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 (beta-TCP) in orthotopic implantation. Seven hundred milligrams of beta-TCP mixed with 1 x 10(6) 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 beta-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 beta-TCP is a promising osteogenic material for repairing bone defects.
Animals ; Biocompatible Materials/metabolism/therapeutic use ; Bone Substitutes/*therapeutic use ; Calcium Phosphates/*therapeutic use ; Dogs ; Fetal Blood/*cytology ; Fracture Fixation/methods/veterinary ; Mesenchymal Stem Cells/*physiology ; Osteogenesis/*physiology ; Tissue Engineering/methods ; Wound Healing/physiology

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 beta-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.
Adipocytes, White/cytology/physiology ; Alkaline Phosphatase/metabolism ; Animals ; Biocompatible Materials/metabolism/*therapeutic use ; Bone Diseases/*therapy ; Bone Marrow Cells/cytology/physiology ; Calcification, Physiologic ; Calcium/metabolism ; Calcium Phosphates/metabolism/therapeutic use ; Cell Proliferation ; Dogs ; Female ; Fetal Blood/cytology/physiology ; Flow Cytometry ; Male ; Mesenchymal Stromal Cells/cytology/*metabolism ; *Osteogenesis ; Polyesters/metabolism/therapeutic use ; Tissue Engineering/*methods ; Vascular Endothelial Growth Factor A/metabolism

The distribution and function of transforming growth factor-beta (TGF-beta) in the region of bone defect repaired by collagen/nano-beta-tricalcium phosphate composite artificial bone (Co/N-TCP) and the ability of Co/N-TCP recruiting osteoblasts to precipitate the repair of bone defect were investigated. Twenty-four domestic rabbits were operated on bilateral cranial bone to create an experimental bone defect of 8.0 mm in diameter through the whole bone. On the left, Co/N-TCP was implanted as experimental group, but on the right, Co/TCP was implanted as control group. At 2nd, 4th, 8th, 12th week after operation, all animals were sacrificed and the implanted materials with surrounding bone were taken out. Immunohistochemical staining was performed for TGF-beta assay by avidin-biotin complex method (SABC). Simultaneously, TGF-beta was quantitatively analyzed by HPIAS-1000 imaging analysis system. The immunohistochemical staining for TGF-beta revealed that osteoblasts and immature osteocytes highly expressed TGF-beta. Diffused TGF-beta positive staining particles appeared in the mesenchymal and fibrous-tissue. There was no significant difference in the TGF-beta positive staining between two groups in the medial region to original osseous beds at different time points (P > 0.05). However, in distal original osseous bed of the defected region, the positive expression of TGF-beta in the Co/N-TCP group was significantly stronger than in the control group (P < 0.05 or 0.01). The Co/N-TCP has good bioactivities and ability of stimulating and conducting TGF-beta to aggregate and precipitate the healing of bone defect.
Animals ; Biocompatible Materials ; therapeutic use ; Calcium Phosphates ; Ceramics ; Collagen ; Fracture Healing ; Implants, Experimental ; Nanotechnology ; Osseointegration ; drug effects ; Osteogenesis ; physiology ; Rabbits ; Skull Fractures ; metabolism ; surgery ; Transforming Growth Factor beta ; analysis ; metabolism

This paper is aimed to assess the efficacy of recombinant human bone morphogenetic protein-2 (rhBMP-2) and bone marrow stromal cell (BMSCs) composited tricalcium phosphate (TCP) in a rat model of posterolateral lumbar intertransverse process fusion. Rat BMSCs were cultured in vitro. Twenty SD rats underwent single-level bilateral intertransverse process spine arthrodesis at L4 and L5. These rats were assigned to two groups according to the graft materials. They received: 10 of the total were treated with the BMSCs with rhBMP-2 and tricalcium phosphate (TCP) as the experimental group, and the other 10 with TCP treatment alone as the control group. All the animals were killed at 4 weeks after surgery and the spine fusion results were assessed by gross inspection, manual palpation, radiography and histology. The fusion rate, the tensile strength and stiffness of the solidly fused levels in the experimental group were statistically higher than that of the controlled group (P < 0.05). These results showed that the spinal fusion could be improved mechanically when rhBMP-2 and BMSCs were added into the TCP.
Animals ; Bone Morphogenetic Protein 2 ; therapeutic use ; Calcium Phosphates ; therapeutic use ; Cells, Cultured ; Combined Modality Therapy ; Female ; Lumbar Vertebrae ; surgery ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Osseointegration ; physiology ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; therapeutic use ; Spinal Fusion ; methods ; Transforming Growth Factor beta ; therapeutic use

Although renal calcium crystal deposits (nephrocalcinosis) may occur in acute phosphate poisoning as well as type 1 renal tubular acidosis (RTA), hyperphosphatemic hypocalcemia is common in the former while normocalcemic hypokalemia is typical in the latter. Here, as a unique coexistence of these two seperated clinical entities, we report a 30-yr-old woman presenting with carpal spasm related to hypocalcemia (ionized calcium of 1.90 mM/L) due to acute phosphate poisoning after oral sodium phosphate bowel preparation, which resolved rapidly after calcium gluconate intravenously. Subsequently, type 1 RTA due to Sjogren's syndrome was unveiled by sustained hypokalemia (3.3 to 3.4 mEq/L), persistent alkaline urine pH (> 6.0) despite metabolic acidosis, and medullary nephrocalcinosis. Through this case report, the differential points of nephrocalcinosis and electrolyte imbalances between them are discussed, and focused more on diagnostic tests and managements of type 1 RTA.
Acidosis, Renal Tubular/*diagnosis/etiology ; Acute Disease ; Adult ; Antibodies, Antinuclear/blood ; Calcium Gluconate/therapeutic use ; Chronic Disease ; Female ; Humans ; Hydrogen-Ion Concentration ; Hypocalcemia/*chemically induced/complications/drug therapy ; Nephrocalcinosis/complications/*diagnosis/ultrasonography ; Parotid Gland/ultrasonography ; Phosphates/*adverse effects ; Salivary Glands/radionuclide imaging ; Sjogren's Syndrome/complications/*diagnosis/metabolism ; Submandibular Gland/ultrasonography