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Research Papers

Hydrostatic Pressure Stimulation of Human Mesenchymal Stem Cells Seeded on Collagen-Based Artificial Extracellular Matrices

[+] Author and Article Information
Ricarda Hess

Institute of Material Science, Max Bergmann Center of Biomaterials, Technische Universität Dresden, 01069 Dresden, Germanyricarda.hess@tu-dresden.de

Timothy Douglas

Department of Oral and Maxillofacial Surgery, University of Kiel, 24105 Kiel, Germanydouglas@mkg.uni-kiel.de

Kenneth A. Myers

McCaig Institute for Bone and Joint Health, University of Calgary, Calgary, AB, T2N 4N1, Canadakmyers@ucalgary.ca

Barbe Rentsch

 Catgut GmbH, 08258 Markneukirchen, Germanycatgut.office@catgut.de

Claudia Rentsch

Trauma and Reconstructive Surgery, University Hospital Carl Gustav Carus Dresden, 01307 Dresden, Germanyclaudia.rentsch@uniklinikum-dresden.de

Hartmut Worch

Institute of Material Science, Max Bergmann Center of Biomaterials, Technische Universität Dresden, 01069 Dresden, Germanyhartmut.worch@tu-dresden.de

Nigel G. Shrive

McCaig Institute for Bone and Joint Health, University of Calgary, Calgary, AB, T2N 4N1, Canadanshrive@ucalgary.ca

David A. Hart

McCaig Institute for Bone and Joint Health, University of Calgary, Calgary, AB, T2N 4N1, Canadahartd@ucalgary.ca

Dieter Scharnweber

Institute of Material Science, Max Bergmann Center of Biomaterials, Technische Universität Dresden, 01069 Dresden, Germanydieter.scharnweber@tu-dresden.de

J Biomech Eng 132(2), 021001 (Dec 28, 2009) (6 pages) doi:10.1115/1.4000194 History: Received March 05, 2009; Revised August 19, 2009; Posted September 10, 2009; Published December 28, 2009; Online December 28, 2009

Human mesenchymal stem cells (hMSCs) from bone marrow are considered a promising cell source for bone tissue engineering applications because of their ability to differentiate into cells of the osteoblastic lineage. Mechanical stimulation is able to promote osteogenic differentiation of hMSC; however, the use of hydrostatic pressure (HP) has not been well studied. Artificial extracellular matrices containing collagen and chondroitin sulfate (CS) have promoted the expression of an osteoblastic phenotype by hMSCs. However, there has been little research into the combined effects of biochemical stimulation by matrices and simultaneous mechanical stimulation. In this study, artificial extracellular matrices generated from collagen and/or CS were coated onto polycaprolactone-co-lactide substrates, seeded with hMSCs and subjected to cyclic HP at various time points during 21 days after cell seeding to investigate the effects of biochemical, mechanical, and combined biochemical and mechanical stimulations. Cell differentiation was assessed by analyzing the expression of alkaline phosphatase (ALP) at the protein- and mRNA levels, as well as for calcium accumulation. The timing of HP stimulation affected hMSC proliferation and expression of ALP activity. HP stimulation after 6 days was most effective at promoting ALP activity. CS-containing matrices promoted the osteogenic differentiation of hMSCs. A combination of both CS-containing matrices and cyclic HP yields optimal effects on osteogenic differentiation of hMSCs on scaffolds compared with individual responses.

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Figures

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Figure 1

(a) Schematic representation of experiments to investigate the effect of cyclic HP on hMSC proliferation seeded on uncoated PCL. Samples were divided into groups, which were stimulated at different time points after seeding. Samples were collected in the course of the experiments at 0 day, 4 days, 7 days, 9 days, 11 days, 15 days, and 21 days after seeding. Experiments were terminated after 21 days. (b) Pressure profile used in experiments. PCL scaffolds with hMSCs were subjected to cyclic HP between 300 kPa and 375 kPa at 0.5 Hz for 1 min every 15 min for a period of 8 h.

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Figure 2

Effect of timing of HP stimulation on hMSC proliferation in the course of 21 days in the case of stimulation at (a) 1 day, (b) 3 days, (c) 6 days, and (d) 13 days after seeding

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Figure 3

Effect of timing of HP stimulation and osteogenic coatings on ALP activity: (a) ALP activity 21 days after seeding and (b) gene expression of ALP and the housekeeping-gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) on samples coated with collagen I and CS 21 days after seeding. (Abbreviations: w/o means without HP stimulation, HP 6 means HP stimulation 6 days after seeding, and HP 13 means HP stimulation 13 days after seeding. All samples were cultured in (DMEM)+10%(FCS)+1/P/S. Significances:  ∗=p<0.05 and  ∗∗=p<0.01.)

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Figure 4

Effect of osteogenic supplements and timing of HP stimulation on calcium accumulation. (Abbreviations: w/o means without HP stimulation, HP 6 means HP stimulation 6 days after seeding, and HP 13 means HP stimulation 13 days after seeding. All samples were cultured in (DMEM)+10%(FCS)+1/P/S. Significance:  ∗=p<0.05.)

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