珍珠质自然涂层钛种植体表面对MC3T3E1成骨样细胞的作用

Effects of Titanium Implant Surface Coated With Natural Nacre on MC3T3E1 Cell Line In vitro

为了研究珍珠质自然涂层钛种植体表面的体外生物相容性,将珍珠质自然涂层的钛片与MC3T3E1成骨样细胞复合培养以观察细胞的生长、增殖和分化.分别以羟基磷灰石涂层钛片和没有涂层的纯钛片作为对照组,以MC3T3E1细胞单纯培养作为空白组,分别培养3天,5天和7天,通过倒置相差显微镜和扫描电镜观察细胞生长情况,流式细胞技术检测细胞增殖活性,金氏比色法检测碱性磷酸酶(ALP)活性以及蛋白质印迹(Western blotting)法测定转化生长因子-β1(TGF-β1)表达水平.结果发现,细胞在珍珠质周围能形成良好附着,在其表面生长丰满.细胞培养第3天,第5天和第7天时,珍珠质表面的细胞增殖指数分别为(35.9±2.5)%、(69.7±3.3)%和(58.2±2.6)%,ALP活性分别为(6.123±2.917)U/g、(17.486±1.986)U/g和(23.987±1.372)U/g.第5天和7天时,实验组的细胞增殖指数、ALP活性和TGF-β1表达水平显著高于对照组和空白组(P<0.05).珍珠质自然涂层钛表面有利于MC3T3E1细胞的生长、增殖和分化,表明了珍珠质涂层能提高种植体表面的生物相容性,有可能会促进种植后的骨整合.

To study the biocompatibility of the titanium dental implant surfaces coated with nacre in vitro, osteoblast-like cells （MC3T3E1） growth on the substrates coated with nacre were compared with that growth on the substrates coated with hydroxyapatite （HA） or nothing and the cells of the blank. MC3T3E1 were incubated on the respective surface for 3days, 5days and 7 days. Cell morphology was estimated by inverted phase contrast microscope and scanning electron microscope （SEM） and cell proliferation was measured by flowcytometry. The King＇s chromometry was used for measuring the activity of ALP in the cell lysate and the expression of TGF-β1 was detected by Western blotting. The results showed that MC3T3E1 cultured on the nacre-coated surface were more spreaded and plumper than the controls on the 3days, 5days and 7days of culture. Meanwhile, the test group showed a superiority to control groups and blank with respects to the cell proliferation index[（35.9±2.5）%, （69.7±3.3）%, （58.2±2.6）%], ALP activity [（6.123±2.917）, （17.486±1.986）, （23.987±1.372） U/g] and TGF-β1 level. In conclusion, the surface coated nacre can promote the proliferation and differentiation of osteoblast, which the mechanism partly may be the increasing expression of TGF-β1.