cerebral ischemia.

Experimental Tracheal Replacement Using 3-dimensional Bioprinted Artificial Trachea with Autologous Epithelial Cells and Chondrocytes.

Various treatment methods for tracheal defects have been attempted, such as artificial implants, allografts, autogenous grafts, and tissue engineering; however, no perfect method has been established. We attempted to create an effective artificial trachea via a tissue engineering method using 3D bio-printing. A multi-layered scaffold was fabricated using a 3D printer. Polycaprolactone (PCL) and hydrogel were used with nasal epithelial and auricular cartilage cells in the printing process. An artificial trachea was transplanted into 15 rabbits and a PCL scaffold without the addition of cells was transplanted into 6 rabbits (controls). All animals were followed up with radiography, CT, and endoscopy at 3, 6, and 12 months. In the control group, 3 out of 6 rabbits died from respiratory symptoms. Surviving rabbits in control group had narrowed tracheas due to the formation of granulation tissue and absence of epithelium regeneration. In the experimental group, 13 of 15 animals survived, and the histologic examination confirmed the regeneration of epithelial cells. Neonatal cartilage was also confirmed at 6 and 12 months. Our artificial trachea was effective in the regeneration of respiratory epithelium, but not in cartilage regeneration. Additional studies are needed to promote cartilage regeneration and improve implant stability.

Development of an improved canine model of percutaneous spinal cord compression injury by balloon catheter.

We developed a minimally invasive canine model of spinal cord injury (SCI). A balloon catheter was inserted into the epidural space via the lumbosacral space, and inflated between L2 and L3 for 30 or 60 min under fluoroscopic guidance. Motor function after SCI was assessed using modified Tarlov scale. All seven dogs showed complete paraplegia after the procedure, neurological problems were evident and the modified Tarlov scores remained at zero after the SCI procedure; no improvement in clinical signs was observed. The dogs underwent 3T MR imaging at 3 days and 1 year after SCI. Histopathologic examinations were conducted at 2 weeks, 12 weeks and 1 year after SCI. In the present study, we described an animal model of minimally invasive spinal cord injury using a balloon catheter without laminectomy under fluoroscopic guidance. And, this percutaneous spinal cord compression injury model has many potential applications. The described percutaneous spinal cord compression injury model offers a new means of administering SCI and has many potential applications.

Hematogenous metastasis of embryonal rhabdomyosarcoma originating from skeletal muscle in a young dog.

An 8-month-old, intact male Golden Retriever with a history of left forelimb lameness for 2 months was presented to the Veterinary Medical Teaching Hospital of Konkuk University (Seoul, Korea). Results of a physical examination revealed a mass in the left axillary region. A thoracic radiography showed an osteolytic lesion in the scapula and the presence of a soft tissue density from the thoracic wall to the scapula. A computerized tomography revealed a mass invading into the scapula, and small nodules in the lung that suggested metastasis. At necropsy, a pale-yellow, irregular, firm, 8 x 10 x 5 cm mass extended from axillary region and destroyed the scapular. In addition, small nodules were noted in the lung. On microscopic examination, the mass consisted of round-to-oval cells, with eccentrically located hyperchromatic nuclei and eosinophilic cytoplasm in fibromyxoid stroma. Tumor cells were observed in blood vessels in the primary mass. Tumor cells strongly expressed vimentin, desmin, and myoglobin. In phosphotungstic acid-hematoxylin staining, cross-striations were detected in rhabdomyoblasts. In periodic acid-Schiff reaction, only a few cells were detected. The diagnosis was primary rhabdomyosarcoma of the appendicular muscle of a young dog. The tumor presumably originated in the skeletal muscle of the limb, invaded into the adjacent scapular bone, and metastasized to the lung.

Transdermal water-in-oil nanocarriers of nitric oxide for triggering penile erection.

Men's sexual health can have significant effects on a man's self-esteem, sexual relationship and male reproductive functions. Although commercially available drugs (e.g., VIAGRA and CIALIS) show effective treatment of erectile dysfunction (ED), patients with severe ED fail to respond to these medicines. Topical nitric-oxide (NO) delivery to penis can be a painless, alternative solution with severe ED because NO triggers erection and diffuses to the trabecular arteries and smooth muscles in the penis. We here develop water-in-oil (W/O) nanoemulsions (NEs) that contain NO and can directly spread on the penis. We optimize NE formation conditions including hydrophilic-lipophilic balance (HLB) and ratio of oil, water and surfactants. Then, by spreading NEs on penis skin of intact middle aged dogs, we verify medication effects and safety of the NEs in vivo. The water-in-oil NEs can be a promising non-invasive medication for ED patients with low response to a phosphodiesterase type 5 (PDE5) inhibitor, thus increasing quality of life in the aging society.

Comparison of articular cartilage with costal cartilage in initial cell yield, degree of dedifferentiation during expansion and redifferentiation capacity.

Costal cartilage has been proposed as an alternative donor of chondrocytes for articular-cartilage repair. In the present study we compared the initial cell yield of chondrocytes from rabbit costal cartilage and their cell expansion rates in monolayer culture with those of articular cartilage. Costal cartilage gave an approx. 2.6-fold higher cell yield than did articular cartilage. During in vitro culture, CCs (costal chondrocytes) grew faster and displayed approx. 3-fold more cell expansion up to P4 (passage 4) than did ACs (articular chondrocytes). In order to match the degree of dedifferentiation during serial cultivation with the cells' expansion rate, type II collagen expression and the emergence of fibroblastic morphology were monitored at each cell passage. Both ACs and CCs gradually lost their chondrocytic phenotype, changed to fibroblast-like cells and displayed a reduced expression of type II collagen. We then also evaluated the redifferentiation capacity of the expanded ACs and CCs by culturing them at high density in collagen gel. Almost fully dedifferentiated CCs at P4 were successfully redifferentiated into hyaline cartilage, which showed the expression of glycosaminoglycan and type II collagen as well as the formation of lacunae and a territorial matrix. In conclusion, costal cartilage may have advantages over articular cartilage as an alternative donor tissue for autologous chondrocytes on the basis of its higher cell yield, higher cell expansion and successful reversion into hyaline cartilage without ossification in vitro. However, although this experiment with a rabbit model gave a better insight into the problem than other experiments have done, it does not answer definitively the question as to which cells are most appropriate for articular cartilage repair in humans.

Primary intrapelvic hemangiosarcoma in a dog.

A 12-year-old, spayed female Schnauzer presented with constipation. A mass was observed in the pelvic cavity, and metastasis was not identified. Mass resection was performed through celiotomy with pubic osteotomy, and hemangiosarcoma was diagnosed. At 10 weeks post-operatively, the patient died of multiple metastasis. Primary intrapelvic hemangiosarcoma is rare in dogs.