Response of MG63 osteoblast-like cells to ordered nanotopographies fabricated using colloidal self-assembly and glancing angle deposition.

Ordered surface nanostructures have attracted much attention in different fields including biomedical engineering because of their potential to study the size effect on cellular response and modulation of cell fate. However, the ability to fabricate large-area ordered nanostructures is typically limited due to high costs and low speed of fabrication. Herein, highly ordered nanostructures with large surface areas (>1.5 × 1.5 cm(2)) were fabricated using a combination of facile techniques including colloidal self-assembly, colloidal lithography, and glancing angle deposition (GLAD). An ordered tantalum (Ta) pattern with 60-nm-height was generated using colloidal lithography. A monolayer of colloidal crystal, i.e., hexagonal close packed 720 nm polystyrene particles, was self-assembled and used as a mask. Ta patterns were subsequently generated by evaporation of Ta through the mask. The feature size was further increased by 100 or 200 nm using GLAD, resulting in the fabrication of four different surfaces (FLAT, Ta60, GLAD100, and GLAD200). Cell adhesion, proliferation, and mineralization of MG63 osteoblast-like cells were investigated on these ordered nanostructures over a 1 week period. Our results showed that cell adhesion, spreading, focal adhesion formation, and filopodia formation of the MG63 osteoblast-like cells were inhibited on the GLAD surfaces, especially the initial (24 h) attachment, resulting in a lower cell density on the GLAD surfaces. After 1 week culture, alkaline phosphatase activity and the amount of Ca was higher on the GLAD surfaces compared with Ta60 and FLAT controls, suggesting that the GLAD surfaces facilitate differentiation of osteoblasts. This study demonstrates that ordered Ta nanotopographies synthesized by combining colloidal lithography with GLAD can improve the mineralization of osteoblast-like cells providing a new platform for biomaterials and bone tissue engineering.

[1]  Joachim P Spatz,et al.  Cooperativity in adhesion cluster formation during initial cell adhesion. , 2008, Biophysical journal.

[2]  W. Tsai,et al.  The roles of RGD and grooved topography in the adhesion, morphology, and differentiation of C2C12 skeletal myoblasts , 2012, Biotechnology and bioengineering.

[3]  Masahiro Ohshima,et al.  Time-lapse observation of cell alignment on nanogrooved patterns , 2009, Journal of The Royal Society Interface.

[4]  C. Wilkinson,et al.  The control of human mesenchymal cell differentiation using nanoscale symmetry and disorder. , 2007, Nature materials.

[5]  Lauren R. Clements,et al.  Screening Mesenchymal Stem Cell Attachment and Differentiation on Porous Silicon Gradients , 2012 .

[6]  Lauren R. Clements,et al.  High-throughput characterisation of osteogenic differentiation of human mesenchymal stem cells using pore size gradients on porous alumina. , 2013, Biomaterials science.

[7]  J. Borges,et al.  An overview of inverted colloidal crystal systems for tissue engineering. , 2014 .

[8]  N. Rosenthal,et al.  Derive and conquer: sourcing and differentiating stem cells for therapeutic applications , 2008, Nature Reviews Drug Discovery.

[9]  R. Tannenbaum,et al.  The effects of combined micron-/submicron-scale surface roughness and nanoscale features on cell proliferation and differentiation. , 2011, Biomaterials.

[10]  F. Guilak,et al.  Control of stem cell fate by physical interactions with the extracellular matrix. , 2009, Cell stem cell.

[11]  Peng-Yuan Wang,et al.  Modulation of alignment and differentiation of skeletal myoblasts by submicron ridges/grooves surface structure , 2010, Biotechnology and bioengineering.

[12]  Ali Khademhosseini,et al.  Engineering microscale topographies to control the cell-substrate interface. , 2012, Biomaterials.

[13]  K. Anselme,et al.  Osteoblast adhesion on biomaterials. , 2000, Biomaterials.

[14]  Jiandong Ding,et al.  Cell–Material Interactions Revealed Via Material Techniques of Surface Patterning , 2013, Advanced materials.

[15]  Nicholas A. Kotov,et al.  A floating self-assembly route to colloidal crystal templates for 3D cell scaffolds , 2005 .

[16]  Massoud Motamedi,et al.  Inverted‐Colloidal‐Crystal Hydrogel Matrices as Three‐Dimensional Cell Scaffolds , 2005 .

[17]  M. Foss,et al.  Low-aspect ratio nanopatterns on bioinert alumina influence the response and morphology of osteoblast-like cells. , 2015, Biomaterials.

[18]  C. Lohmann,et al.  Response of MG63 osteoblast-like cells to titanium and titanium alloy is dependent on surface roughness and composition. , 1998, Biomaterials.

[19]  P. Kingshott,et al.  Self-assembled binary colloidal crystal monolayers as cell culture substrates. , 2015, Journal of materials chemistry. B.

[20]  Cato T Laurencin,et al.  Bone tissue engineering: recent advances and challenges. , 2012, Critical reviews in biomedical engineering.

[21]  Y. Kuo,et al.  Inverted colloidal crystal scaffolds with induced pluripotent stem cells for nerve tissue engineering. , 2013, Colloids and surfaces. B, Biointerfaces.

[22]  A Curtis,et al.  Topographical control of cells. , 1997, Biomaterials.

[23]  Q. Yan,et al.  Colloidal monolayer at the air/water interface: Large-area self-assembly and in-situ annealing , 2013 .

[24]  R. Langer,et al.  Engineering substrate topography at the micro- and nanoscale to control cell function. , 2009, Angewandte Chemie.

[25]  J. Black Biological performance of tantalum. , 1994, Clinical materials.

[26]  W. Tsai,et al.  Modulation of cell attachment and collagen production of anterior cruciate ligament cells via submicron grooves/ridges structures with different cell affinity , 2013, Biotechnology and bioengineering.

[27]  W. Tsai,et al.  Grooved PLGA films incorporated with RGD/YIGSR peptides for potential application on skeletal muscle tissue engineering. , 2013, Colloids and surfaces. B, Biointerfaces.

[28]  W. Tsai,et al.  Modulation of osteogenic, adipogenic and myogenic differentiation of mesenchymal stem cells by submicron grooved topography , 2012, Journal of Materials Science: Materials in Medicine.

[29]  J. Lovmand,et al.  Control of proliferation and osteogenic differentiation of human dental-pulp-derived stem cells by distinct surface structures. , 2014, Acta biomaterialia.

[30]  Megan S. Lord,et al.  Influence of nanoscale surface topography on protein adsorption and cellular response , 2010 .

[31]  Alireza Dolatshahi-Pirouz,et al.  A combinatorial screening of human fibroblast responses on micro-structured surfaces. , 2010, Biomaterials.

[32]  Todd C. McDevitt,et al.  Materials as stem cell regulators. , 2014, Nature materials.

[33]  M. Fedorchak,et al.  Scaling up self-assembly: bottom-up approaches to macroscopic particle organization. , 2015, Soft matter.

[34]  Helmut Thissen,et al.  Screening the attachment and spreading of bone marrow-derived and adipose-derived mesenchymal stem cells on porous silicon gradients , 2012 .

[35]  Peng-Yuan Wang,et al.  Modulation of alignment, elongation and contraction of cardiomyocytes through a combination of nanotopography and rigidity of substrates. , 2011, Acta biomaterialia.

[36]  Yong Yang,et al.  Nanotopography as modulator of human mesenchymal stem cell function. , 2012, Biomaterials.

[37]  V. Sikavitsas,et al.  Biomaterials and bone mechanotransduction. , 2001, Biomaterials.

[38]  M. Foss,et al.  Modulation of human mesenchymal stem cell behavior on ordered tantalum nanotopographies fabricated using colloidal lithography and glancing angle deposition. , 2015, ACS applied materials & interfaces.

[39]  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.

[40]  Lingzhou Zhao,et al.  The role of integrin-linked kinase/β-catenin pathway in the enhanced MG63 differentiation by micro/nano-textured topography. , 2013, Biomaterials.

[41]  A. Yee,et al.  Expression of Oct4 in human embryonic stem cells is dependent on nanotopographical configuration. , 2013, Acta biomaterialia.