Zhang Guangping, Na Zhenwen, Ren Bin, Zhao Xin, Liu Weixian
Department of Oral and Maxillofacial Surgery, Shengjing Hospital of China Medical University Shengyang 110004, Liaoning Province, China.
Department of Prosthetics, School of Stomatology, China Medical University Shenyang 110002, Liaoning Province, China.
Int J Clin Exp Med. 2015 Aug 15;8(8):12172-81. eCollection 2015.
The aim of this study was to investigate the impacts of fluorescent superparamagnetic iron oxide particles (Molday ION Rhodamine B, MIRB) on bioactivities and osteogenetic differentiation of rat bone marrow mesenchymal stem cells (BMSCs). The Cell Counting Kit-8 (CCK-8) method was used to detect the proliferation of superparamagnetic iron oxide (SPIO)-labeled BMSCs and observed the distribution of MIRB in cells; real time -polymerase chain reaction (RT-PCR) method was used to analyze the expressions of such osteogenesis-related genes as bone sialoprotein, alkaline phosphatase (ALP), RUNX2, bonemorphogeneticprotein-2 (BMP-2), type 1 collagen (COL-1) and type 3 collagen (COL-3); ALP-Alizarin red staining and poly-biochemical analyzer were used to qualitatively and quantitatively analyze the osteogenetic metabolites. The labeled MIRB particles distributed in the cytoplasm of BMSCs, the diameter of larger particles could be up to several hundred nanometers, and concentrated around the nuclei, the particles far away from the nuclei were smaller, but the labeled-cells' skeletons and adherent morphology did not change significantly; under the concentration of 25 μg Fe/mL of, MIRB did not affect cellular viabilities of BMSCs, but the gene expressions of bone sialoprotein, ALP, RUNX2 and BMP-2 were decreased, and the secretion amount of ALP and osteocalcin were also declined. MIRB would not affect the proliferation and cell structures of BMSCs, but the SPIO particles aggregated and formed larger granules around the nuclei, which might affect the osteogenesis of BMSCs.
本研究旨在探讨荧光超顺磁性氧化铁颗粒(莫尔戴离子罗丹明B,MIRB)对大鼠骨髓间充质干细胞(BMSCs)生物活性和成骨分化的影响。采用细胞计数试剂盒-8(CCK-8)法检测超顺磁性氧化铁(SPIO)标记的BMSCs的增殖情况,并观察MIRB在细胞内的分布;采用实时聚合酶链反应(RT-PCR)法分析骨唾液蛋白、碱性磷酸酶(ALP)、RUNX2、骨形态发生蛋白-2(BMP-2)、Ⅰ型胶原(COL-1)和Ⅲ型胶原(COL-3)等成骨相关基因的表达;采用ALP-茜素红染色和多功能生化分析仪对成骨代谢产物进行定性和定量分析。标记的MIRB颗粒分布于BMSCs的细胞质中,较大颗粒的直径可达数百纳米,并聚集在细胞核周围,远离细胞核的颗粒较小,但标记细胞的骨架和贴壁形态无明显变化;在25μg Fe/mL的浓度下,MIRB不影响BMSCs的细胞活力,但骨唾液蛋白、ALP、RUNX2和BMP-2的基因表达降低,ALP和骨钙素的分泌量也下降。MIRB不会影响BMSCs的增殖和细胞结构,但SPIO颗粒在细胞核周围聚集形成较大颗粒,可能会影响BMSCs的成骨作用。