无机活性元素支架材料对颌骨缺损骨创面骨祖细胞的生物诱导作用

Bio-inductive Effects of the Inorganic Elements Scaffold Material on Osteogenitor Cell near the Postoperative Jaw Defects

目的:应用颌骨术后缺损骨创面周围骨松质分离培养鉴定成骨细胞,并将其与无机活性元素支架材料在体外复合生长,探讨无机活性元素支架材料对骨祖细胞来源的成骨细胞生物学特性的生物诱导作用。方法:应用组织块贴壁法分离培养出成骨细胞,并用碱性磷酸酶(Burstone偶氮偶联法)染色法、茜素红染色法、透射电镜、流式细胞仪等对其碱性磷酸酶活性、钙结节形成情况、细胞的超微结构、细胞生长周期进行鉴定,然后将体外培养的成骨细胞与无机活性元素支架材料体外复合生长,对复合体进行形态学、扫描电镜、细胞增殖能力、细胞周期变化的测定,评价细胞与材料的相容性。结果:颌骨术后缺损骨创面周围骨松质分离出的成骨细胞具有较强的碱性磷酸酶活性且有大量的钙结节形成,透射电镜观察其具有良好的分泌细胞特性,此成骨细胞与无机活性元素支架材料体外复合培养14 d,分布于支架材料的成骨细胞增殖良好,分泌细胞外基质并形成钙结节,细胞周期未见明显改变。结论:颌骨术后缺损骨创面周围骨松质中含有大量的骨祖细胞,具有较强的向成骨细胞分化和繁殖能力。无机活性元素支架材料均具有较好的细胞相容性,与成骨细胞共同培养有望获得具有三维立体结构的组织工程骨。

Objective： To study the bio-inductive effects of inorganic elements on osteogenitor cell near the postoperative jaw defects.Methods： We isolate and culture osteoblasts from cancellous bone near the postoperative jaw defects and co-combine with cultured inorganic elements in vitro.Alkaline phosphatase（AKP） activity was determined by azo dye method,calcification nodule formed was observed by alizarin red S,the cell ultrastructure was identified by electron microscopy and the growth cycle was evaluated with flow cytometer.Then inorganic elements co-combined with cultured osteoblasts in vitro.The morphological cytoskeleton characters,cell proliferation,mitotic cycle and DNA index were detected.Results： We found that alkaline phosphatase activity and alizarin red S was strongly positive.These cells possess good secreting characteristics.The cell cycle and physiological characteristics did not significantly change after co-cultured 14 days.Conclusion： Cancellous bone near the postoperative jaw defects contain a lot of osteogenitor cells which have osteogenic potential.The inorganic elements scaffold material has good cellular compatibility.It is possible to achieve the tissue engineering bone which possesses three-dimensional stereoscopic structure by co-culturing them.