Osteogenesis of Mesenchymal Stem Cells by Nanoscale Mechanotransduction
Osteogenesis of Mesenchymal Stem Cells by Nanoscale Mechanotransduction
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DOI：
10.1021/nn400202j
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发表时间：
2013-03-01
期刊：
影响因子：
17.1
通讯作者：
Dalby, Matthew J.
中科院分区：
文献类型：
--
作者：
Nikukar, Habib;Reid, Stuart;Dalby, Matthew J.
间充质干细胞很可能会在许多自体再生疗法中找到用途。然而，我们控制细胞干细胞生长和分化的能力目前是有限的，并且这是这些多能细胞的临床应用的主要障碍，特别是当考虑到在培养中不使用可溶性因子或复杂培养基制剂的期望时。此外，临床上需要大量的细胞是目前使用基于材料（刚度，化学，纳米形貌等）的细胞的障碍。培养基在这里，我们给出了使用纳米级正弦机械转导协议（10-14 nm的位移，1 kHz的频率），“nanoking”，以促进成骨细胞在人类间充质干细胞培养的第一个示范。在应用反向压电效应的基础上，我们使用干涉法来开发最佳的干细胞刺激条件，允许在所使用的培养皿的整个表面上传递纳米级线索。免疫荧光，PCR和微阵列的组合，然后被用来证明成骨细胞，和阵列牵连RhoA作为中央成骨细胞分化与材料为基础的战略协议。我们用RhoA激酶的药理学抑制证实了这一点。很容易设想这样的刺激方案被放大以形成大规模的成骨细胞bloreactor，因为标准细胞培养板和孵育器被用于本发明。
It Is likely that mesenchymal stem cells will find use in many autologous regenerative therapies. However, our ability to control cell stem growth and differentiation is presently limited, and this is a major hurdle to the clinical use of these multlpotent cells especially when considering the desire not to use soluble factors or complex media formulations in culture. Also, the large number of cells required to be clinically useful is currently a hurdle to using materials-based (stiffness, chemistry, nanotopography, etc.) culture substrates. Here we give a first demonstration of using nanoscale sinusoidal mechanotransductive protocols (10-14 nm displacements at 1 kHz frequency), "nanokIcking", to promote osteoblastogenesis In human mesenchymal stem cell cultures. On the basis of application of the reverse piezo effect, we use interferometry to develop the optimal stem cell stimulation conditions, allowing delivery of nanoscale cues across the entire surface of the Petri dishes used. A combination of immundluorescence, PCR, and microarray has then been used to demonstrate osteoblastogenesis, and the arrays implicate RhoA as central to osteoblastic differentiation in agreement with materials-based strategies. We validate this with pharmacological inhibition of RhoA klnase. It Is easy to envisage such stimulation protocols being up-scaled to form largescale osteoblast bloreactors as standard cell culture plates and incubators are used In the