Paracrine effect of canine allogenic umbilical cord blood-derived mesenchymal stromal cells mixed with beta-tricalcium phosphate on bone regeneration in ectopic implantations

• Published in: Cytotherapy (Oxford, England) Vol. 12; no. 5; pp. 626 - 636
• Main Authors: Byeon, Ye-Eun, Ryu, Hak-Hyun, Park, Sung Su, Koyama, Yoshihisa, Kikuchi, Masanori, Kim, Wan Hee, Kang, Kyung-Sun, Kweon, Oh-Kyeong
• Format: Journal Article
• Language: English
• Published: England Elsevier Inc 01.09.2010; Informa Healthcare
• Subjects: Acid Phosphatase - metabolism; Advanced Basic Science; Animals; Bone Regeneration; Calcium Phosphates - metabolism; canine umbilical cord blood; Cyclooxygenase 2 - biosynthesis; Cyclooxygenase 2 - genetics; Cytokines - biosynthesis; Cytokines - genetics; Dermatologic Surgical Procedures; Dogs; Fetal Blood - cytology; Isoenzymes - metabolism; Mesenchymal Stem Cell Transplantation; mesenchymal stromal cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - immunology; Mesenchymal Stromal Cells - metabolism; Osteogenesis; Other; Paracrine Communication; paracrine effects; Skin - immunology; Skin - metabolism; Skin - pathology; Tartrate-Resistant Acid Phosphatase; Transplantation, Homologous; Vascular Endothelial Growth Factor A - biosynthesis; Vascular Endothelial Growth Factor A - genetics; β-tricalcium phosphate; bone regeneration; canine umbilical cord blood; β-tricalcium phosphate; paracrine effects; mesenchymal stromal cells
• ISSN: 1465-3249; 1477-2566
• DOI: 10.3109/14653249.2010.481665

Abstract Background aims The aim of this study was to evaluate the paracrine effects of canine umbilical cord blood (cUCB) mesenchymal stromal cells (MSC) mixed with β-tricalcium phosphate (β-TCP) on bone regeneration in ectopic implantation. Methods β-TCP mixed with cUCB MSC (UCB-MSC group), cell lysates (cell lysate group) or a control (control group) were respectively implanted in a subcutaneous pouches in the back of beagle dogs. The implants were harvested 1, 4, 7, 14, 28, 56, 84 days after implantation. Histological findings and stain analyzes of tartrate-resistant acid phosphatase (TRACP) and assays of alkaline phosphatase (ALP) and TRACP were evaluated. The mRNA expression levels of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX-2), vascular endothelial growth factor (VEGF) and transforming growth factor-β (TGF-β) were analyzed using semi-quantitative reverse transcription - polymerase chain reactions (RT-PCR). An enzyme-linked immunosorbent assay (ELISA) was used to confirm the protein expression levels of IL-6, COX-2, VEGF and TGF-β. Results TRACP-positive cells were observed in all groups 7 days after implantation. ALP and TRACP activities in the UCB-MSC group 84 days after implantation were significantly higher than those of the control (P>0.05). Histologic findings after 84 days showed that the osteoid matrix area in the UCB-MSC group was significantly larger than that of the control ( P <0.05). The mRNAs levels of IL-1, IL-6 and VEGF in UCB-MSC and cell lysate groups on day 1 were up-regulated compared with the control. The protein levels of IL-6 and VEGF in the UCB-MSC group at day 1 were significantly higher than that of the other groups ( P <0.05). Conclusions It is suggested that a significant release of cytokines by cUCB MSC, 1 day following implantation, could enhance bone regeneration.