Combination of enzymes and flow perfusion conditions improves osteogenic differentiation of bone marrow stromal cells cultured upon starch/poly(ε-caprolactone) fiber meshes

  1. Ana M. Martins1,2,3,
  2. Anita Saraf3,
  3. Rui A. Sousa1,2,
  4. Catarina M. Alves1,2,
  5. Antonios G. Mikos3,
  6. F. Kurtis Kasper3,
  7. Rui L. Reis1,2,*

Article first published online: 31 MAR 2010

DOI: 10.1002/jbm.a.32785

Issue

Journal of Biomedical Materials Research Part A

Journal of Biomedical Materials Research Part A

Volume 94A, Issue 4, pages 1061–1069, 15 September 2010

Additional Information

How to Cite

Martins, A. M., Saraf, A., Sousa, R. A., Alves, C. M., Mikos, A. G., Kasper, F. K. and Reis, R. L. (2010), Combination of enzymes and flow perfusion conditions improves osteogenic differentiation of bone marrow stromal cells cultured upon starch/poly(ε-caprolactone) fiber meshes. J. Biomed. Mater. Res., 94A: 1061–1069. doi: 10.1002/jbm.a.32785

Author Information

  1. 1

    3B's Research Group—Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Taipas 4806-909, Guimarães, Portugal

  2. 2

    IBB—Institute for Biotechnology and Bioengineering, PT Government Associated Laboratory, Guimarães, Portugal

  3. 3

    Department of Bioengineering, Rice University, 6100 Main Street, Houston, Texas 77005-1892

*3B's Research Group—Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Taipas 4806-909, Guimarães, Portugal

Publication History

  1. Issue published online: 2 AUG 2010
  2. Article first published online: 31 MAR 2010
  3. Manuscript Accepted: 13 JAN 2010
  4. Manuscript Revised: 9 DEC 2009
  5. Manuscript Received: 4 SEP 2009

Funded by

  • Portuguese Foundation for Science and Technology (FCT). Grant Number: SFRH/BPD/26763/2006
  • European NoE EXPERTISSUES. Grant Number: NMP3-CT-2004-500283

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Keywords:

  • starch/poly(ε-polycaprolactone) fiber meshes;
  • α-amylase;
  • lipase;
  • flow perfusion bioreactor;
  • osteogenic differentiation

Abstract

Previous studies have shown that α-amylase and lipase are capable of enhancing the degradation of fiber meshes blends of starch and poly(ε-caprolactone) (SPCL) under dynamic conditions, and consequently to promote the proliferation and osteogenic differentiation of bone marrow stromal cells (MSCs). This study investigated the effect of flow perfusion bioreactor culture in combination with enzymes on the osteogenic differentiation of MSCs. SPCL fiber meshes were seeded with MSCs and cultured with osteogenic medium supplemented with α-amylase, lipase, or a combination of the two for 8 or 16 days using static or flow conditions. Lipase and its combination with α-amylase enhanced cell proliferation after 16 days. In addition, the flow perfusion culture enhanced the infiltration of cells and facilitated greater distribution of extracellular matrix (ECM) throughout the scaffolds in the presence/absence of enzymes. A significant amount of calcium was detected after 16 days in all groups cultured in flow conditions compared with static cultures. Nevertheless, when α-amylase and lipase were included in the flow perfusion cultures, the calcium content was 379 ± 30 μg/scaffold after as few as 8 days. The highest calcium content (1271 ± 32 μg/scaffold) was obtained for SPCL/cell constructs cultured for 16 days in the presence of lipase and flow. Furthermore, von Kossa staining and tetracycline fluorescence of histological sections demonstrated mineral deposition within the scaffolds for all groups cultured for 16 days under flow. However, all the data corroborate that lipase coupled with flow perfusion conditions improve the osteogenic differentiation of MSCs and enhance ECM mineralization. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2010.

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