Comparing the osteogenic potential of canine mesenchymal stem cells derived from adipose tissues, bone marrow, umbilical cord blood, and Wharton’s jelly for treating bone defects

강병재,류학현,박성수,Yoshihisa Koyama,Masanori Kikuchi,우흥명,김완희,권오경 대한수의학회 2012 Journal of Veterinary Science Vol.13 No.3

Alternative sources of mesenchymal stem cells (MSCs) for replacing bone marrow (BM) have been extensively investigated in the field of bone tissue engineering. The purpose of this study was to compare the osteogenic potential of canine MSCs derived from adipose tissue (AT), BM, umbilical cord blood (UCB), and Wharton’s jelly (WJ) using in vitro culture techniques and in vivo orthotopic implantation assays. After canine MSCs were isolated from various tissues, the proliferation and osteogenic potential along with vascular endothelial growth factor (VEGF) production were measured and compared in vitro. For the in vivo assay, MSCs derived from each type of tissue were mixed with β-tricalcium phosphate and implanted into segmental bone defects in dogs. Among the different types of MSCs, AT-MSCs had a higher proliferation potential and BM-MSCs produced the most VEGF. AT-MSCs and UCB-MSCs showed greater in vitro osteogenic potential compared to the other cells. Radiographic and histological analyses showed that all tested MSCs had similar osteogenic capacities, and the level of new bone formation was much higher with implants containing MSCs than cell-free implants. These results indicate that AT-MSCs, UCB-MSCs, and WJ-MSCs can potentially be used in place of BM-MSCs for clinical bone engineering procedures.

Primary Adenocarcinoma of the Minor Duodenal Papilla

Takeru Wakatsuki,Atsushi Irisawa,Tadayuki Takagi,Yoshihisa Koyama,Sayuri Hoshi,Seiichi Takenoshita,Masafumi Abe,Hiromasa Ohira 연세대학교의과대학 2008 Yonsei medical journal Vol.49 No.2

A 70-year-old man was admitted to our institution due to aggravation of blood-sugar level control and because an abdominal CT showed dilatation of the main pancreatic duct. Upper gastrointestinal endoscopy revealed a flat elevated tumor with central ulceration in the second portion of the duodenum. Subsequent duodenoscopy for a more detailed examination showed that the tumor had originated in the minor duodenal papilla. A biopsy specimen showed moderately differentiated adenocarcinoma. Endoscopic retrograde pancreatography via the major duodenal papilla revealed a slightly dilated main pancreatic duct and obstruction of the accessory pancreatic duct. Endoscopic ultrasonography showed a hypoechoic mass in the minor duodenal papilla with retention of the muscularis propria of the duodenum. These findings suggest that the tumor existed only to a limited extent in the minor duodenal papilla, and that the tumor did not infiltrate into the pancreas. For treatment, pylorus-preserving pancreatoduodenectomy was performed, and histological findings revealed a well- differentiated adenocarcinoma that originated in the minor duodenal papilla. Primary adenocarcinoma of the minor duodenal papilla is extremely rare. Our case is the first report of primary adenocarcinoma of the minor duodenal papilla at an early stage with no infiltration into muscularis propria of the duodenum and pancreas.

Implantation of canine umbilical cord blood-derived mesenchymal stem cells mixed with beta-tricalcium phosphate enhances osteogenesis in bone defect model dogs

Byung-Jun Jang,변예은,임지혜,류학현,김완희,Yoshihisa Koyama,Masanori Kikuchi,강경선,권오경 대한수의학회 2008 Journal of Veterinary Science Vol.9 No.4

This study was performed to evaluate the osteogenic effect of allogenic canine umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) mixed with beta-tricalcium phosphate (β-TCP) in orthotopic implantation. Seven hundred milligrams of β-TCP mixed with 1 × 106 UCB-MSCs diluted with 0.5 ml of saline (group CM) and mixed with the same volume of saline as control (group C) were implanted into a 1.5 cm diaphyseal defect and wrapped with PLGC membrane in the radius of Beagle dogs. Radiographs of the antebrachium were made after surgery. The implants were harvested 12 weeks after implantation and specimens were stained with H&E, toluidine blue and Villanueva-Goldner stains for histological examination and histomorphometric analysis of new bone formation. Additionally, UCB-MSCs were applied to a dog with non-union fracture. Radiographically, continuity between implant and host bone was evident at only one of six interfaces in group C by 12 weeks, but in three of six interfaces in group CM. Radiolucency was found only near the bone end in group C at 12 weeks after implantation, but in the entire graft in group CM. Histologically, bone formation was observed around β-TCP in longitudinal sections of implant in both groups. Histomorphometric analysis revealed significantly increased new bone formation in group CM at 12 weeks after implantation (p < 0.05). When applied to the non-union fracture, fracture healing was identified by 6 weeks after injection of UCB-MSCs. The present study indicates that a mixture of UCB-MSCs and β-TCP is a promising osteogenic material for repairing bone defects.

Comparison of Canine Allogenic Umbilical Cord Blood Derived Mesenchymal Stem Cells and Their Lysates Mixed with Beta-Tricalcium Phosphate in Orthotopic Implantation

( Ye Eun Byeon ),( Hak Hyun Ryu ),( Sung Su Park ),( Yoshihisa Koyama ),( Masanori Kikuchi ),( Min Soo Seo ),( Wan Hee Kim ),( Kyung Sun Kang ),( Oh Kyeong Kweon ) 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.4

The objective of this study is to evaluate the effect of mixtures of β-tricalcium phosphate(β-Ca3(PO4)2, β-TCP) granules with canine allogenic umbilical cord blood derived mesenchymal stem cells(UCB-MSC) and their lysates on bone formation. The mixtures were implanted into cylindrical bone defects(7 mm diameter, 10 mm length) made bilaterally by using the trephine in the medial aspect of the distal femur and proximal tibia. Fluorochromes for bone labeling were administered for measuring the rate of mineral apposition at 4, 6 and 8 weeks after implantation. A radiographic evaluation of the radiopacity of the implant in the bone defect was made at 2 weeks intervals. At 10 weeks, postoperatively, bone regeneration and resorption of β-TCP were assessed by micro-computed tomography(CT) in the tibia and by fluorescence microscopy and histomorphological examination in the femur and tibia. In micro CT, cortical bony bridging in the UCB-MSC, cell lysate and control groups was observed in 2, 1 and 0 out of each of the 3 samples, respectively. Bone mineral densities within trabecular bone defects were significantly different among the different groups(UCB-MSC < cell lysate < control) but the mineral apposition rate per day from 4 weeks after implantation was not different. In histomorphology the rates of new bone formation and resorption of the β-TCP in the UCB-MSC group were significantly higher than in the control group of both femur and tibia than in the cell lysate group in the tibia(p<0.05). The rate of new bone formation in the cell lysate group was not different compared to that of the control group in both the femur and the tibia. However, the absorption rate of the β-TCP in the tibia was significantly higher than that of the control group. The UCB-MSC improved the absorption of β-TCP and bone formation without site-dependence; however their lysates showed site-dependent effects. These results suggested that the canine allogenic UCB-MSC mixed with β-TCP had a higher bone healing activity for bone defects than their lysates.

Guided bone regeneration with beta-tricalcium phosphate and poly L- lactide-co-glycolide-co-epsilon-caprolactone membrane in partial defects of canine humerus

권오경,오태훈,Md.Mizanur Rahman,임지혜,김대용,윤정희,김완희,Masanori Kikuchi,Junzo Tanaka,박미선,Yoshihisa Koyama 대한수의학회 2006 Journal of Veterinary Science Vol.7 No.1

This study was performed to evaluate the effect of betatricalcium phosphate and poly L-lactide-co-glycolide-coepsilon- caprolactone (TCP/PLGC) membrane in the repair of partial bone defects in canine proximal humerus. Three adult mixed-breed dogs were used during the experimental period. The length of the defect was quarter of the full length of humerus, and width of the defect was quarter of middle diameter of the lateral aspect of humerus. The humeri of each dog were divided into treatment (TCP/ PLGC) and control groups. The defect was covered with TCP/PLGC membrane in treatment group. To evaluate regeneration of the bone, computerized tomography (CT) and histopathologic examination were performed. The radiopaque lines were appeared at the original defect sites in TCP/PLGC group but below the original site in control at 4th week. Radiopacity and thickness of the defect sites, and radiopaque lines were more increased at 8th week than those of 4th week. Histopathologic findings revealed fibrous connective tissue migration into the defect and the migration inhibited the structure of new cortex to be placed in the original level in control whereas new cortex growth was found in the level of original line in TCP/ PLGC group. However, the new cortical bone in the TCP/ PLGC group was thinner and less organized than the adjacent intact cortex, and the amount of new cancellous bones were also scanty. The result suggested that TCP/ PLGC membrane is a good guided bone regeneration material to restore the original morphology of humerus in partial defect.