Shake Flask Bioreactor System for Dynamic Culture of Mesenchymal Stem Cells on Electrospun PCL-nHA Nanofibrous Scaffolds for Bone Tissue Engineering

Authors
M. Vazirzadeh 1
S. Hashemi-Najafabadi 2
M. Soleimani 3
1 Department of Biomedical Engineering, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran Iran
2 Department of Biomedical Engineering, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran
3 Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Abstract
Objective: In the present study we investigated the effect of a dynamic culture in a shake flask bioreactor (SFB) on the proliferation and differentiation to osteoblasts for human mesenchymal stem cells (hMSCs) cultured on multilayered electrospun PCL-nHA scaffolds.
Methods: First, we prepared PCL-nHA scaffolds by electrospinning. After culturing the hMSCs on the scaffolds in a static state, the seeded scaffolds were divided into two groups (static and SFB culture) and incubated up to 21 days. We assessed biocompatibility and cell differentiation by the MTT, calcium, and alkaline phosphatase (ALP) assays on days 7, 14, and 21.
Results: The MTT assay evaluated hMSCs proliferation rate on the scaffold layers. There was greater cell proliferation (optical density values) on the layers in the bioreactor (OD=2.18) compared to the static state condition (OD=1.68) on day 21. In order to study osteogenic differentiation, we determined the amount of calcium deposition and ALP activity. We observed a 1.6-fold greater level of calcium deposition for the dynamic culture compared to the static culture, which showed increased cell differentiation within the bioreactor on day 21. The ALP results showed that during 14 days, ALP activity within the bioreactor was 1.55-fold higher than the static culture.
Conclusion: The SFB culture displayed a higher proliferation and differentiation of stem cells on PCL-nHA multilayered scaffolds compared to the static state condition.

Keywords

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Pathobiology Reserach
Volume 19, Issue 1
May 2016
Pages 79-91