Effect of substrate stiffness on the osteogenic differentiation of bone marrow stem cells and bone‐derived cells

There is a profound dependence of cell behaviour on the stiffness of its microenvironment. To gain a better understanding of the regulation of cellular differentiation by mechanical cues, we investigated the influence of matrix stiffness (E = 1.46 kPa and E = 26.12 kPa) on differentiated osteogenic cell lineage of bone marrow stem cells (BM‐MSCs) and bone‐derived cells (BDCs) using flexible collagen‐coated polyacrylamide substrates. Differentiation potential was determined by measuring alkaline phosphatase activity, expression of osteoblast‐specific markers including alkaline phosphatase, osteocalcin, Runx2 and collagen type I, as well as assessment of mineralisation (Alizarin Red S staining). We found that osteogenic differentiation can be regulated by the rigidity of the substrate, which may depend on the commitment in multi‐ or uni‐potent targeting cells. Osteogenic differentiation of BM‐MSCs was enhanced on a stiff substrate compared to a soft one, whereas BDCs osteogenic differentiation did not vary depending on the substrate stiffness. The data help in understanding the role of the external mechanical determinants in stem cell differentiation, and can also be useful in translational approach in functional tissue engineering.

[1]  Robert C. Breithaupt,et al.  The influence of stereolithographic scaffold architecture and composition on osteogenic signal expression with rat bone marrow stromal cells. , 2011, Biomaterials.

[2]  F. O'Brien,et al.  Substrate stiffness and contractile behaviour modulate the functional maturation of osteoblasts on a collagen-GAG scaffold. , 2010, Acta biomaterialia.

[3]  P. Janmey,et al.  Tissue Cells Feel and Respond to the Stiffness of Their Substrate , 2005, Science.

[4]  Seema Sharma,et al.  Bone marrow angiogenesis in multiple myeloma and its correlation with clinicopathological factors , 2010, Annals of Hematology.

[5]  Adam J. Engler,et al.  Myotubes differentiate optimally on substrates with tissue-like stiffness , 2004, The Journal of cell biology.

[6]  M. Dembo,et al.  Substrate flexibility regulates growth and apoptosis of normal but not transformed cells. , 2000, American journal of physiology. Cell physiology.

[7]  R. Tuan,et al.  Comparison of Proliferative and Multilineage Differentiation Potential of Human Mesenchymal Stem Cells Derived from Umbilical Cord and Bone Marrow , 2007, Stem cells.

[8]  R. Mahaffy,et al.  Quantitative analysis of the viscoelastic properties of thin regions of fibroblasts using atomic force microscopy. , 2004, Biophysical journal.

[9]  D. Prockop,et al.  Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. , 2006, Cytotherapy.

[10]  W. Grzesik,et al.  Bone matrix RGD glycoproteins: Immunolocalization and interaction with human primary osteoblastic bone cells in vitro , 1994, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[11]  G. Schmalz,et al.  Soft matrix supports osteogenic differentiation of human dental follicle cells. , 2011, Biochemical and biophysical research communications.

[12]  S. Sen,et al.  Matrix Elasticity Directs Stem Cell Lineage Specification , 2006, Cell.

[13]  Y. Wang,et al.  Cell locomotion and focal adhesions are regulated by substrate flexibility. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[14]  G. Karsenty.  Bone formation and factors affecting this process. , 2000, Matrix biology : journal of the International Society for Matrix Biology.

[15]  Xing Wei,et al.  Endothelial differentiation of Wharton's jelly-derived mesenchymal stem cells in comparison with bone marrow-derived mesenchymal stem cells. , 2009, Experimental hematology.

[16]  Małgorzata Witkowska-Zimny.  Transcriptional Control of Osteogenesis , 2012 .

[17]  Y. Wang,et al.  Preparation of a flexible, porous polyacrylamide substrate for mechanical studies of cultured cells. , 1998, Methods in enzymology.

[18]  Jennifer S. Park,et al.  The effect of matrix stiffness on the differentiation of mesenchymal stem cells in response to TGF-β. , 2011, Biomaterials.

[19]  K. Kurzydłowski,et al.  Candidate bone-tissue-engineered product based on human-bone-derived cells and polyurethane scaffold. , 2010, Acta biomaterialia.

[20]  R. Tuan,et al.  Multilineage mesenchymal differentiation potential of human trabecular bone‐derived cells , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[21]  S. Goldenberg,et al.  Dissimilar Differentiation of Mesenchymal Stem Cells from Bone Marrow, Umbilical Cord Blood, and Adipose Tissue , 2008, Experimental biology and medicine.

[22]  R. Gundle,et al.  Isolation and culture of bone-forming cells (osteoblasts) from human bone. , 1996, Methods in molecular medicine.

[23]  M. Lewandowska-Szumieł,et al.  Effect of substrate stiffness on differentiation of umbilical cord stem cells. , 2012, Acta biochimica Polonica.

[24]  Peng-Yuan Wang,et al.  Screening of rat mesenchymal stem cell behaviour on polydimethylsiloxane stiffness gradients. , 2012, Acta biomaterialia.

[25]  D. E. Discher,et al.  Matrix elasticity directs stem cell lineage — Soluble factors that limit osteogenesis , 2009 .

[26]  E. Wróbel,et al.  Perinatal sources of mesenchymal stem cells: Wharton’s jelly, amnion and chorion , 2011, Cellular & Molecular Biology Letters.

[27]  Xing Zhang,et al.  Differentiation of bone marrow mesenchymal stem cells induced by myocardial medium under hypoxic conditions , 2006, Acta Pharmacologica Sinica.

[28]  V. Sogos,et al.  Differentiation of human bone marrow stem cells into cells with a neural phenotype: diverse effects of two specific treatments , 2006, BMC Neuroscience.

[29]  A. Rowlands,et al.  Directing osteogenic and myogenic differentiation of MSCs: interplay of stiffness and adhesive ligand presentation. , 2008, American journal of physiology. Cell physiology.

[30]  Jess G Snedeker,et al.  Biochemical and biomechanical gradients for directed bone marrow stromal cell differentiation toward tendon and bone. , 2010, Biomaterials.

[31]  J. Kozakiewicz,et al.  Moisture-cured silicone-urethanes-candidate materials for tissue engineering: a biocompatibility study in vitro. , 2010, Journal of biomedical materials research. Part A.

[32]  In-Seop Lee,et al.  Enhancement of the ALP activity of C3H10T1/2 cells by the combination of an oxysterol and apatite , 2010, Biomedical materials.