骨髓间充质干细胞来源的外泌体对成骨细胞增殖和分化的影响
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摘要
目的:探讨骨髓间充质干细胞来源的外泌体对成骨细胞增殖和分化的影响。方法:体外分离培养骨髓间充质干细胞,超速离心法收集外泌体,并对成骨细胞进行刺激,细胞计数试剂盒(cell counting kit-8,CCK-8)法检测骨髓间充质干细胞来源的外泌体对成骨细胞增殖的影响,碱性磷酸酶活性检测及茜素红染色观察骨髓间充质干细胞来源的外泌体对成骨细胞矿化的影响,利用实时荧光定量聚合酶链反应(real-time polymerase chain reaction,Real-time PCR)检测成骨相关基因的表达。结果:骨髓间充质干细胞来源的外泌体促进成骨细胞的增殖,上调成骨相关基因的表达,提升碱性磷酸酶活性及促进钙化结节的形成。结论:骨髓间充质干细胞来源的外泌体能够促进成骨细胞的增殖和分化。
Objective: To evaluate the effects of bone marrow mesenchymal stem cells(BMSCs) derived exosomes on the proliferation and differentiation of osteoblasts. Methods: BMSCs were cultured in vitro and exsomes were isolated by ultracentrifugation from supernatants. After treated with BMSCs derived exosomes, the proliferation of osteoblasts was detected by CCK-8 assay. Alkaline phosphatase activity(ALP) analysis and alizarin red staining were adopted to evaluate the effect of BMSCs derived exosomes on mineralization process of osteoblasts. Real-time PCR was utilized to detect the expression of odontogenesis-related genes. Results: BMSCs derived exosomes enhanced the proliferation of osteoblasts and upregulated the expression levels of odontogenesis-related genes. The ALP activity and calcium accumulation of osteoblasts were increased significantly as well. Conclusion: BMSCs derived exosomes could promote the proliferation and differentiation of osteoblasts.
Objective: To evaluate the effects of bone marrow mesenchymal stem cells(BMSCs) derived exosomes on the proliferation and differentiation of osteoblasts. Methods: BMSCs were cultured in vitro and exsomes were isolated by ultracentrifugation from supernatants. After treated with BMSCs derived exosomes, the proliferation of osteoblasts was detected by CCK-8 assay. Alkaline phosphatase activity(ALP) analysis and alizarin red staining were adopted to evaluate the effect of BMSCs derived exosomes on mineralization process of osteoblasts. Real-time PCR was utilized to detect the expression of odontogenesis-related genes. Results: BMSCs derived exosomes enhanced the proliferation of osteoblasts and upregulated the expression levels of odontogenesis-related genes. The ALP activity and calcium accumulation of osteoblasts were increased significantly as well. Conclusion: BMSCs derived exosomes could promote the proliferation and differentiation of osteoblasts.
引文
[1] 李成,周海斌.骨髓间充质干细胞的旁分泌能影响成骨细胞生物学功能[J].中国组织工程研究, 2014, 18(10): 1477-1483.
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[9] Lauzon MA, Drevelle O, Daviau A, et al. Effects of BMP-9 and BMP-2 on the PI3K/Akt pathway in MC3T3-E1 preosteoblasts [J]. Tissue Eng Part A, 2016, 22(17-18): 1075-1085.
[10] Yang D, Okamura H, Nakashima Y, et al. Histone demethylase Jmjd3 regulates osteoblast differentiation via transcription factors Runx2 and osterix [J]. J Biol Chem, 2013, 288(47): 33530-33541.
[11] Tsao YT, Huang YJ, Wu HH, et al. Osteocalcin mediates biomineralization during osteogenic maturation in human mesenchymal stromal cells [J]. Int J Mol Sci, 2017, 18(1). pii: E159.
[2] Yu X, Odenthal M, Fries JW. Exosomes as miRNA carriers: formation-function-future [J]. Int J Mol Sci, 2016, 17(12): 2028.
[3] Xie Y, Chen Y, Zhang L, et al. The roles of bone-derived exosomes and exosomal microRNAs in regulating bone remodelling [J]. J Cell Mol Med, 2017, 21(5): 1033-1041.
[4] 冯晓珂,董岩,李治冶,等.骨髓间充质干细胞复合脱细胞软骨和富血小板血浆的体内成骨实验[J].实用口腔医学杂志, 2017, 33(2): 148-151.
[5] Qin Y, Sun R, Wu C, et al. Exosome: a novel approach to stimulate bone regeneration through regulation of osteogenesis and angiogenesis [J]. Int J Mol Sci, 2016, 17(5). pii: E712.
[6] 何尤夫,石蓓.干细胞源外泌体治疗缺血性心脏病的近况[J].临床与病理杂志,2017,37(2): 422-428.
[7] Osugi M, Katagiri W, Yoshimi R, et al. Conditioned media from mesenchymal stem cells enhanced bone regeneration in rat calvarial bone defects [J]. Tissue Eng Part A, 2012, 18(13-14): 1479-1489.
[8] 张晓晓,李婧,王俊妹,等.银杏叶提取物对成骨细胞分化的影响及相关机制研究[J].口腔医学研究,2017,33(5): 491-495.
[9] Lauzon MA, Drevelle O, Daviau A, et al. Effects of BMP-9 and BMP-2 on the PI3K/Akt pathway in MC3T3-E1 preosteoblasts [J]. Tissue Eng Part A, 2016, 22(17-18): 1075-1085.
[10] Yang D, Okamura H, Nakashima Y, et al. Histone demethylase Jmjd3 regulates osteoblast differentiation via transcription factors Runx2 and osterix [J]. J Biol Chem, 2013, 288(47): 33530-33541.
[11] Tsao YT, Huang YJ, Wu HH, et al. Osteocalcin mediates biomineralization during osteogenic maturation in human mesenchymal stromal cells [J]. Int J Mol Sci, 2017, 18(1). pii: E159.