Koh E T, Torabinejad M, Pitt Ford T R, Brady K, McDonald F
Department of Conservative Dentistry, UMDS, London, England.
J Biomed Mater Res. 1997 Dec 5;37(3):432-9. doi: 10.1002/(sici)1097-4636(19971205)37:3<432::aid-jbm14>3.0.co;2-d.
We report a novel material that appears to stimulate cytokine production in human osteoblasts and allow good adherence of the cells to the material. We have examined cultured osteoblasts (MG-63) in the presence of mineral trioxide aggregate (MTA) as set in moist conditions; secondly, we examined the behavior of these MG-63 cells with respect to cytokine and osteocalcin production and alkaline phosphatase activity. Standard ELISA assays were used for assessment of interleukin (IL)-1 alpha, IL-1 beta, IL-6, macrophage colony stimulating factor (M-CSF), and osteocalcin. Furthermore the levels of alkaline phosphatase were measured to establish the level of differentiation of the cells. Cells without MTA served as controls. Cells also were grown in the presence of polymethylmethacrylate (PMA), the commonly used orthopedic cement. In all dishes cells were seen adhering to the base and MTA at 6 h and had increased to confluence at 144 h. IL-1 alpha (175.1 +/- 32.6 pg/mL), IL-1 beta (154.0 +/- 26.7 pg/mL), and IL-6 (214.7 +/- 21.8 pg/mL) were raised when the cells were grown in the presence of MTA at 144 h, with raised values at all time intervals. M-CSF appeared to be unaffected although the overall value was high (7,045.0 +/- 89.5 pg/mL). In contrast, cells grown in the absence of MTA produced negligible amounts of these cytokines (< pg/mL) as did those cells grown in the presence of PMA. Osteocalcin production increased when cells were grown on MTA from 3.8 +/- 0.87 ng/mL to 19.7 +/- 2.8 ng/mL. No osteocalcin could be detected with PMA. Cells in contact with MTA also appeared to have levels of alkaline phosphatase similar to those reported elsewhere (4.3 +/- 0.21 mumol/mg protein/min). No cells could be found attached to PMA and so no alkaline phosphatase activity could be measured.
我们报告了一种新型材料，该材料似乎能刺激人成骨细胞产生细胞因子，并使细胞能良好地附着于该材料。我们研究了在潮湿条件下培养的成骨细胞（MG - 63）与三氧化矿物凝聚体（MTA）共同存在时的情况；其次，我们研究了这些MG - 63细胞在细胞因子、骨钙素产生及碱性磷酸酶活性方面的行为。采用标准酶联免疫吸附测定（ELISA）法评估白细胞介素（IL）-1α、IL -1β、IL -6、巨噬细胞集落刺激因子（M - CSF）和骨钙素。此外，测量碱性磷酸酶水平以确定细胞的分化程度。未添加MTA的细胞作为对照。细胞也在常用的骨科骨水泥聚甲基丙烯酸甲酯（PMA）存在的情况下生长。在所有培养皿中，细胞在6小时时可见附着于培养皿底部和MTA，在144小时时增殖至汇合。当细胞在144小时与MTA共同培养时，IL -1α（175.1±32.6 pg/mL）、IL -1β（154.0±26.7 pg/mL）和IL -6（214.7±21.8 pg/mL）水平升高，且在所有时间间隔内均升高。尽管总体值较高（7,045.0±89.5 pg/mL），但M - CSF似乎未受影响。相比之下，未添加MTA培养的细胞以及在PMA存在下培养的细胞产生的这些细胞因子量可忽略不计（< pg/mL）。当细胞在MTA上生长时，骨钙素产量从3.8±0.87 ng/mL增加至19.7±2.8 ng/mL。在PMA存在下未检测到骨钙素。与MTA接触的细胞碱性磷酸酶水平似乎与其他地方报道的水平相似（4.3±0.21 μmol/mg蛋白质/分钟）。未发现细胞附着于PMA，因此无法测量碱性磷酸酶活性。
