Ozono Satoru, Fujita Tadahiro, Matsuo Masato, Todoki Kazuo, Ohtomo Takatsune, Negishi Hideyuki, Kawase Toshio
Institute for Frontier Oral Science, Kanagawa Dental College, Yokosuka, Kanagawa, Japan.
Nihon Hotetsu Shika Gakkai Zasshi. 2008 Jul;52(3):366-74. doi: 10.2186/jjps.52.366.
Bone marrow stromal cells (BMSCs) are a promising cell source in applications for tissue engineering and regenerative medicine. Optimization and control of the growth and differentiation of cultivated cells can be achieved by the administration of growth factors and hormones in vitro. This study provided experimental information on the enhancement of the osteogenic potential of rat BMSCs in vitro and in vivo.
Mineralized nodule formation of rat BMSCs in culture for 3 weeks with dexamethasone (Dex)-treated media supplemented with both basic fibroblast growth factor (bFGF) and 17beta -estradiol (E2) was examined by histology. In porous beta-tricalcium phosphate (beta - TCP), proliferation, migration, and differentiation of BMSCs were examined by histology and transmission electron microscopy. After culturing, the composites were subcutaneously implanted into syngeneic rats. The tissues with implants were harvested after 4 weeks and evaluated microscopically by using histological stain.
Dex-treated media supplemented with both bFGF and E2 was the most effective in mineralized nodule formation of BMSCs in vitro. Light and electron microscopy revealed the presence of many cells with developed rough endoplasmic reticulum. Bone formation in the BMSC/beta -TCP composites in cultures in vitro for 3 weeks was observed histologically at 4 weeks after implantation. When BMSC/beta -TCP composites were cultured in Dex-treated media supplemented with both bFGF and E2, the amount of bone formation at implants was substantially greater than that of composites cultured in Dex-treated media supplemented with bFGF.
The combined use of bFGF and E2 could effectively improve the bone-forming ability of BMSCs.
骨髓基质细胞（BMSCs）是组织工程和再生医学应用中一种很有前景的细胞来源。通过在体外施用生长因子和激素，可以实现对培养细胞生长和分化的优化与控制。本研究提供了关于增强大鼠BMSCs体外和体内成骨潜力的实验信息。
通过组织学检查用补充了碱性成纤维细胞生长因子（bFGF）和17β-雌二醇（E2）的地塞米松（Dex）处理的培养基培养3周的大鼠BMSCs的矿化结节形成情况。在多孔β-磷酸三钙（β-TCP）中，通过组织学和透射电子显微镜检查BMSCs的增殖、迁移和分化情况。培养后，将复合材料皮下植入同基因大鼠体内。4周后收获植入物的组织，并使用组织学染色进行显微镜评估。
补充了bFGF和E2的Dex处理培养基在体外对BMSCs的矿化结节形成最有效。光镜和电镜显示存在许多粗面内质网发达的细胞。植入后4周，组织学观察到体外培养3周的BMSC/β-TCP复合材料中有骨形成。当BMSC/β-TCP复合材料在补充了bFGF和E2的Dex处理培养基中培养时，植入物处的骨形成量明显大于在补充了bFGF的Dex处理培养基中培养的复合材料。
bFGF和E2联合使用可有效提高BMSCs的成骨能力。
