title: RhoA-Mediated Functions in C3H10T1/2 Osteoprogenitors Are Substrate Topography Dependent.
abstract: 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.
authors.affiliation: Department of Prosthodontics, School of Dentistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.|Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University, Fukuoka, Japan.
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Ogino
authors.affiliation: Department of Prosthodontics, School of Dentistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors.forename: Ruiwei
authors.initials: R
authors.lastname: Liang
authors.affiliation: Division of Prosthodontics, School of Dentistry, University of Michigan, Ann Arbor, Michigan.
authors.forename: Daniela B S
authors.initials: DB
authors.lastname: Mendonça
authors.affiliation: Division of Prosthodontics, School of Dentistry, University of Michigan, Ann Arbor, Michigan.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Mendonça
authors.affiliation: Department of Prosthodontics, School of Dentistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.|Division of Bio-Prosthodontics, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
authors.forename: Masako
authors.initials: M
authors.lastname: Nagasawa
authors.affiliation: Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University, Fukuoka, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Koyano
authors.affiliation: Department of Prosthodontics, School of Dentistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors.forename: Lyndon F
authors.initials: LF
authors.lastname: Cooper
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D010012:Osteogenesis / Q000502:physiology*; D020741:rho GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein
chemical_list: C526380:RhoA protein, mouse; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein