Ito Takafumi, Kaneko Tomoatsu, Yamanaka Yusuke, Shigetani Yoshimi, Yoshiba Kunihiko, Okiji Takashi
Division of Cariology, Operative Dentistry and Endodontics, Niigata University, Graduate School of Medical and Dental Sciences, Niigata, Japan.
Division of Cariology, Operative Dentistry and Endodontics, Niigata University, Graduate School of Medical and Dental Sciences, Niigata, Japan.
J Endod. 2014 Mar;40(3):379-83. doi: 10.1016/j.joen.2013.11.011. Epub 2013 Dec 19.
This study examined the protein and messenger RNA (mRNA) expression of molecules associated with M2 (wound healing) macrophages in mineral trioxide aggregate (MTA)-implanted rat subcutaneous tissue to elucidate the involvement of M2 macrophages in the connective tissue response to MTA.
Silicone tubes containing freshly mixed MTA or a calcium hydroxide cement (Life; Kerr, Romulus, MI) were subcutaneously implanted into the backs of Wistar rats. Solid silicone rods implanted in different animals served as controls. The specimens were then double immunostained for ED1 (CD68, a general macrophage marker) and ED2 (CD163, an M2 macrophage marker). Immunostaining for CD34 (a marker for vascularization and wound healing) was also performed. Expression levels of CD34, CD163, and mannose receptor c type 1 (an M2 macrophage marker) mRNAs were determined with real-time polymerase chain reaction.
MTA-implanted subcutaneous tissues showed significant increases in the density of ED1+ED2+ macrophages beneath the implantation site and expression levels of CD163 and MMR mRNAs compared with Life-implanted and control tissues. MTA-implanted subcutaneous tissues also showed a significant increase of CD34-immunostained areas and up-regulation of CD34 mRNAs compared with Life-implanted and control tissues.
MTA implantation induced the accumulation of M2 macrophage marker (ED2)-expressing macrophages and enhanced the expression of M2 macrophage marker genes. MTA implantation also enhanced the expression of CD34, suggesting acceleration of the healing/tissue repair process. Taken together, biological connective tissue response to MTA may involve wound healing/tissue repair processes involving M2 macrophages.
本研究检测了三氧化矿物凝聚体（MTA）植入大鼠皮下组织中与M2（伤口愈合）巨噬细胞相关分子的蛋白质和信使核糖核酸（mRNA）表达，以阐明M2巨噬细胞在结缔组织对MTA反应中的作用。
将装有新鲜混合MTA或氢氧化钙水门汀（Life；Kerr，Romulus，密歇根州）的硅胶管皮下植入Wistar大鼠背部。植入不同动物体内的实心硅胶棒作为对照。然后对标本进行ED1（CD68，一种通用巨噬细胞标志物）和ED2（CD163，一种M2巨噬细胞标志物）的双重免疫染色。还进行了CD34（血管生成和伤口愈合标志物）的免疫染色。用实时聚合酶链反应测定CD34、CD163和甘露糖受体c型1（一种M2巨噬细胞标志物）mRNA的表达水平。
与植入Life和对照组织相比，植入MTA的皮下组织在植入部位下方ED1+ED2+巨噬细胞密度以及CD163和MMR mRNA表达水平显著增加。与植入Life和对照组织相比，植入MTA的皮下组织CD34免疫染色区域也显著增加，且CD34 mRNA上调。
MTA植入诱导了表达M2巨噬细胞标志物（ED2）的巨噬细胞积累，并增强了M2巨噬细胞标志物基因的表达。MTA植入还增强了CD34的表达，提示愈合/组织修复过程加速。综上所述，结缔组织对MTA的生物学反应可能涉及M2巨噬细胞参与的伤口愈合/组织修复过程。
