Isolation and Characterization of Extracellular Vesicle from Mesenchymal Stem Cells of the Epidural Fat of the Spine

Methods@#In this study, we isolated the exosomes using the tangential flow filtration (TFF) system with exosome-depleted fetal bovine serum and performed the characterization tests via western blotting, reverse transcription–polymerase chain reaction, nanoparticle tracking analysis (NTA), and transmission electron microscopy. @*Results@#In transmission electron microscopy, the exosome had a diameter of approximately 100–200 nm and had a spherical shape, whereas in the NTA, the exosome had an average diameter of 142.8 nm with a concentration of 1.27×1010 particles/mL. The flow cytometry analysis showed the expression of CD63 and CD81. The western blotting analysis showed the positive markers. @*Conclusions@#These findings showed that isolating the exosomes via TFF resulted in high-quality EF-MSC exosome yield. Further studies with exosomes from EF-MSC are needed to evaluate the function and role of the EF tissue.

A comparative study of the phenotype with kainic acid-induced seizure in DBA/2 mice from three different sources / 한국실험동물학회지

The kainic acid-induced seizure mouse model is widely used in epilepsy research. In this study, we applied kainic acid to the subcutaneous injections of three different sources of DBA/2 mice to compare and evaluate the seizure response. The three mouse sources consisted of DBA/2Kor1 (Korea FDA source), DBA/2A (USA source), and DBA/2 (Japan source), and were purchased from different vendors. To compare the responses of DBA/2 mice to kainic acid injections, we examined the survival rate, seizure phenotype scoring, and behavioral changes. We also evaluated brain lesions using histopathological analysis. Following the administration of kainic acid, almost half of the cohort survived, and the seizure phenotype displayed a moderate level of sensitivity (2 ~ 4 out of 6). In the histopathologic analysis, there was no change in morphological features, and levels of glial fibrillary acidic protein (GFAP) and ionized calcium binding adaptor molecule 1 (Iba-1) increased in the kainic acid-treated groups. However, there was no difference in the neuronal nuclei (NeuN) expression level. All the data showed that the responses in the kainic acid-treated group were similar across the three strains. In conclusion, our results suggest that the three sources of DBA/2 mice (DBA/2Kor1, DBA/2A, and DBA/2B) have similar pathological responses to kainic acid-induced seizures.

A comparative study of the phenotype with kainic acid-induced seizure in DBA/2 mice from three different sources / 한국실험동물학회지

The kainic acid-induced seizure mouse model is widely used in epilepsy research. In this study, we applied kainic acid to the subcutaneous injections of three different sources of DBA/2 mice to compare and evaluate the seizure response. The three mouse sources consisted of DBA/2Kor1 (Korea FDA source), DBA/2A (USA source), and DBA/2 (Japan source), and were purchased from different vendors. To compare the responses of DBA/2 mice to kainic acid injections, we examined the survival rate, seizure phenotype scoring, and behavioral changes. We also evaluated brain lesions using histopathological analysis. Following the administration of kainic acid, almost half of the cohort survived, and the seizure phenotype displayed a moderate level of sensitivity (2 ~ 4 out of 6). In the histopathologic analysis, there was no change in morphological features, and levels of glial fibrillary acidic protein (GFAP) and ionized calcium binding adaptor molecule 1 (Iba-1) increased in the kainic acid-treated groups. However, there was no difference in the neuronal nuclei (NeuN) expression level. All the data showed that the responses in the kainic acid-treated group were similar across the three strains. In conclusion, our results suggest that the three sources of DBA/2 mice (DBA/2Kor1, DBA/2A, and DBA/2B) have similar pathological responses to kainic acid-induced seizures.

Epidural Fat-Derived Mesenchymal Stem Cell: First Report of Epidural Fat-Derived Mesenchymal Stem Cell

STUDY DESIGN: Experimental study. PURPOSE: To determine whether epidural fat (EF) tissue contains mesenchymal stem cells (MSC). OVERVIEW OF LITERATURE: Spine surgeons are unaware of the contents of EF tissue and the reason for its presence between the ligamentum flavum and the dura mater; therefore, EF tissues are routinely eliminated during surgical procedures. However, EF removal causes certain postoperative problems, such as post-laminectomy syndrome. We hypothesized that the EF tissue may play a significant supportive role for the neural structures and other nearby conditions. METHODS: EF tissues were obtained from consenting patients (n=3) during posterior decompression surgery of the lumbar spine. The primary cells were isolated and cultured as per previously described methods with some modifications, and the cell morphology and cumulation were examined. Thereafter, reverse transcription–polymerase chain reaction (RT-PCR), a fluorescence-activated cell sorting (FACS) analysis, and differentiation potency for differentiation into osteoblasts, chondroblasts, and adipocytes were investigated to identify whether the cells derived from EF are MSC. RESULTS: The cells from the EF tissue had a fibroblast or neuron-like morphology that persisted until the senescence at p18. MSC-specific genes, such as OCT4, SOX2, KLF4, MYC, and GAPDH were expressed in the RT-PCR study, while MSC-specific surface markers such as CD105, CD90, and CD73 were exhibited in the FACS analysis. The differentiation properties of EF-MSC for differentiation into the three types of cells (osteoblast, chondroblast, and adipocyte) were also confirmed. CONCLUSIONS: Based on the cell culture, FACS analysis, RT-PCR analysis, and differentiation potent outcomes, all the features of the cells corresponded to MSC. This is the first study to identify EF-MSC derived from the EF tissue.

Comparison study of the response with botulinum toxin muscle injection in the ICR mice from three different sources / 한국실험동물학회지

Botulinum-toxin A (BoNT/A) is a widely used not only for cosmetics but also for various experimental purposes including muscle-related research. In this study, we applied BoNT/A to mouse muscle of three different sources to compare and evaluate the biological and pathological response. The three different mouse sources consist of Korl: ICR (Korea FDA source), A:ICR (USA source) and B:ICR (Japan source) which were purchased from each different vendors. To compare the responses of ICR mice with BoNT/A muscle injection, we examined the body weight, hematological and serum biochemistry analysis. Also, we evaluated the muscle change by histopathological analysis and gene expression patterns of muscle-related target by qPCR. The body weight gain was decreased in the BoNT/ A-treated group compared with the control group. In clinical pathologic analysis and gene expression patterns, the data showed that the responses in the BoNT/A-treated group were similar compared with the control group. Decreased muscle fiber was observed in BoNT/A-treated group compared with control group, while Korl:ICR showed a little low response with the other mouse sources. In conclusion, our results suggest that three different sources ICR mice (Korl:ICR, A:ICR and B:ICR) have a similar biological and pathological responses in BoNT/A muscle injection.

Primary leiomyoma of the ovaries in a dog / 동물의과학연구지

We would like to report a case of leiomyoma of the ovaries in a dog. Leiomyoma is commonly seen in the vagina in dogs. However, it is a very rare neoplasm in the ovaries. As there have only been a few reported cases, this report provides valuable information on veterinary medicine and pathology. Masses found in the ovaries need to be differentiated from other ovarian tumors. Therefore, we describe the gross, histopathological, and immunohistochemical features of a case of ovarian leiomyoma in a 10-year-old female Yorkshire Terrier dog. The mass on the right of the uterus was found accidentally by pelvic ultrasonography. Laparatomy revealed a large multi-nodulated ovarian mass. Grossly, cut surfaces of the mass showed multiple firm whitish nodules in the cortex and bloody loose connective tissue in the medulla. Histopathologically, the cortex of the mass was composed of spindle cells forming interlacing fascicles. The cells had elongated, blunt-ended nuclei and eosinophilic cytoplasm as detected by hematoxylin and eosin staining. Immunohistochemical stained sections were immunoreactive for alpha-smooth muscle actin and desmin but negative for vimentin and S-100. Therefore, differential diagnosis confirmed leiomyoma based on morphology and positive staining for alpha-smooth muscle actin and desmin.

Chronic pyometra due to bilateral ovarian tumors in a South American sea lion / 동물의과학연구지

A 14-year-old female South American sea lion (Otaria byronia) with persistent vaginal secretion and chronic hemorrhagic diarrhea was encountered. During postmortem examination, the uterus was found to resemble a balloon with mucosal congestion and was filled with grayish milky material. The ovaries also had abnormal features, including necrotic surface lesions and multiple whitish foci in the cut section. Hemorrhages and ulcerated changes due to toxemia were observed in other organs, including the liver, spleen, lung, intestines, and lymph nodes. Microscopically, the left ovary contained interlacing fascicles of fibroblast-like cells with blunt-end nuclei showing cytoplasmic positive immunoreactivity against alpha-smooth muscle actin and desmin. The right ovary contained cells with round to cigar-shaped nuclei showing cytoplasmic positive immunoreactivity against vimentin. In conclusion, based on classification of bilateral ovarian tumors as a leiomyoma in the left region and a fibroma in the right region, this sea lion was diagnosed with chronic closed pyometra.

A case of metastatic cholangiocarcinoma in a ferret / 동물의과학연구지

Cholangiocarcinoma is a relatively rare neoplasm in animals and humans. A four-year-old, neutered male ferret presented with depression, anorexia, cachexia, diarrhea, and icterus. Necropsy findings included ascites, multiple white nodules on the surface of the liver, stomach, and duodenum, gross enlargement of the bile duct and right atrium, hemorrhage of the gastric and duodenal mucosa, and icterus of the mesenteric fat. Infiltrative well-differentiated neoplastic biliary epithelial cells forming ducts and acini with a prominent collagenous stroma were observed on microscopic examination of neoplastic lesions within the liver, mesentery, and the serosa of the stomach and duodenum. This is a report on a rare case of obstructive jaundice due to cholangiocarcinoma in a ferret.

Case of Sporadic T-cell Lymphosarcoma in Cattle / 동물의과학연구지

We would like to report a case of bovine lymphosarcoma. Parous cattle from a livestock farmhouse were examined for mutinodular masses in the abdominal cavity after slaughter. For clinical signs, animals presented mild leukemic signs but did not have viral or bacterial infection. Grossly, whitish to yellowish smooth masses similar to fat tissue were covered with a thin membrane. A multilobulated mass formed around the arteri, and there was a large quantity of reddish fluid on the cut surface. Histopathologically, a monomorphic population of lymphocytes was observed along with small amounts of cytoplasm, round nuclei with coarsely granular chromatin, and numerous mitotic figures in the samples. In the tumor lesion, uniformly round cells had invaded with abundant neovascularization. Especially, the immunohistochemical phenotype of tumor cells was positive for anti-CD3 and negative for anti-CD8 and anti-CD20. Therefore, morphological analysis diagnosed the mass as a multinodular bovine lymphosarcoma of T-cell origin without any sign of infection by a viral agent.

The Efficacy of MRI in Tibial Plateau Fractures

PURPOSE: To analysis the efficacy of MRI in assessing fracture configuration and frequency of associated soft tissue injuries in tibial plateau fractures. MATERIALS AND METHODS: In the plain films and MRI of 47 cases with tibial plateau fractures,every fracture was classified by findings on plain film and MRI respectively according to the Schatzker system and they were compared with each other. The degree of displacement and depression of the fracture fragments were measured on plain film and MRI. RESULTS: MRI was more accurate in determining the classification of the fracture and measuring the displacement and depression of fragments. Fracture classifications were changed in 12 cases. Especially in Schatzker VI cases, articular step off over 2 mm and articular fragments over 2X2 cm were revealed in detail by MRI. Evidence of internal derangement of the knee was found in 38 (80.8%) cases. CONCLUSION: Most of the cases with acute tibial plateau fracture were commonly associated with ligamentous and meniscal injuries. MRI can aid in accurate evaluation of tibial plateau fracture patterns and decision of treatment plan.