Mouriès Lucilia Pereira, Almeida Maria-José, Milet Christian, Berland Sophie, Lopez Evelyne
Laboratoire de Physiologie Générale et Comparée, Muséum National d'Histoire Naturelle, Paris, France.
Comp Biochem Physiol B Biochem Mol Biol. 2002 May;132(1):217-29. doi: 10.1016/s1096-4959(01)00524-3.
In vivo and in vitro studies provide strong evidence of the osteogenic activity of nacre obtained from Pinctada maxima. The in vitro studies indicate that diffusible factors from nacre are involved in cell stimulation. The water-soluble matrix (WSM) was extracted from nacre by a non-decalcifying process, and four fractions (SE(1)-SE(4)) were separated by SE-HPLC. Those fractions were tested in vitro on MRC5 fibroblasts. Alkaline phosphatase (ALP) activity was measured as a marker of osteoblastic differentiation. The anti-apoptotic protein Bcl-2 was also immunodetected in cultured osteoblasts from rat calvaria. WSM and fraction SE(4) increased ALP activity. BMP-2 had the same effect on the cells as WSM and SE(4). WSM greatly increased the amount of Bcl-2 in the cytoplasm and nucleus of osteoblasts. These in vitro studies support our initial hypothesis that nacre organic matrix (WSM) of a bivalve mollusk contains signal-molecules that can stimulate the osteogenic pathway in mammalian cells that are targets for bone induction.
体内和体外研究均为取自大珠母贝的珍珠层的成骨活性提供了有力证据。体外研究表明，珍珠层中的可扩散因子参与细胞刺激。通过非脱钙过程从珍珠层中提取水溶性基质（WSM），并通过尺寸排阻高效液相色谱法（SE-HPLC）分离出四个组分（SE(1)-SE(4)）。在体外对MRC5成纤维细胞测试这些组分。测量碱性磷酸酶（ALP）活性作为成骨细胞分化的标志物。还在来自大鼠颅骨的培养成骨细胞中免疫检测抗凋亡蛋白Bcl-2。WSM和组分SE(4)增加了ALP活性。骨形态发生蛋白-2（BMP-2）对细胞的作用与WSM和SE(4)相同。WSM极大地增加了成骨细胞细胞质和细胞核中Bcl-2的量。这些体外研究支持了我们最初的假设，即双壳贝类的珍珠层有机基质（WSM）含有能够刺激作为骨诱导靶点的哺乳动物细胞中的成骨途径的信号分子。
