D Prè, G Ceccarelli, L Visai, L Benedetti, M Imbriani, M G Cusella De Angelis, G Magenes
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引用次数: 29
摘要
为了验证高频振动(HFV)是否促进成体干细胞向骨组织的分化,我们用HFV (30 Hz)每天45分钟的机械刺激骨髓间质细胞(BMSCs),持续21或40天。将细胞植入成骨培养基中,增强向骨组织的分化。采用茜素红试验(Alizarin Red test)、实时荧光定量PCR (real-time PCR)和细胞外基质蛋白含量检测机械处理对分化的影响。此外,我们还分析了机械刺激下BMSC的增殖率和凋亡情况。观察到所有表征分化的参数都有明显的增加。在处理过的样品中,钙的沉积几乎增加了一倍;与后期分化相关的基因表达显著增加,所有成骨蛋白的蛋白含量均较高。最后,增殖结果表明,与对照组相比,受刺激的骨髓间充质干细胞的生长速度下降,但处理和未处理的细胞都不进入凋亡过程。这些发现可以通过改进自体细胞的分化方案和进一步将骨移植物植入患者体内来缩小来自患者细胞的骨替代物的研究与临床应用之间的差距。
High-Frequency Vibration Treatment of Human Bone Marrow Stromal Cells Increases Differentiation toward Bone Tissue.
In order to verify whether differentiation of adult stem cells toward bone tissue is promoted by high-frequency vibration (HFV), bone marrow stromal cells (BMSCs) were mechanically stimulated with HFV (30 Hz) for 45 minutes a day for 21 or 40 days. Cells were seeded in osteogenic medium, which enhances differentiation towards bone tissue. The effects of the mechanical treatment on differentiation were measured by Alizarin Red test, (q) real-time PCR, and protein content of the extracellular matrix. In addition, we analyzed the proliferation rate and apoptosis of BMSC subjected to mechanical stimulation. A strong increase in all parameters characterizing differentiation was observed. Deposition of calcium was almost double in the treated samples; the expression of genes involved in later differentiation was significantly increased and protein content was higher for all osteogenic proteins. Lastly, proliferation results indicated that stimulated BMSCs have a decreased growth rate in comparison with controls, but both treated and untreated cells do not enter the apoptosis process. These findings could reduce the gap between research and clinical application for bone substitutes derived from patient cells by improving the differentiation protocol for autologous cells and a further implant of the bone graft into the patient.