低强度脉冲治疗超声应用于间充质前体细胞不会影响其细胞特性。
Application of low-intensity pulsed therapeutic ultrasound on mesenchymal precursors does not affect their cell properties.
机构信息
Faculty of Biomedical and Health Sciences, Universidad Europea de Madrid, Madrid, Spain.
Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain.
出版信息
PLoS One. 2021 Feb 11;16(2):e0246261. doi: 10.1371/journal.pone.0246261. eCollection 2021.
Ultrasound is considered a safe and non-invasive tool in regenerative medicine and has been used in the clinic for more than twenty years for applications in bone healing after the approval of the Exogen device, also known as low-intensity pulsed ultrasound (LIPUS). Beyond its effects on bone health, LIPUS has also been investigated for wound healing of soft tissues, with positive results for various cell processes including cell proliferation, migration and angiogenesis. As LIPUS has the potential to treat chronic skin wounds, we sought to evaluate the effects produced by a conventional therapeutic ultrasound device at low intensities (also considered LIPUS) on the migration capacity of mouse and human skin mesenchymal precursors (s-MPs). Cells were stimulated for 3 days (20 minutes per day) using a traditional ultrasound device with the following parameters: 100 mW/cm2 with 20% duty cycle and frequency of 3 MHz. At the parameters used, ultrasound failed to affect s-MP proliferation, with no evident changes in morphology or cell groupings, and no changes at the cytoskeletal level. Further, the migration and invasion ability of s-MPs were unaffected by the ultrasound protocol, and no major changes were detected in the gene/protein expression of ROCK1, integrin β1, laminin β1, type I collagen and transforming growth factor β1. Finally, RNA-seq analysis revealed that only 10 genes were differentially expressed after ultrasound stimulation. Among them, 5 encode for small nuclear RNAs and 2 encode for proteins belonging to the nuclear pore complex. Considering the results overall, while the viability of s-MPs was not affected by ultrasound stimulation and no changes were detected in proliferation/migration, RNA-seq analysis would suggest that s-MPs do respond to ultrasound. The use of 100 mW/cm2 intensity or conventional therapeutic ultrasound devices might not be optimal for the stimulation the properties of cell populations. Future studies should investigate the potential application of ultrasound using variations of the tested parameters.
超声被认为是再生医学中一种安全且非侵入性的工具,在 Exogen 设备(也称为低强度脉冲超声,LIPUS)获得批准后,已在临床上用于骨愈合超过 20 年。除了对骨骼健康的影响外,LIPUS 还被用于软组织的伤口愈合研究,对包括细胞增殖、迁移和血管生成在内的各种细胞过程都有积极的结果。由于 LIPUS 有可能治疗慢性皮肤伤口,我们试图评估低强度(也被认为是 LIPUS)的传统治疗性超声设备产生的影响对小鼠和人类皮肤间充质前体细胞(s-MP)的迁移能力的影响。使用传统的超声设备以以下参数刺激细胞 3 天(每天 20 分钟):100 mW/cm2,占空比为 20%,频率为 3 MHz。在使用的参数下,超声未能影响 s-MP 的增殖,细胞形态或细胞群没有明显变化,细胞骨架水平也没有变化。此外,s-MP 的迁移和侵袭能力不受超声方案的影响,ROCK1、整合素β1、层粘连蛋白β1、I 型胶原蛋白和转化生长因子β1 的基因/蛋白表达也没有发生重大变化。最后,RNA-seq 分析显示,超声刺激后只有 10 个基因表达差异。其中,5 个基因编码小核 RNA,2 个基因编码核孔复合体蛋白。综合考虑结果,虽然 s-MP 的活力不受超声刺激的影响,增殖/迁移也没有检测到变化,但 RNA-seq 分析表明 s-MP 确实对超声有反应。使用 100 mW/cm2 强度或传统治疗性超声设备可能不是刺激细胞群体特性的最佳选择。未来的研究应调查使用测试参数变化的超声的潜在应用。
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