The experimental study of constructing tissue engineered bone by compounding zinc-sintered bovine cancellous bone with Marrow stromal cells

Abstract

To study the osteogenic ability of tissue-engineered bone constructed by compounding zinc-sintered bovine cancellous bone with rabbit marrow stromal cells (MSCs) in vivo, the zinc-sintered bovine cancellous bone of beta-tricalcium phosphate (TCP) type was prepared by sintering the fresh calf cancellous bone twice and then loading it with zinc-ion. The rabbit MSCs were cultured, induced and seeded onto the zinc-sintered bovine cancellous bones. The tissue-engineered bones were then implanted into the rabbits' back muscles. The newly formed bone tissues were observed by histological methods and the areas of new osseous tissues were measured at the end of the 4th and 8th week. The zinc-sintered bovine cancellous bones alone were implanted on the other side as control. The osteogenic activity of MSCs was identified by alkaline phosphatase (ALP) staining and calcification nod chinalizarin staining. At the end of 4th week, a small amount of new bone tissues was observed. At the end of 8th week, there were many newly formed bone mature tissues. Moreover, the area of the latter was significantly larger than that of the former (P<0.01), while in the control group there was no new bone formation. The tissue-engineered bone, which was constructed by combining zinc-sintered bovine cancellous bone with MSCs, has satisfactory osteogenic capabilities in vivo.

Keywords

sintered bone(SB) / zinc-ion / marrow stromal cells / tissue engineering

References

[1] Liu S N, Zheng Q X, Zhou Y S. Rabbit's Radius Segmental Bone Defect Repaired with Zinc-sintered Bovine Cancellous Bone. Chin J Exp Surg, 2003, 20(4): 339-340. (in Chinese)

[2] De Groot K. Bioceramics of Calcium Phosphate, 1983 Boca Raton, FL: CRC Press.

[3] Honglian Dai, Xianying Cao, Xiaoxi Li, . The Transformation of Calcium Phosphate Bioceramicsin vivo. Journal of Wuhan University of Technology-Mater. Sci. Ed., 2003, 18(2): 19-22.

[4] Coleman J E. Zinc Protein: Enzymes, Storage Proteins, Transcription Factors, and Replication Proteins. Annu Rev Biomed, 1992, 61: 897-946.

[5] Hashizume M, Yamaguchi M. Stimulatory Effect of β-alanyl-l-histidinato Zinc on Cell Proliferation is Dependent on Protein Synthesis in Osteoblastic MC3T3-E1 Cells. Mol Cell Biochem, 1993, 122: 59-64.

[6] Schroeder LW, Dickens B, Brown WE. Crystallographic Studies of the Role of Mg as a Stabilizing Impurity in β-Ca₃(PO₄)₂. II. Refinement of Mg-containing β-Ca₃ (PO₄)₂. J. Solid State Chem., 1977, 22: 253-262.

[7] Maniatopoulos C, Sodek J, Melcher A H. Bone Formation in vitro by Stromal Cells Obtained Bone Marrow of Young Adult Rats. Cell Tissue Res., 1998, 254(3): 317-317.

[8] Gu Z C, Li Q H, Zhao Y F, . Osteogenic Property of Cultured Rabbit Marrow Stromal Cells under Condition of Bone Induction. Acta Academiae Medicinae Militaris Tertiae, 2002, 24(5): 527-530. (in Chinese)

[9] Kadiyala S, Young R G, Thiede M A. Culture Expanded Canine Mesenchymal Stem Cells Possess Osteochondrogenic Potential in vivo andin vitro. Cell Transplant, 1997, 6(2): 125-134.