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| BMP9 Induced the Osteogenic Differentiation of Human Umbilical Cord Mesenchymal Stem Cells in vitro and vivo |
| LI Ya-sha1, LIU Xing2, BI Yang1, YANG Ke1, ZHAO Li1, GONG Meng-jia1, GUO Qi3 |
1. Department of Pediatric Research Institute, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, Key Laboratory of Pediatrics in Chongqing, Stem Cell Therapy Engineering Technical Center, Chongqing 400014, China;
2. Department of Orthopaedic Surgery, The Children's Hospital of Chongqing Medical University, Chongqing 400014, China;
3. Chongqing Quality Testing Inspection Centrer for Medical Devices, Chongqing 401127, China |
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Abstract Objective:To investigate the effect of Bone morphogenetic proteins 9 on induced osteogenic differentiation of human umbilical cord mesenchymal stem cells in vitro and in vivo.Methods:BMP9 gene-recombinant adenovirus was infected into hUC-MSCs and paralleled with the Ad-GFP as negative control group. Cells in two groups were detected in 3d, 5d, 7d of alkaline phosphatase activity;Immunohistochemical staining was used to calculated the expression of osteocalcin and osteopontin after 14d;Alizarin red staining was used to observe the formation of mineralized nodules after 21d;Then collect both groups of infected hUC-MSCs for the establishment of ectopic osteogenic model through nude mice subcutaneous injection, removed the ectopic induced bone tissue after 4 weeks and present to Micro-CT scan and analysis, H&E staining, Masson Trichrome staining, Alcain Blue staining.Results:It is shown that ALP activity and mineralized nodule formation of BMP9 group was significantly higher than that in control group, Immunohistochemical staining showed positive expression of OCN, OPG in BMP9 group was significantly higher than that of control group; the results of ectopic osteogenic model showed that the control group had no visible subcutaneous mass, Only Ad-BMP9 infected hUC-MSCs can observed the formation of ectopic bone obviously, average value of bone mineral density is 396.05+0.60; H&E staining results showed that the generation of mature bone matrix and bone trabecula in the BMP9 induced ectopic bone tissue, Masson Trichrome staining results showed that BMP9 induced of matrix mineralization, Alcain Blue staining results showed that BMP9 induced endochondral osteogenic effect. Conclusion:The BMP9 significantly induces osteogenic differentiation of hUC-MSCs in vitro and in vivo, to provide the feasibility for clinical cell therapy in bone tissue engineering.
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Received: 29 October 2015
Published: 14 December 2015
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