Porous calcium phosphate cement induces osteogenesis of mesenchymal stem cells under microenvironment

doi:10.3969/j.issn.2095-4344.2016.25.003

Abstract

Abstract:

BACKGROUND: The osteoinduction by biomaterials has been proven in various animal experiments.

OBJECTIVE: To investigate the osteoinduction of porous calcium phosphate cement on bone marrow mesenchymal stem cells.

METHODS: Bone marrow mesenchymal stem cells were isolated from rabbits aged 1 week in vitro and labeled by PKH-67 or PKH-26, respectively. Forty-eight adult New Zealand white rabbits were randomized into four groups and porous calcium phosphate cement was implanted into both sides of gluteus maximus in each rabbit. Then, 1 mL PKH-26-labeled bone barrow mesenchymal stem cells (1×10¹⁰/L) were injected into the superior gluteal artery branch at each side of gluteus maximus near the femur, and 1 mL PKH-67-labeled bone barrow mesenchymal stem cells (1×10¹⁰/L) injected into tissues around the cement (group A); 1 mL PKH-26-labeled bone barrow mesenchymal stem cells (1×10¹⁰/L) were injected into the each side of superior gluteal artery branch (group B); 1 mL PKH-67-labeled bone barrow mesenchymal stem cells (1×10¹⁰/L) were injected into tissues around the cement (group C); the same amount of normal saline was injected into tissues around the cement (group D). At 3, 7 and 12 weeks after implantation, the cement and its surrounding tissues were extracted and detected by fluorescence microscope and Massion staining. Expression of bone morphogenetic protein 2 was analyzed by RT-PCR.

RESULTS AND CONCLUSION: Under fluorescence microscope, PKH-26-labeled bone barrow mesenchymal stem cells attached fast and distributed homogeneously; however, PKH-67-labeled bone barrow mesenchymal stem cells attached slowly and exhibited a gradually homogeneous distribution. Massion staining showed that ectopic new bone formation appeared to have an upward trend in all groups, and the area of new bones in groups A, B and C were larger than that of group D at different time points after implantation. There was a significantly higher expression of bone morphogenetic protein 2 in groups A, B and C compared with the group D at different time points after implantation (P < 0.05), and the expression was the highest in the group A (P < 0.05). In conclusion, the porous calcium phosphate cement can induce bone barrow mesenchymal stem cells chemotaxis and osteogenetic differentiation. Besides, osteoblasts are differentiated from both the surrounding capillaries and body fluid, and capillary-derived mesenchymal stem cells occupy the important position.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Calcium Phosphates, Stem Cells, Tissue Engineering

CLC Number:

R318

Mao Xue-qing, Li Meng, Li Shao-feng, Liu Xing-yan. Porous calcium phosphate cement induces osteogenesis of mesenchymal stem cells under microenvironment[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(25): 3665-3672.

Figures/Tables 5

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