Modulation of alignment and differentiation of skeletal myoblasts by submicron ridges/grooves surface structure

Alignment and fusion of myoblasts into parallel arrays of multinucleated myotubes are critical in skeletal muscle tissue engineering. It is well known that contact guidance by grooves/ridges structures induces myoblasts to align and to migrate along the anisotropic direction. In this study, two series of grooved substrata with different widths (450 and 900 nm) and different depths (100, 350, and 550 nm) were studied on their effects on myoblast adhesion, proliferation, and differentiation into myotubes. We found that C2C12 cells were aligned and elongated along the direction of grooves. Groove depth was more influential on cellular morphology, proliferation, and differentiation than groove width. While cell proliferation was retarded on the grooved surfaces especially on the substrate with 900/550 nm (width/depth), differentiation was also enhanced on the patterned surfaces compared to the flat control. Our results demonstrated the potential of grooved substrata with submicron scale in skeletal muscle tissue engineering. Biotechnol. Bioeng. 2010;106: 285–294. © 2010 Wiley Periodicals, Inc.

[1]  P. Ohara,et al.  Contact guidance in vitro. A light, transmission, and scanning electron microscopic study. , 1979, Experimental cell research.

[2]  C. Murphy,et al.  Responses of human keratocytes to micro- and nanostructured substrates. , 2004, Journal of biomedical materials research. Part A.

[3]  C. Murphy,et al.  Epithelial contact guidance on well-defined micro- and nanostructured substrates , 2003, Journal of Cell Science.

[4]  C. Wilkinson,et al.  Topographical control of cell behaviour: II. Multiple grooved substrata. , 1990, Development.

[5]  C J Murphy,et al.  Effects of synthetic micro- and nano-structured surfaces on cell behavior. , 1999, Biomaterials.

[6]  G. Dunn,et al.  Alignment of fibroblasts on grooved surfaces described by a simple geometric transformation. , 1986, Journal of cell science.

[7]  Matthew J Dalby,et al.  Nucleus alignment and cell signaling in fibroblasts: response to a micro-grooved topography. , 2003, Experimental cell research.

[8]  P Connolly,et al.  Cell guidance by ultrafine topography in vitro. , 1991, Journal of cell science.

[9]  Milan Mrksich,et al.  Micropatterned Surfaces for Control of Cell Shape, Position, and Function , 1998, Biotechnology progress.

[10]  W. Tsai,et al.  Modulation of morphology and functions of human hepatoblastoma cells by nano-grooved substrata. , 2009, Acta biomaterialia.

[11]  William P King,et al.  Myoblast alignment and differentiation on cell culture substrates with microscale topography and model chemistries. , 2007, Biomaterials.

[12]  E. Olson,et al.  Interplay between proliferation and differentiation within the myogenic lineage. , 1992, Developmental biology.

[13]  S. Mohan,et al.  Inhibition of mechanosensitive cation channels inhibits myogenic differentiation by suppressing the expression of myogenic regulatory factors and caspase‐3 activity , 2005, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[14]  Stephen Britland,et al.  Differential response of fetal and neonatal myoblasts to topographical guidance cues in vitro , 1999, Development Genes and Evolution.

[15]  C. Wilkinson,et al.  Role of the cytoskeleton in the reaction of fibroblasts to multiple grooved substrata. , 1995, Cell motility and the cytoskeleton.

[16]  Mathis O. Riehle,et al.  The use of materials patterned on a nano- and micro-metric scale in cellular engineering , 2002 .

[17]  P. Wigmore,et al.  After embryonic day 17, distribution of cells on surface of primary muscle fibres in mouse is non‐random , 1996, Developmental dynamics : an official publication of the American Association of Anatomists.

[18]  Juin-Yih Lai,et al.  Quantitative analysis of osteoblast-like cells (MG63) morphology on nanogrooved substrata with various groove and ridge dimensions. , 2009, Journal of biomedical materials research. Part A.

[19]  P. Clark,et al.  Alignment of myoblasts on ultrafine gratings inhibits fusion in vitro. , 2002, The international journal of biochemistry & cell biology.

[20]  C. S. Chen,et al.  Geometric control of cell life and death. , 1997, Science.

[21]  M. Wakelam The fusion of myoblasts. , 1985, The Biochemical journal.

[22]  Benjamin Chu,et al.  Myotube assembly on nanofibrous and micropatterned polymers. , 2006, Nano letters.

[23]  Shuichi Takayama,et al.  The effect of continuous wavy micropatterns on silicone substrates on the alignment of skeletal muscle myoblasts and myotubes. , 2006, Biomaterials.

[24]  P. Wigmore,et al.  The generation of fiber diversity during myogenesis. , 1998, The International journal of developmental biology.

[25]  Michael Olbrich,et al.  Proliferation of aligned mammalian cells on laser-nanostructured polystyrene. , 2008, Biomaterials.

[26]  Kazunori Shimizu,et al.  Alignment of skeletal muscle myoblasts and myotubes using linear micropatterned surfaces ground with abrasives , 2009, Biotechnology and bioengineering.

[27]  Robert G. Dennis,et al.  Excitability and isometric contractile properties of mammalian skeletal muscle constructs engineered in vitro , 2000, In Vitro Cellular & Developmental Biology - Animal.

[28]  A F von Recum,et al.  Quantitative analysis of fibroblast morphology on microgrooved surfaces with various groove and ridge dimensions. , 1996, Biomaterials.

[29]  Hywel Morgan,et al.  Superhydrophobicity and superhydrophilicity of regular nanopatterns. , 2005, Nano letters.

[30]  Hsuan-Liang Liu,et al.  Fibronectin modulates the morphology of osteoblast-like cells (MG-63) on nano-grooved substrates , 2009, Journal of materials science. Materials in medicine.

[31]  Hansong Zeng,et al.  Fabrication of skeletal muscle constructs by topographic activation of cell alignment , 2009, Biotechnology and bioengineering.