Nagahata Misao, Tsuchiya Toshie, Ishiguro Tatsuya, Matsuda Naoki, Nakatsuchi Yukio, Teramoto Akira, Hachimori Akira, Abe Koji
Division of Medical Devices, National Institute of Health Sciences, Kamiyoga 158-8501, Japan.
Biochem Biophys Res Commun. 2004 Mar 12;315(3):603-11. doi: 10.1016/j.bbrc.2004.01.098.
In this study, we examined the interaction of the osteoblast which forms bone and sulfated hyaluronan (SHya). For the purpose of the creation of a new functional polysaccharide, we introduced a sulfate group in hyaluronan (Hya) of high molecular weight, and SHya of high molecular weight could be obtained for the first time. When rat calvarial osteoblast (rOB) cells were cultured with a high concentration of SHya, they formed aggregated spheroids after 4h and the spheroids grew to about 200microm after 24h. We examined the expression of cell adhesion molecules in order to clarify the mechanism of aggregate formation. The N-cadherin (N-cad) and Connexin43 (Cx43) expression level of rOB cells cultured with SHya remarkably increased after 2h. A difference in the expression of Integrin beta1 (Intbeta1) could not be observed between the SHya addition and control group. The alkaline phosphatase (ALPase) activity of rOB cells cultured with SHya after 8h was significantly enhanced in comparison with control. Therefore, the sulfate group of SHya seems to enhance expression of cell adhesion protein such as N-cad and Cx43, resulting in aggregate formation and further remarkable induction of the ALPase activity of rOB cells.
在本研究中，我们检测了形成骨的成骨细胞与硫酸化透明质酸（SHya）之间的相互作用。为了制备一种新的功能性多糖，我们在高分子量透明质酸（Hya）中引入了硫酸基团，首次获得了高分子量的SHya。当大鼠颅骨成骨细胞（rOB）与高浓度的SHya一起培养时，4小时后它们形成聚集的球体，24小时后球体生长到约200微米。我们检测了细胞粘附分子的表达，以阐明聚集形成的机制。用SHya培养的rOB细胞在2小时后N-钙粘蛋白（N-cad）和连接蛋白43（Cx43）的表达水平显著增加。在添加SHya的组和对照组之间未观察到整合素β1（Intbeta1）表达的差异。与对照组相比，用SHya培养8小时后的rOB细胞的碱性磷酸酶（ALPase）活性显著增强。因此，SHya的硫酸基团似乎增强了N-cad和Cx43等细胞粘附蛋白的表达，导致聚集形成，并进一步显著诱导rOB细胞的ALPase活性。
