RhoA-Mediated Functions in C3H10T1/2 Osteoprogenitors Are Substrate Topography Dependent.
RhoA-Mediated Functions in C3H10T1/2 Osteoprogenitors Are Substrate Topography Dependent.
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DOI：
10.1002/jcp.25100
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发表时间：
2016-03
影响因子：
5.6
通讯作者：
Cooper LF
中科院分区：
文献类型：
--
作者：
Ogino Y;Liang R;Mendonça DB;Mendonça G;Nagasawa M;Koyano K;Cooper LF
表面形貌广泛影响细胞反应。贴壁细胞的活性在一定程度上由RhoA调控，RhoA是GTPases的RhoA家族成员。在这项研究中，我们评估了表面形貌对RhoA活性和相关细胞功能的影响。将小鼠间充质干细胞C3H10T1/2细胞（骨祖细胞）培养在光滑形貌(S)、微形貌(M)和纳米形貌(N)的钛基质上，观察表面形貌对rhoa介导的功能（细胞扩散、粘附、迁移和成骨分化）的影响。利用RhoA药物抑制剂也阐明了RhoA活性对表面形貌的影响。粘附后，M和N贴壁细胞呈多突状，S贴壁细胞呈扁平分布。RhoA抑制剂在所有表面上诱导了明显的更长和更薄的细胞质突起。细胞粘附和成骨分化依赖于S < M和N表面。RhoA抑制增加了S和M表面的粘附，但对N表面没有影响。在伤口愈合实验中，细胞在S、M、N表面的迁移更大，RhoA抑制剂增加了S粘附细胞的迁移，但没有增加N粘附细胞的迁移。RhoA抑制剂对S贴壁细胞的成骨分化有促进作用，而对M和N贴壁细胞没有促进作用。RhoA活性与表面形貌粗糙度相关（S < M， N）。RhoA活性和介导的功能受表面形貌的影响。光滑表面贴壁细胞对RhoA功能表现出高度敏感，而纳米尺度的地形贴壁细胞可能利用其他细胞信号通路来影响贴壁细胞功能，而不管RhoA活性如何。
Surface topography broadly influences cellular responses. Adherent cell activities are regulated, in part, by RhoA, a member of the Rho-family of GTPases. In this study, we evaluated the influence of surface topography on RhoA activity and associated cellular functions. The murine mesenchymal stem cell line C3H10T1/2 cells (osteoprogenitor cells) were cultured on titanium substrates with smooth topography (S), microtopography (M), and nanotopography (N) to evaluate the effect of surface topography on RhoA-mediated functions (cell spreading, adhesion, migration, and osteogenic differentiation). The influence of RhoA activity in the context of surface topography was also elucidated using RhoA pharmacologic inhibitor. Following adhesion, M and N adherent cells developed multiple projections, while S adherent cells had flattened and widespread morphology. RhoA inhibitor induced remarkable longer and thinner cytoplasmic projections on all surfaces. Cell adhesion and osteogenic differentiation was topography dependent with S < M and N surfaces. RhoA inhibition increased adhesion on S and M surfaces, but not N surfaces. Cell migration in a wound healing assay was greater on S versus M versus N surfaces and RhoA inhibitor increased S adherent cell migration, but not N adherent cell migration. RhoA inhibitor enhanced osteogenic differentiation in S adherent cells, but not M or N adherent cells. RhoA activity was surface topography roughness dependent (S < M, N). RhoA activity and -mediated functions are influenced by surface topography. Smooth surface adherent cells appear highly sensitive to RhoA function, while nano-scale topography adherent cell may utilize alternative cellular signaling pathway(s) to influence adherent cellular functions regardless of RhoA activity.