Effects of enamel matrix derivative on proliferation and differentiation of primary osteoblasts
Effects of enamel matrix derivative on proliferation and differentiation of primary osteoblasts
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DOI：
10.1016/j.tripleo.2004.02.063
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发表时间：
2004-08-01
期刊：
影响因子：
--
通讯作者：
Zimmermann, B
中科院分区：
文献类型：
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作者：
Hägewald, S;Pischon, N;Zimmermann, B
Objective.本研究旨在探讨釉基质衍生物（EMD）对小鼠成骨细胞增殖、蛋白质合成和矿化的影响。通过酶消化从小鼠颅骨获得成骨细胞，并与EMD（2-100 μ g/ml）一起单层生长。四唑盐比色法（MTT）和5-溴-2 '-脱氧尿苷（BrdU）掺入法测定代谢活性和细胞增殖。对于分化研究，使用三维类器官培养系统。成骨细胞分化通过碱性磷酸酶（ALP）活性和钙含量来估计。胶原合成通过[H-3]-脯氨酸掺入来评估。电镜观察细胞形态。EMD处理增加代谢细胞活性和BrdU掺入。在类器官培养物中，EMD处理增强了ALP活性和钙积累，但[H-3]-脯氨酸掺入没有。形态学上，发现矿化结节沉积增加。EMD处理增强了原代成骨细胞的细胞活性，并可能支持牙周骨缺损的再生。
Objective. The aim of this study was to investigate the effects of enamel matrix derivative (EMD) on proliferation, protein synthesis, and mineralization in primary mouse osteoblasts.Study design. Osteoblasts were obtained from mouse calvaria by enzymatic digestion and grown in monolayer together with EMD (2-100 mug/ml). Metabolic activity and cell proliferation were determined by tetrazolium salt assay (MTT) and by 5-bromo-2'-deoxyuridine (BrdU) incorporation. For differentiation studies, a 3-dimensional organoid culture system was used. Osteoblastic differentiation was estimated by alkaline phosphatase (ALP) activity and calcium content. Collagen synthesis was assessed by [H-3]-proline incorporation. Morphologic observations were made by electron microscopy.Results. EMD treatments increased metabolic cell activity and BrdU incorporation. In the organoid cultures, ALP activity and calcium accumulation were enhanced by EMD treatment, but [H-3]-proline incorporation was not. Morphologically, an increased deposition of mineralized nodules was found.Conclusions. EMD treatment enhanced cellular activities of primary osteoblasts and might support the regeneration of periodontal bony defects.