Wu Tao, Cheng Ning, Xu Chun, Sun Wei, Yu Chengzhong, Shi Bin
The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, 430079, People's Republic of China.
School of Dentistry, Oral Biology Curriculum, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, 27599.
J Biomed Mater Res A. 2016 Dec;104(12):3004-3014. doi: 10.1002/jbm.a.35841. Epub 2016 Aug 5.
This study evaluated the effect of mesoporous bioglass (MBG) dissolution on the differentiation of bone marrow mesenchymal stem cells (BMSCs) derived from either sham control or ovariectomized (OVX) rats. MBG was fabricated by evaporation-induced self-assembly method. Cell proliferation was tested by Cell Counting Kit-8 assay, and cytoskeletal morphology was observed by fluorescence microscopy. Osteogenic differentiation was evaluated by alkaline phosphatase (ALP) staining and activity, Alizarin Red staining, while adipogenic differentiation was assessed by Oil Red-O staining. Quantitative real-time PCR and Western blot analysis were taken to evaluate the expression of runt-related transcription factor 2 (Runx2) and proliferator-activated receptor-γ (PPARγ). We found that MBG dissolution (0, 25, 50, 100, 200 µg/mL) was nontoxic to BMSCs growth. Sham and OVX BMSCs exhibited the highest ALP activity in 50 µg/mL of MBG osteogenic dissolution, except that sham BMSCs in 100 µg/mL showed the highest ALP activity on day 14. Runx2 was significantly upregulated after 100 µg/mL of MBG stimulation in sham and OVX BMSCs for 7 and 14 days, except that 25 µg/mL showed highest upregulation effect on OVX BMSCs at day 7. PPARγ was downregulated after MBG stimulation. The protein level of Runx2 from the sham BMSCs group was significantly upregulated after lower doses (25 and 50 µg/mL) of MBG stimulation, whereas PPARγ was downregulated in the sham and OVX BMSCs group. Thus, both the osteogenic and adipogenic abilities of BMSCs were damaged under OVX condition. Moreover, lower concentration of MBG dissolution can promote osteogenesis but inhibit adipogenesis of the sham and OVX BMSCs. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3004-3014, 2016.
本研究评估了介孔生物玻璃(MBG)溶解对源自假手术对照或去卵巢(OVX)大鼠的骨髓间充质干细胞(BMSC)分化的影响。MBG采用蒸发诱导自组装法制备。通过细胞计数试剂盒-8法检测细胞增殖,并通过荧光显微镜观察细胞骨架形态。通过碱性磷酸酶(ALP)染色及活性、茜素红染色评估成骨分化,通过油红O染色评估成脂分化。采用定量实时PCR和蛋白质印迹分析评估 runt相关转录因子2(Runx2)和增殖激活受体γ(PPARγ)的表达。我们发现MBG溶解物(0、25、50、100、200μg/mL)对BMSC生长无毒。假手术组和OVX组BMSC在50μg/mL的MBG成骨溶解物中表现出最高的ALP活性,但假手术组BMSC在第14天100μg/mL时显示出最高的ALP活性。在假手术组和OVX组BMSC中,100μg/mL的MBG刺激7天和14天后,Runx2显著上调,但在第7天,25μg/mL对OVX组BMSC显示出最高的上调作用。MBG刺激后PPARγ下调。假手术组BMSC中较低剂量(25和50μg/mL)的MBG刺激后,Runx2的蛋白水平显著上调,而在假手术组和OVX组BMSC中PPARγ下调。因此,在OVX条件下,BMSC的成骨和成脂能力均受损。此外,较低浓度的MBG溶解物可促进假手术组和OVX组BMSC的成骨作用,但抑制其成脂作用。©2016威利期刊公司。《生物医学材料研究杂志》A部分:104A:3004 - 3014,2016年。