Preconditioning Human Mesenchymal Stem Cells with a Low Concentration of BMP2 Stimulates Proliferation and Osteogenic Differentiation In Vitro.
Preconditioning Human Mesenchymal Stem Cells with a Low Concentration of BMP2 Stimulates Proliferation and Osteogenic Differentiation In Vitro.
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DOI：
10.1089/biores.2014.0044
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发表时间：
2014-12-01
影响因子：
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通讯作者：
Bünger C
中科院分区：
文献类型：
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作者：
Lysdahl H;Baatrup A;Foldager CB;Bünger C
使用骨形态发生蛋白-2（BMP-2）进行骨重建的临床试验已显示出有希望的结果。然而，有效所需的相对高的浓度引起了对功效和安全性的担忧。本研究的目的是研究替代治疗策略的成骨作用，其中人骨髓间充质干细胞（hMSCs）在体外用低浓度的BMP 2短时间预处理。将悬浮液中的hMSC用10和20 ng/mL的BMP 2刺激15分钟。去除BMP 2后，将细胞接种并培养在成骨培养基中。从基因和蛋白水平分析预处理对成骨细胞增殖和成骨标志物表达的影响。将结果与来自具有连续BMP 2刺激的培养物的结果进行比较。在预处理和用BMP 2连续刺激的情况下均观察到增殖的显著增加，处理之间没有差异。BMP 2预处理显著增加RUNX 2、COLI、ALP和OC的基因表达以及COLI和ALP的蛋白水平。这在连续刺激中没有发现。通过SMAD 1基因表达增加和SMAD 1/5/8通路中ser 463和ser 465双重磷酸化增加的发现，证实了BMP 2预处理在成骨中的作用。我们的结论是，预处理hMSCs与BMP 2刺激成骨：增殖与基质分泌和基质成熟的hMSCs。这意味着BMP 2预处理可能比连续刺激更有效地诱导hMSCs增殖和成骨分化。BMP 2预处理可以避免大剂量治疗的副作用，有利于BMP 2的临床应用。
Clinical trials using bone morphogenetic protein-2 (BMP2) for bone reconstruction have shown promising results. However, the relatively high concentration needed to be effective raises concerns for efficacy and safety. The aim of this study was to investigate the osteogenic effect of an alternative treatment strategy in which human bone marrow–derived mesenchymal stem cells (hMSCs) are preconditioned with low concentrations of BMP2 for a short time in vitro. hMSCs in suspension were stimulated for 15 min with 10 and 20 ng/mL of BMP2. After the BMP2 was removed, the cells were seeded and cultured in osteogenic medium. The effects of preconditioning were analyzed with regard to proliferation and expression of osteogenic markers at both gene and protein level. The results were compared to those from cultures with continuous BMP2 stimulation. A significant increase in proliferation was seen with both precondition and continuous stimulation with BMP2, with no difference between the treatments. Preconditioning with BMP2 significantly increased gene expression of RUNX2, COLI, ALP, and OC, and protein levels of COLI and ALP. This was not found with continuous stimulation. The role of preconditioning with BMP2 in osteogenesis was validated by findings of increased gene expression of SMAD1 and an increase in dual phosphorylation of ser 463 and ser 465 in the SMAD 1/5/8 pathway. We concluded that preconditioning hMSCs with BMP2 stimulates osteogenesis: proliferation with matrix secretion and matrix maturation of hMSCs. This implies that preconditioning with BMP2 might be more effective at inducing proliferation and osteogenic differentiation of hMSCs than continuous stimulation. Preconditioning with BMP2 could benefit the clinical application of BMP2 since side effects from high-dose treatments could be avoided.