Noshi T, Yoshikawa T, Ikeuchi M, Dohi Y, Ohgushi H, Horiuchi K, Sugimura M, Ichijima K, Yonemasu K
Departments of Oral and Maxillofacial Surgery, Nara Medical University, Kashihara City, Nara 634-8522, Japan.
J Biomed Mater Res. 2000 Dec 15;52(4):621-30. doi: 10.1002/1097-4636(20001215)52:4<621::aid-jbm6>3.0.co;2-a.
A composite of marrow mesenchymal stem cells and porous hydroxyapatite (HA) has in vivo osteogenic potential. To investigate factors enhancing the osteogenic potential of marrow/HA composites, we prepared a bone morphogenetic protein (BMP) fraction from the 4M guanidine extract of bovine bone by heparin-sepharose affinity chromatography. Marrow/HA composites or composites containing marrow mesenchymal stem cells, BMP, and HA (marrow/BMP/HA composites) were implanted subcutaneously in 7-week-old male Fischer rats. BMP/HA composites and HA alone were also implanted. The implants were harvested after 2, 4, or 8 weeks and were prepared for histological and biochemical studies. Histological examination showed obvious de novo bone formation together with active osteoblasts at 2 weeks, as well as more extensive bone formation at 4 and 8 weeks in many pores of the marrow/BMP/HA composites. The marrow/HA composites did not induce bone formation at 2 weeks, but there was moderate bone formation at 4 weeks. At 2 weeks, only marrow/BMP/HA composites resulted in intensive osteogenic activity, judging from alkaline phosphatase and osteocalcin expression at both the protein and gene levels. These results indicate that the combination of marrow mesenchymal stem cells, porous HA, and BMP synergistically enhances osteogenic potential, and may provide a rational basis for their clinical application, although further in vivo experiment is needed.
骨髓间充质干细胞与多孔羟基磷灰石（HA）的复合物具有体内成骨潜力。为了研究增强骨髓/HA复合物成骨潜力的因素，我们通过肝素-琼脂糖亲和层析从牛骨的4M胍提取物中制备了一种骨形态发生蛋白（BMP）组分。将骨髓/HA复合物或含有骨髓间充质干细胞、BMP和HA的复合物（骨髓/BMP/HA复合物）皮下植入7周龄雄性Fischer大鼠体内。同时也植入了BMP/HA复合物和单独的HA。在2、4或8周后取出植入物，准备进行组织学和生化研究。组织学检查显示，在2周时，骨髓/BMP/HA复合物的许多孔隙中有明显的新生骨形成以及活跃的成骨细胞，在4周和8周时骨形成更为广泛。骨髓/HA复合物在2周时未诱导骨形成，但在4周时有中度骨形成。在2周时，从蛋白质和基因水平的碱性磷酸酶和骨钙素表达判断，只有骨髓/BMP/HA复合物产生了强烈的成骨活性。这些结果表明，骨髓间充质干细胞、多孔HA和BMP的组合协同增强了成骨潜力，尽管还需要进一步的体内实验，但可能为它们的临床应用提供合理依据。
