Chen James, Zhang Yan, Mendoza Joseph, Denbesten Pamela
Department of Orofacial Sciences, University of California, San Francisco, CA 94143, USA.
J Exp Zool B Mol Dev Evol. 2009 Jul 15;312B(5):458-64. doi: 10.1002/jez.b.21279.
Calcium is a key component of the mineralized enamel matrix, but may also have a role in ameloblast cell differentiation. In this study we used human ameloblast lineage cells to determine the effect of calcium on cell function. Primary human ameloblast lineage cells were isolated from human fetal tooth buds. Cells were treated with calcium ranging from 0.05 to -1.8 mM. Cell morphology was imaged by phase contrast microscopy, and amelogenin was immunolocalized. Proliferation of cells treated with calcium was measured by BrdU immunoassay. The effect of calcium on mRNA expression of amelogenin, Type 1 collagen, DSPP, amelotin, and KLK-4 was compared by PCR analysis. Von Kossa staining was used to detect mineral formation after cells were pretreated with calcium. Calcium induced cell organization and clustering at 0.1 and 0.3 mM concentrations. Increasing concentrations of calcium significantly reduced ameloblast lineage cell proliferation. The addition of 0.1 mM calcium to the cultures upregulated expression of amelogenin, Type I collagen, and amelotin. After pretreatment with 0.3 mM calcium, the cells could form a mineralized matrix. These studies, which utilized human ameloblast lineage cells grown in vitro, showed that the addition of calcium at 0.1 and 0.3 mM, induced cell differentiation and upregulation of amelogenin Type I collagen and amelotin.
钙是矿化釉质基质的关键成分，但在成釉细胞分化中也可能发挥作用。在本研究中，我们使用人成釉细胞系细胞来确定钙对细胞功能的影响。从人胎儿牙胚中分离出原代人成釉细胞系细胞。用0.05至1.8 mM的钙处理细胞。通过相差显微镜对细胞形态进行成像，并对釉原蛋白进行免疫定位。用BrdU免疫测定法测量经钙处理的细胞的增殖。通过PCR分析比较钙对釉原蛋白、I型胶原、DSPP、釉蛋白和KLK - 4 mRNA表达的影响。在用钙预处理细胞后，使用冯·科萨染色法检测矿化形成。钙在0.1和0.3 mM浓度下诱导细胞组织化和聚集。钙浓度增加显著降低成釉细胞系细胞的增殖。向培养物中添加0.1 mM钙会上调釉原蛋白、I型胶原和釉蛋白的表达。用0.3 mM钙预处理后，细胞能够形成矿化基质。这些利用体外培养的人成釉细胞系细胞进行的研究表明，添加0.1和0.3 mM的钙可诱导细胞分化以及上调釉原蛋白、I型胶原和釉蛋白的表达。
