[Shake flask bioreactor system for dynamic culture of mesenchymal stem cells on electrospun PCL-nHA nanofibrous scaffolds for bone tissue engineering]

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

[Investigation of the dynamic culture medium on proliferation and differentiation of mesenchymal stem cells after culturing on electrospun PCL and PCL-nHA nanofibers]

Objective: More similarity to in vivo may help to increase the proliferation and differentiation of cells at in vitro culture. The present study has investigated the effect of a dynamic culture medium and nano hydroxyapatite [nHA] presence on proliferation and differentiation of mesenchymal stem cells [MSCs] to bone cells using electrospun polycaprolactone [PCL] scaffolds

Methods: We prepared PCL and PCL-nHA scaffolds by electrospinning. After static culturing of the scaffolds with MSCs, the scaffolds, were divided into two groups of static and dynamic cultures. The dynamic culture scaffolds were placed on a shaker. Cell proliferation and differentiation at days 3, 7 and 14 were investigated by MTT, and the calcium and alkaline phosphatase assays

Results: The obtained results from the MTT assay on day 14 showed an increase of 1.1 times in cell proliferation in the dynamic culture compared to the static culture. During this period, the calcium content produced by cells in the dynamic culture at day 14 were 1.23 times higher for PCL scaffolds and 1.46 times higher for PCL-nHA scaffolds compared to the static culture. Alkaline phosphatase levels for the dynamic state PCL scaffold were 1.24 times more and for PCL-nHA scaffolds were 1.28 times more compared to the static culture at day 14

Conclusion: The obtained results from dynamic culture, showed higher proliferation and differentiation of stem cells to bone for both PCL and PCL-nHA scaffolds compared to the static culture. The amount of cell proliferation and differentiation in the scaffolds that contained nHA was more than scaffolds that did not have nHA