Formation of three‐dimensional cell/polymer constructs for bone tissue engineering in a spinner flask and a rotating wall vessel bioreactor
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- 3 July 2002
- journal article
- research article
- Published by Wiley in Journal of Biomedical Materials Research
- Vol. 62 (1) , 136-148
- https://doi.org/10.1002/jbm.10150
Abstract
The aim of this study is to investigate the effect of the cell culture conditions of three‐dimensional polymer scaffolds seeded with rat marrow stromal cells (MSCs) cultured in different bioreactors concerning the ability of these cells to proliferate, differentiate towards the osteoblastic lineage, and generate mineralized extracellular matrix. MSCs harvested from male Sprague–Dawley rats were culture expanded, seeded on three‐dimensional porous 75:25 poly(D,L‐lactic‐co‐glycolic acid) biodegradable scaffolds, and cultured for 21 days under static conditions or in two model bioreactors (a spinner flask and a rotating wall vessel) that enhance mixing of the media and provide better nutrient transport to the seeded cells. The spinner flask culture demonstrated a 60% enhanced proliferation at the end of the first week when compared to static culture. On day 14, all cell/polymer constructs exhibited their maximum alkaline phosphatase activity (AP). Cell/polymer constructs cultured in the spinner flask had 2.4 times higher AP activity than constructs cultured under static conditions on day 14. The total osteocalcin (OC) secretion in the spinner flask culture was 3.5 times higher than the static culture, with a peak OC secretion occurring on day 18. No considerable AP activity and OC secretion were detected in the rotating wall vessel culture throughout the 21‐day culture period. The spinner flask culture had the highest calcium content at day 14. On day 21, the calcium deposition in the spinner flask culture was 6.6 times higher than the static cultured constructs and over 30 times higher than the rotating wall vessel culture. Histological sections showed concentration of cells and mineralization at the exterior of the foams at day 21. This phenomenon may arise from the potential existence of nutrient concentration gradients at the interior of the scaffolds. The better mixing provided in the spinner flask, external to the outer surface of the scaffolds, may explain the accelerated proliferation and differentiation of marrow stromal osteoblasts, and the localization of the enhanced mineralization on the external surface of the scaffolds. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res 62: 136–148, 2002Keywords
This publication has 50 references indexed in Scilit:
- Fluid flow-induced tyrosine phosphorylation and participation of growth factor signaling pathway in osteoblast-like cellsJournal of Cellular Biochemistry, 2000
- Medium Perfusion Enhances Osteogenesis by Murine Osteosarcoma Cells in Three-Dimensional Collagen SpongesJournal of Bone and Mineral Research, 1999
- Marrow‐derived progenitor cell injections enhance new bone formation during distractionJournal of Orthopaedic Research, 1999
- Steady and Transient Fluid Shear Stress Stimulate NO Release in Osteoblasts Through Distinct Biochemical PathwaysJournal of Bone and Mineral Research, 1999
- Nitric Oxide Response to Shear Stress by Human Bone Cell Cultures Is Endothelial Nitric Oxide Synthase DependentBiochemical and Biophysical Research Communications, 1998
- Signal Transduction of Mechanical Stimuli Is Dependent on Microfilament Integrity: Identification of Osteopontin as a Mechanically Induced Gene in OsteoblastsJournal of Bone and Mineral Research, 1997
- Review: Bone tissue engineering: The role of interstitial fluid flowBiotechnology & Bioengineering, 1994
- Measurement of serum osteocalcin with a human-specific two-site immunoradiometric assayJournal of Bone and Mineral Research, 1992
- Morbidity at Bone Graft Donor SitesJournal of Orthopaedic Trauma, 1989
- Tetracycline-based histological analysis of bone remodelingCalcified Tissue International, 1969